evaluation of biocontrol potentlal of endophytic …

i

EVALUATION OF BIOCONTROL POTENTlAL OF ENDOPHYTIC

SPECIES OF PENICILLIUM AGAINST ROOT ROTTING FUNGI

AND ROOT KNOT NEMATODE

FAIZAH UROOJ

(MPhilPhD)

DEPARTEMENT OF BOTANY UNIVERSITY OF KARACHI

KARACHI PAKISTAN

2018

ii

EVALUATION OF BIOCONTROL POTENTIAL OF ENDOPHYTIC SPECIES OF

PENICILLIUM AGAINST ROOT ROTTING FUNGI AND ROOT KNOT

NEMATODE

FAIZAH UROOJ


KARACHI PAKISTAN

2018

iii

EVALUATION OF BIOCONTROL POTENTAIL OF ENDOPHYTIC SPECIES OF


NEMATODE

A Thesis Submitted for the Partial Fulfillment of the Degree of Doctor of Philosophy in

Botany

By

FAIZAH UROOJ


KARACHI PAKISTAN

2018

iv

DEDICATION

Dedicated to my most respected teachers and my beloved

Parents who believe in me and brought out best in me

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LIST OF CONTENTS Page No

1 INTRODUCTION

11 Endophytic fungi

12 Endophytic Penicillium

13 Role of endophytic Penicillium in plant growth

14 Role of endophytic Penicillium as resistance inducers in plant against

biotic and abiotic stresses

15 Soil-borne diseases

16 Soil-borne root rotting fungi and nematode

17 Biological control

2 MATERIALS AND METHODS

21 Collection of sample for the isolation of endophytic Penicillium spp

from different host

22 Isolation and identification of endophytic Penicillium

23 Isolation of the root infecting fungi from soil

231 Soil dilution technique for the isolation of Fusarium spp

232 Baiting technique for the isolation of Rhizoctonia solani

233 Wet sieving and dilution technique for the isolation of

Macrophomina phaseolina

24 In vitro dual culture plate assay for determining the antifungal activity

of Penicillium species

25 Preparation of root knot nematode inoculum

26 Hatching of nematodes

27 Preparation of culture filtrates

28 In vitro antifungal activity of culture filtrates of Penicillium species

29 In vitro antibacterial activity of culture fitrates of Penicillium species

210 In vitro nematicidal activity of culture filtrates of Penicillium species

211 Fractionation of culture filtrates

212 Extraction and fractionation of mycelium of endophytic Penicillium

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213 Spectroscopy of oily fractions eluted from mycelium

212 In vitro antimicrobial activity of fractions of culture filtrates

213 Population of antagonists Colony forming unit (cfu) per ml in

suspension

214 Growth parameter

2141 Physical parameter

2142 Infection percentage of root rot fungi on roots

215 Biochemical parameter

2151 Estimation of polyphenols

2152 Estimation of antioxidant activity

216 Fruit analysis

2161 pH

2162 Moisture content

2163 Tritable acidity (TA)

2164 Total soluble solid (TSS)

2165 Firmness

2166 Total solids

2167 Protein

2168 Carbohydrate

2169 Total polyphenol and antioxidant activity

217 Experimental design

218 Analysis of data

3 EXPERIMENTAL RESULTS

31 Isolation of endophytic Penicillium

32 In vitro fungicidal activity of endophytic Penicillium

33 In vitro fungicidal activity of cell free culture filtrates of endophytic

Penicillium

34 In vitro antibacterial activity of cell free culture filtrates of endophytic

Penicillium

35 In vitro nematicidal activity of cell free culture filtrates of endophytic

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Penicillium spp

36 In-vitro antimicrobial activity of fractions of culture filtrates

361 In-vitro antifungal activity of n-hexane soluble fractions of

culture filtrates

362 In-vitro antibacterial activity of n-hexane soluble fractions of

culture filtrates

363 In-vitro antifungal activity of chloroform soluble fractions of

culture filtrates

364 In-vitro antibacterial activity of chloroform soluble fractions of

culture filtrates

365 Compounds from n-hexane fraction of mycelium of Penicillium

rugulosum

37 Screen house experiments

371 Effect of endophytic Penicillium in soil amended with neem cake

in suppressing the root diseases and growth of sunflower (2016)


in suppressing the root diseases and growth of Sunflower (2017)


in suppressing the root diseases and growth of mung bean

374 Effect of Endophytic Penicillium and Cotton cake in suppressing

the root diseases and growth of Mung Bean

375 Effect of Endophytic Penicillium in suppressing the root diseases

and growth of Mung Bean


in suppressing the root diseases and growth of tomato

377 Effect of endophytic Penicillium in soil amended with cotton

cake in suppressing the root diseases and growth of tomato


in suppressing the root diseases and growth of chickpea

379 Effect of endophytic Penicillium insoil amended with mustard

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cake in suppressing the root diseases and growth of chickpea

3710 Effect of endophytic Penicillium and fungicides in suppressing

the root diseases and growth of sunflower

3711 Effect of endophytic Penicillium as soil drench on growth of

okra plants


tomato plants

38 Field Experiments

381 Effect of Pseudomonas monteilii and endophytic Penicillium as

soil drench on growth of okra plants in soil under field condition


soil drench on growth of tomato plants in soil under field condition

4 DISCUSSION

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NEMATODE

SUMMARY

Endophytes are either bacteria or fungi that reside in the tissues of the plant without causing

any apparent symptoms Some endophytic microorganism may promote growth of plants

help in uptake of nutrients and increase the ability to bear environmental stresses like

salinity drought and reduce biotic stresses During our study plants were collected from

different localities in Karachi Pakistan like Memon Goth Kathor Gadap Gharo Malir and

University of Karachi campus from which endophytic Penicillium were isolated Out of the

eighty samples of the plant 14 isolates of endophytic Penicillium isolated (root stem and

leaves) from wild plants (Achyranthus aspera Atriplex stocksii Euphorbia hirta

Chorchorus tridens) and cultivated plant (Solanum melongena Lycopersicon esculentum

Helianthus annuus Azadirachta indica Abelmoschus esculentus Momordica charantia)

Species of Penicillium identified as P asperum P lilacinum P purpurogenum P

nigricans P rugulosum P restrictum P duclauxi P citrinum P thomii P lividum and P

javanicum Identification of selected isolates of Penicillium was also confirmed by using

molecular biology tools

Antimicrobial activity of 14 endophytic isolates of different species of Penicillium

tested against common fungi (root rotting) viz F oxysporum Fusarium solani

Macrophomina phaseolina and Rhizoctonia solani by dual culture plate assay All EP

isolates showed significant result produced by the inhibition zone Nematicidal potential of

cell free culture filtrates of endophytic Penicillium also has shown significant results After

24 hour 50nematicidal potential showed by Ppurpurogenum (EP-3) while after 48 hours

all other isolates showed 100 mortality

Culture filterates of endophytic Penicillium caused growth suppression of bacteria

Salmonella typhimurium Bacillus subtilis Escherichia coli and Staphylococcus aureus As

concentration increased biocontrol potential of culture filterates of EP increased as well

These outcomes show that endophytic Penicillium could be fullfil the need of discovering of

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new antibiotics Culture filtrates of Penicillium also showed activity of fungicidal against

root rotting fungal pathogens Fsolani Rsolani Mphaseolina Rsolani and Foxysporum

by making inhibitory zone Cuture filterates of 60 microldisc showed more effective results than

20 or 40 microldisc Fractionation of cell free culture filtrates of viable isolates of our

Penicillium (EP) was made in solvents ie chloroform and n-hexane and showed strong

antibacterial and antifungal activity against above described pathogens These results

showed that secondry metabolites having compounds with strong antimicrobial potential

Secondary metabolites producing from endophytic Penicillium spp offer an stimulating

area of investigation for the encounter of novel antimicrobial compounds Hexane fraction

of mycelium of promising isolate EP-5 showed prescence of chemicals

In current research antagonistic potential of Penicillium was assessed against

phytopathogens on sunflower (Helianthus annuus) chickpea (Cicer arietinum) tomato

(Lycopersicon escolentum) mungbean (Vigna radiata) and okra (Abelmoschus esculentus)

in field and screen house experiments Inhibitory affects on Foxysporum Rsolani Fsolani

and Mphaseolina showed by many endophytic Penicillium which causes healthy plant

growth by improving plant length fresh shoot weights in both type of experiments (Screen

house and field) In some experiment polyphenol and antioxidant activity also showed

significant result which might be due to resistance produced by endophytes Endophytic

Penicillium treated plants produced fruits which is better in quality as compared to control

Endophytic Penicillium associated with healthy plants is a source of new bioactive

metabolites which could be exploited in plant protection and also in medicine

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1

1 INTRODUCTION

11 Endophytic fungi

Agricultural production passes through heavy loss due to different abiotic and

biotic stresses Most of the economic areas of the world is agriculture it is the most

eager need of the decade to discover and to create the best approach for sustainable

agriculture and development in crop growth (Rai et al 2014) Endophytes are

microorganisms that live inside the plant tissues for atleast in their life cycle that produce

no visuallized symptoms to the host (Bacon and White 2000) Inside the living host plant

tissues an expensive symptomless plant-microbe association build this phenomena called as

Endophytism(Kusari and Spitteler 2012b) During this complex relationship both partners

can be represented as extremely keen mutualism individual benefits depend on both of them

But their relation might be shift toward parasitism or saprophytism or concerning further

dedicated collaboration with time (Millet et al 2010 Zuccaro et al 2011) Recent studies

proposed endophyte-host plant relations are inconstant and showe a relationship between

mutualistic to antagonistic (Saikkonen et al 1998) Mutual relationship between

photosynthetic organisms and fungi earliest and universal (Berbee 2001 Alexopoulos et

al 1996) Evidence showed the presence of microorganism inside the plant tissues from

the the time of the emergence of higher plant on the earth (Redecker et al 2000) Since

the end of 19th century the inoculum of fungi in symptomless plant has recognized

Guerin (1898) Azevedo (1998) and Endophyte word was first suggested in 1866 de

Bary (1866) Endophytes initially defined in Darnel (Lolium temulentum) Freeman

(1904) they isolated it from wide range of plants from arctic to tropics and from

cultivated to wild ecosystems (Arnold 2007) and so far atleast one endophyte have been

found in all living plants species (Dutta et al 2014)

There have been numerous revisions on the relationship of endophyte and plant

particularly for grasses for instance tall fescue where it has been revealed that

endophytic fungus Neotyphodium coenophialum produce toxins that act as defensive

agent against their predators including insects and other grazing animals (Bultman and

Murphy 2000 Bacon et al 1977) it was found that this fungus could be beneficial for

2

enhancing their host tolerance against stresses of abiotic and biotic (Schardl et al 2004

Saikkonen et al1998) In between other symbiotic associations fungal endophytes are

most commonly competitive (Staniek et al 2008) Fungal endophytes are a very varied

polyphyletic group of microorganism that lives inside host stem leaves and also in roots

Endophytes fungi are present above ground parts of plant which make different from

mycorrhizal fungi but also present in roots Fungi related to rhizosphere and roots of the

plants and had positive effect on the growth of plant and recognized as PGPF (Plant

growth promoting fungi) The significant of PGPF belongs the genus Gliocladium and

Trichoderma (Altomare et al 1999) have proficient of inhabiting the plant roots (Gera Hol

and Cook 2005) Endophytes are considered as avirulent opportunistic plant symbionts

and develop systemic resistance in plants just like rhizobacteria (Harman et al 2004)

Similarly endophytic Acremonium lolii and A coenophialum exposed antibiotic formation

against a variety of fungal plant pathogens in culture (White and Cole 1985) Fungus

Muscodor produced volatile compounds which is mostly used as a fumigants in soil (Ezra et

al 2004 Mercier and Manker 2005) In our previous report endophytic Penicillium spp

isolated from Salvadora species showed noteworthy antimicrobial activity (Korejo et al

2014)

Against numerous diseases many endophytes have capability to produce different

secondry metabolites that have therapeutic effect (Kharwar et al 2011 Kusari and

spiteller 2012b)


In recent search for agricultural and pharmaceutical industries to develop a

effective products Natural products have been recognized as a therapuetic agents and play

a important role in nature So the search is carried out for the production of novel

bioactive metabolites from organisms that reside novel biotopes Endophytic fungi

populate such a biotope (Schulz et al 2002) The genus Penicillium is a group of more

than 200 species inhabiting fibre fruits food items soil marine and various species of

plants (Korejo et al 2014 Gong et al 2012) In same way species of Penicillium

deliberated as soil inhabitant and present as a toxicant on foods materials like fibers

starchy materials and fruits but species of Penicillium have been reported in the form of

3

endophytes and play significant role in plants towards tolerance of stress(Khan and Lee

2013 Waller et al 2005) Fungal endophytes is used as a ironic source of secondry

metabolites for agricultural and medicinal practices (Schulz et al 2002) and lot of exposed

(Huang et al 2008)

Endophytic Penicillium species are the producers of diverse variety of secondary

metabolites (Zhang et al 2006 Schulz and Boyle 2005) ie various penicillins PR-

toxin polyketides xanthoviridicatins E and F chrysogine Chrysogenamide A

sorrentanone xanthocillins secalonic acids sorbicillactones A B sorbivinetone

Ochratoxin A (Hoog et al 2000 Singh et al 2003 Gerhard et al 2005 Vega et al

2006 Lin et al 2008) Penicillium species are known to have antifungal algicidal and

antibiotic activities (Meng et al 2011)

13 Role of endophytic Penicillium in growth of plant

Though current studies have revealed that growth enhancement of plant might be

the reason of the production growth promoting secondary metabolites (gibberellins auxin

cytokinin) from plants due to the prescene of endophytic fungi in the rhizospheric region

(Hamayun et al 2010a) Endophyte and plant relationship have the mojor influence on

plant growth promotion (Hassan et al 2013) though endophytic fungi may be responsible

to enhance the growth of the plant in order to secrete different chemical compounds like

ammonia indole acetic acid (IAA) and phytohormone and (Bal et al 2013) Usually

indole acetic acid acts as growth promoter plants by enhancing cell division and cell

elongation and is necessary for differentiation of tissues of plant (Taghavi et al 2009)

Soil microorganisms have a potential to synthesis a wide range of indole acetic acid that

play a role in plant development (Spaepen and Vanderleyden 2011) on other hand

endophytic fungi isolated from different parts of plants which indicated high amount of

indole acetic acid as compared to those isolates isolated from root-free soil (Spaepen et al

2007) The important role of indole acetic acid in growth of the plant in addition to the

potentail of fungal endophytes to secretes indole acetic acid has increased attention due to

their effectiveness on the concentration and supply of indole acetic acid in tissues of the

plants

4

Endophytic fungi have been considered as producers of phytohormones which act

as strong plant growth enhancer These outcomes proposed that endophytic fungi obtained

in the study produced bioactive metabolites which play magnificent roles in stimulating

growth of the plants (Khan et al 2015) Endophytic Penicillium species produced wide

range of Indole acetic acid and gibberellins thus increases plant growth Gong et al

(2014) reported the effect of Penicillium oxalicum on enhancement of growth of maize

plants where they observed that P oxalicum stimulate the growth of maize plants due to its

phosphate-solubilizing ability

14 Role of endophytic Penicillium as resistance inducers in plant stress

Systemic induced resistance have played a vital role in the survival of the plants to

protect themselves in response to pathogenic organisms (Lim et al 2006) It seems in

almost all plants in response pathogenic attack treated with different organic amendments

and chemicals Phytohormones are present extensively in plant parts Plants secrete an

enormous range of chemicals that are toxic to their predators Phenolic compouds are

bioactive chemicals which are common elements of fruits and vegetables act as defensive

agent against insect and grazing animal (Stevenson et al 1993) In the plants growth

phytochemical compounds which have low molecular weight such as phenolic show a

dynamic part and its production and secretion may be due to both biotic and abiotic factors

(Joachim et al 2007) Phytochemicals protect plants towards abiotic and biotic stresses

and therefore are produced against pathogens attack which are exposed to high energy

radicals like the exposure of UV radiation (Briskin 2000) Due to the significant defensive

roles phenolic phytochemicals have pervasive in most of the plants and find specific place

in most of the groups of foods Cherif et al (1991) reported that phenolic compound play

role in resistance of the plants which are accomplished by the rapid accumulation of at the

infection site resulting in the prevention of the pathogen The function of phenolic

compounds in inhibition of the pathogenic infection which act as a barriers to a

pathogens and develop resistance broadly Imporatant groups of compounds termed as

scavengers of oxygen free radical or antioxidants used to resist the phytopathogen and

protection of the oxidative stress of environment (Conceica et al 2006 Wanas 2006)

Numerous studies demonstrate that soil-borne fungal diseases controlled by antioxidants

5

(Dmitriev 2003) with increasing the phytophenolic compounds which increasing plant

growth development and defense against disease Antioxidants used successfully to

control most of the diseases in plant like Fusarium wilt of chickpea plants(Nighat- Sarwar

et al 2005) in tomato (Mohamed et al 2007) pod rot and peanut root (Elwakil 2003

Mahmoud et al 2006) in pepper damping- off (Rajkumar 2008) faba bean of chocolate

spot (Hassan et al 2006) and in the lupine leaf blight and root rot (Abdel-Monaim 2008)

Antioxidants eg salicylic benzoic acids ascorbic propylgalate in cumin in the form of

seed soaking or in other way such as soil drenching showed protection of diseases

occurred by f spcumini and Fusarium oxysporum (Mostasa 2006) The mechanism of

antioxidants was described in many host-pathogen relations such as a wide range of

enzymes like polyphenol oxidase ascorbate oxidase peroxidase and catalase identified

againsts pathogen infection (Clark et al 2002) or outcomes of most of the treatments with

different antioxidants activity ( El-Khallal 2007 and Abdel-Monaim 2008)

In organic agriculture biocontrol agents have different mode of actions including

production of metabolites against pathogens mycoparasitism competing their place and

their nutrients uptake growth promotion of plants and stimulation of defense mechanim in

most of the plants (Chet et al 1997 Howell 2003) This original biological approach

encourages natural resistances of the plants which leads towards systemic resistance

(Vallad and Goodman 2004) instead of apply effects on the most of the plant pathogens

(Walters and Fountaine 2009) Metabolites produced by biocontrol agents against

pathogenic fungus are main factor to discovering them Many researchers are discovering

bioactive chemicals synthesize by microorganism that control most of the diseases of the

plants (Dowling and OrsquoGara 1994) Induction of systemic resistance through biocontrol

agents changed the certain biochemicals of plant which can consider as resistance markers

(Schonbeck et al 1981) including enzymes accumulation like peroxidase (He et al

2002) It was shown that due to systemic acquired resistance in tomato activation of the

defensive mechanism occurs by the insects (Murugan and Dhandapani 2007) viruses

most of the nematodes bacteria and endophytic fungus (Anfoka and Buchenauer 1997

Laporte et al 2007 Molinari 2008 Vasyukova et al 2007Mandal et al 2009 Hase et

al 2008 Park et al 2008) In the same way Shafique et al (2016) studied that combine

use of the oil cake and P lilacinus and PGPR enhance growth of plant that also suppress

6

the infection of root rotting fungi by improving antioxidant activity and polyphenols

contents of the okra plant

Endophytic microorganisms produce secondary metabolites which are crucially

important as parasiticide insect antifeedent and pathogen inhibitors (Meng et al 2011)

Other benefits for host plant include increased resistance to heavy metals salinity and heat

stress improved drought tolerence protected from grazing animals introduced systemic

resistance to pathogens and promoted growth (Redman et al 2001 Clay and Schardl

2002 Marquez et al 2007 Tejasawi et al 2007) Hence Endophytic fungi increase the

ecological survival of plants by increasing resistance towards abiotic and biotic stress

factors (Schulz and Boyle 2005 Gonthier et al 2006) Hossain et al (2014) reported the

part of Penicillium sp in developing systematic resistance to cucumber infection of leaf

caused by anthracnose phytopathogen Colletotricum orbiculare in the cucumber

Similarly Khan et al (2015) studied the effect of P janthenalum in producing tolerance

against aluminum stress in tomato plants Penicillium endophytes are also help plants to

tolerate stress of salinity by regulating plants hormones (Khan et al 2013 Khan et al

2015) Penicillium strains are safe to environment as they reduces the level of salinity and

increase growth of the plants (Leitao and Enguita 2016)

Furthermost fungal endophyte facilitates induction of systemic acquired resistance

in most of the plants (Bailey et al 2006 Nassimi and Taheri 2017) and play a vital role in

safety and control of infection of plants Endophytic fungi play a chief part in growth

promotion of plant higher production of seed and resist plants against several abiotic

biotic stresses and infections Most of them are produce compounds against pathogenic

microbes phytohormones and different bioactive agrochemicals Eco-friendly and

economically active agricultural products are developed by many potential endophytes

(Rai et al 2014) Penicillum chrysogenum produces hypocrellins B and C which have

strong antifungal activity (Meng et al 2011)


Diseases which are caused by organisms persists in soil and debris on soil surface

are known as soil borne diseases and the organisms which causes such diseases are soil-

7

borne pathogens Soil-borne pathogenic fungi reside for several years in soil in the form of

various dormant structures viz chlamydospores melanized hyphae sclerotia and oospores

and are major cause of lowering yield and quality of plant products (Baysal-Gurel et al

2012 Koike et al 2003) Whereas nematodes survive in soil as free organisms cysts or

eggs (Koike et al 2003) Soil borne pathogens infect belowground along with foliar

tissues of plants The well-known diseases produced by soil-borne fungi are the rots which

effect underground tissues of plants and vascular wilts While some soil-borne pathogens

effect the above ground tissues of plants (Koike et al 2003) Soil-borne diseases are more

harmful under poor soil conditions ie inappropriate drainage system low range of

organic matter low level of fertility poor soil structure and high compaction level of the

soil (Abawi and Widmer 2000)


Among the plant disease causing organisms nematodes which parasitized plant

resulted loss upto 100 billion US$ to the agriculture world annualy and approximately 500

million US$ is wasted on control of nematode (Saifullah et al 2007) Whereas the

infection of root rot caused by Rhizoctonia solani Macrophomina phaseolina Fusarium

species Pythium species and Phytophthora species are most common in the crop plants

producing billions $ losses every year

Infections produced by soil borne pathogens includes damping off root rots and

wilts by Fusarium Phythium and Rhizoctonia Phytophthora verticillium and nematodes

species Fusarium oxysporum and its more than 70 species are known to cause root wilt

and root rot diseases in variety of plants species including tomato plants (Kistler 1997)

Species of Cephaliophora Bipolaris Cephalosporium Corynascus Curvularia

Exerohilum Botryodiplodia Fusarium Melanospora Nigrospora Rhizoctonia

MacrophominaSclerotium and Stemphylium are also potent plant pathogens in Pakistan

(Shahzad and Ghaffar 1995) Root knot nematodes are the members of genus Meloidogyne

(Sharon et al 2001 Taylor and Sasser 1978) Globally 26 of crop losses are resulted by

pathogens (Khan et al 2009) Nematodes alone cause 5 of worlds crop losses (Sasser

and Carter 1975) Soil-borne root infecting fungi and nematodes not only produce diseases

8

in plants but also decrease the biomass of plants and severely decrease the yield of crops

and sometimes even death of plant may occur

Nematodes (Meloidogyne spp) parasitized inside specialized type of feeding cells

into the plant tissues directly and remained inside the plant tissueon the otherhand

parasitic type of fungi also penetrate into the tissues of host and absorbs the nutrients Soil

and rhizosphere microorganisms are difficult to control because of tissues around them So

these endo-parasitic nematode and fungi may be able to control by endophytic

microorganisms colonizing around plant root tissue because they occupies same space and

are come in contact with each other (Hallman et al 1997) Hallman and Sikora (1994

1996) demonstrated that endophytic Fusarium oxysporum isolated from tomato roots had

determental effect on Meloidogyne incognita Colonization of tomato roots by the

endophyte resulted in 60 reduction of Mincognita infestation

Charcoal rot disease produced by Macrophomina phaseolina which is soil

inhabiting fungus having diverse type of distribution and have hazardous to the

production of the crops in most of the arid areas over 500 plant species (Ijaz et al 2012)


Biological control is the management of components of ecosystem in order to

protect plants against pathogens It ensures the preservation of environment by no use of

chemicals (Barea and Jaffries 1995) Most of the fungi used as a biocontrol agents and

have long been studied and various reports are available Such as Perveen et al (1994)

reported the effectiveness of Fusarium oxysporum in order to reduce the infection of the

Macrophomina phaseolina Fusarium solani and Rhizoctonia solani Trichoderma species

have been known for so long as biological control agent of soilborne pathogens and also

act as a symbionts of the plants (Harman and Shoresh 2007) Further they suggest that F

oxysporium is a potential biocontrol agent against these pathogens in tomato and okra

Later Siddiqui and Shaukat (2003) tested Pochonia chlamydospora against Fusarium sp

Rsolani and M phaseolina and found it effective against these pathogens Siddiqui et al

(2000) and Waqas et al (2012) investigated the effects of Penicillium and Phoma

glomerata species on the cucumber in drought and saline stress and reported that these

9

endophytic fungal species increases biomass and growth of economically important crops

Major application in agriculture pharmaceutical and commercial utilization of these

endophytic fungi

The current research focused on the isolation and identification of the endophytic

Penicillium species which is associated with plants which are healthy plants and

evaluation of their antagonistic potential against root rotting fungi using sunflower

munbean tomato chickpean and okra as test crops The report also describes the extraction

and characterization of some new compounds from mycelium of Pregulosum

10


21 Collection of plants for isolation of the endophytic Penicillium spp

Survey of various agricultural fields of Kaarchi and its suburb like Karachi

University campus Memon Goth Kathor Gadap Gharo and Malir were carried out

Healthy wild and cultivated plants alongwith roots were selected collected and were

transported to laboratory and preserved at (4oC) untill Penicillium were isolatedround

about (24) hours


1 g of th sample of the plant either stem root or leaves was separately cleaned

sanitized in 1 bleech for (3) min then with (70) alcohol for (3) min and then washed

with the help of distilled H2o Each sample was chopped in sterilized grinder with 50mL

sterilized water and dilutions of each sample were made upto 1104 and further proceed as

described by Korejo et al (2014) and fungal growth fungi were identified with reference

to Barnett and Hunter (1998) Domsch et al (1980) Dugan (2006) Raper and Thom

(1949) and Visagie et al (2014)

221 Molecular strain typing of promising isolates

The selected endophytic Penicillium isolates P rugulosum (EPAAR5) P

decumbens (EPAIR6) P nigricans (EPSLR4) P asperum (EPHAL10) and P

purpurogenum (EPEHS7) initially identified by morphological characters were further

subjected to molecular identification and strain typing bythe PCR (polymerase chain

reaction) based on molecular techniques recently described (Habiba et al 2018)

Briefly five days old strains grown (1 mL) in broth of YPD at 26degC and cells were

harvested by centrifugation (Hanil Korea) for (14000 rpm) for (10 min) at room

temperature Genomic DNA extraction kit (Norgen biotek Canada) was used for fungi as

per vender instruction while quality and purity of the genomic DNA established in

nanodrop (Nano-Drop 200 Thermo Scientific USA) In case of molecular identification t

rDNA-ITS4 ITS1-58S regions amplified with the help of the primers ITS1 (5acute-

11

TCCGTAGGTGAACCTG CGG-3acute) and ITS4 (5acute-TCCTCCGCTTATTGATATGC-3acute) as

initially described Karimi et al (2015) Reactions of the PCR were performed consisting of

genomic DNA (150 ng) primer set (16 μM each) Dream Taq Master Mix (2x Thermo

Scientific USA) and nuclease free water to a final volume of 20 μL Thermal cycling

carried out in a Master cycler (ProS Eppendorf Germany) with an initial denaturation step

(4 min at 94 ordmC) followed by 40 cycles of denaturation (45 s at 94 ordmC) annealing (45 s at 55

ordmC) and extension (1 min at 72 ordmC) and a final extension at 72 ordmC for 7 min

For genetic variation between the strains Random Amplified Polymorphic DNA

(RAPD) PCR was performed with specific oligonucleotide primer M13 (5acute-GAGGGTGG

CGGTTCT-3acute) as described by Zahid et al (2017) Briefly PCR were performed in a total

volume of 20 microL comprising of genomic DNA (25 microL) primer M13 (16 microM) 2x Dream

Taq PCR mix (10 microL) with additional 1 mM MgCl2 and 10 DMSO (Sigma-Aldrich

USA) Thermal cycling was carried out in a Master cycler (ProS Eppendorf Germany) with

an initial denaturation step (5 min at 95 ordmC) followed by 35 cycles of denaturation (30 s at

90 ordmC) annealing (1min at 40 ordmC) and extension (8 min at 65 ordmC) and a final extension at 68

ordmC for 16 min

PCR products (~10 microL) were subjected to 2 agarose gel electrophoresis

containing ethidium bromide (05 μgmL) 1kb DNA ladder (Fermentas USA) was used to

calibrate the sizes

23 Isolation of the soil borne fungi

231 Soil dilution technique for the iolation of Fusarium species

Fusarium were isolated by soil dilution technique (Nash and Snyder 1962) as

described by (Urooj et al 2018) and identified by Nelson et al (1983) and Booth (1971)

12

232 Baiting technique for the isolation of (Rhizoctonia solani)

Rhizoctonia solani were isolated through baiting technique and identified

(Wilhelm 1955) as described in previous report (Urooj et al 2018)

233 Dilution and wet sieving technique for the isolation of (Macrophomina

phaseolina)

Macrophomina phaseolina were isolated by using techniques (wet sieving and

dilution plating)Sheikh and Ghaffar (1975)

24 In vitro determination of antifungal activity of Penicillium species by dual

culture plate assay

For determination of fungicidal potential of Penicillium spp four common fungi

(root rotting) viz Rhizoctonia solani F oxysporum Macrophomina phaseolina and

Fusarium solani were chosen A disc of the 5 mm of the test and fungi (root rotting) was

inoculated on the opposite side of the Petri dish of 90 mm which was poured with CDA

(Czapeks Dox Agar) pH (72) and incubated (28degC) for (5 days) Inhibition zone was

measured in mm (Korejo et al 2014) Experiment were repeated thrice and replicated four

times

25 Inoculation of the nematode (root knot)

Pure culture of the root knot nematode (Meloidogyne javanica) obtained through

egg masses attached on infected brinjal root Roots were washed under tap water was used

to washed te roots thoroughly stereomicroscope was used to collect egg masses and

transferd in cavity blocks having distilled water and left for the hatching (at room

temperature) after 48 hours juveniles were hatched and proceed for the experiment


Culture filtrates of test Penicillium spp were obtained by growing 5 mm disc of

culture in 100 ml of CDB (Czapekrsquos Dox broth) in (250 ml) flask After (15 days) of the

13

incubation (25-30degC) culture filtrate were collected by filteration and 1-2 drop of

chloroform were added to prevent further growth of any contaminant

28 Determination of antifungal activity of culture filtrates of Penicillium species

in vitro

Culture filtrate were loaded at concentration of 20 40 and 60 microl on thick sterile

filter paper discs and dried and placed in clock wise manner according to concentration in

the plates containing Czapekrsquos Dox Agar Disc of test fungus were inoculated in centre of

plates CDB (Czapekrsquos Dox broth) used as a control and 20 microgdisc carbendazim used as a

positive controlAt 30degC Petri dishes left for (5-7 days) and between test fungus and disc

distance was measured as a inhibition zone Qureshi (2003)


To examine the activity of secondary metabolites of Penicillium spp against

bacteria lawn of test bacterium was prepared in 90mm petri dishes containing Nutrient

Agar medium Culture filtrate of each Penicillium sp at 20 40 and 60 microldisc were loaded

on thick sterile filter paper discs and dried and placed in clock wise manner according to

concentration in the plates having bacterial lawn with nutrient Agar A disc of 5 mm of test

fungus was inoculated in the centre of the plate Discs loaded with sterile broth of

Czapekrsquos Dox served as control whereas penicillin 20microgdisc used as positive control for

the gram positive bacteria and streptomycin 20microgdisc used as a positive control for gram

negative bacteria Petri dishes were kept at 30degC for (2-3 days) The inhibition zone were

measured in mm

14

210 In vitro nematicidal activity of culture filtrate of Penicillium species

To examine the nematicidal potential of the culture filtrate 1 ml of culture filtrate

was filled in a cavity blocks containing 15 picked second stage nematode (Meloidogyne

javanica) larvae As a +ve control distilled H2O water was used 2ml The cavity blocks

were kept at room temperature 25-30C and nematode mortality was recorded after 24-48

hours under stereomicroscope


Culture filtrate was extracted three times with n-hexane and chloroform by shaking

vigorously in a separating funnel The extraction volume of each solvent is approximately

half to that of the filtrate Each solvent layer was allowed to separate out and run off from

the aqueous layer The n-hexane and chloroform fractions were collected pooled

concentrated on a rotary evaporator (Eyela-NE) separately and weighed

28 Determination of antifungal activity of frcations of culture filtrates of

Penicillium species in vitro

Each fraction was re-dissolved in their respective solavent and loaded at

concentration of 20 40 and 60 microl on thick sterile filter paper discs and dried and placed in

clock wise manner according to concentration in the plates containing Czapekrsquos Dox Agar

(CDA) Disc of test fungus were inoculated in centre of plates Czapekrsquos Dox broth (CDB)

used as control and carbendazim at 20 microgdisc used as positive control Petri dishes were

left for 5-7 days at 30degC and distance between test fungus and disc was measured as

inhibition zone (Qureshi 2003)

29 In vitro antibacterial activity of the frcations of culture fitrates of the

Penicillium species

In order to examine the prescence of secondary metabolites of the species of

Penicillium against bacteria lawn of test bacterium was prepared in 90mm petri dishes

containing Nutrient Agar medium Filtrates of cell free culture of the species of Penicillium

species at 20 40 and 60 microldisc were loaded on thick sterile filter paper discs and dried

15

and placed in clock wise manner according to concentration in the plates having bacterial

lawn with nutrient Agar 5 mm disc of test fungus was inoculated in centre of plate Discs

loaded with sterile broth of Czapekrsquos Dox (CDB) used as control whereas penicillin

20microgdisc used as positive control for gram positive bacteria and streptomycin 20microgdisc

served as positive control for gram negative bacteria Petri dishes were kept at (30degC) for

(2-3) days The inhibition zone were measured in mm

212 Extraction and compounds from mycelium of endophytic Penicillium

10 gm mycelium was thoroughly washed with n-hexane solvent to remove excess

water and extraction with (200 mL) n-hexane by Soxhlet extractor for (8 h) The fractions

were evaporated at 40degC through a rotary vacuum evaporator

213 Spectroscopy of oily fractions extrcated from mycelium of Penicillium

regulosum

The oily mass extracted from mycelium and culture filtrate of endophytic fungi

were subjected to GC-MS in order to isolate volatile compound GCMS (Gas

chromatographymass spectrometer) analyzed on High Resolution Mass spectrometer Jeol

HX-110 (Japan) eqquiped with data system DA-5500 with gas chromatograph Hewlett

packard (5890)

213 Determination of colony forming unit (cfu) per ml of suspension

Colony forming unit (cfu) per ml of Penicillium suspension were determined by

dilution plate method Fungi grown on the petri plates added then multiplied by the factor

of the dilutions donated by (cfuml) of the fungi

Cfu ml = Number of colonies of bacteria on plate X Dilution factor

16

214 Growth parameters

2141 Physical growth parameter

On harvesting the experiment physical parameters of the plants which was height

weight of the shoot length and weight of the roots number and weight of fruits were

measured

2142 Percent Infection of fungi (root rot) on roots

To determe of the infection of the root rot fungi method reported by Rahman et al

(2016) was used

215 Biochemical parameters


Dried sample of the leaves crushed in ethanol of 96 vv At 3000rpm for 20min

mixture of the sample centrifuged Supernatants used to anlayse antioxidant Salicylic and

polyphenol activity

Folin-Ciocalteu phenol reagent used for total poly phenol content described

(Chandini et al 2008)


Free radical scavenging assay was determined by DPPH (2 2-Di-phenyl-1-

picrylhydrazyl) used for Antioxidant activity (Zubia et al 2007 Duan et al 2006)

2153 Quantification of salicylic acid (SA)

Salicylic quantification was done by using 01 percent prepared Fecl3 (Ferric Chloride)

described by Warrier et al (2013)

216 analysis of Fruits

17

2161 pH (Power of Hydrogen)

To determine the pH fresh sample of five gram fruit in (10ml) of distilled water

were centrifuged for (20 min) in (3000) rpm Supernatent collected to analyse biochemical

activitySample pH measured as described (AOAC 1990)


To analyse moisture content Fresh fruit determine by the method AOAC (1990)

Fruit moisture content can be calculated as follows

Moisture content= Weight of fresh sample ndash Weight of dried sampletimes 100

-------------------------------------------------------

Weight of fresh sample


Sample of 5-ml titrated against (01 N) NOAH solutions by adding 2-3 drops of

phenolphthalein indicator drops for the persistent of the pink coloration The tritable

acidity was calculated by AOAC (1900)

2164 Total soluble solid

A juice drop transferred on prism surface of the hand refractometer (model

ATAGO) and the brix value was recorded by adjusting the eyepiece which showed TSS in

sucrose

2165 Firmness

Tomato fruit firmness recorded by using a TA-XT (Texture Analyser) with 3mm

diameter of the flat aluminium probe

2166 Total solids

It was determined as described by (James 1995) by subtracting percentage

moisture from 100

18

Total solids () = 100 ndash moisture

2167 Protein

Content of protein measured using (Lowry et al 1951) method

2168 Carbohydrate

Method of Phenol-sulphuric acid used to determine the prescence of carbohydrate

of the fruit sample (Dubios et al 1956)

2169 Antioxidant activity and Total polyphenol

To estimate the polyphenol by Folin-Ciocalteu phenol reagent method used

described as (Chandini et al 2008) To determine the antioxidant activity of fruits

samples used by method described by (Zubia et al 2007 Duan et al 2006)


Complete randomized design or randomized complete block design used as a

ststistical tool in screen house and field conditions experiments


(ANOVA) Analysis of variance included least significant difference (LSD) were

analyse according to experimental design described as Gomez and Gomez (1984) were

used

19



Out of 80 plant samples from both wild and cultivated species (Roots stems and

leaves) 14 samples showed presence of genus Penicillium Endophytic Penicillium spp

isolated (root stem and leaves) from wild plants (Achyranthus aspera Atriplex stocksii

Euphorbia hirta Chorchorus tridens) and cultivated plant (Solanum melongena

Lycopersicon esculentum Helianthus annuus Azadirachta indica Abelmoschus

esculentus Momordica charantia) Fourteen isolates of Penicillium were isolated and

identified on the bases of their morphological feature Species of Penicillium were

identified as P lividum P lilacinum P purpurogenum P nigricans P rugulosum P

restrictum P duclauxi P asperum P thomii P citrinum and P javanicum (Table 1)

32 Molecular Identification of endophytic Penicillium




subjected to molecular identification and strain typing (Habiba et al 2018) PCR

amplification of DNA from endophytic Penicillium strains using a universal genus specific

primer set (ie ITS1 and ITS4) which amplified the product size ranging between 500 to 600

bp for different fungal species while 600bp specific for Penicillium spp All products thus

showing the availability and consistency in size of typical 600bp for Penicillium isolates

(Figure 1A) RAPD-PCR was also performed to established the genotypic variations and

similarities with in the genus Penicillium (Figure 1B) RAPD-PCR is universally used and

based on polymorphism of DNA at the taxonomic level clearly illustrates the discrimination

power at the specie level Moreover the dendrogram of RAPD-PCR analysis revealed the

genetic relatedness between the isolates (Figure 1C) Dendogram represents two distinct

clades in first isolate P rugulosum EPAAR5 and P purpurogenum EPEHS7 were found to

share the same clade (a) whereas P asperum EPHAL10 P nigricans EPSLR4 P

decumbens EPAIR6 and positive control exist together in the second clade (b)

20

21

22

32 In dual culture plate assay antifungal activity of endophytic Penicillium

Fungicidal potential of endophytic species of Penicillium isolates were

examined usually phytopathogens such as Rhizoctonia solani Macrophomina

phaseolina F oxysporum and Fusarium solani using dual culture plate assay The 5mm

diam agar disc of endophytic Penicillium was placed on a 90mm Petri dish poured

with (CDA) Czapekrsquos Dox Agar pH (72) On opposite side of this disc from root

rotting fungi grown in plate a 5mm disc of was cut placed and leave at 28oC and

inhibition zone measured averaged and expressed in mm

All endophytic Penicillium showed best result against common root rot fungi

Maximum inhibition zone (25mm) against Fsolani produced by Ppurpurogenum

then Pdecumbens and P nigricans inhibition zone produced against Rsolani

(Table 1) fig1-7

23

Table 1 Suppression of Macrophomina phaseolina Rhizoctonia solani Fusarium solani and F oxysporum in dual culture plate assay

by the endophytic Penicillium species isolated from different wild and cultivated plants

Fungus Penicillium spp Host name Plant

part MPhaseolina Rsolani Fsolani Foxysporum

Zone of inhibition(mm)

EPSMR1 P citrinum Solanum melongena L

(Solanaceae)

Root 4 4 20 20

EPSMS2 P lilacinum Solanum melongena L (Solanaceae) Stem 6 8 11 14

EPSML3 Ppurpurogenum Solanum melongena L (Solanaceae) leaf 6 5 25 17

EPSLR4 P nigricans Lycopersicon esculentum L

(Solanaceae)

root 5 25 16 21

EPAAR5 P rugulosum Achyranthus aspera L

(Amaranthaceae)

root 3 12 11 20

EPAIR6 P decumbens Azadirachta indica AJuss

(Meliaceae)

root 5 25 13 20

EPEHS7 P purpurogenum Euhorbia hirta L (Euphorbiaceae) stem 6 5 25 17

EPCTS8 P restrictum Chorchorus tridens L (Malvaceae) stem 2 2 5 5

EPASS9 Pduclauxi Atriplex stocksii

(Amaranthaceae)

stem 18 13 11 14

EPHAL10 Pasperum Helianthus annuus L (Asteraceae) leaf 2 2 5 5

EPAER11 P thomii Abelmoschus esculentus L

(Malvaceae)

root 5 8 5 6

EPMCL12 Plividum Momordica charantia L

(Cucurbitaceae)

leaf 18 13 11 14

EPSLR13 Pjavanicum Lycopersicon esculentum L

(Solanaceae)

root 5 24 17 22

EPAER14 Ppurpurogenum Abelmoschus esculentus L

(Malvaceae)

root 5 3 21 12

24

Fig1 Growth inhibition of Foxyspoum by the endophytic Penicillium in dual culture plate

assay

Fig2 Growth inhibition of Fsolani by the endophytic Penicillium in dual culture plate

assay

25


assay

Fig4 Growth inhibition of F solani by the endophytic Penicillium

in dual culture plate assay

26


assay


assay

27


assay

33 In vitro fungicidal potential of culture filtrates of endophytic Penicillium

Penicillium isolates were grown in Czapekrsquos Dox broth pH 72 at 25-30oC for 15

days and through filteration culture filtrate was collected in autoclaved flasks The filtrate of

culture was dropped by chloroform under sterilize conndition to kill fungal propagoles if

any To determine the antifungal activity Disc Diffusion Method was used in which cell free

culture filterates at 20microldisc 40microldisc 60microldisc and control were placed at equal distance

at diferent positions in the petri plates poured with Czapeks Dox Agar pH 72 Water

impregnated disc were used as negative control and carbendazim 20microgdisc were used as

positive control against four root rot fungi viz Rhizoctonia solani Macrophomina

phaseolina F oxysporum and Fusarium solani 5mm disc of each root rot pathogen

Fusarium solani Macrophomina phaseolina F oxysporum and Rhizoctonia solani was

inoculated in the centre of the petri plates were kept 28oC for 5 days Distance between

paper disc and fungal colonies was measured as inhibition zone which were averaged and

showed in mmThe experiment was performed twice and replicated four times

28

Culture filtrate of Penicillium initiated growth suppression of (root rotting) fungi viz R

solani M phaseolina F oxysporum and F solani in vitro M phaseolina was inhibited by

culture filtrates of Plilacinum Pnigricans and Pthomii at 60microldisc by producing

maximum zone of 20mm Plilacinum Pnigricans and Pthomii also showed zone of

inhibition of 15mm at 20microldisc and 17mm at 40microldisc R solani was inhibited by

producing zone of 14mm at 60microldisc from culture filtrates of Plilacinum Ppurpurogenum

(EPSML3) Ppurpurogenum (EPEHS7) Pasperum and Ppurpurogenum (EPAER14)

Pnigricans and Pthomii produced zone of inhibition of 17mm at 60microldisc against F

solani P decumbens P citrinum Ppurpurogenum (EPSML3) EPSLR4 Pregulosum

Ppurpurogenum (EPEHS7) Pduclauxi Pasperum Pthomii Pjavanicum and

Ppurpurogenum (EPAER14) produced zone of inhibition ranging from 12-14mm at

60microldisc(Table 2)

29

Table 2 In vitro growth inhibition of Macrophomina phaseolina Rhizoctonia solani Fusarium solani and Foxysporum by culture

filtrates of endophytic Penicillium species isolated from wild and cultivated plant species

Fungus No Penicillium spp MPhaseolina Rsolani Fsolani Foxysporum


Control 0 0 0 0

+ve Control (Carbendazim 20microgdisc) 8 5 9 7

EPSMR1 P citrinum

20microldisc 8 8 8 10



EPSMS2 Plilacinum




EPSML3 Ppurpurogenum




EPSLR4 P nigricans



30




EPAAR5 P rugulosum




EPAIR6 P decumbens




EPEHS7 Ppurpurogenum




EPCTS8 Prestrictum




EPASS9 P duclauxi


31





EPHAL10 Pasperum




EPAER11 Pthomii




EPMCL12 P lividum




EPSLR13 P javanicum




EPAER14 P purpurogenum

32






33

34 In vitro antibacterial potentail of culture filtrates of endophytic Penicillium

Bacterial lawn of test bacteria was prepared in 90mm Petri dished conating Nutrient

agar and loaded disc of culture filterates at 20microldisc 40microldisc 60microldisc and control were

placed at equal distance in clockwise pattern in according to concentration Water

impregnated disc were used as negative control and Streptomycin 10microgdisc applied as +ve

control for gram +ve bacteria viz Salmonella typhimurium and Escherichia coli and

Penicillin applied as +ve control for gram positive bacteria viz Bacillus subtilus and

Staphlococcus aureus Zones of inhibition produced around the discs after 2-3 days growth

were recorded averaged and showed in millimeter (mm) The performance was conducted

twice and replicated four times

Fourteen isolates of Penicillium species were tested in vitro against four bacterial

species Bacillus subtilus and Staphlococcus aureus (Gram positive) and Salmonella

typhimurium and Escherichia coli (Gram negative)Cell free filtrate of culture of the

Penicillium resulted growth suppression of four bacteria Bsubtilus Saureus S

typhimurium and E coli in vitro Penicillium rugulosum was found to inhibit by Bsubtilus

by producing maximum zone of 9mm at 20microldisc 13mm at 40microldisc and 21mm at

60microldisc P rugulosum was found to inhibit by Saureus by producing maximum zone of

24mm at 20microldisc 30mm at 40microldisc and 30mm at 60microldisc P rugulosum was found to

inhibit S typhimurium by producing maximum zone of 12mm at 20microldisc 20mm at

40microldisc and 20mm at 60microldisc P rugulosum was found to inhibit E coli by producing

maximum zone of 18mm at 20microldisc 22mm at 40microldisc and 22mm at 60microldisc Bsubtilus

was inhibited by P lividum and Plilacinum by producing 16mm and 10mm zone at 20 40

and 60microldisc respectively Saureus was inhibited by P lividum and Plilacinum by

producing zone of inhibition of 18mm at 40 and 60microldisc and 20mm at 60microldisc

respectively E coli was found to inhibit by P decumbens by producing zone of 18mm at all

concentration (Table 3 and Fig 8)

34

Table3 In vitro growth suppression of Bsubtilus Saureus S typhimurium and E coli by culture filtrates of endophytic Penicillium

species

Fungus No Penicillium sp Bsubtilus Saureus S typhimurium E coli

Zone of inhibition mm

Control 0 0 0 0

Streptomycin 20 microgdisc 15 15 15 15

EPSMR1 P citrinum


40 microldisc 6 8 8 6


EPSMS2 Plilacinum








EPSLR4 P nigricans


35





EPAAR5 P rugulosum




EPAIR6 P decumbens








EPCTS8 P restrictum




EPASS9 P duclauxi

36






EPHAL10 Pasperum




EPAER11 Pthomii




EPMCL12 P lividum




EPSLR13 P javanicum




37







38

Fig 8 Growth inhibition of Saureus by the culture filterate of endophytic Penicillium in

disc diffusion method

A=Control B=+ve control C=20microldisc D=40microldisc E=60microldisc

35 In vitro nematicidal potentail of culture filtrates of endophytic Penicillium

spp

Penicillium isolates were grown in CDB (Czapekrsquos Dox broth) pH (72) at (25-

30oC) for 15 days and filtered and culture filtrate was collected in sterile flasks for use

Suspension of 10 juveniles per ml and culture filtrate (1 ml) of Penicillium isolates

shifted in cavity blocks and placed at 26 plusmn5oC These were replicated three times and

mortality rate of juvenile was noticed subsequently 24 and 48 hours

Culture filtrates of endophytic Penicillium exhibited nematicidal effects juveniles

mortality of Meloidogyne javanica occurred at different percentages Out of 14 isolates

tested Ppurpurogenum (EPSML3) initiated 100 killing of juveniles of M javanica in

24 h While 10 isolates initiated 50 or more juveniles mortality in 48 hours (Table 4)

A

B

C

E D

39

Table4 Effect of cell free culture filtrate of endophytic Penicillium spp on juveniles mortality of Meloidogyne javanica after 24 and

48 hours

Treatments Code Juveniles Mortality

24Hours 48Hours

Control(CDA Broth) hellip 0 0

P decumbens EPAIR6 50 76

Pnigricans EPSLR4 10 33

Pregulosum EPAAR5 46 63

P citrinum EPSMR1 36 73

Plilacinum EPSMS2 36 83

Ppurpurogenum EPSML3 100 100

Pduclauxi EPASS9 10 76

Plividum EPMCL12 16 53

Ppurpurogenum EPEHS7 43 76

Prestrictum EPCTS8 76 83

Pthomii EPAER11 43 43

Ppurpurogenum EPAER14 43 76

Pjavanicum EPSLR13 10 33

Pasperum EPHAL10 30 70

40

41

36 In-vitro antimicrobial potentail of solvent fractions of culture filtrtaes of

endophytic Penicillium

In our present study filtrates of culture of each fungus extracted thrice with n-

hexane and then chloroform by shaking vigorously in a separating funnel The extraction

volume of each solvent is approximately half to that of filtrate The n-hexane and

chloroform fractions were collected pooled and finally crude extracts on a rotary vacum

evaporator (Eyela-NE) separately and weighed The dilutions of 15mgml of n-hexane and

chloroform were dissolved in their respective solvents and weighed down on senitized

discs at 20 40 and 60microldisc and dried These are used for antimicrobial test by Disc

Diffusion Method as described for cell free culture filtarates section (Hadacek and Greger

2000) Solvent of respective fractions were served as control streptomycin at 20microgdisc

was used as positive control in determining antibacterial activity against Salmonella

typhimurium Escherichia coli Bacillus subtilus Staphlococcus aureus and Pseudomonas

auroginosa Whereas in antifungal activity carbendazim at 20microgdisc used as positive

control against root rotting fungi Mphaseolina Foxysporum Fsolani and Rsolani

There were four replicates of each treatment

361 In-vitro fungicidal potentail of n-hexane fractions

P rugulosum and Ppurpurogenum (EPEHS7) produced inhibition zones of 20mm

against Mphaseolina whereas P decumbens produced maximum inhibition zones of

25mm against Foxysporum and Fsolani was also inhibited P rugulosum

Ppurpurogenum (EPEHS7) and P nigricans Highest zone of inhibition of 25mm at

60microldisc were produced by P rugulosum against Rsolani (Table 5)

42

Table5 In vitro growth inhibition of M Phaseolina R Solani F solani and F oxysporum by n-Hexane fraction of endophytic

Penicillium species

Fungus No Penicillium sp M phaseolina R solani F solani F oxysporum


Control 0 0 0 0

Carbendazim 20 microgdisc 30 30 30 30

EPSLR4 P nigricans




EPAAR5 P rugulosum




EPAIR6 P decumbens






43


60 microldisc 20 `20 20 0

EPHAL10 Pasperum




44

362 In-vitro antibacterial potentail of n-hexane fractions of culture filtrates of


Pasperum and P rugulosum inhibited Bacillus subtilus by producing inhibition

zones ranging from 12-14mm respectively P rugulosum suppressed the growth of

Staphlococcus aureus by producing inhibition zone 24mm at 60microldisc while P

rugulosum also formed inhibition zones measuring 18mm against Escherichia coli whereas

the inhibition zones of 20mm against Salmonella typhimurium were produced by P

rugulosum Similarly P rugulosum inhibited Pseudomonas auroginosa with zones of

25mm (Table 6 and Fig9-12)

363 In-vitro fungicidal potentail of chloroform fractions of culture filtrates of


P rugulosum produced inhibition zones of 20mm 25mm 20mm and 15mm at

60microldisc against Fsolani Rsolani Mphaseolina Rsolani and Foxysporum (Table 7)

45

Table6 In vitro growth inhibition of Bsubtilus Saureus S typhimurium E coli and Pauroginosa by n-hexane fraction of

endophytic Penicillium species

Penicillium sp Bsubtilus Saureus S typhimurium E coli Pauroginosa


Control 0 0 0 0 0

Streptomycin 20 microgdisc 15 15 15 15 15

EPSLR4 P nigricans

20microldisc 6 10 8 8 8

40 microldisc 9 10 8 8 9

60 microldisc 11 11 9 12 10

EPAAR5 P rugulosum

20microldisc 0 18 18 11 18

40 microldisc 0 21 18 11 22

60 microldisc 0 24 20 18 22

EPAIR6 P decumbens



60 microldisc 0 12 16 0 11




46


EPHAL10 Pasperum


40 microldisc 11 9 6 10 10

60 microldisc 12 11 9 10 12

47

Fig9 Growth inhibition of Pauroginosa by the n-hexane fraction endophytic Penicillium in


Fig10 Growth inhibition of Saureus by the n-Hexane fraction of endophytic Penicillium in


C

+ve C

20microl

60microl

40microl

+veC

20microl

40microl

60microl

C

48

Fig11 Growth inhibition of S typhimurium by the n-Hexane fraction of endophytic

Penicillium in disc diffusion method

Fig12 Growth inhibition of E coli by the n-Hexane fraction of endophytic Penicillium in


C

60microl

40microl

20microl +veC

vCCe

veve

+veC

vCCe

veve

C

60microl

20microl

40microl

49

Table7 In vitro growth suppression of M Phaseolina R Solani F solani and F oxysporum by chloroform fraction of endophytic

Penicillium species

Fungus No Penicillium sp M Phaseolina R Solani F solani F oxysporum


Control 0 0 0 0


EPSLR4 P nigricans




EPAAR5 P rugulosum




EPAIR6 P decumbens







50


EPHAL10 Pasperum




364 In-vitro antibacterial potentail of chloroform fractions of culture filtrates of endophytic Penicillium

P rugulosum inhibited Bacillus subtilus Staphlococcus aureus Salmonella typhimurium and Pseudomonas auroginosa by

producing inhibition zones ranging from 21-18mm P rugulosum while P rugulosum also produced inhibition zones measuring

11mm against Escherichia coli whereas the inhibition zones of 14mm against Escherichia coli were produced by P nigricans

(Table 8 and Fig12)

51

Table8 In vitro growth inhibition of Bsubtilus Saureus S typhimurium E coli and Pauroginosa by chloroform fraction of


Fungus No Penicillium sp Bsubtilus Saureus S typhimurium E coli Pauroginosa


Control 0 0 0 0 0


EPSLR4 P nigricans

20microldisc 16 16 14 14 16

40 microldisc 16 16 14 14 18

60 microldisc 18 16 16 14 20

EPAAR5 P rugulosum

20microldisc 18 18 20 11 20

40 microldisc 18 18 20 11 21

60 microldisc 18 18 20 11 21

EPAIR6 P decumbens






52



EPHAL10 Pasperum




53

4

Fig13 Growth inhibition of S typhimurium by the chloroform fraction of endophytic


C

+ve C

20microl 40microl

60microl

54

3656 Extraction and characterization of compounds from mycelium of endophytic

Penicillium

Czapekrsquos Dox broth of Penicillium regulosum was prepared in (250 ml) conical

flask containing (100 ml) A 5mm disc of test Penicillium was cuttedinoculated and

incubated (25-30degC) and left for 15 days When fungi secreted secondry metabolites then

cell free culture filtrates were obtained by filtering The mycelium was used for the

extraction of compounds


water and extracted with 200 mL n-hexane using a Soxhlet extractor for 8 h The extracts

were filtered and dried at 40degC by using a rotary vacuum evaporator The oily mass

extracted from mycelium of Penicillium regulosum was subjected to GC-MS analysis

GCMS (Gas chromatographymass spectrometer) analyzed on High Resolution Mass

spectrometer Jeol HX-110 (Japan) equipped with data system DA-5500 in combination with

gas chromatograph Hewlett packard (5890)

Total 23 different chemical compounds were obtained from mycelium fraction Volatile

compound such as normal hydrocarbon (akane and alkene) fatty acid alcohol ether

terpenoids and benzene derivatives including cyclohexane and other compounds that were

found among the volatile metabolites were identified by mass spectral data base (Table 9)

55

(1) Nanodecane

(2) Nonadecane

(3) Heptadecane

(4) Heptacosane

(5) Heptacosane

(6) Eicosane

(7) Octadecane

(replib) Nonadecane

50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 2400

50

10057

71

85

99113 127 141 155 169 183 197

(replib) Nonadecane

60 80 100 120 140 160 180 200 220 240 260 2800

50

10057

71

85

99113 127 141 155 169 183 197 268

(replib) Heptadecane

50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 2500

50

10057

71

85

99113 127 141 155 169 182 196 210 240

(replib) Heptacosane

60 80 100 120 140 160 180 200 220 240 260 280 300 320 3400

50

10057

71

85

99113 127 141 155 169 183 197 211 225 239 253 267 281 294 308 322 336


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10057

71

85

99113 127 141 155 169 183 197 211 225 239 253 267 281 294 308 322

(mainlib) Eicosane

60 80 100 120 140 160 180 200 220 240 260 2800

50

10057

71

85

99113

127 141 155 169 183 197 211 225 238 252 282

(replib) Octadecane

60 80 100 120 140 160 180 200 220 240 2600

50

10057

71

85

99113 127 141 155 169 183 197 210 225 254

56

(8) Tetradecanoic acid

(9) Dodecane 2610-trimethyl-

(10) i-Propyl tetradecanoate

(11) i-Propyl 12-methyltetradecanoate

(12) Ethyl 13-methyl-tetradecanoate

(13) Widdrol hydroxyether

(mainlib) Tetradecanoic acid

50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 2400

50

100

55

60

69

73

8397 115

129

138

143157

171

185

199209

228

OH

O

(replib) Dodecane 2610-trimethyl-

60 80 100 120 140 160 180 200 220 240 2600

50

10057

71

85

97

113127

141 155 168183 197 212

(mainlib) i-Propyl tetradecanoate

50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 2500

50

100

60

71

8397

102

111

129

143157

171

185

199

211

228

O

O

(mainlib) i-Propyl 12-methyltetradecanoate

60 80 100 120 140 160 180 200 220 240 2600

50

10057

60

71

83 97

102

111 129

143153 165

177

185

195 213225

242O

O

(mainlib) Ethyl 13-methyl-tetradecanoate

60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

61

70

88

101

115129

143

157

171 185 199 213

227

241 255

270

O

O

(mainlib) Widdrol hydroxyether

60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

69

81

95 109

123

135

140

150

167

177 205223

238

O

OH

57

(14) n-Hexadecanoic acid

(15) Hexadecanoic acid ethyl ester

(16) Oleic Acid

(17) 912-Octadecadienoic acid ethyl ester

(replib) n-Hexadecanoic acid

60 80 100 120 140 160 180 200 220 240 2600

50

10060 73

8397

115

129

143157 171 185

199

213

227 239

256

OH

O

(mainlib) Hexadecanoic acid ethyl ester

60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

61 73

88

101

115129 143

157

171 185 199 213 225239

255 267284

O

O

(mainlib) 912-Octadecadienoic acid ethyl ester

60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

100

55

6781

95

109

123135 150 164 178

192 205 220 234

263

279

308

O

O

(replib) Oleic Acid

60 80 100 120 140 160 180 200 220 240 260 2800

50

10055

69

83

97

111

125137 151 165 180 193 207 222 236

264

282

HO

O

58

(18) Ethyl Oleate

(19) cis-10-Nonadecenoic acid

(20) 2-Propenoic acid 3-(4-methoxyphenyl)- 2-ethylhexyl ester

(21) 12-Benzenedicarboxylic acid diisooctyl ester

(replib) Ethyl Oleate

60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10055

6983

97

111123

137 155180

194 207

222

236

264

281

310

O

O

(mainlib) cis-10-Nonadecenoic acid

60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10055

6983

97

111

125137 151 165 179 194 207 221 236 249 261

278296

HO

O

(mainlib) 2-Propenoic acid 3-(4-methoxyphenyl)- 2-ethylhexyl ester

60 80 100 120 140 160 180 200 220 240 260 280 3000

50

100

55 77 90 103118

133

147

161

178

191 262290

O

O

O

(replib) 12-Benzenedicarboxylic acid diisooctyl ester

60 90 120 150 180 210 240 270 300 330 360 3900

50

100

5770

83 104132

149

167

279

O

O

O

O

(mainlib) Cyclopenta[ad]cycloocten-5-one 1233a456899a1010a-dodecahydro-7-(1-methylethyl)-19a-dimethyl-4-methylene

60 90 120 150 180 210 240 270 300 330 360 3900

50

100

55

69

81

95

107

121

147

173189

215

231

243

258

286

O

59

(22) Cyclopenta[ad]cycloocten-5-one 1233a456899a1010a-dodecahydro-7-(1-

methylethyl)-19a-dimethyl-4-methylene

(23) 2-Aminofluorescein

(mainlib) 2-Aminofluorescein

50 100 150 200 250 300 350 400 450 500 550 600 6500

50

100

63 91

151

189

287

303

318 347

O

O

OHO OH

H2N

60

Table9 GCMS of mycelial fraction of Penicillium regulosum

SNo Scan

No

Systemic Name

(Common Name)

Mol

Formula

Mol

Wt

Ret

Time

Conc

1 2606 Nanodecane C19H40 268 24168 0036

2 2913 Heptadecane C17H36 240 2641 0035

3 2998 Tetradecanoic acid C14H28O2 228 27038 0056

4 3230 Octadecane C18H38 254 28737 0049

5 3264 Dodecane 2610-trimethyl- C15H32 212 28986 0077

6 3331 i-Propyl tetradecanoate C17H34O2 270 29476 0058

7 3381 i-Propyl 12-methyltetradecanoate C18H36O2 284 29842 0097

8 3496 Ethyl 13-methyl-tetradecanoate C17H34O2 270 30684 0054

9 3653 Nonadecane C19H40 268 31834 0064

10 3975 Widdrol hydroxyether C15H26O2 238 34192 0094

11 4096 n-Hexadecanoic acid C16H32O2 256 35078 0079

12 4223 Hexadecanoic acid ethyl ester C18H36O2 284 36007 0094

13 4252 Eicosane C20H42 282 36220 0093

14 5475 Oleic Acid C18H34O2 282 45175 0105

15 5516 912-Octadecadienoic acid ethyl ester C20H36O2 308 45475 0084

16 5546 Ethyl Oleate C20H38O2 310 45694 0065

61

17 5970 cis-10-Nonadecenoic acid C19H36O2 296 48799 0053

18 6023 Heptacosane C27H56 380 49187 0051

19 6072 2-Propenoic acid 3-(4-methoxyphenyl)- 2-ethylhexyl ester C18H26O3 290 49546 0058

20 6281 Heptacosane C27H56 380 51076 0044

21 6591 12-Benzenedicarboxylic acid diisooctyl ester C24H38O4 390 53346 0048

22 6668 Cyclopenta[ad]cycloocten-5-one 1233a456899a1010a-

dodecahydro-7-(1-methylethyl)-19a-dimethyl-4-methylene

C20H30O 286 53910 004

23 8458 2-Aminofluorescein C20H13NO5 347 67016 0135

62


371 Effect of endophytic Penicillium in soil amended with neem cake in inhibition

of the root diseases and growth of sunflower (2016)

Fourteen isolates of endophytic Penicillium viz P duclauxi Plilacinum

Ppurpurogenum (EPSML3) Pnigricans Pregulosum P decumbens Ppurpurogenum

(EPEHS7) P restrictum P citrinum Pasperum Pthomii Ppurpurogenum (EPAER14)

Plividum Pjavanicum and caused growth suppression of four root rotting fungi in vitro A

25ml five-day-old cell suspension of fungal isolates were drench in 1kg soil obtaining from

experimental field of the Department of Botany each clay pots Carbendazim considered as

+ve control against pathogenic fungi Application of endophytic Penicillium and 1 Neem

cake were also applied in another pot set In each pot (6 seeds per pot) seed of sunflower

(Helianthus annuus) were sown and kept four seedlings after germination Treatments were

replicated four times watered daily

After six weeks experiment were harvested to evaluate the potentail of endophytic

Penicillium on the suppression of pathogens and growth of plant and data on height of

plant weight of fresh shoot length of root weight of root were measured and noted The

infection of root rotting fungi roots cleaned with tap water 5 root pieces of 1cm were

sterilized with 1 bleach and placed on plates poured with (Potato Dextrose Agar) PDA

mixed with penicillin (100000 units litre) and streptomycin (02 glitre) After incubation

of 5 day occurrence of root rots were recorded

Plant grown in soil amended with neem cake generally showed less infection of

root rotting fungi related to plant grown in natural soil (un-amended soil) Plant inoculated

with endophytic Penicillium species most of them showed less infection of root rotting

fungi related to control plant Plants grown in pots received Endophytic Pregulosum in

natural soil and also in amended soil with neem cake showed no infection of F oxysporum

Whereas P Pnigricans Pregulosum P citrinum Ppurpurogenum (EPSML3)

Pduclauxi Pthomii Pjavanicum and P decumbens in amended soil with neem cake also

showed no infection of F oxysporum Combine effect of isolates P decumbens

63

Pnigricans P citrinum P lividum Plilacinum Ppurpurogenum (EPSML3) Pduclauxi

Ppurpurogenum (EPEHS7) P restrictum Pthomii Ppurpurogenum (EPAER14)

Pjavanicum and neem cake showed no infection on Fsolani P decumbens Pnigricans

Pregulosum and Pjavanicum also showed no infection of Fsolani when used alone

Plividum alone showed no infection of Mphaseolina on sunflower roots Combine effect

of P decumbens Pnigricans Pregulosum Pthomii and Pjavanicum with neem cake

showed significant reduction on infection of Mphaseolina Application of P decumbens

Pnigricans P citrinum Plividum Ppurpurogenum (EPEHS7) Ppurpurogenum

(EPAER14) and Pjavanicum showed no infection of Rsolani P decumbens

Pregulosum P citrinum Plilacinum Ppurpurogenum (EPSML3) Pduclauxi

Ppurpurogenum (EPEHS7) P restrictum Ppurpurogenum (EPAER14) Pjavanicum

with neem cake showed no infection of Rsolani While Pnigricans Plividum Pthomii

and Pasperum Significantly suppressed the Rsolani infection when applied in neem cake

amended soil (Table 10)

Greater plant height was produced by Ppurpurogenum (EPEHS7) P restrictum

Ppurpurogenum (EPAER14) and Pasperum when applied in neem cake amended soil

However effect of P restrictum and Pasperum with neem cake were significant on fresh

shoot weight (Table 10) Pnigricans Pthomii and Pjavanicum alone showed significant

result of root length and root weight whereas P decumbens and Pduclauxi with neem

cake showed greater root length (Table 11 and Fig13-14)

64

Table10 Effect of endophytic Penicillium and neem cake on the infection of Fusarium solani Foxysporum Rhizoctonia solani and

Macrophomina phaseolina on sunflower roots in green house experiment

Infection

Treatments Code Foxysporum

Fsolani

M phaseolina Rsolani

NS AS NS AS NS AS NS AS

Control hellip 50 187 75 25 75 50 187 125

Carbendazim hellip 25 0 312 62 125 25 125 0

P decumbens EPAIR6 187 0 0 0 25 187 0 0

Pnigricans EPSLR4 62 0 0 0 375 187 0 62

Pregulosum EPAAR5 0 0 0 187 62 187 62 0

P citrinum EPSMR1 375 0 25 0 125 25 0 0

Plilacinum EPSMS2 25 62 187 0 62 50 62 0

Ppurpurogenum EPSML3 50 0 125 0 62 25 62 0

Pduclauxi EPASS9 50 0 62 0 312 312 62 0

Plividum EPMCL12 50 62 50 0 0 50 0 62

Ppurpurogenum EPEHS7 375 187 375 0 50 312 0 0

Prestrictum EPCTS8 50 62 62 0 125 437 62 0

Pthomii EPAER11 62 0 62 0 375 187 62 62

Ppurpurogenum EPAER14 375 187 375 0 50 312 0 0

Pjavanicum EPSLR13 62 0 0 0 375 187 0 0

Pasperum EPHAL10 125 0 25 187 375 312 62 62

LSD005 Treatment=4651 Pathogen=2322 Soil Type=1643

1 Difference greater than LSD values among means in column are significant at plt005

2 Difference greater than LSD values among means in row are significant at plt005

3Mean values in the NS and AS column showing difference greater than LSD value are significantly different at plt005

NS= Natural Soil AS=Amended Soil

65

Table11 Effect of endophytic Penicillium and neem cake on the growth of sunflower in green house experiment

Treatments Code Shoot Length

Shoot Weight

Root Length Root weight

(cm)

(g)

(cm)

(g)


Control 22775 3993 253 535 643 1162 0645 0675

Carbendazim 2585 418 2216 451 742 1287 0715 0622




P citrinum EPSMR1 2599 4681 218 51 94 862 0726 0807



Pduclauxi EPASS9 2541 4487 243 512 1103 1406 077 0786

Plividum EPMCL12 22685 4587 205 539 631 558 0663 0578



Pthomii EPAER11 2824 44 277 527 1221 1218 1005 0645



Pasperum EPHAL10 26186 4918 214 678 918 757 069 0866

LSD005 5141 7881 07911 1821 2551 2821 01951 031



66

372 Effect of endophytic Penicillium with neem cake in inhibition of the root

diseases and growth of Sunflower (2017)

Fourteen isolates of endophytic Penicillium viz P citrinum Plilacinum


(EPEHS7) P restrictum Pduclauxi Pasperum Pthomii Plividum Pjavanicum and

Ppurpurogenum (EPAER14) caused growth suppression of four root rotting fungi in vitro

A 25ml five-day-old cell suspension of fungal isolates were drench in 1kg soil obtaining

from experimental field of the Department of Botany each clay pots Carbendazim

considered as positive control against root rotting fungi Application of endophytic

Penicillium and 1 Neem cake were also applied in another pot set In each pot (6 seeds per

pot) seed of sunflower (Helianthus annuus) were sown and kept four seedlings after

germination Treatments were replicated four times watered daily


Penicillium on the suppression of pathogens and growth of plant and data on plant height

fresh shoot weight root length root weight were measured and noted The infection of

root rotting fungi roots were washed under tap water 5 root pieces of 1cm were sterilized

with 1 bleach and placed on plates poured with Potato Dextrose Agar mixed with

penicillin (100000 units litre) and streptomycin (02 glitre) After incubation of 5 day

occurrence of root rots were recorded

67

68

Fig14 Growth promotion by the endophytic Penicillium in sunflower

Control +veControl EP EP EP

69

Fig14 Growth promotion by the endophytic Penicillium in neem cake amended soil in

sunflower

Control +ve Control EP

+veControl EP

EP

EP EP EP EP

EP

Control

70


root rotting fungi as compared to plant grown in natural soil (un-amended soil) Plant

inoculated with endophytic Penicillium species most of them showed less infection of

root rotting fungi as compared to untreated control Plants grown in pots received

Endophytic Penicillium isolates caused significant reduction except Ppurpurogenum

(EPSML3) and Plividum which caused no reduction as compared to untreated control

on F oxysporum infection Whereas pots received endophytic P citrinum

Ppurpurogenum (EPSML3) Pnigricans Pregulosum P decumbens Pduclauxi

Pthomii Pjavanicum with neem cake showed complete suppression of F oxysporum

Combine effect of isolates Pnigricans P citrinum Plilacinum Plividum P

restrictum Pthomii Pjavanicum and neem cake showed no infection of Fsolani P

decumbens Pnigricans and Pjavanicum also showed complete suppression of

infection of Fsolani while Plividum showed no difference from control when used

alone Plividum alone showed no infection of Mphaseolina on sunflower roots

Combine effect of all treatments with neem cake showed significant reduction on

infection of Mphaseolina Application of P decumbens P citrinum Plividum

Ppurpurogenum (EPEHS7) and Pregulosum showed no infection of Rsolani P

decumbens Pnigricans P citrinum Ppurpurogenum (EPSML3) Pduclauxi

Ppurpurogenum (EPEHS7) P restrictum Ppurpurogenum (EPAER14) and

Pjavanicum with neem cake showed complete suppression of Rsolani (Table 12)

Plant grown in soil amended with neem cake generally showed greater height as

compared to plant grown in natural soil (un-amended soil) Plant inoculated with

endophytic Penicillium species most of them showed larger shoot length as compared to

untreated control Greater plant height was produced by Plilacinum when applied in

neem cake amended soil (Table 13 and Fig 15-17)

71



Infection


Fsolani



Control 50 187 50 25 75 75 187 125

Carbendazim 125 62 312 62 125 25 62 62




P citrinum EPSMR1 375 0 25 0 125 25 0 0



Pduclauxi EPASS9 25 0 62 62 312 187 62 0

Plividum EPMCL12 50 62 50 0 0 50 0 62



Pthomii EPAER11 62 0 62 0 375 187 62 62



Pasperum EPHAL10 125 125 25 187 312 312 62 62






72


Treatments Code

Shoot Length

(cm)

Shoot Weight

(g)


(cm)

(g)


Control 3256 3893 378 642 57 1034 085 131

Carbendazim 3781 4293 452 607 84 1025 124 128




P citrinum EPSMR1 385 6443 373 1425 75 787 088 226



Pduclauxi EPASS9 4412 6275 386 1013 7 768 086 213

Plividum EPMCL12 345 6551 206 1019 706 645 072 161



Pthomii EPAER11 4838 6208 489 953 863 656 096 141



Pasperum EPHAL10 4158 5006 362 818 6102 1275 067 186

LSD005 10331 8971 2271 5521 3021 2171 04581 1071




73

Fig15 Growth promotion by the endophytic Penicillium in soil amended with neem cake

in sunflower

373 Effect of endophytic Penicillium with neem cake in inhibition of root diseases

and mung bean growth

In an experiment a 25 ml cell suspension of five-day-old cultures of Fourteen

isolates of endophytic Penicillium viz P citrinum Plilacinum Ppurpurogenum

(EPSML3) Pnigricans Pregulosum P decumbens Ppurpurogenum (EPEHS7) P

restrictum Pduclauxi Pasperum Pthomii Plividum Pjavanicum and

Ppurpurogenum (EPAER14) were applied in pots filled with 1 Kg soil Endophytic

Penicillium were drench in each pots with 1 neem cake in another pot set Mung bean

(Vigna radiata) seeds were sown pots (6 seeds per pot) Four seedlings were remained in

each pots after germination Treatments were replicated four times and data were noticed

after 45 days

EP

Carbendazim Control

74

No infection of Foxysporum were found Plilacinum Ppurpurogenum (EPSML3)

and Pduclauxi when used in natural soil Whereas infection of Foxysporum was also not

found where Plilacinum Pnigricans and Pduclauxi used in neem cake amended soil

Significant reduction in infection of Fsolani was seen in natural soil by all isolates whereas

in neem cake amended soil all isolates also showed significant reduction other than P

citrinum which showed infection equal to control treatment 75 No infection of

Mphaseolina was showed by P citrinum in both type of soil whereas P restrictum also

showed no infection of Mphaseolina only in natural soil Control showed no infection of

Rsolani in natural soil while Pnigricans Pasperum Pthomii and Pjavanicum in

amended soil showed no infection of Rsolani (Table 14)

Use of endophytic Plividum with neem cake caused a significant increase in

plant height while Pnigricans Plilacinum Ppurpurogenum (EPEHS7) Pasperum

Pthomii Pjavanicum and Ppurpurogenum (EPAER14) showed significant result in

natural soil Ppurpurogenum (EPEHS7) and Ppurpurogenum (EPAER15) showed

significant growth on Shoot weight in natural soil In natural soil greater root length was

showed by Plilacinum whereas in amended soil P restrictum Pasperum Pthomii and

Pjavanicum showed larger root length (Table 15)

75

Table14 Effect of endophytic Penicillium with neem cake on the infection of Fusarium solani Foxysporum Rhizoctonia solani and

Macrophomina phaseolinaon mung bean roots in green house experiment

Infection


Fsolani



Control hellip 50 312 100 75 100 50 0 562





P citrinum EPSMR1 62 62 437 75 0 0 62 62



Pduclauxi EPASS9 0 0 437 375 25 375 62 25

Plividum EPMCL12 62 25 25 687 125 375 62 50



Pthomii EPAER11 62 62 437 25 125 312 0 0



Pasperum EPHAL10 435 125 25 25 25 187 0 0




3 Mean values in the NS and AS column showing difference greater than LSD value are significantly different at plt005


76

Table15 Effect of endophytic Penicillium and neem cake on the growth of mung bean in green house experiment


Shoot Weight


(cm)

(g)

(cm)

(g)


Control hellip 1375 1714 078 08 1531 4652 051 014





P citrinum EPSMR1 1299 1606 059 0617 165 477 039 016



Pduclauxi EPASS9 1187 1916 069 0855 1108 4562 017 016

Plividum EPMCL12 132 2147 061 1358 2252 4785 026 022



Pthomii EPAER11 1463 179 077 1203 1125 7025 031 024



Pasperum EPHAL10 1463 1874 077 1102 1125 702 031 02

LSD005 1611 4011 0191 2141 8421 1151 0171 0071




77

374 Effect of Endophytic Penicillium and cotton cake in inhibition of root

diseases and mung bean growth

A 25 ml five-day-old cell suspension of fourteen isolates of endophytic

Penicillium viz P citrinum Plilacinum Ppurpurogenum (EPSML3) Pnigricans

Pregulosum P decumbens Ppurpurogenum (EPEHS7) P restrictum Pduclauxi

Pasperum Pthomii Plividum Pjavanicum and Ppurpurogenum (EPAER14) were

applied in clay pots filled with 1 Kg soil In similler set endophytic Penicillium were

drench in each pots alongwith 1 cotton cake Seeds of mungbean (Vigna radiata)

were sown Four seedlings were kept in each pot after germination Carbendazim (200

ppm) 25 ml pot considered as positive control

After 45 days data were noted Different Fsolani and Foxysporum infection

showed between plants treated with different isolates was significant Endophytic

Penicillium isolates separete or combine with cotton cake caused significant reduction

M phaseolina infection Plants grown in soil treated with Pnigricans Pregulosum P

decumbens Ppurpurogenum (EPEHS7) Pthomii Plividum Pjavanicum and

Ppurpurogenum (EPAER14) in cotton cake amended soil showed no infection of R

solani (Table 16)

Cotton cake and Pnigricans Pthomii Pjavanicum significant increased root

length and fresh root weight related to control plants While combine use of cotton cake

and P decumbens significantly improved fresh shoot weight (Table 17)

78

Table16 Effect of Endophytic Penicillium and cotton cake on the infection of Fusarium solani Foxysporum Rhizoctonia solani and

Macrophomina phaseolina on mungbean roots in green house experiment

Infection


Fsolani



Control hellip 50 50 100 75 100 75 0 187





P citrinum EPSMR1 62 25 437 437 0 437 62 187



Pduclauxi EPASS9 0 312 437 562 25 562 62 65

Plividum EPMCL12 62 125 25 25 125 25 62 0



Pthomii EPAER11 62 187 437 437 125 375 0 0



Pasperum EPHAL10 437 375 25 312 25 562 0 125






79

Table17 Effect of Endophytic Penicillium and Cotton cake on the growth of mung bean in green house experiment

Treatments Code

Shoot Length

Shoot Weight Root Length Root weight

(cm)

(g)

(cm)

(g)


Control hellip 1375 1364 078 089 1531 613 051 031





P citrinum EPSMR1 1299 1318 059 193 165 961 039 037



Pduclauxi EPASS9 1187 1438 069 13 1108 1178 017 048

Plividum EPMCL12 132 1323 061 107 2252 1024 026 048



Pthomii EPAER11 1463 1435 077 119 1125 1185 031 071



Pasperum EPHAL10 1463 1453 077 128 1125 972 031 046

LSD005 1611 2661 0191 091 8421 271 0171 0291



80

375 Effect of endophytic Penicillium in inhibition of root diseases and

mungbean growth









No infection of Foxysporum was found by Plilacinum and Pduclauxi

treatments Significant reduction in infection of Fsolani was seen by all isolates No

infection of Mphaseolina was showed by P citrinum and P restrictum All treatments

showed significant reduction on infection of Rsolani although Pnigricans P

decumbens Pthomii and Pjavanicum showed 0 infection (Table 18)

Application of Endophytic Pasperum caused a significant increase in plant

height Showed significant result in natural soil P citrinum caused significant growth

on Shoot weight Root length showed non-significant result P decumbens showed

greater fresh root weight (Table 19)

81

Table18 Effect of Endophytic Penicillium on the infection of Fusarium solani Foxysporum Rhizoctonia solani and

Macrophomina phaseolina on mung bean roots in green house experiment

Treatments Code Foxysporum Fsolani M phaseolina Rsolani

Infection

Control --------- 50 100 100 50

Carbendazim --------- 25 50 50 62

P decumbens EPAIR6 125 375 187 0

Pnigricans EPSLR4 62 50 125 0

Pregulosum EPAAR5 125 437 312 62

P citrinum EPSMR1 62 437 0 62

Plilacinum EPSMS2 0 50 312 187

Ppurpurogenum EPSML3 25 25 312 25

Pduclauxi EPASS9 0 437 25 62

Plividum EPMCL12 62 25 125 65

Ppurpurogenum EPEHS7 62 375 187 62

Prestrictum EPCTS8 125 437 0 62

Pthomii EPAER11 62 50 125 0

Ppurpurogenum EPAER14 62 375 187 62

Pjavanicum EPSLR13 62 50 125 0

Pasperum EPHAL10 437 25 25 62

LSD005 Treatment=7601 Pathogen=3802

82

Table19 Effect of endophytic Penicillium on the growth of mung bean in green house experiment

Treatments Code Shoot Lenght Shoot Weight Root Length Root weight

(cm) (g) (cm) (g)

Control ---------- 1475 0522 4972 0098

Carbendazim --------- 1635 0987 3737 009




P citrinum EPSMR1 1344 0987 4617 0137



Pduclauxi EPASS9 1434 0696 4127 0096

Plividum EPMCL12 1639 0752 4147 0121



Pthomii EPAER11 1482 0711 3865 0072



Pasperum EPHAL10 1608 0787 3875 0066

LSD005 2891 0261 0741 0051



83

84

376 Effect of endophytic Penicillium in soil amended with neem cake in

inhibition the root diseases and tomato growth

In this experiment 25 ml of five-day-old cell suspension of fourteen isolates of

endophytic Penicillium viz P citrinum Plilacinum Ppurpurogenum (EPSML3)

Pnigricans Pregulosum P decumbens Ppurpurogenum (EPEHS7) P restrictum

Pduclauxi Pasperum Pthomii Plividum Pjavanicum and Ppurpurogenum

(EPAER14) were applied in each pots filled 1 Kg soil In same other set endophytic

Penicillium were applied in each pots alongwith 10g neem cake per pot Three-week-

old four equal sized tomato (Lycopersicon exculentum) seedlings grown in autoclaved

soil were shifted in pots Carbendazim (200 ppm) 25 ml pot considered as positive

control Treatments were replicated four times and data were noticed after 60 days

Application of endophytic P decumbens P citrinum and Pduclauxi and P

restrictum alone affected a complete suppression of Foxysporum infection Whereas

Pduclauxi was found no infection of Foxysporum when used with neem cake (Table

20) Endophytic Penicillium are found effective against Fsolani in both type of soil

When P decumbens and Pduclauxi were used alone Infection of M phaseolina was

significantly reduced In neem cake amended soil untreated control showed no infection

of M phaseolina Difference in R solani infection among plants received different

treatment was non significant in both type of soil natural and amended (Table 20)

Plants grown in natural soil received P decumbens Pnigricans Pduclauxi

Ppurpurogenum (EPAER14) and Pjavanicum fungal culture showed better growth

than untreated control Pasperum with neem cake showed highly significant plant

height of 24cm (Table 21 and Fig18-20)

85


Macrophomina phaseolina on tomato roots in green house experiment

Infection


Fsolani



Control hellip 437 312 625 625 312 0 312 0





P citrinum EPSMR1 0 50 62 625 625 62 75 0



Pduclauxi EPASS9 0 0 62 25 187 125 50 62

Plividum EPMCL12 50 437 437 562 375 0 687 62



Pthomii EPAER11 187 562 312 562 50 312 562 0



Pasperum EPHAL10 62 312 125 562 25 62 812 0






86

Table 21 Effect of endophytic Penicillium and neem cake on the growth of tomato in green house experiment

Treatments Code Shoot Length Shoot Weight Root Length Root weight

(cm) (g) (cm) (g)


Control hellip 12 1544 18 407 126 333 155 063





P citrinum EPSMR1 1732 1755 297 389 922 1149 064 056



Pduclauxi EPASS9 1672 2255 243 636 1185 597 057 11

Plividum EPMCL12 1307 1303 178 254 1242 529 052 046



Pthomii EPAER11 1328 1375 214 234 148 466 046 055



Pasperum EPHAL10 1328 2412 18 732 1225 775 06 126

LSD005 271 5171 0691 2091 3731 3031 1031 0631



87

Fig18 Growth promotion by the endophytic Penicillium in tomato

EP

88

377 Effect of endophytic Penicillium in soil amended with cotton cake in

inhibition of root diseases and tomato growth






Penicillium were applied in each pots alongwith 10g neem cake per pot Three-week-old

four equal sized tomato (Solanum Lycopersicum) seedlings grown in autoclaved soil

were shifted in pots Carbendazim (200 ppm) 25 ml pot was considered as positive

control Treatments were replicated four times and data were recorded after 60 days

Application of endophytic P decumbens P citrinum Pduclauxi and P

restrictum alone affected a broad inhibition of Foxysporum infection Whereas

Pregulosum was found no infection of Foxysporum when used with cotton cake (Table

22) Endophytic Penicillium are found effective against Fsolani in natural soil In

cotton cake amended soil Pnigricans and Pduclauxi showed significant reduction in

Fsolani infection When P decumbens and Pduclauxi were used alone Infection of M

phaseolina was significantly reduced In cotton cake amended soil Pregulosum P

citrinum Plilacinum Ppurpurogenum (EPSML3) and Plividum showed no infection


treatment was non-significant in natural soil and in cotton cake amended soil no

infection of Rsolani was found (Table 22)

89

Table 22 Effect of endophytic Penicillium and cotton cake on the infection of Fusarium solani Foxysporum Rhizoctonia solani and


Infection


Fsolani



Control hellip 437 50 625 25 312 62 312 0





P citrinum EPSMR1 0 62 62 562 625 0 75 0



Pduclauxi EPASS9 0 562 62 562 187 25 50 0

Plividum EPMCL12 50 187 437 375 375 0 687 0



Pthomii EPAER11 187 312 312 25 50 125 562 0



Pasperum EPHAL10 62 125 125 50 25 62 812 0


1Difference greater than LSD values among means in column are significant at plt005




90



than untreated control P restrictum with cotton cake showed highly significant plant

height Pnigricans and Pjavanicum showed greater fresh shoot weight in amended

soil Root length in both type of soil was non-significant Whereas fresh root weight

was non-significant in natural soil In cotton cake amended soil Pduclauxi showed

significant fresh root weight (Table 23 and Fig21)

378 Effect of endophytic Penicillium with neem cake in inhibition of root

diseases and chickpea growth


Ppurpurogenum (EPSML3) Pnigricans Pregulosum Pdecumbens Ppurpurogenum


Ppurpurogenum (EPAER14) caused suppression of four root rotting fungi in vitro A

25ml cell suspension of five-day-old culture of fungal isolates were drench in each pots

filled with 1kg soil Carbendazim considered as positive control against root rotting

fungi Combine use of endophytic Penicillium and 1 Neem cake were drenched in

another same set Chickpea (Cicer arietinum) seeds were sown in pots (6 seeds per pot)

After one week four seedlings were kept in each pots and extra were detached

Treatments were replicated four times and watered daily Data were recorded after six

weeks

91

Table23 Effect of endophytic Penicillium and cotton cake on the growth of tomato in green house experiment

Treatments Code

Shoot

Length

Shoot

Length

Shoot

Weight

Shoot

Weight

Root

Length

Root

Length

Root

weight

Root

weight

(cm) (cm) (g) (g) (cm) (cm) (g) (g)


Control hellip 12 1633 18 554 126 1757 155 105





P citrinum EPSMR1 1732 1912 297 512 922 9 064 155



Pduclauxi EPASS9 1672 214 243 69 1185 153 057 237

Plividum EPMCL12 1307 2347 178 741 1242 1298 052 156



Pthomii EPAER11 1328 225 214 657 148 155 046 164



Pasperum EPHAL10 1328 2101 18 525 1225 1095 06 135

LSD005 271 4291 0691 3281 3731 5851 1031 091


92

Fig 21 Growth promotion by the endophytic Penicillium in soil amended with cotton

cake in tomato

EP

93

Plants grown in pots received endophytic Penicillium isolates Ppurpurogenum

(EPSML3) and Pthomii in natural soil and in amended soil with neem cake P

decumbens Pnigricans Ppurpurogenum (EPSML3) Ppurpurogenum (EPEHS7)

Pjavanicum and Ppurpurogenum (EPAER14) showed no infection of F oxysporumIn

unamended soil Fsolani was found significantly reduced except isolate Pasperum

Whereas in amended soil infection of Fsolani was non significant In unamended soil

Mphaseolina was found significantly reduced Combine effect of isolates

Ppurpurogenum (EPSML3) Ppurpurogenum (EPEHS7) Ppurpurogenum (EPAER14)

and neem cake showed significant result on Mphaseolina infection Application of

Pregulosum P decumbens P restrictum Pduclauxi Pasperum and Pthomii showed

no infection of Rsolani in natural soil Amended soil with neem cake showed no

infection of Rsolani (Table 24)

Greater plant height was produced by P decumbens Pnigricans Pregulosum

and Pduclauxi when applied in natural soil Effect of P restrictum and P citrinum with

neem cake showed highest plant height Untreated control of amended soil showed

highest value of fresh shoot weight and fresh root weight related to other treatments

whereas fresh shoot weight in natural soil showed significant result in all treatments

except Pthomii P decumbens and Pduclauxi alone showed highest root length and

fresh root weight In amended soil Ppurpurogenum (EPAER14) showed significant

root length (Table 25 and Fig22-27)

94


Macrophomina phaseolina on chickpea roots in green house experiment

Infection


Fsolani



Control hellip 375 0 50 125 437 375 25 0





P citrinum EPSMR1 312 187 187 312 375 50 187 0



Pduclauxi EPASS9 187 375 125 25 375 50 0 0

Plividum EPMCL12 62 62 437 125 62 625 25 0



Pthomii EPAER11 0 187 437 187 62 25 0 0



Pasperum EPHAL10 125 62 50 125 125 812 0 0






95

Table25 Effect of endophytic Penicillium and neem cake on the growth of chickpea in green house experiment


(cm) (g) (cm) (g)


Control hellip 2369 2225 274 837 274 975 211 303





P citrinum EPSMR1 267 3384 313 668 2397 975 394 098



Pduclauxi EPASS9 2925 2911 376 388 3037 1293 522 116

Plividum EPMCL12 2768 2801 31 698 2155 1132 35 109



Pthomii EPAER11 239 30 296 799 2416 1062 427 125



Pasperum EPHAL10 2856 2891 344 763 1921 1352 306 13

LSD005 471 4931 0941 3331 7321 5451 1611 11071



96

Fig22 Growth promotion by the endophytic Penicillium in chickpea


EP

S

EP

97


EP

EP

98


in chickpea

Fig 26 Growth promotion by the endophytic Penicillium in soil amended with neem cake

in chickpea

EP

EP

99


in chickpea

379 Effect of endophytic Penicillium with mustard cake in suppressing the root

diseases and growth of chickpea







fungi Combine use of endophytic Penicillium and 1 mustared cake were drenched in




weeks

Root rot fungi infection was less in amended soil as compared to unamended

soil No infection of Foxysporum was found in Ppurpurogenum (EPSML3) and

Pthomii in unamended soil P citrinum Ppurpurogenum (EPSML3) Pnigricans

Pregulosum P decumbens Ppurpurogenum (EPEHS7) Pduclauxi Pjavanicum and

Ppurpurogenum (EPAER14) with mustard cake amendment showed complete

suppression of Foxysporum P decumbens and Ppurpurogenum (EPSML3) in

amended soil showed less infection of Fsolani while Plividum showed 100 infection

of Fsolani in amended soil Infection of M phaseolina in unamended soil was

significant whereas in amended soil untreated control showed no infection of M

phaseolina Treatment of Pthomii and Ppurpurogenum (EPAER14) in mustard cake

amended soil showed less infection of R solani while P citrinum Pnigricans

Pregulosum Pduclauxi Pjavanicum and Plividum showed non-significant result

(Table 26)

100

Natural soil showed greater plant height as compared to mustard cake amended

soil Pnigricans showed greater plant length as compared to other treatments In

amended soil plant height was non-significant statisticaly (Table 27)

101

Table 26 Effect of endophytic Penicillium and mustard cake on the infection of Fusarium solani Foxysporum Rhizoctonia solani and


Infection



Control hellip 375 125 50 312 437 0 25 187





P citrinum EPSMR1 312 0 187 625 375 187 187 312



Pduclauxi EPASS9 187 0 125 625 375 62 0 312

Plividum EPMCL12 62 62 437 100 62 25 25 312



Pthomii EPAER11 0 62 437 125 62 62 0 62



Pasperum EPHAL10 125 0 50 187 125 0 0 0






102

Table 27 Effect of endophytic Penicillium and mustard cake on the growth of chickpea in green house experiment


(cm) (g) (cm) (g)


Control hellip 2369 2188 274 406 274 692 211 58





P citrinum EPSMR1 267 1916 313 556 2397 756 394 1172



Pduclauxi EPASS9 2925 192 376 561 3037 1115 522 819

Plividum EPMCL12 2768 1929 31 417 2155 946 35 383



Pthomii EPAER11 239 2305 296 626 2416 9 427 931



Pasperum EPHAL10 2856 1662 344 582 1921 925 306 834

LSD005 471 6131 0941 3011 7321 2921 1611 6151



NS=NaturalSoilAS=AmendedSoil

103

3710 Effect of endophytic Penicillium and fungicides in inhibition of root infection

and sunflower growth

Four isolates of endophytic Penicillium viz P citrinum (EPSMR1) Pnigricans

(EPSLR4) P decumbens (EPAIR6) and Pasperum (EPHAL10) caused suppression of

four root rotting fungi in vitro and revealed significant growth in in vivo were selected to

evaluate the combine effect with three different fungicides (Feast-M Carbendazim and

Topsin-M) A 25ml five-day-old cell suspension of fungal isolates were applied in pots

filled with 1kg soil In same other set pots were also applied combine application of

endophytic Penicillium and fungicides Each fungicide were also drench 25ml of 200ppm

in each pot Sunflower (Helianthus annuus) seeds were sown in pot (6 seeds per pot)

After one week four seedlings were kept in pots and extra were detached Treatments were

replicated four times and watered according to requirement Data were recorded after six

weeks

All three fungicides alone showed no infection of F oxysporum Plants grown in pots

received endophytic Penicillium isolate P decumbens and Pasperum with Feast-M showed

no infection of infection of F oxysporum Plants grown in pots received endophytic

Penicillium isolate Pnigricans with carbendazim and Pnigricans and P citrinum with

Topsin-M showed complete suppression of infection of F oxysporum All treatments

showed less infection of Fsolani as compared to control All treatments showed less

infection of Mphaseolina as compared to untreated control except P citrinum Pnigricans

alone and P decumbens Pasperum combine with Topsin-M showed 100 Mphaseolina

infection on sunflower roots Combine effect of Pasperum with Topsin-M and P citrinum

alone showed no infection of Rsolani Feast-M+ Pasperum and carbendazim showed no

difference from untreated control (Table 28)

Greater plant height was produced by carbendazim+ Pnigricans However greater

fresh shoot weight was produced by Feast-M alone (Table 29)

104

Table 28 Effect of endophytic Penicillium and fungicides on the infection of Fusarium solani Foxysporum Rhizoctonia solani and


Infection

Treatments Foxysporum Fsolani M phaseolina Rsolani

Control 75 100 100 75

Feast-M 0 37 687 625

Feast-M+ P citrinum 62 75 625 687

Feast-M+ Pnigricans 187 812 687 687

Feast-M+ P decumbens 0 312 50 625

Feast-M+ Pasperum 0 50 81 75

Carbendazim 0 812 75 75

Carbendazim+P citrinum 62 562 87 687

Carbendazim+ Pnigricans 0 75 625 187

Carbendazim+P decumbens 62 812 812 687

Carbendazim+ Pasperum 187 562 75 312

Topsin-M 0 437 812 62

Topsin-M+ P citrinum 0 812 437 125

Topsin-M+ Pnigricans 0 75 312 437

Topsin-M+P decumbens 687 687 100 25

Topsin-M+ Pasperum 875 25 100 0

P citrinum 437 687 100 0

Pnigricans 125 812 100 62

P decumbens 187 50 437 187

Pasperum 125 50 562 125




105

Table 29 Effect of endophytic Penicillium and fungicides on the growth of sunflower in green house experiment

Treatments ShootLength ShootWeight Root Length Root weight

Control 3197 339 288 288

Feast-M 4269 451 526 526





Carbendazim 3675 319 398 398

Carbendazim+ P citrinum 3933 326 464 464


Carbendazim+ P decumbens 3807 315 527 527


Topsin-M 3935 314 383 383



Topsin-M+ P decumbens 337 229 409 409


P citrinum 3268 249 432 432

Pnigricans 2788 201 401 401

P decumbens 3421 3007 446 446

Pasperum 3262 229 363 363

LSD005 5751 0811 1041 1041



106

3711 Effect of endophytic Penicillium on okra growth

In this experiment six seeds of okra (Abelmoschus esculentus) were sown in

earthen pots filled with 5 kg garden soil and watered watered daily to gained the 50

WHC (Keen and Raczkowiski 1921) P nigricans (EPSLR4) P rugulosum (EPAAR5)

and P decumbens (EPAIR6) (8x107 cfumL) used as soil drench in each pot and four

seedlings were kept after germination Treatments were replicated four times in screen

house Carbendazim was considered as a positive control and data were recorded after 90

days of germination

Treatments showed significant (Plt005) reduction of F solani and R solani

related to control (Table 30)

Application of P rugulosum resulted maximum plant height highest shoot weight

and root length while maximum root weight produced due to the treatment of carbendazim

and P decumbens Maximum number of fruits produced by Pnigricans and P decumbens

resulted highest fresh fruit weight(Table 31)

Highest polyphenol content resulted by Pnigricans and highest antioxidant activity

determined due to the drenching of Pnigricans after 1 minute and after 30 minute

Application of P rugulosum resulted maximum production of salicylic acid (Table 31)

Application of antagonist showed significant outcome on okra fruits Highest pH

showed by Pnigricans Application of P decumbens resulted highest tritable acidity value

then in Pnigricans and P rugulosum (Table 33) Application of carbendazim resulted

highest moisture content then in P rugulosum in fruits Maximum protein resulted by P

rugulosum then in P decumbens while highest carbohydrate caused by P decumbens

then in Pnigricans All the treatments showed significant (Plt005) Increased polyphenol

content showed by all treatments as compared to control (Table 34) P decumbens

resulted highest polyphenol followed by P rugulosum as compared to untreated plants P

rugulosum resulted significant improve in antioxidant potentail(Fig28)

107

Table30 Effect of endophytic Penicillium as soil drench on the infection of Macrophomina phaseolina Rhizoctonia solani Fusarium

solani and F oxysporum in garden soil

Infection


Control 0 50 625 50


P decumbens 0 0 625 312


P rugulosum 0 187 562 25




108

Table31 Effect of endophytic Penicillium as soil drench on growth of okra plants in garden soil

Treatments Shoot Length Shoot Weight Root Length Root Weight Number of

Fruits Fruit weight

(cm) (g) (cm) (g)

Control 3831 1058 1596 305 023c 708c

Carbendazim 3421 832 1659 546 045b 683c

P decumbens 4523 1167 1756 438 052a 1106a

Pnigricans 4265 1172 1794 188 054a 894b

P rugulosum 4592 1295 1967 2405 025c 533d

LSD005 511 4281 3431 581 00261 04841


109

Table32 Effect of endophytic Penicillium as soil drench on polyphenol salicylic acid and antioxidant activity of okra plants in garden

soil

Treatments Polyphenol Antioxidant () Salicylic Acid

microgml After 1 minute After 30 minutes microgml

Control 137e 2711e 2878e 0053d

Carbendazim 172d 4608d 4908d 0048e

P decumbens 308c 4974c 5256c 0093c

Pnigricans 424a 5744a 6229a 0116b

P rugulosum 364b 5393b 5859b 0161a

LSD005 00311 01361 04211 00041


110

Table33 Effect of endophytic Penicillium as soil drench on biochemical parameters of ok ra fruits

Treatments pH Tritable acidity Moisture content Total solids Total Soluble Solid

Sucrose

Control 587c 0087c 8668d 1353b 245d

Carbendazim 585c 013b 9175a 803e 257c

P decumbens 59c 0194a 8434e 1559a 31a

Pnigricans 629a 0128b 8715c 1287c 28b

P rugulosum 605b 0128b 8808b 1185d 317a

LSD005 0121 000571 0211 01031 0121


111

Table 34 Effect of endophytic Penicillium as soil drench on polyphenol antioxidant activity protein and carbohydrates of okra fruits

in garden soil

Treatments Antioxidant Polyphenol Protein Carbohydrates

microgml microgml microgml

Control 2647e 665e 13e 69d

Carbendazim 3575d 734d 27d 86c

P decumbens 4906c 1613a 5263b 1033a

Pnigricans 5115b 96c 39c 99b

P rugulosum 5631a 122b 5566a 9833b

LSD005 10591 01441 21941 3711


112

3712 Effect of endophytic Penicillium on the growth root rotting fungi and

induction of systemic resistance in tomato

Filled earthen pots with 5 kg of soil and watered according to requirement to

maintain 50 WHC (Keen and Raczkowiski 1921) P nigricans (EPSLR4) P

rugulosum (EPAAR5) and P decumbens (EPAIR6) (8x107 cfumL) used as soil drench

Four equal sized seedlings of tomato were transfered in pots Treatments were four time

replicated Carbendazim was considered as a positive control and data were recorded

after 90 days

Most of the treatment showed significant (Plt005) results of R solani F solani

and M phaseolina as relation to control plants (Table 35)

Application of Pnigricans showed highest plant height shoot weight by P

decumbens Maximum number of fruits produced by Pnigricans and P decumbens


P rugulosum showed improved polyphenol as compare to control plants

Highest antioxidant activity resulted by P decumbens and carbendazim after 1 minute

and after 30 minute P rugulosum showed highest antioxidant activity Application of

Pnigricans and P decumbens resulted maximum production of salicylic acid (Table

37)

Application of endophytic Penicillium showed significant effect on tomato

fruits Highest pH noticed when soil treated with Pnigricans and P decumbens

Maximun tritable acidity produced by P decumbens (Table 38) Highest protein

produced by P rugulosum then in P decumbens while carbohydrate resulted by

Pnigricans followed by P decumbens All the treatments showed increase polyphenol

content as compare to control (Table 39) Pnigricans showed significant enhancment in

antioxidant activity related to control

113



Infection


Control 312 100 937 562





LSD005 Treatment1=1455 Pathogen2=1302



114

Table36 Effect of endophytic Penicillium as soil drench on growth of tomato plants in garden soil

Treatments Shoot Length Shoot Weight Root Length Root Weight Number of Fruits Fruit weight

(cm) (g) (cm) (g)

Control 52 1974 1816 35 30a 5801b

Carbendazim 4646 1322 1629 237 20c 4083a

P decumbens 443 2161 1283 418 2133c 995a

Pnigricans 55 1892 1561 315 32a 4286d

P rugulosum 5197 1695 1205 334 256b 4779c

LSD005 1481 18611 5391 4011 3781 0131


115

Table 37 Effect of endophytic Penicillium as soil drench on polyphenol salicylic acid and antioxidant activity of tomato plants in

garden soil



Control 090a 40a 139a 014a

Carbendazim 019a 49a 127a 018a

P decumbens 0076a 44a 131a 019a

Pnigricans 0076a 33a 103a 019a

P rugulosum 0108a 33a 292a 017a

LSD005 01081 01671 0301 00791


116

Table 38 Effect of endophytic Penicillium as soil drench on biochemical parameters of tomato fruits

Treatments pH Tritable acidity Firmness Total Soluble Solid

N Sucrose

Control 411c 023c 34a 323c

Carbendazim 418b 027bc 143b 806a

P decumbens 43a 034a 076b 676ab

Pnigricans 43a 030ab 126bc 613b

P rugulosum 418b 030ab 086bc 686ab

LSD005 00621 00541 0211 1311


117

Table 39 Effect of endophytic Penicillium as soil drench on polyphenol antioxidant activity protein and carbohydrates of tomato

fruits in garden soil



Control 1966c 573e 16d 63a

Carbendazim 333b 756d 28c 78a


Pnigricans 52a 1026c 41b 97a

P rugulosum 496a 125b 52a 96a

LSD005 5591 0471 5771 2391


118

38 FIELD EXPERIMENTS

381 Effect of Pseudomonas monteilii and endophytic Penicillium on okra growth in

field condition

The experiment carried out in 2 times 2 meter field and replicated four times Cell

suspension of endophytic Penicillium (8x107 cfumL) were drench at 200-ml per meter row

alone and in combination with Pseudomonas monteilii 20 seeds of okra were seeded in

rows Topsin-M at 200 ppm were also used alone as a positive control On the basis upon

the requirement plants were watered with difference of 2-3 days The field had infestation

of 2080 cfug of soil of a diverse population of F solani and F oxysporum 10-22

sclerotia of M phaseolina g of soil and 8-17 colonization of R solani on sorghum

seeds used as baits naturally To evaluate the potential of Pseudomonas monteilii and

endophytic Penicillium plants were harvested (form each row 4 plants took) after 45 and

90 days of germination Incidence of root rotting fungi plant physical parameters and

resistance biomarkers were recorded

Significant (Plt005) inhibition of F oxysporum showed by most of treatments as

compere to control except P rugulosum P decumbens + Pseudomonas monteilii and

Topsin-M after 45 days (Table 40) Maximum reduction of Fsolani were observed in

plants treated with Pseudomonas monteilii and Pnigricans + Pseudomonas monteilii after

45 days While maximum reduction of M phaseolina observed in application of P

rugulosum+ Pseudomonas monteilii after 45 days Application of P rugulosum+

Pseudomonas monteilii and Pnigricans showed maximum reduction of Rsolani after 45

days

Highest length of shoot and weight of shoot were observed in plants Maximum

plant hieght were observed after 45 and 90 days intervals with mixed application of

Pnigricans with Pseudomonas monteilii Highest weight of shoot were also observed in

combine application of Pnigricans with Pseudomonas monteilii after 45 and 90 days

while application of Pseudomonas monteilii resulted maximum length of root after 45

days Significant increase in root length produced after 90 days from combine application

of Pnigricans with Pseudomonas monteilii Highest root weight resulted from combine

119

application of Pnigricans with Pseudomonas monteilii after 45 and 90 days Combine

application of P decumbens with Pseudomonas monteilii resulted highest number and

weight of fruits produced after 90 days (Table 41)

After 45 days most of the treatments shown significantly high phenols except

Topsin-M Most of the treatments shown maximum antioxidant activity significantly

except P rugulosum after 1 minute whereas maximum antioxidant activity showed by

Pseudomonas monteilii after 30 minutes P decumbens showed maximum production of

salicylic acid after 45 days (Table 42)

All the treatment showed significant effect on phenolic content except Topsin-M

and P decumbens whereas all the treatment showed significant effect on antioxidant

activity except Topsin-M and P decumbens with Pseudomonas monteilii after 1 and 30

minutes after 90 days Maximum production of salicylic acid showed in combine treatment

of Pnigricans with Pseudomonas monteilii after 90 days (Table 43)

In this experiment combine application of Pseudomonas monteilii and endophytic

Penicillium showed significant increase in physiobiochemical of okra fruits Combine

activity of Pnigricans + Pseudomonas monteilii resulted highest antioxidant activity in

fruits followed by Pseudomonas monteilii alone Highest polyphenol content resulted due

to the application of Pseudomonas monteilii followed by combine application of P

rugulosum with Pseudomonas monteilii Protein were showed maximum in combine

application of P decumbens with Pseudomonas monteilii and Pseudomonas monteilii

alone (Table 44) On the other side carbohydrate content observed highest in combine

application of P rugulosum with Pseudomonas monteilii Application of Pseudomonas

monteilii resulted maximum of total solids whereas combination of P rugulosum with

Pseudomonas monteilii produced highest of moisture Significant increase in pH showed

by Topsin-M followed by combination of Pnigricans with Pseudomonas monteilii and

maximum tritable acidity was showed by P decumbens (Table 45)

120

Table 40 Effect of Pseudomonas monteilii and endophytic Penicillium as soil drench on the infection of M phaseolina Rsolani F

solani and F oxysporum in soil under field condition

Infection


45 90 45 90 45 90 45 90

Control 375 0 562 312 937 100 562 0

Topsin-M 375 0 625 25 937 100 687 0

Pseudomonas monteilii 25 62 25 312 875 100 625 0

P decumbens 62 0 50 375 68 100 375 0

Pnigricans 125 187 562 687 875 100 312 0

P rugulosum 312 62 562 375 812 100 437 0

P rugulosum + Pseudomonas monteilii 187 12 312 50 625 937 312 0

P decumbens + Pseudomonas monteilii 312 62 437 25 812 687 562 0

Pnigricans + Pseudomonas monteilii 62 125 25 375 687 625 75 0

LSD005 Treatments1= 8931 Pathogens2=5952 Treatments1=13341 Pathogens2=8 892



121

Table 41 Effect of Pseudomonas monteilii and endophytic Penicillium as soil drench on growth of okra plants under the field

condition

Treatments Shoot Length

(cm)

Shoot Weight

(g)

Root Length

(cm)

Root Weight

(g)

Number

of Fruits

Fruit

weight

Control 45 90 45 90 45 90 45 90 90 90

Topsin-M 4178 6192 2228 4325 1368 2426 204 823 086g 246i

Pmontelii 422 6375 1765 4731 1267 2377 133 98 12f 31h

Penicillium decumbens 477 6861 2271 507 1839 2684 255 1056 246b 456d

P nigricans 4233 6617 1971 4887 1486 2578 167 1003 143e 1146a

Prugulosum 4866 7083 1635 5095 1378 2311 172 967 176d 331g

P rugulosum 4373 7026 2063 2051 1371 2464 169 709 123f 35f

P rugulosum + P monteilii 5768 8658 3164 5518 1167 3008 207 1208 143e 42e

P decumbens + P monteilii 5553 9499 1867 5897 1409 2938 187 1217 277a 661b

Pnigricans + P monteilii 5907 9867 4043 6095 14 3188 296 1923 22c 623c

LSD005 961 1321 131 1181 3551 1371 0831 2961 0111 0111


122

Table 42 Effect of Pseudomonas monteilii and endophytic Penicillium as soil drench on polyphenol salicylic acid and antioxidant

activity of okra plants in soil under field condition after 45 days

Treatments

Polyphenol

microgml

Antioxidant () Salicylic Acid

microgml After 1 minute After 30 minutes

Control 183h 7314e 7721e 007f

Topsin-M 146i 9119a 9886a 0113d

Pseudomonas monteilii 321f 784d 8466d 0144c

P decumbens 245g 6639g 6858g 0168a

Pnigricans 573c 8044c 8852c 0084e

P rugulosum 474d 7074f 7643f 0154bc

P rugulosum + P monteilii 336e 5045i 6038h 0105d

P decumbens + P monteilii 713b 5186h 5779i 0086e

Pnigricans + P monteilii 773a 8356b 8992b 0165ab

LSD005 00721 10191 06531 00121



123


activity of okra plants in soil under the field condition after 90 days

Treatments Polyphenol

microgml



Control 25def 6656e 7135f 0038g

Topsin-M 183f 4922f 5575g 0074bc

Pseudomonas monteilii 326cde 8345a 8885a 0052e

P decumbens 226ef 7804b 8539b 0072c

Pnigricans 52b 7726c 8233c 0066d

P rugulosum 41c 7165d 7851d 0042f

P rugulosum + P monteilii 343cd 7744c 8241c 0066d

P decumbens + P monteilii 683a 3254g 4917h 0077b

Pnigricans + P monteilii 74a 6852e 7604e 0105a

LSD005 10061 05191 04731 0003081


124

Table44 Effect of Pseudomonas monteilii and endophytic Penicillium as soil drench on polyphenol antioxidant activity protein and

carbohydrates of okra fruits in soil under field condition

Treatments Antioxidant Polyphenol Protein Carbohydrate


Control 5102g 646g 1466g 5966f

Topsin-M 5514f 716f 2566f 67e

Pseudomonas monteilii 6662b 136a 6766a 126a

P decumbens 5933d 976d 56d 101b

Pnigricans 5838d 816e 43e 92d

P rugulosum 6521c 114c 59c 96c

P rugulosum + P monteilii 5659e 124b 66b 102b

P decumbens + P monteilii 6616bc 11c 6766a 100b

Pnigricans + P monteilii 6909a 86e 56d 97c

LSD005 10451 06241 14081 2471


125

Table 45 Effect of Pseudomonas monteilii and endophytic Penicillium as soil drench on

biochemical parameters of okra fruits under field condition

Treatments pH

Tritable

acidity

Moisture

content

Total

solids

Total Soluble

Solid

Sucrose

Control 624a 0102c 8774b 1222f 1425e

Topsin-M 619ab 0126b 8653e 1339b 1475e

Pseudomonas monteilii 615b 0124b 8458f 1522a 2975d

P decumbens 606d 0185a 8632e 1355b 3125cd

Pnigricans 613bc 0127b 8752bcd 1249de 33bc

P rugulosum 607cd 0124b 8735cd 1256d 302d

P rugulosum + P monteilii 606d 0123b 8842a 117g 375a

P decumbens + Pmonteilii 603d 0122b 876bc 1233ef 342b

Pnigricans + P monteilii 616b 0125b 8723d 128c 305d

LSD005 00641 00041 03021 0171 02221


126

127

128

4 DISCUSSION

Microbes and Higher plants are the rich source of novel drugs In last 50 years

numerous effective drugs primarily extracted from fungi have been discoverd

(Smedsgaard and Nielsen 2005) Among them many bioactive compounds have been

produced from endophytes also known as an exceptional source as its capability to

inhabitate the plants in every environmental condition (Strobel and Daisy 2003) In

current study 14 endophytic Penicillium isolates were isolated (root stem and leaves)

from wild plants (Achyranthus aspera Atriplex stocksii Euphorbia hirta Chorchorus

tridens) and cultivated plant (Solanum melongena Lycopersicon esculentum

Helianthus annuus Azadirachta indica Abelmoschus esculentus Momordica

charantia) collected from different parts of Sindh province These findings is an

agreement to the earlier reports about the existence of Penicillium as endophyte

(Korejo et al 2014) Similar as (Ravindran et al 2012) A flavus from

mangrovesreported as an endophytes also

The microbes exist inter andor intra celluler of plant called ldquoendophytesrdquo

Endophytes gives variety of advantages to the host with vast applications in agriculture

and medicine (Clay and Rudgers 2005 Alvarez-Loayza 2011) Endophytes reside

inside the plant effects on plant health and survival They give strenght against abiotic

and biotic stresses and take nourishment from the plant Almost all vascular plants

studied till date have endophytic fungi in parts of their life cycle Plant pathogens and

pests are comparatively less attacked medicinal plants therefore endophytic micro-biota

can be of boundless significance in protecting plants from pests (Kaushik 2012)

Several studies on synthesis of secondry metabolites isolated from endophytic

fungi have found Among them some compounds used to discover new therapeutic

drugs (Strobel et al 2004) About 300000 plant species presented on land having

atleast one or more of fungi From many different plants including trees like yew and

pine and fodders like sorghum clover alfalfa and vegetables like tomatoes carrot

radish sweet potatoes lettuce and soybean fruits like citrus pineapple banana

pineapple and cereal grains like wheatrice and maizeand other crops like sugarcane

129

coffee and marigold have been examined for endophytes (Rosenblueth and Romero

2006) Several plants of medicinal importance such as Actinidia macrosperma (wild

kiwifruit) Ricinus communisTectona grandis Samanea saman Garcinia Picrorhiza

kurroa Cannabis sativa Withania somnifera Rauwolfia serpentine Cedrus deodara

Abies pindrow Pinus roxburgii Nothapodytes nimmoniana Platanus orientalis

Artemisia annua Brucea javanica M sieboldii and Calotropis procera have been

studied for endophytes Species of Alternaria Colletotrichum Aspergillus Fusarium

Gliocladium Cunninghamella Phomopsis Alternaria Fusarium Chaetomium

Nigrospora Cladosporium Alternaria Fusarium Aspergillus Curvularia

Cladosporium sp Aspergillus sp Nigrospora sp Fusarium sp Trichoderma sp

Chaetomium sp Alternaria sp Paecilomyces sp and Phyllostica are frequently

isolated from many agricultural and native plant species as endophytic fungi (Rubini et

al 2005 Guo et al 2008 Veja et al 2008 Gazis and Chaverri 2010 Kurose et al

2012 Parsa et al 2016) and Penicillium (H Kim 2014 Hassan 2017 Gautam 2013

Meng 2011 Peterson 2005 Qader 2015 Devi 2014 Shoeb 2014 Yin Lu et al 2011

Sandhu et al 2014 Phongpaichit et al 2006ukanyanee et al 2006 Qadri et al

2013 Liang 2014Cai and Wang 2012 Sandhu et al 2014b Cai 2012 Qadri 2013

In current study most of the endophytic Penicillium isolated Endophytic fungi

identified according to Domsch et al (1980) Dugan (2006) Raper and Thom (1949)

Barnett and Hunter (1998) and Visagie et al (2014) Identification of the promising

isolates was done through PCR amplification

Endophytic Penicillium isolated and tested for vitro and vivo activity in current

report most of the isolates showed inhibitory potential for fungi (root rotting) Fungal

endophytes that have useful impact on plant growth as biocontrol agents because their

effect against disease by inhabiting internal tissues of plants (Yuan et al 2017

Amatuzzi 2017) Similar biological position as pathogenic microorganism Berg et al

(2005) But in difference to plant pathogens they do not cause injury to host plant and

go inside plants for taking nourishment (Kobayashi and Palumbo 2000) Various

research are existing regarding the valuable function of fungal endophytes like act as

antagonist to phytopathogens and enhance growth of several crops (Waqas et al 2015

130

Veja et al 2008 Bahar et al 2011 Mendoza and Sikora 2009) Moreover

commercial application of Aspergillus spp Penicillium spp and Chaetomium spp for

the making of bioactive compounds that reveal antimicrobial and fungicidal activities (

Wang et al 2012 Jouda et al 2014)

In crop plants fungal endophytes are slightly recognized to play a role in the

production of gibberellins and resistance to stress abiotically Abiotic stressors like

drought heat and salinity symbiotic fungi can help plants to minimize the effect of

these stresses (Rodriguez et al 2008) In coastal plants fungal strains of P

funiculosum and P janthinellum are produced resistance against salt stress (Khan et al

2011 2013) Endophytic P citrinum produced gibberellins for their plant host (Khan et

al 2008) For plant growing stages with leaf enlargement pollen growth seed

sprouting stem elongation gibberellins are essential (Achard et al 2009) and influence

the growth of plant and adjustment throughout the early stages Thus endophytic fungi

possibly support their host plant to take nutrients and also stimulate hosts

growth The Trichoderma spp as considered to a giver of resistance facilitating plant

protection (Rubini et al 2005 Verma et al 2007 Bailey et al 2009 Kurose et al

2012) In this report cell free filtrates of culture and their fractions of endophytic

Penicillium exposed significant Escherichia coli Staphylococcus aureus Salmonella

typhimurium antibacterial activity against Bacillus subtilis Staphylococcus aureus and

Pseudomonas aeruginosa by forming inhibition zone in disc diffusion method

Endophytic Penicillium are also effective against bacterial pathogens with root rotting

fungi (Manmeet and Thind 2002) assessed antagonistic activity of Bacillus subtilis

Pseudomonas aeruginosa Trichoderma harzianum and Penicillium notatum against

causative agent of the bacterial blight of rice caused by Xanthomonas oryzae pv

oryzae in vitro and results showed that B subtilis P fluorescens and T harzianum

stop the growth of pathogen Our findings are an agreement to (Korejo et al 2014)

They reported that cell free filtrates of culture of endophytic Penicillium spp revealed

antifungal and antibacterial potentail Against a humen pathogen Vibriocholerae

(MCM B-322) produced desease cholera the cell free culture of P

chrysogenum revealed significant potential (Devi et al 2012) Many fungal endophytes

are the main source to secrete bioactive compounds (Stinson et al 2003 Corrado and

131

Rodrigues 2004 Ezra et al 2004 Kim et al 2004 Liu et al 2004 Wiyakrutta et al

2004 Atmosukarto et al 2005 Chomchoen et al 2005 Li et al 2005) Among them

seven isolates such as Hypocreales sp PSU-ES26 isolated

by C serrulata Trichoderma spp PSU-ES8 and PSU-ES38 isolated by H ovalis

and Penicillium sp PSU-ES43 Fusarium sp PSU-ES73 Stephanonectriasp PSU-

ES172 and an unidentified endophyte PSU-ES190 isolated by T hemprichii revealed

strong antimicrobial potential against human pathogens (Supaphon et al 2013) There

is eager requirement to discover novel drugs because of infectious diseases and drug

resistance microbes developing day by day Endophytic Penicillium could be a new

origin of treatments for the diseases caused by pathogens

In infectious plants fungal endophytes released the biotic stress with time

duration of 3 6 and 12 day after treatment by lowering the concentration of jasmonic

acid and salicylic acid as compare to control diseased plants Moreover these findings

reported the Penicillium citrinum (LWL4) relationship had a improved helpful impact

on plants of sunflower than Aspergillus terreus LWL5(Waqas 2015) Endophyte

naturally occurring in plants provide defense to plants by different way of mechanisms

such as the secretion of toxicant for pathogens and occasionally to disrupt the cell

membrane causing cell death of the pathogen (Ganley et al 2008 Shittu et al 2009)

Researche reported the justification of the pathogenic infections through the application

of fungal endophytes in plants like F verticillioides (Lee et al 2009) non-pathogenic

mutants of Colletotrichum magna (Redman et al 1999) Xylaria sp (Arnold et al

2003) Colletotrichum specie Fusarium nectria specie and Colletotrichum

gloeosporioides Clonostachys rosea and Acremonium zeae (Poling et al 2008)

Botryosphaeria ribis and (Mejıacutea et al 2008) In current research we assumed that the

application of endophytic Penicillium in plants might protect plants from adverse

effects of the soil born root-rotting fungi The inoculation of endophytic fungi may

inhibit the development of initial infection and prevent disease in this way not only

disease severity decreased but enhanced growth of the plant and yield (Mei and Flinn

2010) Our reseach shows that during pathogenic infection and mutual associations of

the endophytes lower the incidence of disease and improved the yield and biomass of

the plants Promotion of the host plant growth and inhibition of plant pathogen

132

infection may be increase the absorbance of nutrient which causes improved biomass of

plant and growth (Muthukumarasamy et al 2002) In the current study endophytic

Penicillium limited root-rot disease and also promote the health of the plants as

compare to control plants These are the comparision of the results as described by

Serfling et al (2007) The results similar to earlier findings on the plant growth

enhancement by endophytic fungi (Hamayun et al 2010 Khan et al 2011 2012

2013)

Endophytic P cyclopium Penicillium corylophilum P funiculosum are

recognized as GA-producers (Hasan 2002 Khan et al 2011) P citrinum (Khan et al

2008) Penicillium specie (Hamayun et al 2010) Resistance against insect attack and

pathogens enhanced by GA-producing endophytes which alter defense hormones such

as JA and SA In terms of abiotic stress (drought heat stress and salinity) these

endophytes may change the level of abscisic acid and induce resistance Endophytes

may have influencial role 0n the production of biochemicals and alter antioxidant

activities which is the main cause of improving growth of the plants(Waller et al

2005 Hossain et al 2007 Khan et al 2012 Waqas et al 2012 Khan et al 2013)

Chemical fertilizer showed negative impact on plants status The wide

applications of these inorganic fertilizers also causes deterioration to the soil fertility

by losing physiochemical and biological features of soil (Altuhaish et al 2014) In

addition a harmful effect on environment the chemical fertilizers have low level of

efficacy which may reduce nutrients uptake by the plants (Adesemoye et al 2009)

Application of organic amendments is sound known for inhibition of soil-borne

infections improving crops and yield (Ehteshamul-Haque et al 1996 Ikram and Dawar

2015 Sultana et al 2011 Lazarovits 2001 Stone et al 2003) Organic amendments

showed significant effects on crop health and production not only as a result of inhibiting

inoculum of soil pathogens but improve soil quality (Bailey and Lazarovits 2003)

Organic amendments including green manure peats and composts animal manure has

been proposed to sustain and improve fertility of soil and also soil structure for

conventional biological systems of agriculture (Cavigelli and Thien 2003 Magid et al

2001 Conklin et al 2002) and reduce occurrence level of the infections due to soil

133

containg plant pathogens (Noble and Coventry 2005 Litterick et al 2004) It is exposed

that organic amendments can be active against damages produced by fungal pathogens

such as Verticillium dahliae (Lazarovits et al 1999) Rhizoctonia solani (Diab et al 2003)

Phytophthora spp (Szczech and Smolinacuteska 2001) Pythium spp (Veeken et al 2005

MCKellar and Nelson 2003)Sclerotinia spp (Lumsden et al 1983 Boulter et al 2002)

Thielaviopsis basicola (Papavizas 1968) and) Fusarium spp (Szczech 1999) In current

research use of organic amendments like neem cake cotton cake and mustered cake

alone or with combine application of Penicillium spp significantly (plt005) increase

plant growth and cause growth reduction of root rotting fungi as compared to carbendazim

Population of total fungi and bacteria increased by organic soil amendment

which inhibit pathogens growth due to loss of ability to compete with beneficial

microbes (Gilbert et aI 1968) In our study a positive influence of numerous oil cakes

such as cake of neem and mustard on growth of plant was observed which is as

simillar as the findings of the Pandey et al (2005) and Goswami et al (2006) who

reported the use of different oil cakes such as neem and mustards in soil which showed

positive effects on growth of plant

Mixtures of Penicillium with various organic amendments applied in our study

resulted increasing the effectiveness of beneficial microobes for suppressing the fungi

causing the root rots in the present study This is same as the results of (Van Gundy

1965 Oka 2010) who described the combine effect of oil cakes and Pesturia penetrans

which change the soil features might be due to affect on nematode behaviours

(hatching movement and survival) Soil amendment resulting the decrease of the

occurrence of root knot nematodes and Fusarium spp on mung bean plants

(Ehtashamul-Haq et al 1993) Decomposition process of organic amendment released

sunbtances which produced antagonists and resistance too (Lumsden et al 1983)

which promote the inhibition of pathogen T harzianum used as a biocontrol agent with

neem cake showed significant infection on the reduction of Fusarium spp and

improved the development of plants (Nand 2002) Combine application of organic

amendment and PGPR might be resulted reduction of root-rot infections and fungal

pathogens with improved soyabean production (Inam-ul-Haq et al 2012)

134

Among agricultural fertilizer such as neem (Azadirachta indica) and its

products broadly described as a potential fertilizer (Gajalakshmi and Abbasi 2004) and

fungal diseases controlled by them (Dubey et al 2009 Amadioha 2000) insect pests

(Schmutterer 1995Ascher 1993) nematodes which parasitized by plant (Akhtar and

Mahmood 1995) bacteria (Abbasi et al 2003)) Some Studies have been revealed the

surprising potentail of neem products like neem seed oil against R solani M

phaseolina F moniliforme and (Niaz et al 2008) neem seed kernel extract against

Alternaria alternate Trichothecium roseum Monilinia fructicola Penicillium

expansum and Monilinia fructicola (Wang et al 2010) neem seeds and neem leaves

extract for control of F oxysporum Sclerotinia sclerotiorum and R solani (Moslem

and El-Kholie 2009) In our study neem cake mustard cake and cotton cake separate

or within combination of endophytic Penicillium which significantly (plt005) inhibit

the root rotting fungi and increasing the growth of plant Reduction in pre and post

emergence mortality of cotton and in the occurrence of R solani M phaseolina showed

by neem cake which is commonly used as a natural pesticide(Vyas et al 1990 Jeyara-

Jan et al 1987) Multiple nutrients which are having capacity to improve soil

characteristics are found in organic materials (Orrell and Bennett 2013) They also

provide organic substances like acids that help to breakdown soil nutrients and make

them easily accessible for the plants (Husson 2013)

Use of pesticides for reduction of root rotting fungi and plant parasites is costly

approach and resulting destruction of soil environment (Sukul 2001) Use of

bantagonist is an efficient way to overcome root rotting fungi and lethal nematodes

(root knot) (Whapham et al 1994 Ehteshamul-Haque et al 1995 1996) Usually

suppression of the plant pathogens occured by the direct secretion of toxicant such as

phenolic compounds and indirectly enhancing soil microbes by the application of soil

amendments (Shaukat et al 2001Ali et al 2001) In the present report selected

isolates of endophytic Penicillium separate or mixed use with Carbendazim Feast-M

and Topsin-M not only significantly inhibited the infection of root rooting fungi and

enhanced the growth of sunflower but mixed application also produced additional

defense against pathogen penetration and promote growth Plant centered toxicant

within organic amendments revealed promising outcomes in the management of root

135

infecting fungi present in soil (Ghaffar 1995) Organis amendments give better

environment to soil by providing energy and nutrients which support microbes and

plants to grow and survive successfully (Drinkwater et al 1995) Combination of

beneficial microbes by means of various plant colonizing forms with organic

amendment may be convenient for the inhibition of diseases by using different

biocontrol mechanisms for phytopathogens Combine application of different strains of

PGPR resulted significant inhibition of cucumber pathogens consistently (Raupach and

Kloepper 1998)

For crop protection one of the most favorable alternative approach is activation

of resistance within plant among current strategies (Walters and Fountaine 2009

Anderson et al 2006 Walters et al 2005) These alternative stratigies does not kill

phytopathogen directly (Walters and Fountaine 2009) but encouragement of natural

defence system of plant which introduces systemic acquired resistance (Vallad and

Goodman 2004) In case of abiotic and biotic stress a broad series of bioactive

compounds are release by the plant in natural environment that are injurious to

pathogens and grazing animals Phenolic phytochemicals are basic constituents of fruits

and vegetable of bioactive compounds that function as a resistant against insect and

herbivores (Stevenson et al 1993) Due to their significant protective biological role

phenolic compounds are pervasive in all plants so found in all nutrients In plants

resistant reaction of phenols resulting in the separation of phytopathogens which are

categorized due to the quick and early accumulation of phenolics at the infection site

(Cheacuterif et al 1991)

Phenolic compounds are impotant bioactive metabolites can act as antioxidants

against oxidative stress which leads many benefits to plants (Urquiaga and Leighton

2000 Grassmann et al 2000) also termed as free radical- scavengers Phenolic

compounds and antioxidants have close relation (Kumar et al 2008) Phenolic and

lycopene compounds are carotenoids a big source of antioxidants present in tomatoes

richly (Pinela et al 2011 Sahlin et al 2004 Ilahy et al 2001 George 2004)

Organic tomatoes are economically important with relation to conventional tomatoes

(Kapoulas et al 2011) due to their improved quality and ecofriendly nature Phenolic

136

compounds gives better taste as compared to conventional fruits (Benbrook 2005) In

our research better quality of okra and tomato fruits are produced by endophytic

Penicillium as compared to chemical fungicides and control in both screen house and field

condition

In the present study endophytic Penicillium not affected pH of fruit juice of

okra and tomato compared to untreated plant fruits Our findings were in line with (Oke

et al 2005 Carrijo and Hochmuth 2000) who described that pH of tomato fruit juice

not changed by phosphorus use Combine use of endophytic Penicillium with

Psuedomonas montellii improved TSS (total soluble solids) and tritable Acidity of okra

fruit Total soluble solids consist of acids sugars and other constituents existing in THE

fruits of the tomato (Balibrea et al 2006) Instead of inorganic fertilizer application of

biocontrol agents significantly increased brix content in tomato (Oke et al 2005)

The improved quality of fruit Ash content due to the high utilization of the nutrients

of the soil (Mauromicale et al 2011) The variation present in total soluble solids might

be due to the variability of the gene(Riahi et al 2009) In addition of chemical fertilizer

to soil had a significant function in food safety but however made soil harder that

resulted destruction in soil quality (Lai et al 2002) and the soil mineral absorption

decreased through roots Similarly from the soil availability or absorption of mineral

nutrients due to greater moisture content that improved prescence of mineral in soil

(Van veen and Kuikman 1990)

In the present research application of endophytic Penicillium significantly

impoved the carbohydrate protein antioxidant and polyphenol contents of the tomato

and okra fruits The increment of root surface area ultimately increased water

absorption and nutrient uptake due to endophytic Penicillium increased the above

contents These findings are an agreement with Rashed (2002) who described that

antagonistic microbes improved nutrient uptake (El-Ghadban et al 2002)

The biofertilizers impact positively on okra fruits was confirmed by previous

studies described by (Adediran et al 2001 Adejumo et al 2010) The photosynthetic

activity will also be improved as a consequence of improved interception of light when

137

all nutrient is in the right proportion (Subbarao and Ravi 2001) which ultimately

improves vegetative growth and efficient transport of photosynthetic product from

source to sink

Therapeutic effects of active compounds from fungal source have been noticed

from several years and new drugs have exposed and obtained extracted from the

endophytic fungi (Teakahashi and Lucas 2008 Hormazabol et al 2005) A new

endophytic fungus Muscodor albus was isolated from cinnamon tree (Cinnamomum

zeylanicum) formed volatile compunds that executes fungi causing disases (Strobel et

al 2001 Strobel 2006) (Liu et al 2013 Raghunath et al 2012) has discoverd two

new compouds named as nigerasterols A 6 8 (14) 22-hexadehydro-5α9 α-epidioxy-

315-dihydroxy sterols and B from endophytic fungi (Aspergillus niger)

23 compounds were isolated from endophytic Penicillium regulosum mycelia

Hexane fraction of mycelium were characterized by GCMS to identify the chemical

compounds most of them are hydrocarbon fatty acid alcohol and benzene derivatives

Some compounds were characterized from our isolate such as Widdrol hydroxyether

Eicosane Oleic acid Ethyl Oleate and 2-Aminofluorescein Because of the prescence of

these chemical compounds this fungus might have a capability to act against pathogenic

bacteria and fungi and showed a promising result against both type of bacteria such as

gram-ve and gram +ve

Adametizine A produced by Penicillium sp having antibacterial activity against

Aeromonas hydrophila Vibrio harveyi Staphyloccocus aureus Vibrio parahaemolyticus

and antifungal activity against Gaeumannomyces graminis (Liu et al 2015) Arisugacin

K produced by Penicillium sp having antibacterial activity against Escherichia coli (Li et

al 2014) Cillifuranone produced by Penicillium sp having antibacterial activity against

Xanthomonas campestris and antifungal activity againsts Septoria tritici (Wiese et al

2011) Comazaphilones produced by Penicillium sp having antibacterial activity against

S aureus Pseudomonas fluorescens Bacillus subtilis (Gao et al 2011) Communol A

FndashG produced by Penicillium sp having antibacterial activity against Enterobacter

aerogenes E coli (Wang et al 2012) Conidiogenone B produced by Penicillium sp

138

having antibacterial activity against Pseudomonas fluorescens Pseudomonas aeruginosa

Staphylococcus epidermidis S aureus mr and antifungal activity against Candida

albicans (Gao et al 2011) Dictyosphaeric acid A produced by Penicillium sp having

antibacterial activity against S aureus Enterococcus faecium S aureus mr and

antifungal activity against C albicans (Bugni et al 2004) Isocyclocitrinols produced by

Penicillium sp having antibacterial activity against Enterococcus durans S epidermidis

(Amagata et al 2003) Peniciadametizines produced by Penicillium sp having antifungal

activity against Alternaria brassicae (Liu et al 2015) Penicifuran A produced by

Penicillium sp having antibacterial activity against Bacillus cereus Staphylococcus

albus (Qi et al 2013) Penicilactone produced by Penicillium sp having antibacterial

activity against S aureus mr (Trisuwan et al 2009) Penicimonoterpene produced by

Penicillium sp having antibacterial activity against E coli A hydrophila S aureus

Micrococcus luteus V parahaemolyticus and V harveyi (Zhao et al 2014) and

antifungal activity against A brassicae Aspergillus niger Fusarium graminearum (Gao

et al 2011 and Zhao JC et al 2014) Penicisteroid A which is produced by Penicillium

sp having strong antifungal activity in response to A brassicae A niger (Gao et al

2011) Penicitide A which is produced by Penicillium sp having stronge antifungal

activity in response to A brassicae A niger (Gao et al 2011) Penicyclones AndashE islated

from Penicillium sp having antibacterial activity against S aureus (Guo et al 2015)

Perinadine A which is produced by Penicillium sp having antibacterial activity against

B subtilis M luteus (Sasaki et al 2005) Pinodiketopiperazine A produced by

Penicillium sp having antibacterial activity against E coli (Wang et al 2013)

Scalusamide A produced by Penicillium sp having antibacterial activity against M luteus

and antifungal activity against Cryptococcus neoformans (Tsuda et al 2005) Terretrione

D produced by Penicillium sp having antifungal activity againsts C albicans (Shaala

LA et al 2015) and Xestodecalactone B produced by Penicillium sp having antifungal

activity againsts C albicans (Edrada et al 2002) These references supports our results

that our isolate have antimicrobial activity It also have showen a positive result on the

growth of the by enhancing the plant growth and also suppressing infection of root rot

fungi almost in all crops which are experimented

Conclusion

139

There is eager need for natural (environment friendly) chemotherapeutic and

agrochemical agents instead of synthetic toxic chemicals Natural products produced by

endophytes have been tested against infectious agents against plant pathogens One of the

single greatest challenge is control of soil-borne pathogens including parasitic nematodes

facing recent agriculture worldwide Soil-borne fungi and fungi like organisms

including Macrophomina phaseolina Fusarium species Phytophthora spp

Rhizoctonia solani and root knot nematodes commonly (Meloidogyne species) result

severe economic damages both in greenhouse and field production system In

agricultural and pharmaceteucal industry application of endophytes with their related

benefits has now been new approach in rescent years Despite the assistances related to

endophytic bacteria and fungi in plant disease management they are still largely

unexplored Genus Penicilium has been familiar for their significant secretion of

secondry metabolites among them and was also found to play important function in

plants against stress tolerance Penicilium spp secrete a variety of pharmaceutically

vital compounds with antibacterial antifungal insecticidal and nematicidal activities

In this study endophytic Penicillium isolated from healthy plants revealed

significant potential against root infecting fungi both in field condition and screen house

Although endophytes are now widely used in other different fields

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ii

EVALUATION OF BIOCONTROL POTENTIAL OF ENDOPHYTIC SPECIES OF


NEMATODE

FAIZAH UROOJ


KARACHI PAKISTAN

2018

iii



NEMATODE


Botany

By

FAIZAH UROOJ


KARACHI PAKISTAN

2018

iv

DEDICATION



v


1 INTRODUCTION

11 Endophytic fungi










from different host

















vi




suspension







216 Fruit analysis

2161 pH




2165 Firmness

2166 Total solids

2167 Protein

2168 Carbohydrate








Penicillium


Penicillium


vii

Penicillium spp



culture filtrates


culture filtrates


culture filtrates


culture filtrates


rugulosum



















viii





okra plants


tomato plants






4 DISCUSSION

ix



NEMATODE

SUMMARY


























x






















xi

xii

1

1 INTRODUCTION

11 Endophytic fungi




























2

















2014)



spiteller 2012b)











3




(Huang et al 2008)

























plants

4































5
































6






























7






























8






























9



endophytic fungi






10







about (24) hours





















11
















ordmC for 16 min



calibrate the sizes





12





phaseolina)




culture plate assay







times










13




in vitro

















measured in mm

14























Penicillium species





15












regulosum





packard (5890)






16





measured



(2016) was used





polyphenol activity










17









-------------------------------------------------------









sucrose

2165 Firmness



2166 Total solids


moisture from 100

18


2167 Protein


2168 Carbohydrate













used

19





























20

21

22












(Table 1) fig1-7

23







(Solanaceae)

Root 4 4 20 20




(Solanaceae)

root 5 25 16 21


(Amaranthaceae)

root 3 12 11 20


(Meliaceae)

root 5 25 13 20




(Amaranthaceae)

stem 18 13 11 14



(Malvaceae)

root 5 8 5 6


(Cucurbitaceae)

leaf 18 13 11 14


(Solanaceae)

root 5 24 17 22


(Malvaceae)

root 5 3 21 12

24


assay


assay

25


assay



26


assay


assay

27


assay















28













29





Control 0 0 0 0


EPSMR1 P citrinum




EPSMS2 Plilacinum








EPSLR4 P nigricans



30




EPAAR5 P rugulosum




EPAIR6 P decumbens








EPCTS8 Prestrictum




EPASS9 P duclauxi


31





EPHAL10 Pasperum




EPAER11 Pthomii




EPMCL12 P lividum




EPSLR13 P javanicum





32






33



























34


species



Control 0 0 0 0


EPSMR1 P citrinum




EPSMS2 Plilacinum








EPSLR4 P nigricans


35





EPAAR5 P rugulosum




EPAIR6 P decumbens








EPCTS8 P restrictum




EPASS9 P duclauxi

36






EPHAL10 Pasperum




EPAER11 Pthomii




EPMCL12 P lividum




EPSLR13 P javanicum




37







38





spp










A

B

C

E D

39


48 hours


24Hours 48Hours
















40

41























42


Penicillium species



Control 0 0 0 0


EPSLR4 P nigricans




EPAAR5 P rugulosum




EPAIR6 P decumbens






43



EPHAL10 Pasperum




44














45





Control 0 0 0 0 0


EPSLR4 P nigricans



60 microldisc 11 11 9 12 10

EPAAR5 P rugulosum

20microldisc 0 18 18 11 18

40 microldisc 0 21 18 11 22

60 microldisc 0 24 20 18 22

EPAIR6 P decumbens



60 microldisc 0 12 16 0 11




46


EPHAL10 Pasperum


40 microldisc 11 9 6 10 10

60 microldisc 12 11 9 10 12

47





C

+ve C

20microl

60microl

40microl

+veC

20microl

40microl

60microl

C

48





C

60microl

40microl

20microl +veC

vCCe

veve

+veC

vCCe

veve

C

60microl

20microl

40microl

49


Penicillium species



Control 0 0 0 0


EPSLR4 P nigricans




EPAAR5 P rugulosum




EPAIR6 P decumbens







50


EPHAL10 Pasperum








(Table 8 and Fig12)

51





Control 0 0 0 0 0


EPSLR4 P nigricans

20microldisc 16 16 14 14 16

40 microldisc 16 16 14 14 18

60 microldisc 18 16 16 14 20

EPAAR5 P rugulosum

20microldisc 18 18 20 11 20

40 microldisc 18 18 20 11 21

60 microldisc 18 18 20 11 21

EPAIR6 P decumbens






52



EPHAL10 Pasperum




53

4



C

+ve C

20microl 40microl

60microl

54


Penicillium

















55

(1) Nanodecane

(2) Nonadecane

(3) Heptadecane

(4) Heptacosane

(5) Heptacosane

(6) Eicosane

(7) Octadecane

(replib) Nonadecane

50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 2400

50

10057

71

85

99113 127 141 155 169 183 197

(replib) Nonadecane

60 80 100 120 140 160 180 200 220 240 260 2800

50

10057

71

85

99113 127 141 155 169 183 197 268


50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 2500

50

10057

71

85

99113 127 141 155 169 182 196 210 240


60 80 100 120 140 160 180 200 220 240 260 280 300 320 3400

50

10057

71

85

99113 127 141 155 169 183 197 211 225 239 253 267 281 294 308 322 336


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10057

71

85

99113 127 141 155 169 183 197 211 225 239 253 267 281 294 308 322

(mainlib) Eicosane

60 80 100 120 140 160 180 200 220 240 260 2800

50

10057

71

85

99113

127 141 155 169 183 197 211 225 238 252 282

(replib) Octadecane

60 80 100 120 140 160 180 200 220 240 2600

50

10057

71

85

99113 127 141 155 169 183 197 210 225 254

56








50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 2400

50

100

55

60

69

73

8397 115

129

138

143157

171

185

199209

228

OH

O


60 80 100 120 140 160 180 200 220 240 2600

50

10057

71

85

97

113127

141 155 168183 197 212


50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 2500

50

100

60

71

8397

102

111

129

143157

171

185

199

211

228

O

O


60 80 100 120 140 160 180 200 220 240 2600

50

10057

60

71

83 97

102

111 129

143153 165

177

185

195 213225

242O

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

61

70

88

101

115129

143

157

171 185 199 213

227

241 255

270

O

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

69

81

95 109

123

135

140

150

167

177 205223

238

O

OH

57



(16) Oleic Acid



60 80 100 120 140 160 180 200 220 240 2600

50

10060 73

8397

115

129

143157 171 185

199

213

227 239

256

OH

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

61 73

88

101

115129 143

157

171 185 199 213 225239

255 267284

O

O


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

100

55

6781

95

109

123135 150 164 178

192 205 220 234

263

279

308

O

O

(replib) Oleic Acid

60 80 100 120 140 160 180 200 220 240 260 2800

50

10055

69

83

97

111

125137 151 165 180 193 207 222 236

264

282

HO

O

58

(18) Ethyl Oleate





60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10055

6983

97

111123

137 155180

194 207

222

236

264

281

310

O

O


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10055

6983

97

111

125137 151 165 179 194 207 221 236 249 261

278296

HO

O


60 80 100 120 140 160 180 200 220 240 260 280 3000

50

100

55 77 90 103118

133

147

161

178

191 262290

O

O

O


60 90 120 150 180 210 240 270 300 330 360 3900

50

100

5770

83 104132

149

167

279

O

O

O

O


60 90 120 150 180 210 240 270 300 330 360 3900

50

100

55

69

81

95

107

121

147

173189

215

231

243

258

286

O

59





50 100 150 200 250 300 350 400 450 500 550 600 6500

50

100

63 91

151

189

287

303

318 347

O

O

OHO OH

H2N

60


SNo Scan

No

Systemic Name

(Common Name)

Mol

Formula

Mol

Wt

Ret

Time

Conc

1 2606 Nanodecane C19H40 268 24168 0036

2 2913 Heptadecane C17H36 240 2641 0035


4 3230 Octadecane C18H38 254 28737 0049





9 3653 Nonadecane C19H40 268 31834 0064




13 4252 Eicosane C20H42 282 36220 0093

14 5475 Oleic Acid C18H34O2 282 45175 0105


16 5546 Ethyl Oleate C20H38O2 310 45694 0065

61


18 6023 Heptacosane C27H56 380 49187 0051


20 6281 Heptacosane C27H56 380 51076 0044




C20H30O 286 53910 004


62





























63





















64



Infection


Fsolani



Control hellip 50 187 75 25 75 50 187 125





P citrinum EPSMR1 375 0 25 0 125 25 0 0



Pduclauxi EPASS9 50 0 62 0 312 312 62 0

Plividum EPMCL12 50 62 50 0 0 50 0 62



Pthomii EPAER11 62 0 62 0 375 187 62 62



Pasperum EPHAL10 125 0 25 187 375 312 62 62






65



Shoot Weight


(cm)

(g)

(cm)

(g)


Control 22775 3993 253 535 643 1162 0645 0675

Carbendazim 2585 418 2216 451 742 1287 0715 0622




P citrinum EPSMR1 2599 4681 218 51 94 862 0726 0807



Pduclauxi EPASS9 2541 4487 243 512 1103 1406 077 0786

Plividum EPMCL12 22685 4587 205 539 631 558 0663 0578



Pthomii EPAER11 2824 44 277 527 1221 1218 1005 0645



Pasperum EPHAL10 26186 4918 214 678 918 757 069 0866

LSD005 5141 7881 07911 1821 2551 2821 01951 031



66




















67

68



69


sunflower


+veControl EP

EP

EP EP EP EP

EP

Control

70


























71



Infection


Fsolani



Control 50 187 50 25 75 75 187 125

Carbendazim 125 62 312 62 125 25 62 62




P citrinum EPSMR1 375 0 25 0 125 25 0 0



Pduclauxi EPASS9 25 0 62 62 312 187 62 0

Plividum EPMCL12 50 62 50 0 0 50 0 62



Pthomii EPAER11 62 0 62 0 375 187 62 62



Pasperum EPHAL10 125 125 25 187 312 312 62 62






72


Treatments Code

Shoot Length

(cm)

Shoot Weight

(g)


(cm)

(g)


Control 3256 3893 378 642 57 1034 085 131

Carbendazim 3781 4293 452 607 84 1025 124 128




P citrinum EPSMR1 385 6443 373 1425 75 787 088 226



Pduclauxi EPASS9 4412 6275 386 1013 7 768 086 213

Plividum EPMCL12 345 6551 206 1019 706 645 072 161



Pthomii EPAER11 4838 6208 489 953 863 656 096 141



Pasperum EPHAL10 4158 5006 362 818 6102 1275 067 186

LSD005 10331 8971 2271 5521 3021 2171 04581 1071




73


in sunflower











after 45 days

EP

Carbendazim Control

74


















75



Infection


Fsolani



Control hellip 50 312 100 75 100 50 0 562





P citrinum EPSMR1 62 62 437 75 0 0 62 62



Pduclauxi EPASS9 0 0 437 375 25 375 62 25

Plividum EPMCL12 62 25 25 687 125 375 62 50



Pthomii EPAER11 62 62 437 25 125 312 0 0



Pasperum EPHAL10 435 125 25 25 25 187 0 0






76



Shoot Weight


(cm)

(g)

(cm)

(g)


Control hellip 1375 1714 078 08 1531 4652 051 014





P citrinum EPSMR1 1299 1606 059 0617 165 477 039 016



Pduclauxi EPASS9 1187 1916 069 0855 1108 4562 017 016

Plividum EPMCL12 132 2147 061 1358 2252 4785 026 022



Pthomii EPAER11 1463 179 077 1203 1125 7025 031 024



Pasperum EPHAL10 1463 1874 077 1102 1125 702 031 02

LSD005 1611 4011 0191 2141 8421 1151 0171 0071




77

















solani (Table 16)




78



Infection


Fsolani



Control hellip 50 50 100 75 100 75 0 187





P citrinum EPSMR1 62 25 437 437 0 437 62 187



Pduclauxi EPASS9 0 312 437 562 25 562 62 65

Plividum EPMCL12 62 125 25 25 125 25 62 0



Pthomii EPAER11 62 187 437 437 125 375 0 0



Pasperum EPHAL10 437 375 25 312 25 562 0 125






79


Treatments Code

Shoot Length


(cm)

(g)

(cm)

(g)


Control hellip 1375 1364 078 089 1531 613 051 031





P citrinum EPSMR1 1299 1318 059 193 165 961 039 037



Pduclauxi EPASS9 1187 1438 069 13 1108 1178 017 048

Plividum EPMCL12 132 1323 061 107 2252 1024 026 048



Pthomii EPAER11 1463 1435 077 119 1125 1185 031 071



Pasperum EPHAL10 1463 1453 077 128 1125 972 031 046

LSD005 1611 2661 0191 091 8421 271 0171 0291



80


mungbean growth


















81




Infection

Control --------- 50 100 100 50

Carbendazim --------- 25 50 50 62
















82



(cm) (g) (cm) (g)

Control ---------- 1475 0522 4972 0098

Carbendazim --------- 1635 0987 3737 009




P citrinum EPSMR1 1344 0987 4617 0137



Pduclauxi EPASS9 1434 0696 4127 0096

Plividum EPMCL12 1639 0752 4147 0121



Pthomii EPAER11 1482 0711 3865 0072



Pasperum EPHAL10 1608 0787 3875 0066

LSD005 2891 0261 0741 0051



83

84
























85



Infection


Fsolani



Control hellip 437 312 625 625 312 0 312 0





P citrinum EPSMR1 0 50 62 625 625 62 75 0



Pduclauxi EPASS9 0 0 62 25 187 125 50 62

Plividum EPMCL12 50 437 437 562 375 0 687 62



Pthomii EPAER11 187 562 312 562 50 312 562 0



Pasperum EPHAL10 62 312 125 562 25 62 812 0






86



(cm) (g) (cm) (g)


Control hellip 12 1544 18 407 126 333 155 063





P citrinum EPSMR1 1732 1755 297 389 922 1149 064 056



Pduclauxi EPASS9 1672 2255 243 636 1185 597 057 11

Plividum EPMCL12 1307 1303 178 254 1242 529 052 046



Pthomii EPAER11 1328 1375 214 234 148 466 046 055



Pasperum EPHAL10 1328 2412 18 732 1225 775 06 126

LSD005 271 5171 0691 2091 3731 3031 1031 0631



87


EP

88























89



Infection


Fsolani



Control hellip 437 50 625 25 312 62 312 0





P citrinum EPSMR1 0 62 62 562 625 0 75 0



Pduclauxi EPASS9 0 562 62 562 187 25 50 0

Plividum EPMCL12 50 187 437 375 375 0 687 0



Pthomii EPAER11 187 312 312 25 50 125 562 0



Pasperum EPHAL10 62 125 125 50 25 62 812 0






90




















weeks

91


Treatments Code

Shoot

Length

Shoot

Length

Shoot

Weight

Shoot

Weight

Root

Length

Root

Length

Root

weight

Root

weight

(cm) (cm) (g) (g) (cm) (cm) (g) (g)


Control hellip 12 1633 18 554 126 1757 155 105





P citrinum EPSMR1 1732 1912 297 512 922 9 064 155



Pduclauxi EPASS9 1672 214 243 69 1185 153 057 237

Plividum EPMCL12 1307 2347 178 741 1242 1298 052 156



Pthomii EPAER11 1328 225 214 657 148 155 046 164



Pasperum EPHAL10 1328 2101 18 525 1225 1095 06 135

LSD005 271 4291 0691 3281 3731 5851 1031 091


92


cake in tomato

EP

93





















94



Infection


Fsolani



Control hellip 375 0 50 125 437 375 25 0





P citrinum EPSMR1 312 187 187 312 375 50 187 0



Pduclauxi EPASS9 187 375 125 25 375 50 0 0

Plividum EPMCL12 62 62 437 125 62 625 25 0



Pthomii EPAER11 0 187 437 187 62 25 0 0



Pasperum EPHAL10 125 62 50 125 125 812 0 0






95



(cm) (g) (cm) (g)


Control hellip 2369 2225 274 837 274 975 211 303





P citrinum EPSMR1 267 3384 313 668 2397 975 394 098



Pduclauxi EPASS9 2925 2911 376 388 3037 1293 522 116

Plividum EPMCL12 2768 2801 31 698 2155 1132 35 109



Pthomii EPAER11 239 30 296 799 2416 1062 427 125



Pasperum EPHAL10 2856 2891 344 763 1921 1352 306 13

LSD005 471 4931 0941 3331 7321 5451 1611 11071



96



EP

S

EP

97


EP

EP

98


in chickpea


in chickpea

EP

EP

99


in chickpea













weeks













(Table 26)

100




101



Infection



Control hellip 375 125 50 312 437 0 25 187





P citrinum EPSMR1 312 0 187 625 375 187 187 312



Pduclauxi EPASS9 187 0 125 625 375 62 0 312

Plividum EPMCL12 62 62 437 100 62 25 25 312



Pthomii EPAER11 0 62 437 125 62 62 0 62



Pasperum EPHAL10 125 0 50 187 125 0 0 0






102



(cm) (g) (cm) (g)


Control hellip 2369 2188 274 406 274 692 211 58





P citrinum EPSMR1 267 1916 313 556 2397 756 394 1172



Pduclauxi EPASS9 2925 192 376 561 3037 1115 522 819

Plividum EPMCL12 2768 1929 31 417 2155 946 35 383



Pthomii EPAER11 239 2305 296 626 2416 9 427 931



Pasperum EPHAL10 2856 1662 344 582 1921 925 306 834

LSD005 471 6131 0941 3011 7321 2921 1611 6151




103













weeks














104



Infection


Control 75 100 100 75

Feast-M 0 37 687 625










Topsin-M 0 437 812 62





P citrinum 437 687 100 0


P decumbens 187 50 437 187

Pasperum 125 50 562 125




105



Control 3197 339 288 288

Feast-M 4269 451 526 526





Carbendazim 3675 319 398 398





Topsin-M 3935 314 383 383





P citrinum 3268 249 432 432

Pnigricans 2788 201 401 401

P decumbens 3421 3007 446 446

Pasperum 3262 229 363 363

LSD005 5751 0811 1041 1041



106








days of germination



















107



Infection


Control 0 50 625 50








108



Fruits Fruit weight

(cm) (g) (cm) (g)

Control 3831 1058 1596 305 023c 708c

Carbendazim 3421 832 1659 546 045b 683c

P decumbens 4523 1167 1756 438 052a 1106a

Pnigricans 4265 1172 1794 188 054a 894b

P rugulosum 4592 1295 1967 2405 025c 533d

LSD005 511 4281 3431 581 00261 04841


109


soil



Control 137e 2711e 2878e 0053d





LSD005 00311 01361 04211 00041


110



Sucrose

Control 587c 0087c 8668d 1353b 245d





LSD005 0121 000571 0211 01031 0121


111


in garden soil








LSD005 10591 01441 21941 3711


112








after 90 days










37)








113



Infection


Control 312 100 937 562








114



(cm) (g) (cm) (g)

Control 52 1974 1816 35 30a 5801b

Carbendazim 4646 1322 1629 237 20c 4083a

P decumbens 443 2161 1283 418 2133c 995a

Pnigricans 55 1892 1561 315 32a 4286d

P rugulosum 5197 1695 1205 334 256b 4779c

LSD005 1481 18611 5391 4011 3781 0131


115


garden soil








LSD005 01081 01671 0301 00791


116



N Sucrose






LSD005 00621 00541 0211 1311


117










LSD005 5591 0471 5771 2391


118



field condition



















days








119



























120



Infection


45 90 45 90 45 90 45 90

Control 375 0 562 312 937 100 562 0

Topsin-M 375 0 625 25 937 100 687 0


P decumbens 62 0 50 375 68 100 375 0

Pnigricans 125 187 562 687 875 100 312 0

P rugulosum 312 62 562 375 812 100 437 0







121


condition


(cm)

Shoot Weight

(g)

Root Length

(cm)

Root Weight

(g)

Number

of Fruits

Fruit

weight

Control 45 90 45 90 45 90 45 90 90 90

Topsin-M 4178 6192 2228 4325 1368 2426 204 823 086g 246i

Pmontelii 422 6375 1765 4731 1267 2377 133 98 12f 31h


P nigricans 4233 6617 1971 4887 1486 2578 167 1003 143e 1146a

Prugulosum 4866 7083 1635 5095 1378 2311 172 967 176d 331g

P rugulosum 4373 7026 2063 2051 1371 2464 169 709 123f 35f




LSD005 961 1321 131 1181 3551 1371 0831 2961 0111 0111


122



Treatments

Polyphenol

microgml



Control 183h 7314e 7721e 007f

Topsin-M 146i 9119a 9886a 0113d








LSD005 00721 10191 06531 00121



123




microgml




Topsin-M 183f 4922f 5575g 0074bc








LSD005 10061 05191 04731 0003081


124





Control 5102g 646g 1466g 5966f

Topsin-M 5514f 716f 2566f 67e








LSD005 10451 06241 14081 2471


125



Treatments pH

Tritable

acidity

Moisture

content

Total

solids

Total Soluble

Solid

Sucrose

Control 624a 0102c 8774b 1222f 1425e

Topsin-M 619ab 0126b 8653e 1339b 1475e








LSD005 00641 00041 03021 0171 02221


126

127

128

4 DISCUSSION





























129































130
































131
































132







2013)
























133































134
































135








Kloepper 1998)























136




condition


























137



source to sink



























138































Conclusion

139



















REFERENCES

140



Eco 15(1) 37-47






University












Ecol 58 921-929








99-102



141


2933



















78(3) 429-435




New York NY






142























21(12) pp 1387-1397









143











Engelmann










Ecol 51(2) 215-229











144





Hortic Sci 35 67-72
























6(5) 162-166



145





Acad of Sci 102(35) 12465-12470










Pathol 111 101-112






41-48



4(1) 34-43







146



185(11) 3458-3468













5 17-24



356



Press)









147





Trop Sci 35 294-299






75270 Pakistan












717-732



Pathol J 2 75-79





148








255(7) 2751-2760







21 2894-2897









Protect 46 627-635








149



Phytopathol 58 1051






















Prod 78 2699-2703





150









207













43



106




Pathol 57 870-876

151



Resour 8(2) 1634-1645






8(6) 687-695
















63(4) 391-400



87 4-10




152



S00218596120005-12



Pak J Bot 47(4) 1547-1551




261







York Pages 86













209-214

153



York pp 660-662






April 1996



Lett 100(8) 084102














Plant Biol 13(1) 86





154





















BMC Microbiol 13 51





70-78




155

















289-295














88

156



Pathol 23 1-7






299(1) 31-37








553-558














157
































158


228 7382





Newslett 5 32-34



Pl Nutrit 22(4-5) 835-853





649



513-515 doi 101126science1136237



129 189-196











159





and Ecol 39(2) 163-165

















1048


Minia Univ







160



62















40 403-408













161



doi 103389fpls00432




32-34






516-522


















162


Toxicol 50(3-4) 829-834







1041722167-0412100022 5 of 7 Volume 5bull Issue 1bull 1000225 Med














3 342-351







163



















Mar 55 269-279












164















647






319-343










165








York 326-365




686



106(9) 996-1004





Mycol Res 106(9) 996-1004






13(4) 1698-1709





166






Biol Sci 4 1240- 1242












1623




16(4) 403-406










167





Interact 3(2) 75-93




Ecol 19 2917-2933







1037-1042





2950










168







43(1) 1-6














82










169























43-68







Sci 416 382-385

170




Bioch 37 21312140



Mycologia 98 31-42


















Sci 102(38) 13386-13391



95 1368-1373



147 523- 535

171





Res 4 1100-1104






Drugs 11 2230-2238




Interact 10(1) 280-287









3(1-2) 90-97






489

172




585




36(1) 82


















January 20 2017






173



449-458





iii



NEMATODE


Botany

By

FAIZAH UROOJ


KARACHI PAKISTAN

2018

iv

DEDICATION



v


1 INTRODUCTION

11 Endophytic fungi










from different host

















vi




suspension







216 Fruit analysis

2161 pH




2165 Firmness

2166 Total solids

2167 Protein

2168 Carbohydrate








Penicillium


Penicillium


vii

Penicillium spp



culture filtrates


culture filtrates


culture filtrates


culture filtrates


rugulosum



















viii





okra plants


tomato plants






4 DISCUSSION

ix



NEMATODE

SUMMARY


























x






















xi

xii

1

1 INTRODUCTION

11 Endophytic fungi




























2

















2014)



spiteller 2012b)











3




(Huang et al 2008)

























plants

4































5
































6






























7






























8






























9



endophytic fungi






10







about (24) hours





















11
















ordmC for 16 min



calibrate the sizes





12





phaseolina)




culture plate assay







times










13




in vitro

















measured in mm

14























Penicillium species





15












regulosum





packard (5890)






16





measured



(2016) was used





polyphenol activity










17









-------------------------------------------------------









sucrose

2165 Firmness



2166 Total solids


moisture from 100

18


2167 Protein


2168 Carbohydrate













used

19





























20

21

22












(Table 1) fig1-7

23







(Solanaceae)

Root 4 4 20 20




(Solanaceae)

root 5 25 16 21


(Amaranthaceae)

root 3 12 11 20


(Meliaceae)

root 5 25 13 20




(Amaranthaceae)

stem 18 13 11 14



(Malvaceae)

root 5 8 5 6


(Cucurbitaceae)

leaf 18 13 11 14


(Solanaceae)

root 5 24 17 22


(Malvaceae)

root 5 3 21 12

24


assay


assay

25


assay



26


assay


assay

27


assay















28













29





Control 0 0 0 0


EPSMR1 P citrinum




EPSMS2 Plilacinum








EPSLR4 P nigricans



30




EPAAR5 P rugulosum




EPAIR6 P decumbens








EPCTS8 Prestrictum




EPASS9 P duclauxi


31





EPHAL10 Pasperum




EPAER11 Pthomii




EPMCL12 P lividum




EPSLR13 P javanicum





32






33



























34


species



Control 0 0 0 0


EPSMR1 P citrinum




EPSMS2 Plilacinum








EPSLR4 P nigricans


35





EPAAR5 P rugulosum




EPAIR6 P decumbens








EPCTS8 P restrictum




EPASS9 P duclauxi

36






EPHAL10 Pasperum




EPAER11 Pthomii




EPMCL12 P lividum




EPSLR13 P javanicum




37







38





spp










A

B

C

E D

39


48 hours


24Hours 48Hours
















40

41























42


Penicillium species



Control 0 0 0 0


EPSLR4 P nigricans




EPAAR5 P rugulosum




EPAIR6 P decumbens






43



EPHAL10 Pasperum




44














45





Control 0 0 0 0 0


EPSLR4 P nigricans



60 microldisc 11 11 9 12 10

EPAAR5 P rugulosum

20microldisc 0 18 18 11 18

40 microldisc 0 21 18 11 22

60 microldisc 0 24 20 18 22

EPAIR6 P decumbens



60 microldisc 0 12 16 0 11




46


EPHAL10 Pasperum


40 microldisc 11 9 6 10 10

60 microldisc 12 11 9 10 12

47





C

+ve C

20microl

60microl

40microl

+veC

20microl

40microl

60microl

C

48





C

60microl

40microl

20microl +veC

vCCe

veve

+veC

vCCe

veve

C

60microl

20microl

40microl

49


Penicillium species



Control 0 0 0 0


EPSLR4 P nigricans




EPAAR5 P rugulosum




EPAIR6 P decumbens







50


EPHAL10 Pasperum








(Table 8 and Fig12)

51





Control 0 0 0 0 0


EPSLR4 P nigricans

20microldisc 16 16 14 14 16

40 microldisc 16 16 14 14 18

60 microldisc 18 16 16 14 20

EPAAR5 P rugulosum

20microldisc 18 18 20 11 20

40 microldisc 18 18 20 11 21

60 microldisc 18 18 20 11 21

EPAIR6 P decumbens






52



EPHAL10 Pasperum




53

4



C

+ve C

20microl 40microl

60microl

54


Penicillium

















55

(1) Nanodecane

(2) Nonadecane

(3) Heptadecane

(4) Heptacosane

(5) Heptacosane

(6) Eicosane

(7) Octadecane

(replib) Nonadecane

50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 2400

50

10057

71

85

99113 127 141 155 169 183 197

(replib) Nonadecane

60 80 100 120 140 160 180 200 220 240 260 2800

50

10057

71

85

99113 127 141 155 169 183 197 268


50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 2500

50

10057

71

85

99113 127 141 155 169 182 196 210 240


60 80 100 120 140 160 180 200 220 240 260 280 300 320 3400

50

10057

71

85

99113 127 141 155 169 183 197 211 225 239 253 267 281 294 308 322 336


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10057

71

85

99113 127 141 155 169 183 197 211 225 239 253 267 281 294 308 322

(mainlib) Eicosane

60 80 100 120 140 160 180 200 220 240 260 2800

50

10057

71

85

99113

127 141 155 169 183 197 211 225 238 252 282

(replib) Octadecane

60 80 100 120 140 160 180 200 220 240 2600

50

10057

71

85

99113 127 141 155 169 183 197 210 225 254

56








50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 2400

50

100

55

60

69

73

8397 115

129

138

143157

171

185

199209

228

OH

O


60 80 100 120 140 160 180 200 220 240 2600

50

10057

71

85

97

113127

141 155 168183 197 212


50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 2500

50

100

60

71

8397

102

111

129

143157

171

185

199

211

228

O

O


60 80 100 120 140 160 180 200 220 240 2600

50

10057

60

71

83 97

102

111 129

143153 165

177

185

195 213225

242O

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

61

70

88

101

115129

143

157

171 185 199 213

227

241 255

270

O

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

69

81

95 109

123

135

140

150

167

177 205223

238

O

OH

57



(16) Oleic Acid



60 80 100 120 140 160 180 200 220 240 2600

50

10060 73

8397

115

129

143157 171 185

199

213

227 239

256

OH

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

61 73

88

101

115129 143

157

171 185 199 213 225239

255 267284

O

O


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

100

55

6781

95

109

123135 150 164 178

192 205 220 234

263

279

308

O

O

(replib) Oleic Acid

60 80 100 120 140 160 180 200 220 240 260 2800

50

10055

69

83

97

111

125137 151 165 180 193 207 222 236

264

282

HO

O

58

(18) Ethyl Oleate





60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10055

6983

97

111123

137 155180

194 207

222

236

264

281

310

O

O


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10055

6983

97

111

125137 151 165 179 194 207 221 236 249 261

278296

HO

O


60 80 100 120 140 160 180 200 220 240 260 280 3000

50

100

55 77 90 103118

133

147

161

178

191 262290

O

O

O


60 90 120 150 180 210 240 270 300 330 360 3900

50

100

5770

83 104132

149

167

279

O

O

O

O


60 90 120 150 180 210 240 270 300 330 360 3900

50

100

55

69

81

95

107

121

147

173189

215

231

243

258

286

O

59





50 100 150 200 250 300 350 400 450 500 550 600 6500

50

100

63 91

151

189

287

303

318 347

O

O

OHO OH

H2N

60


SNo Scan

No

Systemic Name

(Common Name)

Mol

Formula

Mol

Wt

Ret

Time

Conc

1 2606 Nanodecane C19H40 268 24168 0036

2 2913 Heptadecane C17H36 240 2641 0035


4 3230 Octadecane C18H38 254 28737 0049





9 3653 Nonadecane C19H40 268 31834 0064




13 4252 Eicosane C20H42 282 36220 0093

14 5475 Oleic Acid C18H34O2 282 45175 0105


16 5546 Ethyl Oleate C20H38O2 310 45694 0065

61


18 6023 Heptacosane C27H56 380 49187 0051


20 6281 Heptacosane C27H56 380 51076 0044




C20H30O 286 53910 004


62





























63





















64



Infection


Fsolani



Control hellip 50 187 75 25 75 50 187 125





P citrinum EPSMR1 375 0 25 0 125 25 0 0



Pduclauxi EPASS9 50 0 62 0 312 312 62 0

Plividum EPMCL12 50 62 50 0 0 50 0 62



Pthomii EPAER11 62 0 62 0 375 187 62 62



Pasperum EPHAL10 125 0 25 187 375 312 62 62






65



Shoot Weight


(cm)

(g)

(cm)

(g)


Control 22775 3993 253 535 643 1162 0645 0675

Carbendazim 2585 418 2216 451 742 1287 0715 0622




P citrinum EPSMR1 2599 4681 218 51 94 862 0726 0807



Pduclauxi EPASS9 2541 4487 243 512 1103 1406 077 0786

Plividum EPMCL12 22685 4587 205 539 631 558 0663 0578



Pthomii EPAER11 2824 44 277 527 1221 1218 1005 0645



Pasperum EPHAL10 26186 4918 214 678 918 757 069 0866

LSD005 5141 7881 07911 1821 2551 2821 01951 031



66




















67

68



69


sunflower


+veControl EP

EP

EP EP EP EP

EP

Control

70


























71



Infection


Fsolani



Control 50 187 50 25 75 75 187 125

Carbendazim 125 62 312 62 125 25 62 62




P citrinum EPSMR1 375 0 25 0 125 25 0 0



Pduclauxi EPASS9 25 0 62 62 312 187 62 0

Plividum EPMCL12 50 62 50 0 0 50 0 62



Pthomii EPAER11 62 0 62 0 375 187 62 62



Pasperum EPHAL10 125 125 25 187 312 312 62 62






72


Treatments Code

Shoot Length

(cm)

Shoot Weight

(g)


(cm)

(g)


Control 3256 3893 378 642 57 1034 085 131

Carbendazim 3781 4293 452 607 84 1025 124 128




P citrinum EPSMR1 385 6443 373 1425 75 787 088 226



Pduclauxi EPASS9 4412 6275 386 1013 7 768 086 213

Plividum EPMCL12 345 6551 206 1019 706 645 072 161



Pthomii EPAER11 4838 6208 489 953 863 656 096 141



Pasperum EPHAL10 4158 5006 362 818 6102 1275 067 186

LSD005 10331 8971 2271 5521 3021 2171 04581 1071




73


in sunflower











after 45 days

EP

Carbendazim Control

74


















75



Infection


Fsolani



Control hellip 50 312 100 75 100 50 0 562





P citrinum EPSMR1 62 62 437 75 0 0 62 62



Pduclauxi EPASS9 0 0 437 375 25 375 62 25

Plividum EPMCL12 62 25 25 687 125 375 62 50



Pthomii EPAER11 62 62 437 25 125 312 0 0



Pasperum EPHAL10 435 125 25 25 25 187 0 0






76



Shoot Weight


(cm)

(g)

(cm)

(g)


Control hellip 1375 1714 078 08 1531 4652 051 014





P citrinum EPSMR1 1299 1606 059 0617 165 477 039 016



Pduclauxi EPASS9 1187 1916 069 0855 1108 4562 017 016

Plividum EPMCL12 132 2147 061 1358 2252 4785 026 022



Pthomii EPAER11 1463 179 077 1203 1125 7025 031 024



Pasperum EPHAL10 1463 1874 077 1102 1125 702 031 02

LSD005 1611 4011 0191 2141 8421 1151 0171 0071




77

















solani (Table 16)




78



Infection


Fsolani



Control hellip 50 50 100 75 100 75 0 187





P citrinum EPSMR1 62 25 437 437 0 437 62 187



Pduclauxi EPASS9 0 312 437 562 25 562 62 65

Plividum EPMCL12 62 125 25 25 125 25 62 0



Pthomii EPAER11 62 187 437 437 125 375 0 0



Pasperum EPHAL10 437 375 25 312 25 562 0 125






79


Treatments Code

Shoot Length


(cm)

(g)

(cm)

(g)


Control hellip 1375 1364 078 089 1531 613 051 031





P citrinum EPSMR1 1299 1318 059 193 165 961 039 037



Pduclauxi EPASS9 1187 1438 069 13 1108 1178 017 048

Plividum EPMCL12 132 1323 061 107 2252 1024 026 048



Pthomii EPAER11 1463 1435 077 119 1125 1185 031 071



Pasperum EPHAL10 1463 1453 077 128 1125 972 031 046

LSD005 1611 2661 0191 091 8421 271 0171 0291



80


mungbean growth


















81




Infection

Control --------- 50 100 100 50

Carbendazim --------- 25 50 50 62
















82



(cm) (g) (cm) (g)

Control ---------- 1475 0522 4972 0098

Carbendazim --------- 1635 0987 3737 009




P citrinum EPSMR1 1344 0987 4617 0137



Pduclauxi EPASS9 1434 0696 4127 0096

Plividum EPMCL12 1639 0752 4147 0121



Pthomii EPAER11 1482 0711 3865 0072



Pasperum EPHAL10 1608 0787 3875 0066

LSD005 2891 0261 0741 0051



83

84
























85



Infection


Fsolani



Control hellip 437 312 625 625 312 0 312 0





P citrinum EPSMR1 0 50 62 625 625 62 75 0



Pduclauxi EPASS9 0 0 62 25 187 125 50 62

Plividum EPMCL12 50 437 437 562 375 0 687 62



Pthomii EPAER11 187 562 312 562 50 312 562 0



Pasperum EPHAL10 62 312 125 562 25 62 812 0






86



(cm) (g) (cm) (g)


Control hellip 12 1544 18 407 126 333 155 063





P citrinum EPSMR1 1732 1755 297 389 922 1149 064 056



Pduclauxi EPASS9 1672 2255 243 636 1185 597 057 11

Plividum EPMCL12 1307 1303 178 254 1242 529 052 046



Pthomii EPAER11 1328 1375 214 234 148 466 046 055



Pasperum EPHAL10 1328 2412 18 732 1225 775 06 126

LSD005 271 5171 0691 2091 3731 3031 1031 0631



87


EP

88























89



Infection


Fsolani



Control hellip 437 50 625 25 312 62 312 0





P citrinum EPSMR1 0 62 62 562 625 0 75 0



Pduclauxi EPASS9 0 562 62 562 187 25 50 0

Plividum EPMCL12 50 187 437 375 375 0 687 0



Pthomii EPAER11 187 312 312 25 50 125 562 0



Pasperum EPHAL10 62 125 125 50 25 62 812 0






90




















weeks

91


Treatments Code

Shoot

Length

Shoot

Length

Shoot

Weight

Shoot

Weight

Root

Length

Root

Length

Root

weight

Root

weight

(cm) (cm) (g) (g) (cm) (cm) (g) (g)


Control hellip 12 1633 18 554 126 1757 155 105





P citrinum EPSMR1 1732 1912 297 512 922 9 064 155



Pduclauxi EPASS9 1672 214 243 69 1185 153 057 237

Plividum EPMCL12 1307 2347 178 741 1242 1298 052 156



Pthomii EPAER11 1328 225 214 657 148 155 046 164



Pasperum EPHAL10 1328 2101 18 525 1225 1095 06 135

LSD005 271 4291 0691 3281 3731 5851 1031 091


92


cake in tomato

EP

93





















94



Infection


Fsolani



Control hellip 375 0 50 125 437 375 25 0





P citrinum EPSMR1 312 187 187 312 375 50 187 0



Pduclauxi EPASS9 187 375 125 25 375 50 0 0

Plividum EPMCL12 62 62 437 125 62 625 25 0



Pthomii EPAER11 0 187 437 187 62 25 0 0



Pasperum EPHAL10 125 62 50 125 125 812 0 0






95



(cm) (g) (cm) (g)


Control hellip 2369 2225 274 837 274 975 211 303





P citrinum EPSMR1 267 3384 313 668 2397 975 394 098



Pduclauxi EPASS9 2925 2911 376 388 3037 1293 522 116

Plividum EPMCL12 2768 2801 31 698 2155 1132 35 109



Pthomii EPAER11 239 30 296 799 2416 1062 427 125



Pasperum EPHAL10 2856 2891 344 763 1921 1352 306 13

LSD005 471 4931 0941 3331 7321 5451 1611 11071



96



EP

S

EP

97


EP

EP

98


in chickpea


in chickpea

EP

EP

99


in chickpea













weeks













(Table 26)

100




101



Infection



Control hellip 375 125 50 312 437 0 25 187





P citrinum EPSMR1 312 0 187 625 375 187 187 312



Pduclauxi EPASS9 187 0 125 625 375 62 0 312

Plividum EPMCL12 62 62 437 100 62 25 25 312



Pthomii EPAER11 0 62 437 125 62 62 0 62



Pasperum EPHAL10 125 0 50 187 125 0 0 0






102



(cm) (g) (cm) (g)


Control hellip 2369 2188 274 406 274 692 211 58





P citrinum EPSMR1 267 1916 313 556 2397 756 394 1172



Pduclauxi EPASS9 2925 192 376 561 3037 1115 522 819

Plividum EPMCL12 2768 1929 31 417 2155 946 35 383



Pthomii EPAER11 239 2305 296 626 2416 9 427 931



Pasperum EPHAL10 2856 1662 344 582 1921 925 306 834

LSD005 471 6131 0941 3011 7321 2921 1611 6151




103













weeks














104



Infection


Control 75 100 100 75

Feast-M 0 37 687 625










Topsin-M 0 437 812 62





P citrinum 437 687 100 0


P decumbens 187 50 437 187

Pasperum 125 50 562 125




105



Control 3197 339 288 288

Feast-M 4269 451 526 526





Carbendazim 3675 319 398 398





Topsin-M 3935 314 383 383





P citrinum 3268 249 432 432

Pnigricans 2788 201 401 401

P decumbens 3421 3007 446 446

Pasperum 3262 229 363 363

LSD005 5751 0811 1041 1041



106








days of germination



















107



Infection


Control 0 50 625 50








108



Fruits Fruit weight

(cm) (g) (cm) (g)

Control 3831 1058 1596 305 023c 708c

Carbendazim 3421 832 1659 546 045b 683c

P decumbens 4523 1167 1756 438 052a 1106a

Pnigricans 4265 1172 1794 188 054a 894b

P rugulosum 4592 1295 1967 2405 025c 533d

LSD005 511 4281 3431 581 00261 04841


109


soil



Control 137e 2711e 2878e 0053d





LSD005 00311 01361 04211 00041


110



Sucrose

Control 587c 0087c 8668d 1353b 245d





LSD005 0121 000571 0211 01031 0121


111


in garden soil








LSD005 10591 01441 21941 3711


112








after 90 days










37)








113



Infection


Control 312 100 937 562








114



(cm) (g) (cm) (g)

Control 52 1974 1816 35 30a 5801b

Carbendazim 4646 1322 1629 237 20c 4083a

P decumbens 443 2161 1283 418 2133c 995a

Pnigricans 55 1892 1561 315 32a 4286d

P rugulosum 5197 1695 1205 334 256b 4779c

LSD005 1481 18611 5391 4011 3781 0131


115


garden soil








LSD005 01081 01671 0301 00791


116



N Sucrose






LSD005 00621 00541 0211 1311


117










LSD005 5591 0471 5771 2391


118



field condition



















days








119



























120



Infection


45 90 45 90 45 90 45 90

Control 375 0 562 312 937 100 562 0

Topsin-M 375 0 625 25 937 100 687 0


P decumbens 62 0 50 375 68 100 375 0

Pnigricans 125 187 562 687 875 100 312 0

P rugulosum 312 62 562 375 812 100 437 0







121


condition


(cm)

Shoot Weight

(g)

Root Length

(cm)

Root Weight

(g)

Number

of Fruits

Fruit

weight

Control 45 90 45 90 45 90 45 90 90 90

Topsin-M 4178 6192 2228 4325 1368 2426 204 823 086g 246i

Pmontelii 422 6375 1765 4731 1267 2377 133 98 12f 31h


P nigricans 4233 6617 1971 4887 1486 2578 167 1003 143e 1146a

Prugulosum 4866 7083 1635 5095 1378 2311 172 967 176d 331g

P rugulosum 4373 7026 2063 2051 1371 2464 169 709 123f 35f




LSD005 961 1321 131 1181 3551 1371 0831 2961 0111 0111


122



Treatments

Polyphenol

microgml



Control 183h 7314e 7721e 007f

Topsin-M 146i 9119a 9886a 0113d








LSD005 00721 10191 06531 00121



123




microgml




Topsin-M 183f 4922f 5575g 0074bc








LSD005 10061 05191 04731 0003081


124





Control 5102g 646g 1466g 5966f

Topsin-M 5514f 716f 2566f 67e








LSD005 10451 06241 14081 2471


125



Treatments pH

Tritable

acidity

Moisture

content

Total

solids

Total Soluble

Solid

Sucrose

Control 624a 0102c 8774b 1222f 1425e

Topsin-M 619ab 0126b 8653e 1339b 1475e








LSD005 00641 00041 03021 0171 02221


126

127

128

4 DISCUSSION





























129































130
































131
































132







2013)
























133































134
































135








Kloepper 1998)























136




condition


























137



source to sink



























138































Conclusion

139



















REFERENCES

140



Eco 15(1) 37-47






University












Ecol 58 921-929








99-102



141


2933



















78(3) 429-435




New York NY






142























21(12) pp 1387-1397









143











Engelmann










Ecol 51(2) 215-229











144





Hortic Sci 35 67-72
























6(5) 162-166



145





Acad of Sci 102(35) 12465-12470










Pathol 111 101-112






41-48



4(1) 34-43







146



185(11) 3458-3468













5 17-24



356



Press)









147





Trop Sci 35 294-299






75270 Pakistan












717-732



Pathol J 2 75-79





148








255(7) 2751-2760







21 2894-2897









Protect 46 627-635








149



Phytopathol 58 1051






















Prod 78 2699-2703





150









207













43



106




Pathol 57 870-876

151



Resour 8(2) 1634-1645






8(6) 687-695
















63(4) 391-400



87 4-10




152



S00218596120005-12



Pak J Bot 47(4) 1547-1551




261







York Pages 86













209-214

153



York pp 660-662






April 1996



Lett 100(8) 084102














Plant Biol 13(1) 86





154





















BMC Microbiol 13 51





70-78




155

















289-295














88

156



Pathol 23 1-7






299(1) 31-37








553-558














157
































158


228 7382





Newslett 5 32-34



Pl Nutrit 22(4-5) 835-853





649



513-515 doi 101126science1136237



129 189-196











159





and Ecol 39(2) 163-165

















1048


Minia Univ







160



62















40 403-408













161



doi 103389fpls00432




32-34






516-522


















162


Toxicol 50(3-4) 829-834





















3 342-351







163



















Mar 55 269-279












164















647






319-343










165








York 326-365




686



106(9) 996-1004





Mycol Res 106(9) 996-1004






13(4) 1698-1709





166






Biol Sci 4 1240- 1242












1623




16(4) 403-406










167





Interact 3(2) 75-93




Ecol 19 2917-2933







1037-1042





2950










168







43(1) 1-6














82










169























43-68







Sci 416 382-385

170




Bioch 37 21312140



Mycologia 98 31-42


















Sci 102(38) 13386-13391



95 1368-1373



147 523- 535

171





Res 4 1100-1104






Drugs 11 2230-2238




Interact 10(1) 280-287









3(1-2) 90-97






489

172




585




36(1) 82


















January 20 2017






173



449-458





iv

DEDICATION



v


1 INTRODUCTION

11 Endophytic fungi










from different host

















vi




suspension







216 Fruit analysis

2161 pH




2165 Firmness

2166 Total solids

2167 Protein

2168 Carbohydrate








Penicillium


Penicillium


vii

Penicillium spp



culture filtrates


culture filtrates


culture filtrates


culture filtrates


rugulosum



















viii





okra plants


tomato plants






4 DISCUSSION

ix



NEMATODE

SUMMARY


























x






















xi

xii

1

1 INTRODUCTION

11 Endophytic fungi




























2

















2014)



spiteller 2012b)











3




(Huang et al 2008)

























plants

4































5
































6






























7






























8






























9



endophytic fungi






10







about (24) hours





















11
















ordmC for 16 min



calibrate the sizes





12





phaseolina)




culture plate assay







times










13




in vitro

















measured in mm

14























Penicillium species





15












regulosum





packard (5890)






16





measured



(2016) was used





polyphenol activity










17









-------------------------------------------------------









sucrose

2165 Firmness



2166 Total solids


moisture from 100

18


2167 Protein


2168 Carbohydrate













used

19





























20

21

22












(Table 1) fig1-7

23







(Solanaceae)

Root 4 4 20 20




(Solanaceae)

root 5 25 16 21


(Amaranthaceae)

root 3 12 11 20


(Meliaceae)

root 5 25 13 20




(Amaranthaceae)

stem 18 13 11 14



(Malvaceae)

root 5 8 5 6


(Cucurbitaceae)

leaf 18 13 11 14


(Solanaceae)

root 5 24 17 22


(Malvaceae)

root 5 3 21 12

24


assay


assay

25


assay



26


assay


assay

27


assay















28













29





Control 0 0 0 0


EPSMR1 P citrinum




EPSMS2 Plilacinum








EPSLR4 P nigricans



30




EPAAR5 P rugulosum




EPAIR6 P decumbens








EPCTS8 Prestrictum




EPASS9 P duclauxi


31





EPHAL10 Pasperum




EPAER11 Pthomii




EPMCL12 P lividum




EPSLR13 P javanicum





32






33



























34


species



Control 0 0 0 0


EPSMR1 P citrinum




EPSMS2 Plilacinum








EPSLR4 P nigricans


35





EPAAR5 P rugulosum




EPAIR6 P decumbens








EPCTS8 P restrictum




EPASS9 P duclauxi

36






EPHAL10 Pasperum




EPAER11 Pthomii




EPMCL12 P lividum




EPSLR13 P javanicum




37







38





spp










A

B

C

E D

39


48 hours


24Hours 48Hours
















40

41























42


Penicillium species



Control 0 0 0 0


EPSLR4 P nigricans




EPAAR5 P rugulosum




EPAIR6 P decumbens






43



EPHAL10 Pasperum




44














45





Control 0 0 0 0 0


EPSLR4 P nigricans



60 microldisc 11 11 9 12 10

EPAAR5 P rugulosum

20microldisc 0 18 18 11 18

40 microldisc 0 21 18 11 22

60 microldisc 0 24 20 18 22

EPAIR6 P decumbens



60 microldisc 0 12 16 0 11




46


EPHAL10 Pasperum


40 microldisc 11 9 6 10 10

60 microldisc 12 11 9 10 12

47





C

+ve C

20microl

60microl

40microl

+veC

20microl

40microl

60microl

C

48





C

60microl

40microl

20microl +veC

vCCe

veve

+veC

vCCe

veve

C

60microl

20microl

40microl

49


Penicillium species



Control 0 0 0 0


EPSLR4 P nigricans




EPAAR5 P rugulosum




EPAIR6 P decumbens







50


EPHAL10 Pasperum








(Table 8 and Fig12)

51





Control 0 0 0 0 0


EPSLR4 P nigricans

20microldisc 16 16 14 14 16

40 microldisc 16 16 14 14 18

60 microldisc 18 16 16 14 20

EPAAR5 P rugulosum

20microldisc 18 18 20 11 20

40 microldisc 18 18 20 11 21

60 microldisc 18 18 20 11 21

EPAIR6 P decumbens






52



EPHAL10 Pasperum




53

4



C

+ve C

20microl 40microl

60microl

54


Penicillium

















55

(1) Nanodecane

(2) Nonadecane

(3) Heptadecane

(4) Heptacosane

(5) Heptacosane

(6) Eicosane

(7) Octadecane

(replib) Nonadecane

50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 2400

50

10057

71

85

99113 127 141 155 169 183 197

(replib) Nonadecane

60 80 100 120 140 160 180 200 220 240 260 2800

50

10057

71

85

99113 127 141 155 169 183 197 268


50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 2500

50

10057

71

85

99113 127 141 155 169 182 196 210 240


60 80 100 120 140 160 180 200 220 240 260 280 300 320 3400

50

10057

71

85

99113 127 141 155 169 183 197 211 225 239 253 267 281 294 308 322 336


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10057

71

85

99113 127 141 155 169 183 197 211 225 239 253 267 281 294 308 322

(mainlib) Eicosane

60 80 100 120 140 160 180 200 220 240 260 2800

50

10057

71

85

99113

127 141 155 169 183 197 211 225 238 252 282

(replib) Octadecane

60 80 100 120 140 160 180 200 220 240 2600

50

10057

71

85

99113 127 141 155 169 183 197 210 225 254

56








50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 2400

50

100

55

60

69

73

8397 115

129

138

143157

171

185

199209

228

OH

O


60 80 100 120 140 160 180 200 220 240 2600

50

10057

71

85

97

113127

141 155 168183 197 212


50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 2500

50

100

60

71

8397

102

111

129

143157

171

185

199

211

228

O

O


60 80 100 120 140 160 180 200 220 240 2600

50

10057

60

71

83 97

102

111 129

143153 165

177

185

195 213225

242O

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

61

70

88

101

115129

143

157

171 185 199 213

227

241 255

270

O

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

69

81

95 109

123

135

140

150

167

177 205223

238

O

OH

57



(16) Oleic Acid



60 80 100 120 140 160 180 200 220 240 2600

50

10060 73

8397

115

129

143157 171 185

199

213

227 239

256

OH

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

61 73

88

101

115129 143

157

171 185 199 213 225239

255 267284

O

O


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

100

55

6781

95

109

123135 150 164 178

192 205 220 234

263

279

308

O

O

(replib) Oleic Acid

60 80 100 120 140 160 180 200 220 240 260 2800

50

10055

69

83

97

111

125137 151 165 180 193 207 222 236

264

282

HO

O

58

(18) Ethyl Oleate





60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10055

6983

97

111123

137 155180

194 207

222

236

264

281

310

O

O


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10055

6983

97

111

125137 151 165 179 194 207 221 236 249 261

278296

HO

O


60 80 100 120 140 160 180 200 220 240 260 280 3000

50

100

55 77 90 103118

133

147

161

178

191 262290

O

O

O


60 90 120 150 180 210 240 270 300 330 360 3900

50

100

5770

83 104132

149

167

279

O

O

O

O


60 90 120 150 180 210 240 270 300 330 360 3900

50

100

55

69

81

95

107

121

147

173189

215

231

243

258

286

O

59





50 100 150 200 250 300 350 400 450 500 550 600 6500

50

100

63 91

151

189

287

303

318 347

O

O

OHO OH

H2N

60


SNo Scan

No

Systemic Name

(Common Name)

Mol

Formula

Mol

Wt

Ret

Time

Conc

1 2606 Nanodecane C19H40 268 24168 0036

2 2913 Heptadecane C17H36 240 2641 0035


4 3230 Octadecane C18H38 254 28737 0049





9 3653 Nonadecane C19H40 268 31834 0064




13 4252 Eicosane C20H42 282 36220 0093

14 5475 Oleic Acid C18H34O2 282 45175 0105


16 5546 Ethyl Oleate C20H38O2 310 45694 0065

61


18 6023 Heptacosane C27H56 380 49187 0051


20 6281 Heptacosane C27H56 380 51076 0044




C20H30O 286 53910 004


62





























63





















64



Infection


Fsolani



Control hellip 50 187 75 25 75 50 187 125





P citrinum EPSMR1 375 0 25 0 125 25 0 0



Pduclauxi EPASS9 50 0 62 0 312 312 62 0

Plividum EPMCL12 50 62 50 0 0 50 0 62



Pthomii EPAER11 62 0 62 0 375 187 62 62



Pasperum EPHAL10 125 0 25 187 375 312 62 62






65



Shoot Weight


(cm)

(g)

(cm)

(g)


Control 22775 3993 253 535 643 1162 0645 0675

Carbendazim 2585 418 2216 451 742 1287 0715 0622




P citrinum EPSMR1 2599 4681 218 51 94 862 0726 0807



Pduclauxi EPASS9 2541 4487 243 512 1103 1406 077 0786

Plividum EPMCL12 22685 4587 205 539 631 558 0663 0578



Pthomii EPAER11 2824 44 277 527 1221 1218 1005 0645



Pasperum EPHAL10 26186 4918 214 678 918 757 069 0866

LSD005 5141 7881 07911 1821 2551 2821 01951 031



66




















67

68



69


sunflower


+veControl EP

EP

EP EP EP EP

EP

Control

70


























71



Infection


Fsolani



Control 50 187 50 25 75 75 187 125

Carbendazim 125 62 312 62 125 25 62 62




P citrinum EPSMR1 375 0 25 0 125 25 0 0



Pduclauxi EPASS9 25 0 62 62 312 187 62 0

Plividum EPMCL12 50 62 50 0 0 50 0 62



Pthomii EPAER11 62 0 62 0 375 187 62 62



Pasperum EPHAL10 125 125 25 187 312 312 62 62






72


Treatments Code

Shoot Length

(cm)

Shoot Weight

(g)


(cm)

(g)


Control 3256 3893 378 642 57 1034 085 131

Carbendazim 3781 4293 452 607 84 1025 124 128




P citrinum EPSMR1 385 6443 373 1425 75 787 088 226



Pduclauxi EPASS9 4412 6275 386 1013 7 768 086 213

Plividum EPMCL12 345 6551 206 1019 706 645 072 161



Pthomii EPAER11 4838 6208 489 953 863 656 096 141



Pasperum EPHAL10 4158 5006 362 818 6102 1275 067 186

LSD005 10331 8971 2271 5521 3021 2171 04581 1071




73


in sunflower











after 45 days

EP

Carbendazim Control

74


















75



Infection


Fsolani



Control hellip 50 312 100 75 100 50 0 562





P citrinum EPSMR1 62 62 437 75 0 0 62 62



Pduclauxi EPASS9 0 0 437 375 25 375 62 25

Plividum EPMCL12 62 25 25 687 125 375 62 50



Pthomii EPAER11 62 62 437 25 125 312 0 0



Pasperum EPHAL10 435 125 25 25 25 187 0 0






76



Shoot Weight


(cm)

(g)

(cm)

(g)


Control hellip 1375 1714 078 08 1531 4652 051 014





P citrinum EPSMR1 1299 1606 059 0617 165 477 039 016



Pduclauxi EPASS9 1187 1916 069 0855 1108 4562 017 016

Plividum EPMCL12 132 2147 061 1358 2252 4785 026 022



Pthomii EPAER11 1463 179 077 1203 1125 7025 031 024



Pasperum EPHAL10 1463 1874 077 1102 1125 702 031 02

LSD005 1611 4011 0191 2141 8421 1151 0171 0071




77

















solani (Table 16)




78



Infection


Fsolani



Control hellip 50 50 100 75 100 75 0 187





P citrinum EPSMR1 62 25 437 437 0 437 62 187



Pduclauxi EPASS9 0 312 437 562 25 562 62 65

Plividum EPMCL12 62 125 25 25 125 25 62 0



Pthomii EPAER11 62 187 437 437 125 375 0 0



Pasperum EPHAL10 437 375 25 312 25 562 0 125






79


Treatments Code

Shoot Length


(cm)

(g)

(cm)

(g)


Control hellip 1375 1364 078 089 1531 613 051 031





P citrinum EPSMR1 1299 1318 059 193 165 961 039 037



Pduclauxi EPASS9 1187 1438 069 13 1108 1178 017 048

Plividum EPMCL12 132 1323 061 107 2252 1024 026 048



Pthomii EPAER11 1463 1435 077 119 1125 1185 031 071



Pasperum EPHAL10 1463 1453 077 128 1125 972 031 046

LSD005 1611 2661 0191 091 8421 271 0171 0291



80


mungbean growth


















81




Infection

Control --------- 50 100 100 50

Carbendazim --------- 25 50 50 62
















82



(cm) (g) (cm) (g)

Control ---------- 1475 0522 4972 0098

Carbendazim --------- 1635 0987 3737 009




P citrinum EPSMR1 1344 0987 4617 0137



Pduclauxi EPASS9 1434 0696 4127 0096

Plividum EPMCL12 1639 0752 4147 0121



Pthomii EPAER11 1482 0711 3865 0072



Pasperum EPHAL10 1608 0787 3875 0066

LSD005 2891 0261 0741 0051



83

84
























85



Infection


Fsolani



Control hellip 437 312 625 625 312 0 312 0





P citrinum EPSMR1 0 50 62 625 625 62 75 0



Pduclauxi EPASS9 0 0 62 25 187 125 50 62

Plividum EPMCL12 50 437 437 562 375 0 687 62



Pthomii EPAER11 187 562 312 562 50 312 562 0



Pasperum EPHAL10 62 312 125 562 25 62 812 0






86



(cm) (g) (cm) (g)


Control hellip 12 1544 18 407 126 333 155 063





P citrinum EPSMR1 1732 1755 297 389 922 1149 064 056



Pduclauxi EPASS9 1672 2255 243 636 1185 597 057 11

Plividum EPMCL12 1307 1303 178 254 1242 529 052 046



Pthomii EPAER11 1328 1375 214 234 148 466 046 055



Pasperum EPHAL10 1328 2412 18 732 1225 775 06 126

LSD005 271 5171 0691 2091 3731 3031 1031 0631



87


EP

88























89



Infection


Fsolani



Control hellip 437 50 625 25 312 62 312 0





P citrinum EPSMR1 0 62 62 562 625 0 75 0



Pduclauxi EPASS9 0 562 62 562 187 25 50 0

Plividum EPMCL12 50 187 437 375 375 0 687 0



Pthomii EPAER11 187 312 312 25 50 125 562 0



Pasperum EPHAL10 62 125 125 50 25 62 812 0






90




















weeks

91


Treatments Code

Shoot

Length

Shoot

Length

Shoot

Weight

Shoot

Weight

Root

Length

Root

Length

Root

weight

Root

weight

(cm) (cm) (g) (g) (cm) (cm) (g) (g)


Control hellip 12 1633 18 554 126 1757 155 105





P citrinum EPSMR1 1732 1912 297 512 922 9 064 155



Pduclauxi EPASS9 1672 214 243 69 1185 153 057 237

Plividum EPMCL12 1307 2347 178 741 1242 1298 052 156



Pthomii EPAER11 1328 225 214 657 148 155 046 164



Pasperum EPHAL10 1328 2101 18 525 1225 1095 06 135

LSD005 271 4291 0691 3281 3731 5851 1031 091


92


cake in tomato

EP

93





















94



Infection


Fsolani



Control hellip 375 0 50 125 437 375 25 0





P citrinum EPSMR1 312 187 187 312 375 50 187 0



Pduclauxi EPASS9 187 375 125 25 375 50 0 0

Plividum EPMCL12 62 62 437 125 62 625 25 0



Pthomii EPAER11 0 187 437 187 62 25 0 0



Pasperum EPHAL10 125 62 50 125 125 812 0 0






95



(cm) (g) (cm) (g)


Control hellip 2369 2225 274 837 274 975 211 303





P citrinum EPSMR1 267 3384 313 668 2397 975 394 098



Pduclauxi EPASS9 2925 2911 376 388 3037 1293 522 116

Plividum EPMCL12 2768 2801 31 698 2155 1132 35 109



Pthomii EPAER11 239 30 296 799 2416 1062 427 125



Pasperum EPHAL10 2856 2891 344 763 1921 1352 306 13

LSD005 471 4931 0941 3331 7321 5451 1611 11071



96



EP

S

EP

97


EP

EP

98


in chickpea


in chickpea

EP

EP

99


in chickpea













weeks













(Table 26)

100




101



Infection



Control hellip 375 125 50 312 437 0 25 187





P citrinum EPSMR1 312 0 187 625 375 187 187 312



Pduclauxi EPASS9 187 0 125 625 375 62 0 312

Plividum EPMCL12 62 62 437 100 62 25 25 312



Pthomii EPAER11 0 62 437 125 62 62 0 62



Pasperum EPHAL10 125 0 50 187 125 0 0 0






102



(cm) (g) (cm) (g)


Control hellip 2369 2188 274 406 274 692 211 58





P citrinum EPSMR1 267 1916 313 556 2397 756 394 1172



Pduclauxi EPASS9 2925 192 376 561 3037 1115 522 819

Plividum EPMCL12 2768 1929 31 417 2155 946 35 383



Pthomii EPAER11 239 2305 296 626 2416 9 427 931



Pasperum EPHAL10 2856 1662 344 582 1921 925 306 834

LSD005 471 6131 0941 3011 7321 2921 1611 6151




103













weeks














104



Infection


Control 75 100 100 75

Feast-M 0 37 687 625










Topsin-M 0 437 812 62





P citrinum 437 687 100 0


P decumbens 187 50 437 187

Pasperum 125 50 562 125




105



Control 3197 339 288 288

Feast-M 4269 451 526 526





Carbendazim 3675 319 398 398





Topsin-M 3935 314 383 383





P citrinum 3268 249 432 432

Pnigricans 2788 201 401 401

P decumbens 3421 3007 446 446

Pasperum 3262 229 363 363

LSD005 5751 0811 1041 1041



106








days of germination



















107



Infection


Control 0 50 625 50








108



Fruits Fruit weight

(cm) (g) (cm) (g)

Control 3831 1058 1596 305 023c 708c

Carbendazim 3421 832 1659 546 045b 683c

P decumbens 4523 1167 1756 438 052a 1106a

Pnigricans 4265 1172 1794 188 054a 894b

P rugulosum 4592 1295 1967 2405 025c 533d

LSD005 511 4281 3431 581 00261 04841


109


soil



Control 137e 2711e 2878e 0053d





LSD005 00311 01361 04211 00041


110



Sucrose

Control 587c 0087c 8668d 1353b 245d





LSD005 0121 000571 0211 01031 0121


111


in garden soil








LSD005 10591 01441 21941 3711


112








after 90 days










37)








113



Infection


Control 312 100 937 562








114



(cm) (g) (cm) (g)

Control 52 1974 1816 35 30a 5801b

Carbendazim 4646 1322 1629 237 20c 4083a

P decumbens 443 2161 1283 418 2133c 995a

Pnigricans 55 1892 1561 315 32a 4286d

P rugulosum 5197 1695 1205 334 256b 4779c

LSD005 1481 18611 5391 4011 3781 0131


115


garden soil








LSD005 01081 01671 0301 00791


116



N Sucrose






LSD005 00621 00541 0211 1311


117










LSD005 5591 0471 5771 2391


118



field condition



















days








119



























120



Infection


45 90 45 90 45 90 45 90

Control 375 0 562 312 937 100 562 0

Topsin-M 375 0 625 25 937 100 687 0


P decumbens 62 0 50 375 68 100 375 0

Pnigricans 125 187 562 687 875 100 312 0

P rugulosum 312 62 562 375 812 100 437 0







121


condition


(cm)

Shoot Weight

(g)

Root Length

(cm)

Root Weight

(g)

Number

of Fruits

Fruit

weight

Control 45 90 45 90 45 90 45 90 90 90

Topsin-M 4178 6192 2228 4325 1368 2426 204 823 086g 246i

Pmontelii 422 6375 1765 4731 1267 2377 133 98 12f 31h


P nigricans 4233 6617 1971 4887 1486 2578 167 1003 143e 1146a

Prugulosum 4866 7083 1635 5095 1378 2311 172 967 176d 331g

P rugulosum 4373 7026 2063 2051 1371 2464 169 709 123f 35f




LSD005 961 1321 131 1181 3551 1371 0831 2961 0111 0111


122



Treatments

Polyphenol

microgml



Control 183h 7314e 7721e 007f

Topsin-M 146i 9119a 9886a 0113d








LSD005 00721 10191 06531 00121



123




microgml




Topsin-M 183f 4922f 5575g 0074bc








LSD005 10061 05191 04731 0003081


124





Control 5102g 646g 1466g 5966f

Topsin-M 5514f 716f 2566f 67e








LSD005 10451 06241 14081 2471


125



Treatments pH

Tritable

acidity

Moisture

content

Total

solids

Total Soluble

Solid

Sucrose

Control 624a 0102c 8774b 1222f 1425e

Topsin-M 619ab 0126b 8653e 1339b 1475e








LSD005 00641 00041 03021 0171 02221


126

127

128

4 DISCUSSION





























129































130
































131
































132







2013)
























133































134
































135








Kloepper 1998)























136




condition


























137



source to sink



























138































Conclusion

139



















REFERENCES

140



Eco 15(1) 37-47






University












Ecol 58 921-929








99-102



141


2933



















78(3) 429-435




New York NY






142























21(12) pp 1387-1397









143











Engelmann










Ecol 51(2) 215-229











144





Hortic Sci 35 67-72
























6(5) 162-166



145





Acad of Sci 102(35) 12465-12470










Pathol 111 101-112






41-48



4(1) 34-43







146



185(11) 3458-3468













5 17-24



356



Press)









147





Trop Sci 35 294-299






75270 Pakistan












717-732



Pathol J 2 75-79





148








255(7) 2751-2760







21 2894-2897









Protect 46 627-635








149



Phytopathol 58 1051






















Prod 78 2699-2703





150









207













43



106




Pathol 57 870-876

151



Resour 8(2) 1634-1645






8(6) 687-695
















63(4) 391-400



87 4-10




152



S00218596120005-12



Pak J Bot 47(4) 1547-1551




261







York Pages 86













209-214

153



York pp 660-662






April 1996



Lett 100(8) 084102














Plant Biol 13(1) 86





154





















BMC Microbiol 13 51





70-78




155

















289-295














88

156



Pathol 23 1-7






299(1) 31-37








553-558














157
































158


228 7382





Newslett 5 32-34



Pl Nutrit 22(4-5) 835-853





649



513-515 doi 101126science1136237



129 189-196











159





and Ecol 39(2) 163-165

















1048


Minia Univ







160



62















40 403-408













161



doi 103389fpls00432




32-34






516-522


















162


Toxicol 50(3-4) 829-834





















3 342-351







163



















Mar 55 269-279












164















647






319-343










165








York 326-365




686



106(9) 996-1004





Mycol Res 106(9) 996-1004






13(4) 1698-1709





166






Biol Sci 4 1240- 1242












1623




16(4) 403-406










167





Interact 3(2) 75-93




Ecol 19 2917-2933







1037-1042





2950










168







43(1) 1-6














82










169























43-68







Sci 416 382-385

170




Bioch 37 21312140



Mycologia 98 31-42


















Sci 102(38) 13386-13391



95 1368-1373



147 523- 535

171





Res 4 1100-1104






Drugs 11 2230-2238




Interact 10(1) 280-287









3(1-2) 90-97






489

172




585




36(1) 82


















January 20 2017






173



449-458





v


1 INTRODUCTION

11 Endophytic fungi










from different host

















vi




suspension







216 Fruit analysis

2161 pH




2165 Firmness

2166 Total solids

2167 Protein

2168 Carbohydrate








Penicillium


Penicillium


vii

Penicillium spp



culture filtrates


culture filtrates


culture filtrates


culture filtrates


rugulosum



















viii





okra plants


tomato plants






4 DISCUSSION

ix



NEMATODE

SUMMARY


























x






















xi

xii

1

1 INTRODUCTION

11 Endophytic fungi




























2

















2014)



spiteller 2012b)











3




(Huang et al 2008)

























plants

4































5
































6






























7






























8






























9



endophytic fungi






10







about (24) hours





















11
















ordmC for 16 min



calibrate the sizes





12





phaseolina)




culture plate assay







times










13




in vitro

















measured in mm

14























Penicillium species





15












regulosum





packard (5890)






16





measured



(2016) was used





polyphenol activity










17









-------------------------------------------------------









sucrose

2165 Firmness



2166 Total solids


moisture from 100

18


2167 Protein


2168 Carbohydrate













used

19





























20

21

22












(Table 1) fig1-7

23







(Solanaceae)

Root 4 4 20 20




(Solanaceae)

root 5 25 16 21


(Amaranthaceae)

root 3 12 11 20


(Meliaceae)

root 5 25 13 20




(Amaranthaceae)

stem 18 13 11 14



(Malvaceae)

root 5 8 5 6


(Cucurbitaceae)

leaf 18 13 11 14


(Solanaceae)

root 5 24 17 22


(Malvaceae)

root 5 3 21 12

24


assay


assay

25


assay



26


assay


assay

27


assay















28













29





Control 0 0 0 0


EPSMR1 P citrinum




EPSMS2 Plilacinum








EPSLR4 P nigricans



30




EPAAR5 P rugulosum




EPAIR6 P decumbens








EPCTS8 Prestrictum




EPASS9 P duclauxi


31





EPHAL10 Pasperum




EPAER11 Pthomii




EPMCL12 P lividum




EPSLR13 P javanicum





32






33



























34


species



Control 0 0 0 0


EPSMR1 P citrinum




EPSMS2 Plilacinum








EPSLR4 P nigricans


35





EPAAR5 P rugulosum




EPAIR6 P decumbens








EPCTS8 P restrictum




EPASS9 P duclauxi

36






EPHAL10 Pasperum




EPAER11 Pthomii




EPMCL12 P lividum




EPSLR13 P javanicum




37







38





spp










A

B

C

E D

39


48 hours


24Hours 48Hours
















40

41























42


Penicillium species



Control 0 0 0 0


EPSLR4 P nigricans




EPAAR5 P rugulosum




EPAIR6 P decumbens






43



EPHAL10 Pasperum




44














45





Control 0 0 0 0 0


EPSLR4 P nigricans



60 microldisc 11 11 9 12 10

EPAAR5 P rugulosum

20microldisc 0 18 18 11 18

40 microldisc 0 21 18 11 22

60 microldisc 0 24 20 18 22

EPAIR6 P decumbens



60 microldisc 0 12 16 0 11




46


EPHAL10 Pasperum


40 microldisc 11 9 6 10 10

60 microldisc 12 11 9 10 12

47





C

+ve C

20microl

60microl

40microl

+veC

20microl

40microl

60microl

C

48





C

60microl

40microl

20microl +veC

vCCe

veve

+veC

vCCe

veve

C

60microl

20microl

40microl

49


Penicillium species



Control 0 0 0 0


EPSLR4 P nigricans




EPAAR5 P rugulosum




EPAIR6 P decumbens







50


EPHAL10 Pasperum








(Table 8 and Fig12)

51





Control 0 0 0 0 0


EPSLR4 P nigricans

20microldisc 16 16 14 14 16

40 microldisc 16 16 14 14 18

60 microldisc 18 16 16 14 20

EPAAR5 P rugulosum

20microldisc 18 18 20 11 20

40 microldisc 18 18 20 11 21

60 microldisc 18 18 20 11 21

EPAIR6 P decumbens






52



EPHAL10 Pasperum




53

4



C

+ve C

20microl 40microl

60microl

54


Penicillium

















55

(1) Nanodecane

(2) Nonadecane

(3) Heptadecane

(4) Heptacosane

(5) Heptacosane

(6) Eicosane

(7) Octadecane

(replib) Nonadecane

50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 2400

50

10057

71

85

99113 127 141 155 169 183 197

(replib) Nonadecane

60 80 100 120 140 160 180 200 220 240 260 2800

50

10057

71

85

99113 127 141 155 169 183 197 268


50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 2500

50

10057

71

85

99113 127 141 155 169 182 196 210 240


60 80 100 120 140 160 180 200 220 240 260 280 300 320 3400

50

10057

71

85

99113 127 141 155 169 183 197 211 225 239 253 267 281 294 308 322 336


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10057

71

85

99113 127 141 155 169 183 197 211 225 239 253 267 281 294 308 322

(mainlib) Eicosane

60 80 100 120 140 160 180 200 220 240 260 2800

50

10057

71

85

99113

127 141 155 169 183 197 211 225 238 252 282

(replib) Octadecane

60 80 100 120 140 160 180 200 220 240 2600

50

10057

71

85

99113 127 141 155 169 183 197 210 225 254

56








50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 2400

50

100

55

60

69

73

8397 115

129

138

143157

171

185

199209

228

OH

O


60 80 100 120 140 160 180 200 220 240 2600

50

10057

71

85

97

113127

141 155 168183 197 212


50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 2500

50

100

60

71

8397

102

111

129

143157

171

185

199

211

228

O

O


60 80 100 120 140 160 180 200 220 240 2600

50

10057

60

71

83 97

102

111 129

143153 165

177

185

195 213225

242O

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

61

70

88

101

115129

143

157

171 185 199 213

227

241 255

270

O

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

69

81

95 109

123

135

140

150

167

177 205223

238

O

OH

57



(16) Oleic Acid



60 80 100 120 140 160 180 200 220 240 2600

50

10060 73

8397

115

129

143157 171 185

199

213

227 239

256

OH

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

61 73

88

101

115129 143

157

171 185 199 213 225239

255 267284

O

O


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

100

55

6781

95

109

123135 150 164 178

192 205 220 234

263

279

308

O

O

(replib) Oleic Acid

60 80 100 120 140 160 180 200 220 240 260 2800

50

10055

69

83

97

111

125137 151 165 180 193 207 222 236

264

282

HO

O

58

(18) Ethyl Oleate





60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10055

6983

97

111123

137 155180

194 207

222

236

264

281

310

O

O


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10055

6983

97

111

125137 151 165 179 194 207 221 236 249 261

278296

HO

O


60 80 100 120 140 160 180 200 220 240 260 280 3000

50

100

55 77 90 103118

133

147

161

178

191 262290

O

O

O


60 90 120 150 180 210 240 270 300 330 360 3900

50

100

5770

83 104132

149

167

279

O

O

O

O


60 90 120 150 180 210 240 270 300 330 360 3900

50

100

55

69

81

95

107

121

147

173189

215

231

243

258

286

O

59





50 100 150 200 250 300 350 400 450 500 550 600 6500

50

100

63 91

151

189

287

303

318 347

O

O

OHO OH

H2N

60


SNo Scan

No

Systemic Name

(Common Name)

Mol

Formula

Mol

Wt

Ret

Time

Conc

1 2606 Nanodecane C19H40 268 24168 0036

2 2913 Heptadecane C17H36 240 2641 0035


4 3230 Octadecane C18H38 254 28737 0049





9 3653 Nonadecane C19H40 268 31834 0064




13 4252 Eicosane C20H42 282 36220 0093

14 5475 Oleic Acid C18H34O2 282 45175 0105


16 5546 Ethyl Oleate C20H38O2 310 45694 0065

61


18 6023 Heptacosane C27H56 380 49187 0051


20 6281 Heptacosane C27H56 380 51076 0044




C20H30O 286 53910 004


62





























63





















64



Infection


Fsolani



Control hellip 50 187 75 25 75 50 187 125





P citrinum EPSMR1 375 0 25 0 125 25 0 0



Pduclauxi EPASS9 50 0 62 0 312 312 62 0

Plividum EPMCL12 50 62 50 0 0 50 0 62



Pthomii EPAER11 62 0 62 0 375 187 62 62



Pasperum EPHAL10 125 0 25 187 375 312 62 62






65



Shoot Weight


(cm)

(g)

(cm)

(g)


Control 22775 3993 253 535 643 1162 0645 0675

Carbendazim 2585 418 2216 451 742 1287 0715 0622




P citrinum EPSMR1 2599 4681 218 51 94 862 0726 0807



Pduclauxi EPASS9 2541 4487 243 512 1103 1406 077 0786

Plividum EPMCL12 22685 4587 205 539 631 558 0663 0578



Pthomii EPAER11 2824 44 277 527 1221 1218 1005 0645



Pasperum EPHAL10 26186 4918 214 678 918 757 069 0866

LSD005 5141 7881 07911 1821 2551 2821 01951 031



66




















67

68



69


sunflower


+veControl EP

EP

EP EP EP EP

EP

Control

70


























71



Infection


Fsolani



Control 50 187 50 25 75 75 187 125

Carbendazim 125 62 312 62 125 25 62 62




P citrinum EPSMR1 375 0 25 0 125 25 0 0



Pduclauxi EPASS9 25 0 62 62 312 187 62 0

Plividum EPMCL12 50 62 50 0 0 50 0 62



Pthomii EPAER11 62 0 62 0 375 187 62 62



Pasperum EPHAL10 125 125 25 187 312 312 62 62






72


Treatments Code

Shoot Length

(cm)

Shoot Weight

(g)


(cm)

(g)


Control 3256 3893 378 642 57 1034 085 131

Carbendazim 3781 4293 452 607 84 1025 124 128




P citrinum EPSMR1 385 6443 373 1425 75 787 088 226



Pduclauxi EPASS9 4412 6275 386 1013 7 768 086 213

Plividum EPMCL12 345 6551 206 1019 706 645 072 161



Pthomii EPAER11 4838 6208 489 953 863 656 096 141



Pasperum EPHAL10 4158 5006 362 818 6102 1275 067 186

LSD005 10331 8971 2271 5521 3021 2171 04581 1071




73


in sunflower











after 45 days

EP

Carbendazim Control

74


















75



Infection


Fsolani



Control hellip 50 312 100 75 100 50 0 562





P citrinum EPSMR1 62 62 437 75 0 0 62 62



Pduclauxi EPASS9 0 0 437 375 25 375 62 25

Plividum EPMCL12 62 25 25 687 125 375 62 50



Pthomii EPAER11 62 62 437 25 125 312 0 0



Pasperum EPHAL10 435 125 25 25 25 187 0 0






76



Shoot Weight


(cm)

(g)

(cm)

(g)


Control hellip 1375 1714 078 08 1531 4652 051 014





P citrinum EPSMR1 1299 1606 059 0617 165 477 039 016



Pduclauxi EPASS9 1187 1916 069 0855 1108 4562 017 016

Plividum EPMCL12 132 2147 061 1358 2252 4785 026 022



Pthomii EPAER11 1463 179 077 1203 1125 7025 031 024



Pasperum EPHAL10 1463 1874 077 1102 1125 702 031 02

LSD005 1611 4011 0191 2141 8421 1151 0171 0071




77

















solani (Table 16)




78



Infection


Fsolani



Control hellip 50 50 100 75 100 75 0 187





P citrinum EPSMR1 62 25 437 437 0 437 62 187



Pduclauxi EPASS9 0 312 437 562 25 562 62 65

Plividum EPMCL12 62 125 25 25 125 25 62 0



Pthomii EPAER11 62 187 437 437 125 375 0 0



Pasperum EPHAL10 437 375 25 312 25 562 0 125






79


Treatments Code

Shoot Length


(cm)

(g)

(cm)

(g)


Control hellip 1375 1364 078 089 1531 613 051 031





P citrinum EPSMR1 1299 1318 059 193 165 961 039 037



Pduclauxi EPASS9 1187 1438 069 13 1108 1178 017 048

Plividum EPMCL12 132 1323 061 107 2252 1024 026 048



Pthomii EPAER11 1463 1435 077 119 1125 1185 031 071



Pasperum EPHAL10 1463 1453 077 128 1125 972 031 046

LSD005 1611 2661 0191 091 8421 271 0171 0291



80


mungbean growth


















81




Infection

Control --------- 50 100 100 50

Carbendazim --------- 25 50 50 62
















82



(cm) (g) (cm) (g)

Control ---------- 1475 0522 4972 0098

Carbendazim --------- 1635 0987 3737 009




P citrinum EPSMR1 1344 0987 4617 0137



Pduclauxi EPASS9 1434 0696 4127 0096

Plividum EPMCL12 1639 0752 4147 0121



Pthomii EPAER11 1482 0711 3865 0072



Pasperum EPHAL10 1608 0787 3875 0066

LSD005 2891 0261 0741 0051



83

84
























85



Infection


Fsolani



Control hellip 437 312 625 625 312 0 312 0





P citrinum EPSMR1 0 50 62 625 625 62 75 0



Pduclauxi EPASS9 0 0 62 25 187 125 50 62

Plividum EPMCL12 50 437 437 562 375 0 687 62



Pthomii EPAER11 187 562 312 562 50 312 562 0



Pasperum EPHAL10 62 312 125 562 25 62 812 0






86



(cm) (g) (cm) (g)


Control hellip 12 1544 18 407 126 333 155 063





P citrinum EPSMR1 1732 1755 297 389 922 1149 064 056



Pduclauxi EPASS9 1672 2255 243 636 1185 597 057 11

Plividum EPMCL12 1307 1303 178 254 1242 529 052 046



Pthomii EPAER11 1328 1375 214 234 148 466 046 055



Pasperum EPHAL10 1328 2412 18 732 1225 775 06 126

LSD005 271 5171 0691 2091 3731 3031 1031 0631



87


EP

88























89



Infection


Fsolani



Control hellip 437 50 625 25 312 62 312 0





P citrinum EPSMR1 0 62 62 562 625 0 75 0



Pduclauxi EPASS9 0 562 62 562 187 25 50 0

Plividum EPMCL12 50 187 437 375 375 0 687 0



Pthomii EPAER11 187 312 312 25 50 125 562 0



Pasperum EPHAL10 62 125 125 50 25 62 812 0






90




















weeks

91


Treatments Code

Shoot

Length

Shoot

Length

Shoot

Weight

Shoot

Weight

Root

Length

Root

Length

Root

weight

Root

weight

(cm) (cm) (g) (g) (cm) (cm) (g) (g)


Control hellip 12 1633 18 554 126 1757 155 105





P citrinum EPSMR1 1732 1912 297 512 922 9 064 155



Pduclauxi EPASS9 1672 214 243 69 1185 153 057 237

Plividum EPMCL12 1307 2347 178 741 1242 1298 052 156



Pthomii EPAER11 1328 225 214 657 148 155 046 164



Pasperum EPHAL10 1328 2101 18 525 1225 1095 06 135

LSD005 271 4291 0691 3281 3731 5851 1031 091


92


cake in tomato

EP

93





















94



Infection


Fsolani



Control hellip 375 0 50 125 437 375 25 0





P citrinum EPSMR1 312 187 187 312 375 50 187 0



Pduclauxi EPASS9 187 375 125 25 375 50 0 0

Plividum EPMCL12 62 62 437 125 62 625 25 0



Pthomii EPAER11 0 187 437 187 62 25 0 0



Pasperum EPHAL10 125 62 50 125 125 812 0 0






95



(cm) (g) (cm) (g)


Control hellip 2369 2225 274 837 274 975 211 303





P citrinum EPSMR1 267 3384 313 668 2397 975 394 098



Pduclauxi EPASS9 2925 2911 376 388 3037 1293 522 116

Plividum EPMCL12 2768 2801 31 698 2155 1132 35 109



Pthomii EPAER11 239 30 296 799 2416 1062 427 125



Pasperum EPHAL10 2856 2891 344 763 1921 1352 306 13

LSD005 471 4931 0941 3331 7321 5451 1611 11071



96



EP

S

EP

97


EP

EP

98


in chickpea


in chickpea

EP

EP

99


in chickpea













weeks













(Table 26)

100




101



Infection



Control hellip 375 125 50 312 437 0 25 187





P citrinum EPSMR1 312 0 187 625 375 187 187 312



Pduclauxi EPASS9 187 0 125 625 375 62 0 312

Plividum EPMCL12 62 62 437 100 62 25 25 312



Pthomii EPAER11 0 62 437 125 62 62 0 62



Pasperum EPHAL10 125 0 50 187 125 0 0 0






102



(cm) (g) (cm) (g)


Control hellip 2369 2188 274 406 274 692 211 58





P citrinum EPSMR1 267 1916 313 556 2397 756 394 1172



Pduclauxi EPASS9 2925 192 376 561 3037 1115 522 819

Plividum EPMCL12 2768 1929 31 417 2155 946 35 383



Pthomii EPAER11 239 2305 296 626 2416 9 427 931



Pasperum EPHAL10 2856 1662 344 582 1921 925 306 834

LSD005 471 6131 0941 3011 7321 2921 1611 6151




103













weeks














104



Infection


Control 75 100 100 75

Feast-M 0 37 687 625










Topsin-M 0 437 812 62





P citrinum 437 687 100 0


P decumbens 187 50 437 187

Pasperum 125 50 562 125




105



Control 3197 339 288 288

Feast-M 4269 451 526 526





Carbendazim 3675 319 398 398





Topsin-M 3935 314 383 383





P citrinum 3268 249 432 432

Pnigricans 2788 201 401 401

P decumbens 3421 3007 446 446

Pasperum 3262 229 363 363

LSD005 5751 0811 1041 1041



106








days of germination



















107



Infection


Control 0 50 625 50








108



Fruits Fruit weight

(cm) (g) (cm) (g)

Control 3831 1058 1596 305 023c 708c

Carbendazim 3421 832 1659 546 045b 683c

P decumbens 4523 1167 1756 438 052a 1106a

Pnigricans 4265 1172 1794 188 054a 894b

P rugulosum 4592 1295 1967 2405 025c 533d

LSD005 511 4281 3431 581 00261 04841


109


soil



Control 137e 2711e 2878e 0053d





LSD005 00311 01361 04211 00041


110



Sucrose

Control 587c 0087c 8668d 1353b 245d





LSD005 0121 000571 0211 01031 0121


111


in garden soil








LSD005 10591 01441 21941 3711


112








after 90 days










37)








113



Infection


Control 312 100 937 562








114



(cm) (g) (cm) (g)

Control 52 1974 1816 35 30a 5801b

Carbendazim 4646 1322 1629 237 20c 4083a

P decumbens 443 2161 1283 418 2133c 995a

Pnigricans 55 1892 1561 315 32a 4286d

P rugulosum 5197 1695 1205 334 256b 4779c

LSD005 1481 18611 5391 4011 3781 0131


115


garden soil








LSD005 01081 01671 0301 00791


116



N Sucrose






LSD005 00621 00541 0211 1311


117










LSD005 5591 0471 5771 2391


118



field condition



















days








119



























120



Infection


45 90 45 90 45 90 45 90

Control 375 0 562 312 937 100 562 0

Topsin-M 375 0 625 25 937 100 687 0


P decumbens 62 0 50 375 68 100 375 0

Pnigricans 125 187 562 687 875 100 312 0

P rugulosum 312 62 562 375 812 100 437 0







121


condition


(cm)

Shoot Weight

(g)

Root Length

(cm)

Root Weight

(g)

Number

of Fruits

Fruit

weight

Control 45 90 45 90 45 90 45 90 90 90

Topsin-M 4178 6192 2228 4325 1368 2426 204 823 086g 246i

Pmontelii 422 6375 1765 4731 1267 2377 133 98 12f 31h


P nigricans 4233 6617 1971 4887 1486 2578 167 1003 143e 1146a

Prugulosum 4866 7083 1635 5095 1378 2311 172 967 176d 331g

P rugulosum 4373 7026 2063 2051 1371 2464 169 709 123f 35f




LSD005 961 1321 131 1181 3551 1371 0831 2961 0111 0111


122



Treatments

Polyphenol

microgml



Control 183h 7314e 7721e 007f

Topsin-M 146i 9119a 9886a 0113d








LSD005 00721 10191 06531 00121



123




microgml




Topsin-M 183f 4922f 5575g 0074bc








LSD005 10061 05191 04731 0003081


124





Control 5102g 646g 1466g 5966f

Topsin-M 5514f 716f 2566f 67e








LSD005 10451 06241 14081 2471


125



Treatments pH

Tritable

acidity

Moisture

content

Total

solids

Total Soluble

Solid

Sucrose

Control 624a 0102c 8774b 1222f 1425e

Topsin-M 619ab 0126b 8653e 1339b 1475e








LSD005 00641 00041 03021 0171 02221


126

127

128

4 DISCUSSION





























129































130
































131
































132







2013)
























133































134
































135








Kloepper 1998)























136




condition


























137



source to sink



























138































Conclusion

139



















REFERENCES

140



Eco 15(1) 37-47






University












Ecol 58 921-929








99-102



141


2933



















78(3) 429-435




New York NY






142























21(12) pp 1387-1397









143











Engelmann










Ecol 51(2) 215-229











144





Hortic Sci 35 67-72
























6(5) 162-166



145





Acad of Sci 102(35) 12465-12470










Pathol 111 101-112






41-48



4(1) 34-43







146



185(11) 3458-3468













5 17-24



356



Press)









147





Trop Sci 35 294-299






75270 Pakistan












717-732



Pathol J 2 75-79





148








255(7) 2751-2760







21 2894-2897









Protect 46 627-635








149



Phytopathol 58 1051






















Prod 78 2699-2703





150









207













43



106




Pathol 57 870-876

151



Resour 8(2) 1634-1645






8(6) 687-695
















63(4) 391-400



87 4-10




152



S00218596120005-12



Pak J Bot 47(4) 1547-1551




261







York Pages 86













209-214

153



York pp 660-662






April 1996



Lett 100(8) 084102














Plant Biol 13(1) 86





154





















BMC Microbiol 13 51





70-78




155

















289-295














88

156



Pathol 23 1-7






299(1) 31-37








553-558














157
































158


228 7382





Newslett 5 32-34



Pl Nutrit 22(4-5) 835-853





649



513-515 doi 101126science1136237



129 189-196











159





and Ecol 39(2) 163-165

















1048


Minia Univ







160



62















40 403-408













161



doi 103389fpls00432




32-34






516-522


















162


Toxicol 50(3-4) 829-834





















3 342-351







163



















Mar 55 269-279












164















647






319-343










165








York 326-365




686



106(9) 996-1004





Mycol Res 106(9) 996-1004






13(4) 1698-1709





166






Biol Sci 4 1240- 1242












1623




16(4) 403-406










167





Interact 3(2) 75-93




Ecol 19 2917-2933







1037-1042





2950










168







43(1) 1-6














82










169























43-68







Sci 416 382-385

170




Bioch 37 21312140



Mycologia 98 31-42


















Sci 102(38) 13386-13391



95 1368-1373



147 523- 535

171





Res 4 1100-1104






Drugs 11 2230-2238




Interact 10(1) 280-287









3(1-2) 90-97






489

172




585




36(1) 82


















January 20 2017






173



449-458





vi




suspension







216 Fruit analysis

2161 pH




2165 Firmness

2166 Total solids

2167 Protein

2168 Carbohydrate








Penicillium


Penicillium


vii

Penicillium spp



culture filtrates


culture filtrates


culture filtrates


culture filtrates


rugulosum



















viii





okra plants


tomato plants






4 DISCUSSION

ix



NEMATODE

SUMMARY


























x






















xi

xii

1

1 INTRODUCTION

11 Endophytic fungi




























2

















2014)



spiteller 2012b)











3




(Huang et al 2008)

























plants

4































5
































6






























7






























8






























9



endophytic fungi






10







about (24) hours





















11
















ordmC for 16 min



calibrate the sizes





12





phaseolina)




culture plate assay







times










13




in vitro

















measured in mm

14























Penicillium species





15












regulosum





packard (5890)






16





measured



(2016) was used





polyphenol activity










17









-------------------------------------------------------









sucrose

2165 Firmness



2166 Total solids


moisture from 100

18


2167 Protein


2168 Carbohydrate













used

19





























20

21

22












(Table 1) fig1-7

23







(Solanaceae)

Root 4 4 20 20




(Solanaceae)

root 5 25 16 21


(Amaranthaceae)

root 3 12 11 20


(Meliaceae)

root 5 25 13 20




(Amaranthaceae)

stem 18 13 11 14



(Malvaceae)

root 5 8 5 6


(Cucurbitaceae)

leaf 18 13 11 14


(Solanaceae)

root 5 24 17 22


(Malvaceae)

root 5 3 21 12

24


assay


assay

25


assay



26


assay


assay

27


assay















28













29





Control 0 0 0 0


EPSMR1 P citrinum




EPSMS2 Plilacinum








EPSLR4 P nigricans



30




EPAAR5 P rugulosum




EPAIR6 P decumbens








EPCTS8 Prestrictum




EPASS9 P duclauxi


31





EPHAL10 Pasperum




EPAER11 Pthomii




EPMCL12 P lividum




EPSLR13 P javanicum





32






33



























34


species



Control 0 0 0 0


EPSMR1 P citrinum




EPSMS2 Plilacinum








EPSLR4 P nigricans


35





EPAAR5 P rugulosum




EPAIR6 P decumbens








EPCTS8 P restrictum




EPASS9 P duclauxi

36






EPHAL10 Pasperum




EPAER11 Pthomii




EPMCL12 P lividum




EPSLR13 P javanicum




37







38





spp










A

B

C

E D

39


48 hours


24Hours 48Hours
















40

41























42


Penicillium species



Control 0 0 0 0


EPSLR4 P nigricans




EPAAR5 P rugulosum




EPAIR6 P decumbens






43



EPHAL10 Pasperum




44














45





Control 0 0 0 0 0


EPSLR4 P nigricans



60 microldisc 11 11 9 12 10

EPAAR5 P rugulosum

20microldisc 0 18 18 11 18

40 microldisc 0 21 18 11 22

60 microldisc 0 24 20 18 22

EPAIR6 P decumbens



60 microldisc 0 12 16 0 11




46


EPHAL10 Pasperum


40 microldisc 11 9 6 10 10

60 microldisc 12 11 9 10 12

47





C

+ve C

20microl

60microl

40microl

+veC

20microl

40microl

60microl

C

48





C

60microl

40microl

20microl +veC

vCCe

veve

+veC

vCCe

veve

C

60microl

20microl

40microl

49


Penicillium species



Control 0 0 0 0


EPSLR4 P nigricans




EPAAR5 P rugulosum




EPAIR6 P decumbens







50


EPHAL10 Pasperum








(Table 8 and Fig12)

51





Control 0 0 0 0 0


EPSLR4 P nigricans

20microldisc 16 16 14 14 16

40 microldisc 16 16 14 14 18

60 microldisc 18 16 16 14 20

EPAAR5 P rugulosum

20microldisc 18 18 20 11 20

40 microldisc 18 18 20 11 21

60 microldisc 18 18 20 11 21

EPAIR6 P decumbens






52



EPHAL10 Pasperum




53

4



C

+ve C

20microl 40microl

60microl

54


Penicillium

















55

(1) Nanodecane

(2) Nonadecane

(3) Heptadecane

(4) Heptacosane

(5) Heptacosane

(6) Eicosane

(7) Octadecane

(replib) Nonadecane

50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 2400

50

10057

71

85

99113 127 141 155 169 183 197

(replib) Nonadecane

60 80 100 120 140 160 180 200 220 240 260 2800

50

10057

71

85

99113 127 141 155 169 183 197 268


50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 2500

50

10057

71

85

99113 127 141 155 169 182 196 210 240


60 80 100 120 140 160 180 200 220 240 260 280 300 320 3400

50

10057

71

85

99113 127 141 155 169 183 197 211 225 239 253 267 281 294 308 322 336


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10057

71

85

99113 127 141 155 169 183 197 211 225 239 253 267 281 294 308 322

(mainlib) Eicosane

60 80 100 120 140 160 180 200 220 240 260 2800

50

10057

71

85

99113

127 141 155 169 183 197 211 225 238 252 282

(replib) Octadecane

60 80 100 120 140 160 180 200 220 240 2600

50

10057

71

85

99113 127 141 155 169 183 197 210 225 254

56








50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 2400

50

100

55

60

69

73

8397 115

129

138

143157

171

185

199209

228

OH

O


60 80 100 120 140 160 180 200 220 240 2600

50

10057

71

85

97

113127

141 155 168183 197 212


50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 2500

50

100

60

71

8397

102

111

129

143157

171

185

199

211

228

O

O


60 80 100 120 140 160 180 200 220 240 2600

50

10057

60

71

83 97

102

111 129

143153 165

177

185

195 213225

242O

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

61

70

88

101

115129

143

157

171 185 199 213

227

241 255

270

O

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

69

81

95 109

123

135

140

150

167

177 205223

238

O

OH

57



(16) Oleic Acid



60 80 100 120 140 160 180 200 220 240 2600

50

10060 73

8397

115

129

143157 171 185

199

213

227 239

256

OH

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

61 73

88

101

115129 143

157

171 185 199 213 225239

255 267284

O

O


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

100

55

6781

95

109

123135 150 164 178

192 205 220 234

263

279

308

O

O

(replib) Oleic Acid

60 80 100 120 140 160 180 200 220 240 260 2800

50

10055

69

83

97

111

125137 151 165 180 193 207 222 236

264

282

HO

O

58

(18) Ethyl Oleate





60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10055

6983

97

111123

137 155180

194 207

222

236

264

281

310

O

O


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10055

6983

97

111

125137 151 165 179 194 207 221 236 249 261

278296

HO

O


60 80 100 120 140 160 180 200 220 240 260 280 3000

50

100

55 77 90 103118

133

147

161

178

191 262290

O

O

O


60 90 120 150 180 210 240 270 300 330 360 3900

50

100

5770

83 104132

149

167

279

O

O

O

O


60 90 120 150 180 210 240 270 300 330 360 3900

50

100

55

69

81

95

107

121

147

173189

215

231

243

258

286

O

59





50 100 150 200 250 300 350 400 450 500 550 600 6500

50

100

63 91

151

189

287

303

318 347

O

O

OHO OH

H2N

60


SNo Scan

No

Systemic Name

(Common Name)

Mol

Formula

Mol

Wt

Ret

Time

Conc

1 2606 Nanodecane C19H40 268 24168 0036

2 2913 Heptadecane C17H36 240 2641 0035


4 3230 Octadecane C18H38 254 28737 0049





9 3653 Nonadecane C19H40 268 31834 0064




13 4252 Eicosane C20H42 282 36220 0093

14 5475 Oleic Acid C18H34O2 282 45175 0105


16 5546 Ethyl Oleate C20H38O2 310 45694 0065

61


18 6023 Heptacosane C27H56 380 49187 0051


20 6281 Heptacosane C27H56 380 51076 0044




C20H30O 286 53910 004


62





























63





















64



Infection


Fsolani



Control hellip 50 187 75 25 75 50 187 125





P citrinum EPSMR1 375 0 25 0 125 25 0 0



Pduclauxi EPASS9 50 0 62 0 312 312 62 0

Plividum EPMCL12 50 62 50 0 0 50 0 62



Pthomii EPAER11 62 0 62 0 375 187 62 62



Pasperum EPHAL10 125 0 25 187 375 312 62 62






65



Shoot Weight


(cm)

(g)

(cm)

(g)


Control 22775 3993 253 535 643 1162 0645 0675

Carbendazim 2585 418 2216 451 742 1287 0715 0622




P citrinum EPSMR1 2599 4681 218 51 94 862 0726 0807



Pduclauxi EPASS9 2541 4487 243 512 1103 1406 077 0786

Plividum EPMCL12 22685 4587 205 539 631 558 0663 0578



Pthomii EPAER11 2824 44 277 527 1221 1218 1005 0645



Pasperum EPHAL10 26186 4918 214 678 918 757 069 0866

LSD005 5141 7881 07911 1821 2551 2821 01951 031



66




















67

68



69


sunflower


+veControl EP

EP

EP EP EP EP

EP

Control

70


























71



Infection


Fsolani



Control 50 187 50 25 75 75 187 125

Carbendazim 125 62 312 62 125 25 62 62




P citrinum EPSMR1 375 0 25 0 125 25 0 0



Pduclauxi EPASS9 25 0 62 62 312 187 62 0

Plividum EPMCL12 50 62 50 0 0 50 0 62



Pthomii EPAER11 62 0 62 0 375 187 62 62



Pasperum EPHAL10 125 125 25 187 312 312 62 62






72


Treatments Code

Shoot Length

(cm)

Shoot Weight

(g)


(cm)

(g)


Control 3256 3893 378 642 57 1034 085 131

Carbendazim 3781 4293 452 607 84 1025 124 128




P citrinum EPSMR1 385 6443 373 1425 75 787 088 226



Pduclauxi EPASS9 4412 6275 386 1013 7 768 086 213

Plividum EPMCL12 345 6551 206 1019 706 645 072 161



Pthomii EPAER11 4838 6208 489 953 863 656 096 141



Pasperum EPHAL10 4158 5006 362 818 6102 1275 067 186

LSD005 10331 8971 2271 5521 3021 2171 04581 1071




73


in sunflower











after 45 days

EP

Carbendazim Control

74


















75



Infection


Fsolani



Control hellip 50 312 100 75 100 50 0 562





P citrinum EPSMR1 62 62 437 75 0 0 62 62



Pduclauxi EPASS9 0 0 437 375 25 375 62 25

Plividum EPMCL12 62 25 25 687 125 375 62 50



Pthomii EPAER11 62 62 437 25 125 312 0 0



Pasperum EPHAL10 435 125 25 25 25 187 0 0






76



Shoot Weight


(cm)

(g)

(cm)

(g)


Control hellip 1375 1714 078 08 1531 4652 051 014





P citrinum EPSMR1 1299 1606 059 0617 165 477 039 016



Pduclauxi EPASS9 1187 1916 069 0855 1108 4562 017 016

Plividum EPMCL12 132 2147 061 1358 2252 4785 026 022



Pthomii EPAER11 1463 179 077 1203 1125 7025 031 024



Pasperum EPHAL10 1463 1874 077 1102 1125 702 031 02

LSD005 1611 4011 0191 2141 8421 1151 0171 0071




77

















solani (Table 16)




78



Infection


Fsolani



Control hellip 50 50 100 75 100 75 0 187





P citrinum EPSMR1 62 25 437 437 0 437 62 187



Pduclauxi EPASS9 0 312 437 562 25 562 62 65

Plividum EPMCL12 62 125 25 25 125 25 62 0



Pthomii EPAER11 62 187 437 437 125 375 0 0



Pasperum EPHAL10 437 375 25 312 25 562 0 125






79


Treatments Code

Shoot Length


(cm)

(g)

(cm)

(g)


Control hellip 1375 1364 078 089 1531 613 051 031





P citrinum EPSMR1 1299 1318 059 193 165 961 039 037



Pduclauxi EPASS9 1187 1438 069 13 1108 1178 017 048

Plividum EPMCL12 132 1323 061 107 2252 1024 026 048



Pthomii EPAER11 1463 1435 077 119 1125 1185 031 071



Pasperum EPHAL10 1463 1453 077 128 1125 972 031 046

LSD005 1611 2661 0191 091 8421 271 0171 0291



80


mungbean growth


















81




Infection

Control --------- 50 100 100 50

Carbendazim --------- 25 50 50 62
















82



(cm) (g) (cm) (g)

Control ---------- 1475 0522 4972 0098

Carbendazim --------- 1635 0987 3737 009




P citrinum EPSMR1 1344 0987 4617 0137



Pduclauxi EPASS9 1434 0696 4127 0096

Plividum EPMCL12 1639 0752 4147 0121



Pthomii EPAER11 1482 0711 3865 0072



Pasperum EPHAL10 1608 0787 3875 0066

LSD005 2891 0261 0741 0051



83

84
























85



Infection


Fsolani



Control hellip 437 312 625 625 312 0 312 0





P citrinum EPSMR1 0 50 62 625 625 62 75 0



Pduclauxi EPASS9 0 0 62 25 187 125 50 62

Plividum EPMCL12 50 437 437 562 375 0 687 62



Pthomii EPAER11 187 562 312 562 50 312 562 0



Pasperum EPHAL10 62 312 125 562 25 62 812 0






86



(cm) (g) (cm) (g)


Control hellip 12 1544 18 407 126 333 155 063





P citrinum EPSMR1 1732 1755 297 389 922 1149 064 056



Pduclauxi EPASS9 1672 2255 243 636 1185 597 057 11

Plividum EPMCL12 1307 1303 178 254 1242 529 052 046



Pthomii EPAER11 1328 1375 214 234 148 466 046 055



Pasperum EPHAL10 1328 2412 18 732 1225 775 06 126

LSD005 271 5171 0691 2091 3731 3031 1031 0631



87


EP

88























89



Infection


Fsolani



Control hellip 437 50 625 25 312 62 312 0





P citrinum EPSMR1 0 62 62 562 625 0 75 0



Pduclauxi EPASS9 0 562 62 562 187 25 50 0

Plividum EPMCL12 50 187 437 375 375 0 687 0



Pthomii EPAER11 187 312 312 25 50 125 562 0



Pasperum EPHAL10 62 125 125 50 25 62 812 0






90




















weeks

91


Treatments Code

Shoot

Length

Shoot

Length

Shoot

Weight

Shoot

Weight

Root

Length

Root

Length

Root

weight

Root

weight

(cm) (cm) (g) (g) (cm) (cm) (g) (g)


Control hellip 12 1633 18 554 126 1757 155 105





P citrinum EPSMR1 1732 1912 297 512 922 9 064 155



Pduclauxi EPASS9 1672 214 243 69 1185 153 057 237

Plividum EPMCL12 1307 2347 178 741 1242 1298 052 156



Pthomii EPAER11 1328 225 214 657 148 155 046 164



Pasperum EPHAL10 1328 2101 18 525 1225 1095 06 135

LSD005 271 4291 0691 3281 3731 5851 1031 091


92


cake in tomato

EP

93





















94



Infection


Fsolani



Control hellip 375 0 50 125 437 375 25 0





P citrinum EPSMR1 312 187 187 312 375 50 187 0



Pduclauxi EPASS9 187 375 125 25 375 50 0 0

Plividum EPMCL12 62 62 437 125 62 625 25 0



Pthomii EPAER11 0 187 437 187 62 25 0 0



Pasperum EPHAL10 125 62 50 125 125 812 0 0






95



(cm) (g) (cm) (g)


Control hellip 2369 2225 274 837 274 975 211 303





P citrinum EPSMR1 267 3384 313 668 2397 975 394 098



Pduclauxi EPASS9 2925 2911 376 388 3037 1293 522 116

Plividum EPMCL12 2768 2801 31 698 2155 1132 35 109



Pthomii EPAER11 239 30 296 799 2416 1062 427 125



Pasperum EPHAL10 2856 2891 344 763 1921 1352 306 13

LSD005 471 4931 0941 3331 7321 5451 1611 11071



96



EP

S

EP

97


EP

EP

98


in chickpea


in chickpea

EP

EP

99


in chickpea













weeks













(Table 26)

100




101



Infection



Control hellip 375 125 50 312 437 0 25 187





P citrinum EPSMR1 312 0 187 625 375 187 187 312



Pduclauxi EPASS9 187 0 125 625 375 62 0 312

Plividum EPMCL12 62 62 437 100 62 25 25 312



Pthomii EPAER11 0 62 437 125 62 62 0 62



Pasperum EPHAL10 125 0 50 187 125 0 0 0






102



(cm) (g) (cm) (g)


Control hellip 2369 2188 274 406 274 692 211 58





P citrinum EPSMR1 267 1916 313 556 2397 756 394 1172



Pduclauxi EPASS9 2925 192 376 561 3037 1115 522 819

Plividum EPMCL12 2768 1929 31 417 2155 946 35 383



Pthomii EPAER11 239 2305 296 626 2416 9 427 931



Pasperum EPHAL10 2856 1662 344 582 1921 925 306 834

LSD005 471 6131 0941 3011 7321 2921 1611 6151




103













weeks














104



Infection


Control 75 100 100 75

Feast-M 0 37 687 625










Topsin-M 0 437 812 62





P citrinum 437 687 100 0


P decumbens 187 50 437 187

Pasperum 125 50 562 125




105



Control 3197 339 288 288

Feast-M 4269 451 526 526





Carbendazim 3675 319 398 398





Topsin-M 3935 314 383 383





P citrinum 3268 249 432 432

Pnigricans 2788 201 401 401

P decumbens 3421 3007 446 446

Pasperum 3262 229 363 363

LSD005 5751 0811 1041 1041



106








days of germination



















107



Infection


Control 0 50 625 50








108



Fruits Fruit weight

(cm) (g) (cm) (g)

Control 3831 1058 1596 305 023c 708c

Carbendazim 3421 832 1659 546 045b 683c

P decumbens 4523 1167 1756 438 052a 1106a

Pnigricans 4265 1172 1794 188 054a 894b

P rugulosum 4592 1295 1967 2405 025c 533d

LSD005 511 4281 3431 581 00261 04841


109


soil



Control 137e 2711e 2878e 0053d





LSD005 00311 01361 04211 00041


110



Sucrose

Control 587c 0087c 8668d 1353b 245d





LSD005 0121 000571 0211 01031 0121


111


in garden soil








LSD005 10591 01441 21941 3711


112








after 90 days










37)








113



Infection


Control 312 100 937 562








114



(cm) (g) (cm) (g)

Control 52 1974 1816 35 30a 5801b

Carbendazim 4646 1322 1629 237 20c 4083a

P decumbens 443 2161 1283 418 2133c 995a

Pnigricans 55 1892 1561 315 32a 4286d

P rugulosum 5197 1695 1205 334 256b 4779c

LSD005 1481 18611 5391 4011 3781 0131


115


garden soil








LSD005 01081 01671 0301 00791


116



N Sucrose






LSD005 00621 00541 0211 1311


117










LSD005 5591 0471 5771 2391


118



field condition



















days








119



























120



Infection


45 90 45 90 45 90 45 90

Control 375 0 562 312 937 100 562 0

Topsin-M 375 0 625 25 937 100 687 0


P decumbens 62 0 50 375 68 100 375 0

Pnigricans 125 187 562 687 875 100 312 0

P rugulosum 312 62 562 375 812 100 437 0







121


condition


(cm)

Shoot Weight

(g)

Root Length

(cm)

Root Weight

(g)

Number

of Fruits

Fruit

weight

Control 45 90 45 90 45 90 45 90 90 90

Topsin-M 4178 6192 2228 4325 1368 2426 204 823 086g 246i

Pmontelii 422 6375 1765 4731 1267 2377 133 98 12f 31h


P nigricans 4233 6617 1971 4887 1486 2578 167 1003 143e 1146a

Prugulosum 4866 7083 1635 5095 1378 2311 172 967 176d 331g

P rugulosum 4373 7026 2063 2051 1371 2464 169 709 123f 35f




LSD005 961 1321 131 1181 3551 1371 0831 2961 0111 0111


122



Treatments

Polyphenol

microgml



Control 183h 7314e 7721e 007f

Topsin-M 146i 9119a 9886a 0113d








LSD005 00721 10191 06531 00121



123




microgml




Topsin-M 183f 4922f 5575g 0074bc








LSD005 10061 05191 04731 0003081


124





Control 5102g 646g 1466g 5966f

Topsin-M 5514f 716f 2566f 67e








LSD005 10451 06241 14081 2471


125



Treatments pH

Tritable

acidity

Moisture

content

Total

solids

Total Soluble

Solid

Sucrose

Control 624a 0102c 8774b 1222f 1425e

Topsin-M 619ab 0126b 8653e 1339b 1475e








LSD005 00641 00041 03021 0171 02221


126

127

128

4 DISCUSSION





























129































130
































131
































132







2013)
























133































134
































135








Kloepper 1998)























136




condition


























137



source to sink



























138































Conclusion

139



















REFERENCES

140



Eco 15(1) 37-47






University












Ecol 58 921-929








99-102



141


2933



















78(3) 429-435




New York NY






142























21(12) pp 1387-1397









143











Engelmann










Ecol 51(2) 215-229











144





Hortic Sci 35 67-72
























6(5) 162-166



145





Acad of Sci 102(35) 12465-12470










Pathol 111 101-112






41-48



4(1) 34-43







146



185(11) 3458-3468













5 17-24



356



Press)









147





Trop Sci 35 294-299






75270 Pakistan












717-732



Pathol J 2 75-79





148








255(7) 2751-2760







21 2894-2897









Protect 46 627-635








149



Phytopathol 58 1051






















Prod 78 2699-2703





150









207













43



106




Pathol 57 870-876

151



Resour 8(2) 1634-1645






8(6) 687-695
















63(4) 391-400



87 4-10




152



S00218596120005-12



Pak J Bot 47(4) 1547-1551




261







York Pages 86













209-214

153



York pp 660-662






April 1996



Lett 100(8) 084102














Plant Biol 13(1) 86





154





















BMC Microbiol 13 51





70-78




155

















289-295














88

156



Pathol 23 1-7






299(1) 31-37








553-558














157
































158


228 7382





Newslett 5 32-34



Pl Nutrit 22(4-5) 835-853





649



513-515 doi 101126science1136237



129 189-196











159





and Ecol 39(2) 163-165

















1048


Minia Univ







160



62















40 403-408













161



doi 103389fpls00432




32-34






516-522


















162


Toxicol 50(3-4) 829-834





















3 342-351







163



















Mar 55 269-279












164















647






319-343










165








York 326-365




686



106(9) 996-1004





Mycol Res 106(9) 996-1004






13(4) 1698-1709





166






Biol Sci 4 1240- 1242












1623




16(4) 403-406










167





Interact 3(2) 75-93




Ecol 19 2917-2933







1037-1042





2950










168







43(1) 1-6














82










169























43-68







Sci 416 382-385

170




Bioch 37 21312140



Mycologia 98 31-42


















Sci 102(38) 13386-13391



95 1368-1373



147 523- 535

171





Res 4 1100-1104






Drugs 11 2230-2238




Interact 10(1) 280-287









3(1-2) 90-97






489

172




585




36(1) 82


















January 20 2017






173



449-458





vii

Penicillium spp



culture filtrates


culture filtrates


culture filtrates


culture filtrates


rugulosum



















viii





okra plants


tomato plants






4 DISCUSSION

ix



NEMATODE

SUMMARY


























x






















xi

xii

1

1 INTRODUCTION

11 Endophytic fungi




























2

















2014)



spiteller 2012b)











3




(Huang et al 2008)

























plants

4































5
































6






























7






























8






























9



endophytic fungi






10







about (24) hours





















11
















ordmC for 16 min



calibrate the sizes





12





phaseolina)




culture plate assay







times










13




in vitro

















measured in mm

14























Penicillium species





15












regulosum





packard (5890)






16





measured



(2016) was used





polyphenol activity










17









-------------------------------------------------------









sucrose

2165 Firmness



2166 Total solids


moisture from 100

18


2167 Protein


2168 Carbohydrate













used

19





























20

21

22












(Table 1) fig1-7

23







(Solanaceae)

Root 4 4 20 20




(Solanaceae)

root 5 25 16 21


(Amaranthaceae)

root 3 12 11 20


(Meliaceae)

root 5 25 13 20




(Amaranthaceae)

stem 18 13 11 14



(Malvaceae)

root 5 8 5 6


(Cucurbitaceae)

leaf 18 13 11 14


(Solanaceae)

root 5 24 17 22


(Malvaceae)

root 5 3 21 12

24


assay


assay

25


assay



26


assay


assay

27


assay















28













29





Control 0 0 0 0


EPSMR1 P citrinum




EPSMS2 Plilacinum








EPSLR4 P nigricans



30




EPAAR5 P rugulosum




EPAIR6 P decumbens








EPCTS8 Prestrictum




EPASS9 P duclauxi


31





EPHAL10 Pasperum




EPAER11 Pthomii




EPMCL12 P lividum




EPSLR13 P javanicum





32






33



























34


species



Control 0 0 0 0


EPSMR1 P citrinum




EPSMS2 Plilacinum








EPSLR4 P nigricans


35





EPAAR5 P rugulosum




EPAIR6 P decumbens








EPCTS8 P restrictum




EPASS9 P duclauxi

36






EPHAL10 Pasperum




EPAER11 Pthomii




EPMCL12 P lividum




EPSLR13 P javanicum




37







38





spp










A

B

C

E D

39


48 hours


24Hours 48Hours
















40

41























42


Penicillium species



Control 0 0 0 0


EPSLR4 P nigricans




EPAAR5 P rugulosum




EPAIR6 P decumbens






43



EPHAL10 Pasperum




44














45





Control 0 0 0 0 0


EPSLR4 P nigricans



60 microldisc 11 11 9 12 10

EPAAR5 P rugulosum

20microldisc 0 18 18 11 18

40 microldisc 0 21 18 11 22

60 microldisc 0 24 20 18 22

EPAIR6 P decumbens



60 microldisc 0 12 16 0 11




46


EPHAL10 Pasperum


40 microldisc 11 9 6 10 10

60 microldisc 12 11 9 10 12

47





C

+ve C

20microl

60microl

40microl

+veC

20microl

40microl

60microl

C

48





C

60microl

40microl

20microl +veC

vCCe

veve

+veC

vCCe

veve

C

60microl

20microl

40microl

49


Penicillium species



Control 0 0 0 0


EPSLR4 P nigricans




EPAAR5 P rugulosum




EPAIR6 P decumbens







50


EPHAL10 Pasperum








(Table 8 and Fig12)

51





Control 0 0 0 0 0


EPSLR4 P nigricans

20microldisc 16 16 14 14 16

40 microldisc 16 16 14 14 18

60 microldisc 18 16 16 14 20

EPAAR5 P rugulosum

20microldisc 18 18 20 11 20

40 microldisc 18 18 20 11 21

60 microldisc 18 18 20 11 21

EPAIR6 P decumbens






52



EPHAL10 Pasperum




53

4



C

+ve C

20microl 40microl

60microl

54


Penicillium

















55

(1) Nanodecane

(2) Nonadecane

(3) Heptadecane

(4) Heptacosane

(5) Heptacosane

(6) Eicosane

(7) Octadecane

(replib) Nonadecane

50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 2400

50

10057

71

85

99113 127 141 155 169 183 197

(replib) Nonadecane

60 80 100 120 140 160 180 200 220 240 260 2800

50

10057

71

85

99113 127 141 155 169 183 197 268


50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 2500

50

10057

71

85

99113 127 141 155 169 182 196 210 240


60 80 100 120 140 160 180 200 220 240 260 280 300 320 3400

50

10057

71

85

99113 127 141 155 169 183 197 211 225 239 253 267 281 294 308 322 336


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10057

71

85

99113 127 141 155 169 183 197 211 225 239 253 267 281 294 308 322

(mainlib) Eicosane

60 80 100 120 140 160 180 200 220 240 260 2800

50

10057

71

85

99113

127 141 155 169 183 197 211 225 238 252 282

(replib) Octadecane

60 80 100 120 140 160 180 200 220 240 2600

50

10057

71

85

99113 127 141 155 169 183 197 210 225 254

56








50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 2400

50

100

55

60

69

73

8397 115

129

138

143157

171

185

199209

228

OH

O


60 80 100 120 140 160 180 200 220 240 2600

50

10057

71

85

97

113127

141 155 168183 197 212


50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 2500

50

100

60

71

8397

102

111

129

143157

171

185

199

211

228

O

O


60 80 100 120 140 160 180 200 220 240 2600

50

10057

60

71

83 97

102

111 129

143153 165

177

185

195 213225

242O

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

61

70

88

101

115129

143

157

171 185 199 213

227

241 255

270

O

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

69

81

95 109

123

135

140

150

167

177 205223

238

O

OH

57



(16) Oleic Acid



60 80 100 120 140 160 180 200 220 240 2600

50

10060 73

8397

115

129

143157 171 185

199

213

227 239

256

OH

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

61 73

88

101

115129 143

157

171 185 199 213 225239

255 267284

O

O


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

100

55

6781

95

109

123135 150 164 178

192 205 220 234

263

279

308

O

O

(replib) Oleic Acid

60 80 100 120 140 160 180 200 220 240 260 2800

50

10055

69

83

97

111

125137 151 165 180 193 207 222 236

264

282

HO

O

58

(18) Ethyl Oleate





60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10055

6983

97

111123

137 155180

194 207

222

236

264

281

310

O

O


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10055

6983

97

111

125137 151 165 179 194 207 221 236 249 261

278296

HO

O


60 80 100 120 140 160 180 200 220 240 260 280 3000

50

100

55 77 90 103118

133

147

161

178

191 262290

O

O

O


60 90 120 150 180 210 240 270 300 330 360 3900

50

100

5770

83 104132

149

167

279

O

O

O

O


60 90 120 150 180 210 240 270 300 330 360 3900

50

100

55

69

81

95

107

121

147

173189

215

231

243

258

286

O

59





50 100 150 200 250 300 350 400 450 500 550 600 6500

50

100

63 91

151

189

287

303

318 347

O

O

OHO OH

H2N

60


SNo Scan

No

Systemic Name

(Common Name)

Mol

Formula

Mol

Wt

Ret

Time

Conc

1 2606 Nanodecane C19H40 268 24168 0036

2 2913 Heptadecane C17H36 240 2641 0035


4 3230 Octadecane C18H38 254 28737 0049





9 3653 Nonadecane C19H40 268 31834 0064




13 4252 Eicosane C20H42 282 36220 0093

14 5475 Oleic Acid C18H34O2 282 45175 0105


16 5546 Ethyl Oleate C20H38O2 310 45694 0065

61


18 6023 Heptacosane C27H56 380 49187 0051


20 6281 Heptacosane C27H56 380 51076 0044




C20H30O 286 53910 004


62





























63





















64



Infection


Fsolani



Control hellip 50 187 75 25 75 50 187 125





P citrinum EPSMR1 375 0 25 0 125 25 0 0



Pduclauxi EPASS9 50 0 62 0 312 312 62 0

Plividum EPMCL12 50 62 50 0 0 50 0 62



Pthomii EPAER11 62 0 62 0 375 187 62 62



Pasperum EPHAL10 125 0 25 187 375 312 62 62






65



Shoot Weight


(cm)

(g)

(cm)

(g)


Control 22775 3993 253 535 643 1162 0645 0675

Carbendazim 2585 418 2216 451 742 1287 0715 0622




P citrinum EPSMR1 2599 4681 218 51 94 862 0726 0807



Pduclauxi EPASS9 2541 4487 243 512 1103 1406 077 0786

Plividum EPMCL12 22685 4587 205 539 631 558 0663 0578



Pthomii EPAER11 2824 44 277 527 1221 1218 1005 0645



Pasperum EPHAL10 26186 4918 214 678 918 757 069 0866

LSD005 5141 7881 07911 1821 2551 2821 01951 031



66




















67

68



69


sunflower


+veControl EP

EP

EP EP EP EP

EP

Control

70


























71



Infection


Fsolani



Control 50 187 50 25 75 75 187 125

Carbendazim 125 62 312 62 125 25 62 62




P citrinum EPSMR1 375 0 25 0 125 25 0 0



Pduclauxi EPASS9 25 0 62 62 312 187 62 0

Plividum EPMCL12 50 62 50 0 0 50 0 62



Pthomii EPAER11 62 0 62 0 375 187 62 62



Pasperum EPHAL10 125 125 25 187 312 312 62 62






72


Treatments Code

Shoot Length

(cm)

Shoot Weight

(g)


(cm)

(g)


Control 3256 3893 378 642 57 1034 085 131

Carbendazim 3781 4293 452 607 84 1025 124 128




P citrinum EPSMR1 385 6443 373 1425 75 787 088 226



Pduclauxi EPASS9 4412 6275 386 1013 7 768 086 213

Plividum EPMCL12 345 6551 206 1019 706 645 072 161



Pthomii EPAER11 4838 6208 489 953 863 656 096 141



Pasperum EPHAL10 4158 5006 362 818 6102 1275 067 186

LSD005 10331 8971 2271 5521 3021 2171 04581 1071




73


in sunflower











after 45 days

EP

Carbendazim Control

74


















75



Infection


Fsolani



Control hellip 50 312 100 75 100 50 0 562





P citrinum EPSMR1 62 62 437 75 0 0 62 62



Pduclauxi EPASS9 0 0 437 375 25 375 62 25

Plividum EPMCL12 62 25 25 687 125 375 62 50



Pthomii EPAER11 62 62 437 25 125 312 0 0



Pasperum EPHAL10 435 125 25 25 25 187 0 0






76



Shoot Weight


(cm)

(g)

(cm)

(g)


Control hellip 1375 1714 078 08 1531 4652 051 014





P citrinum EPSMR1 1299 1606 059 0617 165 477 039 016



Pduclauxi EPASS9 1187 1916 069 0855 1108 4562 017 016

Plividum EPMCL12 132 2147 061 1358 2252 4785 026 022



Pthomii EPAER11 1463 179 077 1203 1125 7025 031 024



Pasperum EPHAL10 1463 1874 077 1102 1125 702 031 02

LSD005 1611 4011 0191 2141 8421 1151 0171 0071




77

















solani (Table 16)




78



Infection


Fsolani



Control hellip 50 50 100 75 100 75 0 187





P citrinum EPSMR1 62 25 437 437 0 437 62 187



Pduclauxi EPASS9 0 312 437 562 25 562 62 65

Plividum EPMCL12 62 125 25 25 125 25 62 0



Pthomii EPAER11 62 187 437 437 125 375 0 0



Pasperum EPHAL10 437 375 25 312 25 562 0 125






79


Treatments Code

Shoot Length


(cm)

(g)

(cm)

(g)


Control hellip 1375 1364 078 089 1531 613 051 031





P citrinum EPSMR1 1299 1318 059 193 165 961 039 037



Pduclauxi EPASS9 1187 1438 069 13 1108 1178 017 048

Plividum EPMCL12 132 1323 061 107 2252 1024 026 048



Pthomii EPAER11 1463 1435 077 119 1125 1185 031 071



Pasperum EPHAL10 1463 1453 077 128 1125 972 031 046

LSD005 1611 2661 0191 091 8421 271 0171 0291



80


mungbean growth


















81




Infection

Control --------- 50 100 100 50

Carbendazim --------- 25 50 50 62
















82



(cm) (g) (cm) (g)

Control ---------- 1475 0522 4972 0098

Carbendazim --------- 1635 0987 3737 009




P citrinum EPSMR1 1344 0987 4617 0137



Pduclauxi EPASS9 1434 0696 4127 0096

Plividum EPMCL12 1639 0752 4147 0121



Pthomii EPAER11 1482 0711 3865 0072



Pasperum EPHAL10 1608 0787 3875 0066

LSD005 2891 0261 0741 0051



83

84
























85



Infection


Fsolani



Control hellip 437 312 625 625 312 0 312 0





P citrinum EPSMR1 0 50 62 625 625 62 75 0



Pduclauxi EPASS9 0 0 62 25 187 125 50 62

Plividum EPMCL12 50 437 437 562 375 0 687 62



Pthomii EPAER11 187 562 312 562 50 312 562 0



Pasperum EPHAL10 62 312 125 562 25 62 812 0






86



(cm) (g) (cm) (g)


Control hellip 12 1544 18 407 126 333 155 063





P citrinum EPSMR1 1732 1755 297 389 922 1149 064 056



Pduclauxi EPASS9 1672 2255 243 636 1185 597 057 11

Plividum EPMCL12 1307 1303 178 254 1242 529 052 046



Pthomii EPAER11 1328 1375 214 234 148 466 046 055



Pasperum EPHAL10 1328 2412 18 732 1225 775 06 126

LSD005 271 5171 0691 2091 3731 3031 1031 0631



87


EP

88























89



Infection


Fsolani



Control hellip 437 50 625 25 312 62 312 0





P citrinum EPSMR1 0 62 62 562 625 0 75 0



Pduclauxi EPASS9 0 562 62 562 187 25 50 0

Plividum EPMCL12 50 187 437 375 375 0 687 0



Pthomii EPAER11 187 312 312 25 50 125 562 0



Pasperum EPHAL10 62 125 125 50 25 62 812 0






90




















weeks

91


Treatments Code

Shoot

Length

Shoot

Length

Shoot

Weight

Shoot

Weight

Root

Length

Root

Length

Root

weight

Root

weight

(cm) (cm) (g) (g) (cm) (cm) (g) (g)


Control hellip 12 1633 18 554 126 1757 155 105





P citrinum EPSMR1 1732 1912 297 512 922 9 064 155



Pduclauxi EPASS9 1672 214 243 69 1185 153 057 237

Plividum EPMCL12 1307 2347 178 741 1242 1298 052 156



Pthomii EPAER11 1328 225 214 657 148 155 046 164



Pasperum EPHAL10 1328 2101 18 525 1225 1095 06 135

LSD005 271 4291 0691 3281 3731 5851 1031 091


92


cake in tomato

EP

93





















94



Infection


Fsolani



Control hellip 375 0 50 125 437 375 25 0





P citrinum EPSMR1 312 187 187 312 375 50 187 0



Pduclauxi EPASS9 187 375 125 25 375 50 0 0

Plividum EPMCL12 62 62 437 125 62 625 25 0



Pthomii EPAER11 0 187 437 187 62 25 0 0



Pasperum EPHAL10 125 62 50 125 125 812 0 0






95



(cm) (g) (cm) (g)


Control hellip 2369 2225 274 837 274 975 211 303





P citrinum EPSMR1 267 3384 313 668 2397 975 394 098



Pduclauxi EPASS9 2925 2911 376 388 3037 1293 522 116

Plividum EPMCL12 2768 2801 31 698 2155 1132 35 109



Pthomii EPAER11 239 30 296 799 2416 1062 427 125



Pasperum EPHAL10 2856 2891 344 763 1921 1352 306 13

LSD005 471 4931 0941 3331 7321 5451 1611 11071



96



EP

S

EP

97


EP

EP

98


in chickpea


in chickpea

EP

EP

99


in chickpea













weeks













(Table 26)

100




101



Infection



Control hellip 375 125 50 312 437 0 25 187





P citrinum EPSMR1 312 0 187 625 375 187 187 312



Pduclauxi EPASS9 187 0 125 625 375 62 0 312

Plividum EPMCL12 62 62 437 100 62 25 25 312



Pthomii EPAER11 0 62 437 125 62 62 0 62



Pasperum EPHAL10 125 0 50 187 125 0 0 0






102



(cm) (g) (cm) (g)


Control hellip 2369 2188 274 406 274 692 211 58





P citrinum EPSMR1 267 1916 313 556 2397 756 394 1172



Pduclauxi EPASS9 2925 192 376 561 3037 1115 522 819

Plividum EPMCL12 2768 1929 31 417 2155 946 35 383



Pthomii EPAER11 239 2305 296 626 2416 9 427 931



Pasperum EPHAL10 2856 1662 344 582 1921 925 306 834

LSD005 471 6131 0941 3011 7321 2921 1611 6151




103













weeks














104



Infection


Control 75 100 100 75

Feast-M 0 37 687 625










Topsin-M 0 437 812 62





P citrinum 437 687 100 0


P decumbens 187 50 437 187

Pasperum 125 50 562 125




105



Control 3197 339 288 288

Feast-M 4269 451 526 526





Carbendazim 3675 319 398 398





Topsin-M 3935 314 383 383





P citrinum 3268 249 432 432

Pnigricans 2788 201 401 401

P decumbens 3421 3007 446 446

Pasperum 3262 229 363 363

LSD005 5751 0811 1041 1041



106








days of germination



















107



Infection


Control 0 50 625 50








108



Fruits Fruit weight

(cm) (g) (cm) (g)

Control 3831 1058 1596 305 023c 708c

Carbendazim 3421 832 1659 546 045b 683c

P decumbens 4523 1167 1756 438 052a 1106a

Pnigricans 4265 1172 1794 188 054a 894b

P rugulosum 4592 1295 1967 2405 025c 533d

LSD005 511 4281 3431 581 00261 04841


109


soil



Control 137e 2711e 2878e 0053d





LSD005 00311 01361 04211 00041


110



Sucrose

Control 587c 0087c 8668d 1353b 245d





LSD005 0121 000571 0211 01031 0121


111


in garden soil








LSD005 10591 01441 21941 3711


112








after 90 days










37)








113



Infection


Control 312 100 937 562








114



(cm) (g) (cm) (g)

Control 52 1974 1816 35 30a 5801b

Carbendazim 4646 1322 1629 237 20c 4083a

P decumbens 443 2161 1283 418 2133c 995a

Pnigricans 55 1892 1561 315 32a 4286d

P rugulosum 5197 1695 1205 334 256b 4779c

LSD005 1481 18611 5391 4011 3781 0131


115


garden soil








LSD005 01081 01671 0301 00791


116



N Sucrose






LSD005 00621 00541 0211 1311


117










LSD005 5591 0471 5771 2391


118



field condition



















days








119



























120



Infection


45 90 45 90 45 90 45 90

Control 375 0 562 312 937 100 562 0

Topsin-M 375 0 625 25 937 100 687 0


P decumbens 62 0 50 375 68 100 375 0

Pnigricans 125 187 562 687 875 100 312 0

P rugulosum 312 62 562 375 812 100 437 0







121


condition


(cm)

Shoot Weight

(g)

Root Length

(cm)

Root Weight

(g)

Number

of Fruits

Fruit

weight

Control 45 90 45 90 45 90 45 90 90 90

Topsin-M 4178 6192 2228 4325 1368 2426 204 823 086g 246i

Pmontelii 422 6375 1765 4731 1267 2377 133 98 12f 31h


P nigricans 4233 6617 1971 4887 1486 2578 167 1003 143e 1146a

Prugulosum 4866 7083 1635 5095 1378 2311 172 967 176d 331g

P rugulosum 4373 7026 2063 2051 1371 2464 169 709 123f 35f




LSD005 961 1321 131 1181 3551 1371 0831 2961 0111 0111


122



Treatments

Polyphenol

microgml



Control 183h 7314e 7721e 007f

Topsin-M 146i 9119a 9886a 0113d








LSD005 00721 10191 06531 00121



123




microgml




Topsin-M 183f 4922f 5575g 0074bc








LSD005 10061 05191 04731 0003081


124





Control 5102g 646g 1466g 5966f

Topsin-M 5514f 716f 2566f 67e








LSD005 10451 06241 14081 2471


125



Treatments pH

Tritable

acidity

Moisture

content

Total

solids

Total Soluble

Solid

Sucrose

Control 624a 0102c 8774b 1222f 1425e

Topsin-M 619ab 0126b 8653e 1339b 1475e








LSD005 00641 00041 03021 0171 02221


126

127

128

4 DISCUSSION





























129































130
































131
































132







2013)
























133































134
































135








Kloepper 1998)























136




condition


























137



source to sink



























138































Conclusion

139



















REFERENCES

140



Eco 15(1) 37-47






University












Ecol 58 921-929








99-102



141


2933



















78(3) 429-435




New York NY






142























21(12) pp 1387-1397









143











Engelmann










Ecol 51(2) 215-229











144





Hortic Sci 35 67-72
























6(5) 162-166



145





Acad of Sci 102(35) 12465-12470










Pathol 111 101-112






41-48



4(1) 34-43







146



185(11) 3458-3468













5 17-24



356



Press)









147





Trop Sci 35 294-299






75270 Pakistan












717-732



Pathol J 2 75-79





148








255(7) 2751-2760







21 2894-2897









Protect 46 627-635








149



Phytopathol 58 1051






















Prod 78 2699-2703





150









207













43



106




Pathol 57 870-876

151



Resour 8(2) 1634-1645






8(6) 687-695
















63(4) 391-400



87 4-10




152



S00218596120005-12



Pak J Bot 47(4) 1547-1551




261







York Pages 86













209-214

153



York pp 660-662






April 1996



Lett 100(8) 084102














Plant Biol 13(1) 86





154





















BMC Microbiol 13 51





70-78




155

















289-295














88

156



Pathol 23 1-7






299(1) 31-37








553-558














157
































158


228 7382





Newslett 5 32-34



Pl Nutrit 22(4-5) 835-853





649



513-515 doi 101126science1136237



129 189-196











159





and Ecol 39(2) 163-165

















1048


Minia Univ







160



62















40 403-408













161



doi 103389fpls00432




32-34






516-522


















162


Toxicol 50(3-4) 829-834





















3 342-351







163



















Mar 55 269-279












164















647






319-343










165








York 326-365




686



106(9) 996-1004





Mycol Res 106(9) 996-1004






13(4) 1698-1709





166






Biol Sci 4 1240- 1242












1623




16(4) 403-406










167





Interact 3(2) 75-93




Ecol 19 2917-2933







1037-1042





2950










168







43(1) 1-6














82










169























43-68







Sci 416 382-385

170




Bioch 37 21312140



Mycologia 98 31-42


















Sci 102(38) 13386-13391



95 1368-1373



147 523- 535

171





Res 4 1100-1104






Drugs 11 2230-2238




Interact 10(1) 280-287









3(1-2) 90-97






489

172




585




36(1) 82


















January 20 2017






173



449-458





viii





okra plants


tomato plants






4 DISCUSSION

ix



NEMATODE

SUMMARY


























x






















xi

xii

1

1 INTRODUCTION

11 Endophytic fungi




























2

















2014)



spiteller 2012b)











3




(Huang et al 2008)

























plants

4































5
































6






























7






























8






























9



endophytic fungi






10







about (24) hours





















11
















ordmC for 16 min



calibrate the sizes





12





phaseolina)




culture plate assay







times










13




in vitro

















measured in mm

14























Penicillium species





15












regulosum





packard (5890)






16





measured



(2016) was used





polyphenol activity










17









-------------------------------------------------------









sucrose

2165 Firmness



2166 Total solids


moisture from 100

18


2167 Protein


2168 Carbohydrate













used

19





























20

21

22












(Table 1) fig1-7

23







(Solanaceae)

Root 4 4 20 20




(Solanaceae)

root 5 25 16 21


(Amaranthaceae)

root 3 12 11 20


(Meliaceae)

root 5 25 13 20




(Amaranthaceae)

stem 18 13 11 14



(Malvaceae)

root 5 8 5 6


(Cucurbitaceae)

leaf 18 13 11 14


(Solanaceae)

root 5 24 17 22


(Malvaceae)

root 5 3 21 12

24


assay


assay

25


assay



26


assay


assay

27


assay















28













29





Control 0 0 0 0


EPSMR1 P citrinum




EPSMS2 Plilacinum








EPSLR4 P nigricans



30




EPAAR5 P rugulosum




EPAIR6 P decumbens








EPCTS8 Prestrictum




EPASS9 P duclauxi


31





EPHAL10 Pasperum




EPAER11 Pthomii




EPMCL12 P lividum




EPSLR13 P javanicum





32






33



























34


species



Control 0 0 0 0


EPSMR1 P citrinum




EPSMS2 Plilacinum








EPSLR4 P nigricans


35





EPAAR5 P rugulosum




EPAIR6 P decumbens








EPCTS8 P restrictum




EPASS9 P duclauxi

36






EPHAL10 Pasperum




EPAER11 Pthomii




EPMCL12 P lividum




EPSLR13 P javanicum




37







38





spp










A

B

C

E D

39


48 hours


24Hours 48Hours
















40

41























42


Penicillium species



Control 0 0 0 0


EPSLR4 P nigricans




EPAAR5 P rugulosum




EPAIR6 P decumbens






43



EPHAL10 Pasperum




44














45





Control 0 0 0 0 0


EPSLR4 P nigricans



60 microldisc 11 11 9 12 10

EPAAR5 P rugulosum

20microldisc 0 18 18 11 18

40 microldisc 0 21 18 11 22

60 microldisc 0 24 20 18 22

EPAIR6 P decumbens



60 microldisc 0 12 16 0 11




46


EPHAL10 Pasperum


40 microldisc 11 9 6 10 10

60 microldisc 12 11 9 10 12

47





C

+ve C

20microl

60microl

40microl

+veC

20microl

40microl

60microl

C

48





C

60microl

40microl

20microl +veC

vCCe

veve

+veC

vCCe

veve

C

60microl

20microl

40microl

49


Penicillium species



Control 0 0 0 0


EPSLR4 P nigricans




EPAAR5 P rugulosum




EPAIR6 P decumbens







50


EPHAL10 Pasperum








(Table 8 and Fig12)

51





Control 0 0 0 0 0


EPSLR4 P nigricans

20microldisc 16 16 14 14 16

40 microldisc 16 16 14 14 18

60 microldisc 18 16 16 14 20

EPAAR5 P rugulosum

20microldisc 18 18 20 11 20

40 microldisc 18 18 20 11 21

60 microldisc 18 18 20 11 21

EPAIR6 P decumbens






52



EPHAL10 Pasperum




53

4



C

+ve C

20microl 40microl

60microl

54


Penicillium

















55

(1) Nanodecane

(2) Nonadecane

(3) Heptadecane

(4) Heptacosane

(5) Heptacosane

(6) Eicosane

(7) Octadecane

(replib) Nonadecane

50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 2400

50

10057

71

85

99113 127 141 155 169 183 197

(replib) Nonadecane

60 80 100 120 140 160 180 200 220 240 260 2800

50

10057

71

85

99113 127 141 155 169 183 197 268


50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 2500

50

10057

71

85

99113 127 141 155 169 182 196 210 240


60 80 100 120 140 160 180 200 220 240 260 280 300 320 3400

50

10057

71

85

99113 127 141 155 169 183 197 211 225 239 253 267 281 294 308 322 336


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10057

71

85

99113 127 141 155 169 183 197 211 225 239 253 267 281 294 308 322

(mainlib) Eicosane

60 80 100 120 140 160 180 200 220 240 260 2800

50

10057

71

85

99113

127 141 155 169 183 197 211 225 238 252 282

(replib) Octadecane

60 80 100 120 140 160 180 200 220 240 2600

50

10057

71

85

99113 127 141 155 169 183 197 210 225 254

56








50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 2400

50

100

55

60

69

73

8397 115

129

138

143157

171

185

199209

228

OH

O


60 80 100 120 140 160 180 200 220 240 2600

50

10057

71

85

97

113127

141 155 168183 197 212


50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 2500

50

100

60

71

8397

102

111

129

143157

171

185

199

211

228

O

O


60 80 100 120 140 160 180 200 220 240 2600

50

10057

60

71

83 97

102

111 129

143153 165

177

185

195 213225

242O

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

61

70

88

101

115129

143

157

171 185 199 213

227

241 255

270

O

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

69

81

95 109

123

135

140

150

167

177 205223

238

O

OH

57



(16) Oleic Acid



60 80 100 120 140 160 180 200 220 240 2600

50

10060 73

8397

115

129

143157 171 185

199

213

227 239

256

OH

O


60 80 100 120 140 160 180 200 220 240 260 2800

50

100

55

61 73

88

101

115129 143

157

171 185 199 213 225239

255 267284

O

O


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

100

55

6781

95

109

123135 150 164 178

192 205 220 234

263

279

308

O

O

(replib) Oleic Acid

60 80 100 120 140 160 180 200 220 240 260 2800

50

10055

69

83

97

111

125137 151 165 180 193 207 222 236

264

282

HO

O

58

(18) Ethyl Oleate





60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10055

6983

97

111123

137 155180

194 207

222

236

264

281

310

O

O


60 80 100 120 140 160 180 200 220 240 260 280 300 3200

50

10055

6983

97

111

125137 151 165 179 194 207 221 236 249 261

278296

HO

O


60 80 100 120 140 160 180 200 220 240 260 280 3000

50

100

55 77 90 103118

133

147

161

178

191 262290

O

O

O


60 90 120 150 180 210 240 270 300 330 360 3900

50

100

5770

83 104132

149

167

279

O

O

O

O


60 90 120 150 180 210 240 270 300 330 360 3900

50

100

55

69

81

95

107

121

147

173189

215

231

243

258

286

O

59





50 100 150 200 250 300 350 400 450 500 550 600 6500

50

100

63 91

151

189

287

303

318 347

O

O

OHO OH

H2N

60


SNo Scan

No

Systemic Name

(Common Name)

Mol

Formula

Mol

Wt

Ret

Time

Conc

1 2606 Nanodecane C19H40 268 24168 0036

2 2913 Heptadecane C17H36 240 2641 0035


4 3230 Octadecane C18H38 254 28737 0049





9 3653 Nonadecane C19H40 268 31834 0064




13 4252 Eicosane C20H42 282 36220 0093

14 5475 Oleic Acid C18H34O2 282 45175 0105


16 5546 Ethyl Oleate C20H38O2 310 45694 0065

61


18 6023 Heptacosane C27H56 380 49187 0051


20 6281 Heptacosane C27H56 380 51076 0044




C20H30O 286 53910 004


62





























63





















64



Infection


Fsolani



Control hellip 50 187 75 25 75 50 187 125





P citrinum EPSMR1 375 0 25 0 125 25 0 0



Pduclauxi EPASS9 50 0 62 0 312 312 62 0

Plividum EPMCL12 50 62 50 0 0 50 0 62



Pthomii EPAER11 62 0 62 0 375 187 62 62



Pasperum EPHAL10 125 0 25 187 375 312 62 62






65



Shoot Weight


(cm)

(g)

(cm)

(g)


Control 22775 3993 253 535 643 1162 0645 0675

Carbendazim 2585 418 2216 451 742 1287 0715 0622




P citrinum EPSMR1 2599 4681 218 51 94 862 0726 0807



Pduclauxi EPASS9 2541 4487 243 512 1103 1406 077 0786

Plividum EPMCL12 22685 4587 205 539 631 558 0663 0578



Pthomii EPAER11 2824 44 277 527 1221 1218 1005 0645



Pasperum EPHAL10 26186 4918 214 678 918 757 069 0866

LSD005 5141 7881 07911 1821 2551 2821 01951 031



66




















67

68



69


sunflower


+veControl EP

EP

EP EP EP EP

EP

Control

70


























71



Infection


Fsolani



Control 50 187 50 25 75 75 187 125

Carbendazim 125 62 312 62 125 25 62 62




P citrinum EPSMR1 375 0 25 0 125 25 0 0



Pduclauxi EPASS9 25 0 62 62 312 187 62 0

Plividum EPMCL12 50 62 50 0 0 50 0 62



Pthomii EPAER11 62 0 62 0 375 187 62 62



Pasperum EPHAL10 125 125 25 187 312 312 62 62






72


Treatments Code

Shoot Length

(cm)

Shoot Weight

(g)


(cm)

(g)


Control 3256 3893 378 642 57 1034 085 131

Carbendazim 3781 4293 452 607 84 1025 124 128




P citrinum EPSMR1 385 6443 373 1425 75 787 088 226



Pduclauxi EPASS9 4412 6275 386 1013 7 768 086 213

Plividum EPMCL12 345 6551 206 1019 706 645 072 161



Pthomii EPAER11 4838 6208 489 953 863 656 096 141



Pasperum EPHAL10 4158 5006 362 818 6102 1275 067 186

LSD005 10331 8971 2271 5521 3021 2171 04581 1071




73


in sunflower











after 45 days

EP

Carbendazim Control

74


















75



Infection


Fsolani



Control hellip 50 312 100 75 100 50 0 562





P citrinum EPSMR1 62 62 437 75 0 0 62 62



Pduclauxi EPASS9 0 0 437 375 25 375 62 25

Plividum EPMCL12 62 25 25 687 125 375 62 50



Pthomii EPAER11 62 62 437 25 125 312 0 0



Pasperum EPHAL10 435 125 25 25 25 187 0 0






76



Shoot Weight


(cm)

(g)

(cm)

(g)


Control hellip 1375 1714 078 08 1531 4652 051 014





P citrinum EPSMR1 1299 1606 059 0617 165 477 039 016



Pduclauxi EPASS9 1187 1916 069 0855 1108 4562 017 016

Plividum EPMCL12 132 2147 061 1358 2252 4785 026 022



Pthomii EPAER11 1463 179 077 1203 1125 7025 031 024



Pasperum EPHAL10 1463 1874 077 1102 1125 702 031 02

LSD005 1611 4011 0191 2141 8421 1151 0171 0071




77

















solani (Table 16)




78



Infection


Fsolani



Control hellip 50 50 100 75 100 75 0 187





P citrinum EPSMR1 62 25 437 437 0 437 62 187



Pduclauxi EPASS9 0 312 437 562 25 562 62 65

Plividum EPMCL12 62 125 25 25 125 25 62 0



Pthomii EPAER11 62 187 437 437 125 375 0 0



Pasperum EPHAL10 437 375 25 312 25 562 0 125






79


Treatments Code

Shoot Length


(cm)

(g)

(cm)

(g)


Control hellip 1375 1364 078 089 1531 613 051 031





P citrinum EPSMR1 1299 1318 059 193 165 961 039 037



Pduclauxi EPASS9 1187 1438 069 13 1108 1178 017 048

Plividum EPMCL12 132 1323 061 107 2252 1024 026 048



Pthomii EPAER11 1463 1435 077 119 1125 1185 031 071



Pasperum EPHAL10 1463 1453 077 128 1125 972 031 046

LSD005 1611 2661 0191 091 8421 271 0171 0291



80


mungbean growth


















81




Infection

Control --------- 50 100 100 50

Carbendazim --------- 25 50 50 62
















82



(cm) (g) (cm) (g)

Control ---------- 1475 0522 4972 0098

Carbendazim --------- 1635 0987 3737 009




P citrinum EPSMR1 1344 0987 4617 0137



Pduclauxi EPASS9 1434 0696 4127 0096

Plividum EPMCL12 1639 0752 4147 0121



Pthomii EPAER11 1482 0711 3865 0072



Pasperum EPHAL10 1608 0787 3875 0066

LSD005 2891 0261 0741 0051



83

84
























85



Infection


Fsolani



Control hellip 437 312 625 625 312 0 312 0





P citrinum EPSMR1 0 50 62 625 625 62 75 0



Pduclauxi EPASS9 0 0 62 25 187 125 50 62

Plividum EPMCL12 50 437 437 562 375 0 687 62



Pthomii EPAER11 187 562 312 562 50 312 562 0



Pasperum EPHAL10 62 312 125 562 25 62 812 0






86



(cm) (g) (cm) (g)


Control hellip 12 1544 18 407 126 333 155 063





P citrinum EPSMR1 1732 1755 297 389 922 1149 064 056



Pduclauxi EPASS9 1672 2255 243 636 1185 597 057 11

Plividum EPMCL12 1307 1303 178 254 1242 529 052 046



Pthomii EPAER11 1328 1375 214 234 148 466 046 055



Pasperum EPHAL10 1328 2412 18 732 1225 775 06 126

LSD005 271 5171 0691 2091 3731 3031 1031 0631



87


EP

88























89



Infection


Fsolani



Control hellip 437 50 625 25 312 62 312 0





P citrinum EPSMR1 0 62 62 562 625 0 75 0



Pduclauxi EPASS9 0 562 62 562 187 25 50 0

Plividum EPMCL12 50 187 437 375 375 0 687 0



Pthomii EPAER11 187 312 312 25 50 125 562 0



Pasperum EPHAL10 62 125 125 50 25 62 812 0






90




















weeks

91


Treatments Code

Shoot

Length

Shoot

Length

Shoot

Weight

Shoot

Weight

Root

Length

Root

Length

Root

weight

Root

weight

(cm) (cm) (g) (g) (cm) (cm) (g) (g)


Control hellip 12 1633 18 554 126 1757 155 105





P citrinum EPSMR1 1732 1912 297 512 922 9 064 155



Pduclauxi EPASS9 1672 214 243 69 1185 153 057 237

Plividum EPMCL12 1307 2347 178 741 1242 1298 052 156



Pthomii EPAER11 1328 225 214 657 148 155 046 164



Pasperum EPHAL10 1328 2101 18 525 1225 1095 06 135

LSD005 271 4291 0691 3281 3731 5851 1031 091


92


cake in tomato

EP

93





















94



Infection


Fsolani



Control hellip 375 0 50 125 437 375 25 0





P citrinum EPSMR1 312 187 187 312 375 50 187 0



Pduclauxi EPASS9 187 375 125 25 375 50 0 0

Plividum EPMCL12 62 62 437 125 62 625 25 0



Pthomii EPAER11 0 187 437 187 62 25 0 0



Pasperum EPHAL10 125 62 50 125 125 812 0 0






95



(cm) (g) (cm) (g)


Control hellip 2369 2225 274 837 274 975 211 303





P citrinum EPSMR1 267 3384 313 668 2397 975 394 098



Pduclauxi EPASS9 2925 2911 376 388 3037 1293 522 116

Plividum EPMCL12 2768 2801 31 698 2155 1132 35 109



Pthomii EPAER11 239 30 296 799 2416 1062 427 125



Pasperum EPHAL10 2856 2891 344 763 1921 1352 306 13

LSD005 471 4931 0941 3331 7321 5451 1611 11071



96



EP

S

EP

97


EP

EP

98


in chickpea


in chickpea

EP

EP

99


in chickpea













weeks













(Table 26)

100




101



Infection



Control hellip 375 125 50 312 437 0 25 187





P citrinum EPSMR1 312 0 187 625 375 187 187 312



Pduclauxi EPASS9 187 0 125 625 375 62 0 312

Plividum EPMCL12 62 62 437 100 62 25 25 312



Pthomii EPAER11 0 62 437 125 62 62 0 62



Pasperum EPHAL10 125 0 50 187 125 0 0 0






102



(cm) (g) (cm) (g)


Control hellip 2369 2188 274 406 274 692 211 58





P citrinum EPSMR1 267 1916 313 556 2397 756 394 1172



Pduclauxi EPASS9 2925 192 376 561 3037 1115 522 819

Plividum EPMCL12 2768 1929 31 417 2155 946 35 383



Pthomii EPAER11 239 2305 296 626 2416 9 427 931



Pasperum EPHAL10 2856 1662 344 582 1921 925 306 834

LSD005 471 6131 0941 3011 7321 2921 1611 6151




103













weeks














104



Infection


Control 75 100 100 75

Feast-M 0 37 687 625










Topsin-M 0 437 812 62





P citrinum 437 687 100 0


P decumbens 187 50 437 187

Pasperum 125 50 562 125




105



Control 3197 339 288 288

Feast-M 4269 451 526 526





Carbendazim 3675 319 398 398





Topsin-M 3935 314 383 383





P citrinum 3268 249 432 432

Pnigricans 2788 201 401 401

P decumbens 3421 3007 446 446

Pasperum 3262 229 363 363

LSD005 5751 0811 1041 1041



106








days of germination



















107



Infection


Control 0 50 625 50








108



Fruits Fruit weight

(cm) (g) (cm) (g)

Control 3831 1058 1596 305 023c 708c

Carbendazim 3421 832 1659 546 045b 683c

P decumbens 4523 1167 1756 438 052a 1106a

Pnigricans 4265 1172 1794 188 054a 894b

P rugulosum 4592 1295 1967 2405 025c 533d

LSD005 511 4281 3431 581 00261 04841


109


soil



Control 137e 2711e 2878e 0053d





LSD005 00311 01361 04211 00041


110



Sucrose

Control 587c 0087c 8668d 1353b 245d





LSD005 0121 000571 0211 01031 0121


111


in garden soil








LSD005 10591 01441 21941 3711


112








after 90 days










37)








113



Infection


Control 312 100 937 562








114



(cm) (g) (cm) (g)

Control 52 1974 1816 35 30a 5801b

Carbendazim 4646 1322 1629 237 20c 4083a

P decumbens 443 2161 1283 418 2133c 995a

Pnigricans 55 1892 1561 315 32a 4286d

P rugulosum 5197 1695 1205 334 256b 4779c

LSD005 1481 18611 5391 4011 3781 0131


115


garden soil








LSD005 01081 01671 0301 00791


116



N Sucrose






LSD005 00621 00541 0211 1311


117










LSD005 5591 0471 5771 2391


118



field condition



















days








119



























120



Infection


45 90 45 90 45 90 45 90

Control 375 0 562 312 937 100 562 0

Topsin-M 375 0 625 25 937 100 687 0


P decumbens 62 0 50 375 68 100 375 0

Pnigricans 125 187 562 687 875 100 312 0

P rugulosum 312 62 562 375 812 100 437 0







121


condition


(cm)

Shoot Weight

(g)

Root Length

(cm)

Root Weight

(g)

Number

of Fruits

Fruit

weight

Control 45 90 45 90 45 90 45 90 90 90

Topsin-M 4178 6192 2228 4325 1368 2426 204 823 086g 246i

Pmontelii 422 6375 1765 4731 1267 2377 133 98 12f 31h


P nigricans 4233 6617 1971 4887 1486 2578 167 1003 143e 1146a

Prugulosum 4866 7083 1635 5095 1378 2311 172 967 176d 331g

P rugulosum 4373 7026 2063 2051 1371 2464 169 709 123f 35f




LSD005 961 1321 131 1181 3551 1371 0831 2961 0111 0111


122



Treatments

Polyphenol

microgml



Control 183h 7314e 7721e 007f

Topsin-M 146i 9119a 9886a 0113d








LSD005 00721 10191 06531 00121



123




microgml




Topsin-M 183f 4922f 5575g 0074bc








LSD005 10061 05191 04731 0003081


124





Control 5102g 646g 1466g 5966f

Topsin-M 5514f 716f 2566f 67e








LSD005 10451 06241 14081 2471


125



Treatments pH

Tritable

acidity

Moisture

content

Total

solids

Total Soluble

Solid

Sucrose

Control 624a 0102c 8774b 1222f 1425e

Topsin-M 619ab 0126b 8653e 1339b 1475e








LSD005 00641 00041 03021 0171 02221


126

127

128

4 DISCUSSION





























129































130
































131
































132







2013)
























133































134
































135








Kloepper 1998)























136




condition


























137



source to sink



























138































Conclusion

139



















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140



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January 20 2017






173



449-458





evaluation of biocontrol potentlal of endophytic …

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