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EX-SITU MACROPROPAGATION STUDY IN BANANA

(Musa paradisiaca L.) cv.GRAND NAINE UNDER SOUTH

GUJARAT CONDITION

A

THESIS

SUBMITTED TO THE

NAVSARI AGRICULTURAL UNIVERSITY

NAVSARI

IN PARTIAL FULFILMENT OF THE REQUIREMENTS

FOR

THE AWARD OF THE DEGREE

OF

MASTER OF SCIENCE

(HORTICULTURE)

IN

FRUIT SCIENCE

BY

RAVANI DINESHKUMAR GANESHBHAI

B.Sc. (Agri.)

DEPARTMENT OF FRUIT SCIENCE ASPEE COLLEGE OF HORTICULTURE AND FORESTRY

NAVSARI AGRICULTURAL UNIVERSITY NAVSARI – 396 450

APRIL - 2013

Registration No.: 04-0876-2010

ABBREVIATIONS

SR. NO. ABBREVIATION MEANING

1 % Per cent

2 BAP 6-Banzylaminopurine

3 NAA α-Naphthalene Acetic Acid

4 IBA Indole Butyric Acid

5 C.D. Critical difference

6 @ At the rate of

7 DAP Day after planting

8 C.V. Co-efficient of variance

9 cv. Cultivar

10 et al. Et alii (and others)

11 Fig. Figure

12 l Litre (Unit of volume)

13 S.Em. Standard error of mean

14 µM Micromol

15 Var. Variety

16 cm Centimeter

17 i.e. That is

18 PGR Plant Growth Regulator

19 T Treatment

20 MT Metric tonne

21 Mg milligram

22 ppm Parts per million

23 Ha Hectare

24 ME Malt extract

25 WPM Woody plant medium

26 BCR Benefit Cost Ratio

DEDICATED

TO

MY BELOVED

PARENTS

AND

GURU

ABSTRACT

“EX-SITU MACROPROPAGATION STUDY IN BANANA

(Musa paradisiaca L.) cv. GRAND NAINE UNDER SOUTH

GUJARAT CONDITION”

Student Major Advisor

Ravani Dineshkumar G. Dr. A. N. Patel

DEPARTMENT OF FRUIT SCIENCE

ASPEE COLLEGE OF HORTICULTURE AND FORESTRY

NAVSARI AGRICULTURAL UNIVERSITY, NAVSARI

ABSTRACT

The present investigation entitled “Ex-situ

macropropagation study in banana (Musa paradisiaca L.)

cv. Grand Naine under South Gujarat condition” was

carried out during 2011-2012 at Fruit Research Station, Navsari

Agricultural University, Gandevi. The experiment was laid out in

a Completely Randomized Design (CRD). In experiment, sword

suckers were used as propagation material and decortication

technique adopted in corms of sword suckers. During ex-situ

decortication of sword suckers, they were pared partially and

then growing point was excised out and suckers treated with

nine treatments of different level of BAP and NAA (2ppm and

4ppm) alone, combintion of both and control. All treatments were

replicated thrice and 20 suckers were taken for each treatment

uniformly. Effects of these treatments on growth parameter were

recorded.

Results revealed that the application of growth

regulators to the propagation material resulted in better

vegetative growth, early bud initiation in suckers of banana.

The minimum days (9.21) required to bud initiation

was observed in suckers which were treated with 2 ppm BAP + 4

ppm NAA (T7) and maximum number of shoots per sucker (5.93)

also observed in the same treatment. The longest shoot (14.45,

21.27 and 24.68 cm) and maximum shoot girth (8.13, 9.28 and

10.26 cm) at 30, 60 and 90 DAP, were produced by the treatment

combination of 2 ppm BAP + 4 ppm NAA. The maximum number

of leaves (4.66, 6.33 and 7.33 leaves/explant at 30, 60 and 90

DAP) were produced in the same treatment and it also recorded

maximum leaf area (170.93, 280.37 and 370.35 cm2), number of

roots (6.40, 8.20 and 10.60), root length (16.83, 21.40 and

28.19 cm) and survival percentage (93.33, 88.33 and 79.17

%) at 30, 60 and 90 DAP, respectively.

There was no any contamination, browning and insect-

pest incidence observed due to any of the growth regulator

treatments and their combinations.

Based on the observation recorded it can be concluded

that, the early sprouting of buds, maximum number of shoots,

shoot length, shoot girth, number of leaves per shoot, leaf area,

number of roots per shoot and root length were recorded with the

treatment 2 ppm BAP + 4 ppm NAA at all growth stages. The

survival percentage (73.52 %) was recorded the highest after

bagging in polythene bag at 90 days. Other combinations of PGR

had also positive response on growth and survival as compared

to control. After 90 days, four to five healthy plants of banana

can be obtained from each sucker in the above said treatment.

These plants can be used as healthy planting material. This

method of propagation was also found economically beneficial.

Thus it can be inferred that macropropagation

provides cheap, simple and relatively rapid technique for

vegetative multiplication of Musa species that could be amenable

to the resource poor, unskilled, small and marginal farmers.

Dr. A. N. Patel Associate Research Scientist (Fruit),

Fruit Research Station,

Navsari Agricultural University,

Gandevi - 396360

CERTIFICATE

This is to certify that the thesis entitled “EX-SITU

MACROPROPAGATION STUDY IN BANANA (Musa

paradisiaca L.) cv.GRAND NAINE UNDER SOUTH

GUJARAT CONDITION.” submitted by RAVANI

DINESHKUMAR GANESHBHAI in partial fulfilment of the

requirement for the award of degree of MASTER OF SCIENCE

(HORTICULTURE) IN FRUIT SCIENCE is a record of bonafide

research work carried out by him under my guidance and

supervision and the thesis has not previously formed on the basis

for the award of any degree, diploma or other similar title.

Place: Navsari (A. N. Patel)

Date: 5th April 2013 Major Advisor

DECLARATION

This is to certify that the whole of the research work

reported in this thesis in partial fulfilment of requirements for the

award of the degree of Master of Science in Horticulture in the

subject of Fruit Science is the result of investigations done by

undersigned under the direct guidance and supervision of

Dr. A. N. Patel (Major Advisor), Associate Research Scientist, Fruit

Research Station, Navsari Agricultural University, Gandevi and no

part of the research work has been submitted for any other degree

so far.

Place: Navsari

Date: 5th April 2013

(Ravani Dineshkumar G.)

Countersigned by

(A. N. Patel) Major Advisor &

Associate Research Scientist (Fruit), Fruit Research Station,

Navsari Agricultural University, Gandevi - 396360

AC K N O W L E D G E ME N T

At this gratifying moment of completion of my research

problem, I feel obliged to record my gratitude to those who have helped me.

Indeed the words at my command are not adequate to convey

the depth of my feeling and gratitude to my major advisor Dr. A. N.

Patel sir , Associate Research Scientist (Fruit) , Fruit Research Station,

N.A.U., Gandevi for his most valuable and inspiring guidance with his

friendly nature, love and affection, for his attention and magnanimous

attitude right from the first day, constant encouragement, enormous help

and constructive criticism throughout the course of this investigation and

preparation of this manuscript.

I feel a great pleasure in getting this proud privilege offering

my sincerest and devoted thanks to my minor advisor Dr. M. M. Patel ,

Assistant Professor (AICRP on Palms), ASPEE College of Horticulture

and Forestry , Navsari Agricultural University , Navsari; and other

members of my advisory committee Dr. B. V. Padhiar , Professor

(Horticulture), ASPEE College of Horticulture and Forestry, Navsari

Agricultural University, Navsari and Dr. B. K. Bhatt , Assosiate

Professor (Agricultural Statistics), ASPEE College of Horticulture and

Forestry, Navsari Agricultural University, Navsari for their worthy

suggestions, ever willing help and unbiased attitude throughout the course

of this investigation.

I acknowledge with thanks for the facilities provided by

Dr. A. N. Sabalpara , the Director of Research, Navsari Agricultural

University , Navsari , Dr. N. L. Patel , Dean and Principal, ASPEE

College of Horticulture and Forestry, Navsari, during the course of my

studies.

It is my great pleasure to express special thanks to Dr. B. N.

Kolambe sir and Dr. Taslim Ahmed sir and Amrutbhai, Denybhai,

Umeshbhai, Babukaka, Chandubhai and all staff member of Gandevi farm

who helped me to do this task and made it easy for me.

I am highly thankful to academic staff member Janakbhai,

Nilamben and all the staff members from ASPEE College of Horticultu re

and Forestry and Library staff, Central Library , Navsari Agricultural

University for their valuable guidance and co -operation throughout the

course of my studies.

Though thank is a taboo in friendship, my conscience does not

permit to refrain myself from expressing my heartfelt feeling towards my

beloved friends Piyush, Priyesh, Suresh, Chandu, Ajay, Mahesh,

Manoj, Hitesh, Haimil, Harshad, Sachin, Mihir, Mukeshbhai,

Niravbhai, Rushabhbhai, Jilenbhai, Pareshbhai, Ketanbhai,

Yagnesh, Nitin, Bhupesh and all PG friends who gave direct and

indirect sympathetic touch for completion of my research work and their

jolly company made my life richer.

My vocabulary fails to get words to express deep sense of

gratitude and indebtedness to my parents, my father Ganeshbhai , mother

Vimlaben , grandfather Laljibhai , grandmother Nanbaiben and my

Brother Rohit , Jignesh, Suresh, Bharat, Haresh, Nitin and Ankit

and my nephew Shubh, Chintan, Dhruvit, Ishan, Meet and Tirth for

their everlasting love, constant encouragement, p rayer support and

sacrifice , without which this dream could not have become a reality. They

are there every step of the way; they hold my hand throughout the long

journey, they are truly the wind beneath my wings.

Last but not least, I am writing this acknowledgement

because of Gods bless.

Place: Navsari

Date: 5thApril 2013 (Ravani Dineshkumar G.)

CONTENT

CHEPTER

NO. TITLE PAGE NO.

I. INTRODUCTION

II. REVIEW OF LITERATURE

III. MATERIALS AND METHODS

IV. EXPERIMENTAL RESULTS

V. DISCUSSION

VI. SUMMARY AND CONCLUSION

REFERENCES

APPENDIX

LIST OF TABLES

TABLE

NO. TITLE PAGE

NO.

4.1 Effect of growth regulators on bud initiation

days of banana cv. Grand Naine.

4.2

Effect of growth regulators on no. of shoots

per sucker of banana cv. Grand Naine. (30

days after treatment)

4.3

Effect of growth regulators on shoot length

(cm) at different stage of growth of banana cv.

Grand Naine.

4.4

Effect of growth regulators on shoot girth


Grand Naine.

4.5

Effect of growth regulators on no. of leaves at

different stage of growth of banana cv. Grand

Naine.

4.6

Effect of growth regulators on leaf area (cm 2 )

at different stage of growth of banana cv.

Grand Naine.

4.7

Effect of growth regulators on no. of roots at

different stage of growth of banana cv. Grand

Naine.

4.8

Effect of growth regulators on root length


Grand Naine.

4.9

Effect of growth regulators on survival

percent at different stage of growth of banana

cv. Grand Naine.

4.10 Effect of growth regulators on pest and

disease incidence of banana cv. Grand Naine.

4.11 Economics of different macropropagation

treatments.

LIST OF FIGURES

FIGURE

NO. TITLE

AFTER

PAGE

1.

Effect of growth regulators on bud

initiation days of banana cv. Grand

Naine.

2.

Effect of growth regulators on no. of

shoots per sucker of banana cv. Grand

Naine. (30 days after treatment)

3.

Effect of growth regulators on shoot

length (cm) at different stage of growth of

banana cv. Grand Naine.

4.

Effect of growth regulators on shoot girth

(cm) at different stage of growth of


5.

Effect of growth regulators on no. of

leaves at different stage of growth of


6.

Effect of growth regulators on leaf area

(cm2) at different stage of growth of


7.

Effect of growth regulators on no. of roots

at different stage of growth of banana cv.

Grand Naine.

8.

Effect of growth regulators on root length

(cm) at different stage of growth of


9.

Effect of growth regulators on survival

percent at different stage of growth of


LIST OF PLATES

PLATE

NO. TITLE

AFTER

PAGE

1. Preparation of bed 20

2. The macropropagation technique 21

3. Laying out of experiment 22

4. Bud initiation 29

5. Removing of secondary shoot 29

6. No. of shoots per sucker (30 days after treatment) 30

7. General view of experiment at 30, 60 and 90 days 34

8. Survival plants 41

INTRODUCTION

I. INTRODUCTION

Banana (Musa paradisiaca L.) is a large

herbaceous perennial monocotyledonous and monocarpic

plant, which belongs to family musaceae in order

scitamineae. Banana, one of the earl iest crops cultivated

by man, remains to be one of the most important fruit

crops, especially of the tropics. The term „banana‟ was

introduced from the Guinea Coast of West Africa by the

Portuguese, while the term „plantain' (for cooking bananas)

was derived from „plantano‟ of the Spaniards. The term

“banana” includes all edible varieties eaten as ripe fruits

or as cooked food. The great historian Disraeli mentioned

banana as the most delicious thing in the world.

Bananas were called “figa” in Europe in the 10 th

century A.D. and it is still so in the West Indies. In India,

banana is commonly called as “Kela” in the northern

states. In South India it is called „Arati‟, „Anati‟ in Andhra

Pradesh, „Bale‟ in Karnataka, „Vazhai‟ in Tamil Nadu and

„Vazha‟ in Kera la. In the Sanskrit literature it is often

referred to as „Kadali‟ or „Rambha‟.

Banana and plantain are widely grown in India

with great socio-economic significance and is interwoven

in the cultural heritage of the nation. Banana is known as

„Apple of the Paradise‟. Banana is one of the largest

photosynthetic units in the plant kingdom. Banana is a

source of food, fodder, fibers, beverages, fermentable

sugars, medicines, flavorings, cooked foods, silage,

fragrance, rope, cordage, garlands, shelter, clothing,

smoking material, wrapping/parcelization, making house

roofs and wall linings and has numerous religious as well

as industrial uses like in making resin/gum/glue/latex,

dye and tanning. Owing to these multifaceted uses, it is

referred as „Kalpatharu‟.

In India, banana is fourth important food crops

in terms of gross value and is exceeded only by paddy,

wheat and milk products. It is also a dessert fruit for

millions, apart from a staple food owing to its rich and

easily digestible carbohydrates with a ca lorific value of 67-

137/100 g per fruit; it is a good source of vitamin A (190

IU per 100 g of edible portion) and vitamin-C (100 mg/

100g) and fair source of vitamin B and B2. Fruits are also

rich source of minerals like magnesium, sodium,

potassium, phosphorus and a fair source of calcium and

iron. It makes healthy and salt free balanced diet than

many fruits. One hectare of banana yields 37.5 million

calories of energy as compared to 2.5 million calories from

wheat and multifarious uses. About 24 bananas each

weighing around 100 g would provide the energy

requirement (2400 calories per day) of a man (Singh,

2002).

Table No: 1.The nutritive value of ripe banana fruit.

Sr. No. Composition Per cent/ppm

1 Miosture 70.0 %

2 Phosphorus 290.0 ppm

3 Carbohydrates 27.0 %

4 Calcium 80.0 ppm

5 Crude fiber 0.5 %

6 Iron 0.6 ppm

7 Protein 1.2 %

8 Carotene 0.5 ppm

9 Fat 0.3 %

10 Riboflavin 0.5 ppm

11 Ash 0.9 %

12 Niacin 7.0 ppm

13 Ascorbic acid 120.0 ppm

Medicinally banana act as preventive for heart

ailments. Use of pseudostem core has been well recognized

as medicine in dissolving kidney stones. Stalks are

mashed and used as poultice for sprains or broken bones,

root sap from certain varieties is used as a medicine to

thrush a child‟s mouth irritation and to treat skin warts,

recently pounded banana peels have been found to contain

antibiotic properties.

India is the largest banana producer and

consumer country in the world contributing about 15 per

cent of the total world production. Among the fruits,

banana holds first position in production and productivity

in India. It ranks second in area after mango. After the

introduction of Grand Naine (Musa AAA) it is gaining

popularity and approximately 80-90 % basrai cultivation

has been replaced. In India, annual production of banana

is 297.80 lakh tonnes from an area of 8.3 lakh ha spread

all over the country (Anon. 2011a).

Banana covers 12 per cent of the total area under

fruits, contributing nearly one third of total fruit

production in the country. In India, Tamil Nadu,

Maharashtra, Karnataka, Kerala, Assam and Gujarat are

the leading banana producing states. The highest

productivity was noted 65.8 tones/ha in Tamilnadu

followed by Gujarat (61.5 t/ha) in the year 2010-2011.

(Anon. 2011b).

In Gujarat state, area under total fruit crops 3.49

lakh ha and production 72.45 lakh MT of which banana

crop occupies 64,700 ha area with annual production of

39,78,000 MT. State wise area, production and

productivity of banana fruit crop in the year 2010-2011 is

given in appendix-I.

The largest area is covered under Bharuch

district with an area of 0.14 lakh ha with the highest

production of 8.82 lakh metric tonnes. Navsari district

covers an area of 440 ha with a production of 24,220 MT

annually. Banana is cultivated in the districts of Surat,

Anand, Narmada, Junagadh, Vadodara and Valsad because

of favourable agro-climatic conditions and abundant

supply of irrigation water through well and canal. District

wise area and production of banana fruit crop in the year

2010-11 is given in appendix-II.

Natural regeneration is very slow in banana due

to hormone-mediated apical dominance of mother plants.

Wild bananas have polyarchic architecture and they

produce relatively a large number of shoots from the base

of mother plants while in commercial varieties production

of new suckers follows a hierarchical pattern due to a

strong apical dominance exerted by main plants. Thus

natural regeneration is very slow in banana leading to

production of only 5-20 suckers in its life time depending

on variety.

Looking to the area under banana cultivation in

India, requiring 0.125 billion plants per annum. This is an

enormous requirement in a vegetatively propagated crop

like banana. Presently, 15-20 viable companies are

involved in a production of tissue-cultured banana plants

with an annual production of 2-5 million plantlets. Thus,

only 4.0 per cent is being produced through tissue culture,

while the rest 96 per cent of planting material requirement

is being catered as suckers (Uma et al., 2008).

Plant growth regulators are known to regulate

and modify various physiological processes within the

plant, which ultimately affect the growth, yield and quality

of plants, thereby they influence on morphological

characters and yield. The growth regulators are absorbed

by growing plant both ways by vegetative and floral parts,

when they are applied at appropriate concentration.

No research work has been done using growth

regulators for macropropagation in banana under sou th

Gujarat agro - climatic condition. Therefore, in the present

investigation an attempt has been made on the “ex-situ

macropropagation studies in banana cv. Grand Naine

under South Gujarat conditions” to achieve to following

objectives:

1) To find out the suitable growth regulator and it‟s level

of concentration for macropropagation in banana.

2) To assess the effect of plant growth regulators in

macropropagation in banana individually and in

various combinations.

3) To produce quality planting material for small and

marginal farmers.

REVIEW OF

LITERATURE

II. REVIEW OF LITERATURE

The macropropagation in banana is recently developed

techniques in southern part of India. Non availability of literature

on specific technology, the literature on micropropagation on

banana and other crops like Guava, Papaya, Pomegranate and

Citrus are reviewed here in support of present research.

2.1 Banana

Bhaskar et al. (1993) experimented on micropropagation

studies in banana and it was seen that the treatment combination

of NAA 0.5 ppm + BAP 3.0 ppm was found to be the best as all the

three explants tried took the minimum number of days for the

establishment of culture. In the trial conducted to study the effect

of phytohormones on multiple shoot production using shoot tip

explants, each explant contained 11.0 axillary shoots when the

basal proliferation medium contained NAA 1.0 ppm + BA 10.0 ppm

or BA 10.0 ppm alone.

*Bekheet and Saker (1999) studied rapid mass

micropropagation of banana. They observed that the highest

number of proliferated shoots of the three cultivars was recorded

with 6 mg/l BAP. Among different types of auxins used for rooting

of banana shoots, Napthalene Acetic Acid was more effective on

root formation than Indole Acetic Acid (IAA) Indole -3- Butyric Acid

(IBA).

Malik et al. (2000) studied in vitro multiplication of

banana cv. Desi. It was observed that 6- benzylaminopurine (BAP)

at 5 mg/l gave maximum (4 to 5) plantlets.

*Acharjee et al. (2004) employed in vitro shoot tip culture

technique for clonal propagation of indigenous banana cultivars

viz., 'Bhimkhol', 'Malbhog', 'Kanchkhol' and 'Jahaji'. They noted

that shoot elongation in a medium supplemented with lower (2.5

mg/l) concentration of BA. They also observed that rooted plantlets

were best within 10-20 days in a medium containing 0.2 mg/l NAA.

Gubbuk and Pekmezcl (2004) revealed that

supplementation of 20 µM BAP on MS medium produced the best

multiplication and elongation in all 3 bananas Alanya 5, Anamur

10 and Bozyazi 14. The highest stem diameter was recorded for all

banana types with 1 µM NAA, followed by active charcoal.

Rahman et al. (2004) reported that among the different

levels of BAP, better response was found with 5.0 mg/l BAP in

respect of survivability (91.67%), single shoot formation rate

(62.50%) and formation of hard meristamatic ball like structure

(29.17%). Maximum shoot multiplication (4.52 /explants) with

highest shoot length (3.62cm) was achieved on MS medium

containing 4.0 mg/l BAP + 1.5 mg/l NAA treatment in Banana

cv.BARI-I.

Akbar and Roy (2006) reported that the explants were

cultured the explants on MS medium supplemented with 0.5 mg/l

each of BA, KN and NAA. They observed a large number of shoots

developement and shoot proliferation with the progression of the

number of subcultures.

An investigation carried out by Bhor and Gujar (2007)

revealed that the maximum numbers of shoots (7.3) were recorded

on MS medium supplemented with BAP (2 mg/l) in Banana

cv.Grand Naine.

The result obtained by Kalimuthu et al. (2007)

revealed that the shoot proliferation (95 % ) was best in

BAP + NAA (3.0 + 0.2 mg/l), followed by (75 % ) the next

combination of BAP + NAA (4.0 + 0.2 mg/l) in Banana cv.

Dwarf Cavendish. The poor response of shoot initiation

ability was noticed both in first (0.5 + 0.2 mg/l of BAP +

NAA) and last (5.0 + 0.2 mg/l BAP + NAA) combinations of

the growth regulators.

Muhammad et al. (2007) recorded the maximum

number of shoots regenerated from a single shoot tip with

liquid MS medium containing 4.0 mg/l BAP.

Viehmannova et al. (2007) studied the influence

of growth regulators on root induction In Vitro of the Musa

genus and reported that the largest number of roots has

been found with the variant 5.4 µM NAA (4.79 roots per

plant).

Al-amin et al. (2009) observed highest shoot prolifiration

of shoots per explants and largest shoot at 10, 20 and 30 days

after inoculation with the treatment combination of 7.5 mg/l BAP +

0.5 mg/l NAA.

Karim et al. (2009) reported that among different

concentration of BAP + NAA, better response found with 7.5 mg/l

BAP + 0.5 mg/l NAA in respect of shoot proliferation (0.75,

2.75 and 6.25 shoots per explant), shoot length (1.03, 2.45 and

3.38 cm) and number of leaves (2.50, 3.25 and 7.00 leaves per

explant) at 10, 20 and 30 days after inoculation.

Azam et al. (2010) recorded average shoot length of 3.1

cm, shoot proliferation and elongation response was the strongest

in the MS medium enriched with BAP at 2.0 mg/l. It was observed

that BAP concentration below 1.5 mg/l did not improve the shoot

regeneration. BAP raised shoot proliferation profiency from 10 to

87 %, when its concentration was increased from 0.5 to 2.0 mg/l.

Buah et al. (2010) reported that media supplemented

with 4.5 mg/l BAP induced the highest number of shoots after

eight weeks of culture. Mean shoot height was higher (39.5 cm) in

Apantu pa cultured on 4.5 mg/l BAP while in Oniaba, the highest

shoot height was achived on medium supplemented with BAP 7.5

mg/l.

A study was carried out by Ali et al. (2011) on shoot

formation response from shoot apical meristem. They reported that

MS medium containing 1.0 mg/l BAP showed best response for

shoot formation.

Bhosale et al. (2011) studied in vitro shoot multiplication

in different species of banana and reported that among the various

levels of BAP in three different varieties of banana viz. Ardhapuri,

Basrai, Shrimanti, 7 mg/l BAP shows increased average no. of

shoots.

Jafari et al. (2011) reported that BAP at 22 µM induced

the highest number of normal and elongated shoots, although

more shoots proliferated on initiation medium supplemented with

BAP at 33 µM.

Mukunthakumar et al. (2011) observed that the best

result of shoot multiplication was obtained from the explants in the

media containing 6.0 mg/l BAP.

2.2 Guava

Ali et al. (2003) reported that the highest concentration

(BAP 2 mg/l) induced the maximum number of shoots (3.72) in

green house grown plants. They also noticed that the shoots

derived from green house raised explants were the tallest (3.0 cm)

at 0.5 mg/l BAP in Guava.

Zamir et al. (2003) reported that the highest number of

shoots (43) was developed into plantlets when MS medium was

supplemented with BAP 1.0 mg/l combined with 250 mg/l L -

glutamine.

Rai et al. (2009) studied on shoot multiplication and

plant regeneration of guava (Psidium guajava L.) from nodal

explants of in vitro raised plantlets and reported that among

various concentration of BAP, the highest number of shoots per an

explant was observed on medium containing 1 mg/l BAP.

2.3 Papaya

Rahaman et al. (1992) reported that the maximum

frequency of survived explants and resumed growth on Murashige

and Skoog medium was observed with 10 mg BAP and 1 mg NAA

per litre in papaya.

Naik and Shah (1996) reported that establishment of the

seedling explant was maximum on MS medium supplemented with

either BAP alone at 0.5 mg/l or with BAP 0.5 mg/l and NAA 0.01

mg/l, however the multiplication rate was the highest when NAA

was incorporated alongwith BAP 0.5 mg/l in the medium.

Kabir et al. (2007) observed that shoot proliferation was

best in MS medium containing BAP 1.0 mg/l + KIN 0.5 mg/l and

BAP 1.0 mg/l + NAA 0.5 mg/l. The maximum number of shoot per

culture (28.2) and length of the largest shoot (1.7 cm) were also

observed with MS medium supplemented with BAP 1.0 mg/l + KIN

0.5 mg/l.

2.4 Pomegranate

An in-vitro regeneration studies in pomegranate

cv. Ganesh was conducted by Murkute et al. (2004). They

reported that the maximum shoot height was attained up

to 3.19 and 3.11 cm from shoot tip and nodal segment

respectively, in the treatment MS + BAP 1.0 mg/l + NAA

0.5 mg/l. while the maximum rooting percentage (66.4 % )

within 8 to 9 days were recorded with treatment MS + NAA

0.5 mg/l.

Upadhyay and Badyal (2007) stated that the

cuttings treated with IBA 2000 ppm gave the highest

percentage survival of cuttings i.e. 81.33 % followed by

76.85 % in NAA 100 ppm + IBA 2000 ppm in pomegranate.

Patil et al. (2011) showed that the highest

average growth response (99 %) was recorded on MS

medium containing BAP 1.8 mg/l, whereas the highest

average maximum leaf number (15-20) was recorded on MS

medium containing 0.4 mg/l BAP and 0.3 mg/l NAA.

2.5 Citrus

Desai et al. (1996) studied in vitro propagation of acid

lime (Citrus aurantifolia Swingle) var. 'Kagzi lime'. Maximum

number of shoots (2.63) was obtained with 0.25 mg/l BAP and 200

mg/l ME. In treatment 0.1 mg/l NAA, recorded cent percent

rooting within 10-30 days of incubation.

Al-Khayri and Al-Bahrany (2001) reported that the best

results for multiple shoot formation (8 shoots per node) were

obtained with 1 mg/l BAP and 0.5 mg/l Kinetin.

Krishan Kumar et al. (2001) reported that the maximum

number of shoot (2.06) and leaves (4.56) were found when treated

with (1.0 mg/l) BAP, whereas, maximum shoot length (1.30 cm)

was observed at low concentration of BAP (0.25 mg/l) in Sweet

orange cv. Mosambi.

Krishan Kumar et al. (2001) reported that the maximum

rooting (63.33 %) and regenerated shoots of kinnow mandarin were

observed in IBA (0.5 mg/l) + NAA (0.5 mg/l). They also observed

that the different level of BAP evaluated, maximum mean shoot

proliferation, shoot number, shoot length and leaf number were

observed in BAP @ 1.0 mg/l.

Begum et al. (2003) reported that maximum percentage

of shoot regeneration was obtained on half strength MS medium in

the presence of 1.0 mg/l BAP from callus in three varieties of

pummelo. While different concentrations of NAA, IBA or IAA were

used in half strength of MS medium for root formation, 100 %

regenerated shoots initiated roots on half strength MS + 0.1 mg/l

NAA.

Mukhtar et al. (2005) observed that the highest

percentage of shoot induction was obtained from the shoot which

were cultured on MS media supplemented with 2.0 mg/l of

benzylaminopurine (BAP) in both species i.e. Mosambi and Lemon,

while the highest rooting percentage in Mosambi was obtained at

concentration of 1.5 mg/l NAA. However, in Lemon highest

response of rooting was obtained at high level of NAA i.e. 2.0 mg/l.

Investigation was conducted by Murkute et al.

(2008) to develop a rapid in vitro regeneration protocol for

Citrus jambhiri and C. karna. They observed that the

treatment containing 1mg/l BAP, 0.5 mg/l NAA and 40

mg/l adenine sulphate significantly improved culture

responses including percentage shoot bud formation, days

required for shoot initiation, shoot length and number of

resultant shoots per explant.

Jajoo (2010) experimented on in vitro propagation

of Citrus limonia Osbeck through nucellar embryo culture

and she noted that 6- benzylaminopurine at a

concentration of 2.22 mM induced highest numbers of

multiple shoots (18.26) per explant.

Savita et al. (2010) observed that the

regeneration response (71.89 %) was better from nodal

segment derived callus cultures on MS medium

supplemented with BA (3 mg/l) and NAA (0.5 mg/l). While,

maximum rooting response (71 % ) was observed on MS

medium supplemented with NAA at 0.5 mg/l.

Haripyaree et al. (2011) observed that the

maximum numbers of shoots were induced on medium

containing 0.25 mg/l BA together with 0.50 mg/l NAA or 1

mg/l BA with 0.50 mg/l Kinetin. The highest numbers of

roots were produced with 2 mg/l NAA.

2.6 Other fruits

Sontakke et al. (1996) have taken a propagation trial on

fig cv. Daultabad involving two types of cutting, two growth

regulators IAA and NAA in two concentration 150 and 300 ppm

and 25 and 50 ppm, respectively. The results showed that the

treatment involving two bud cutting treated with 25 ppm NAA

recorded good result for parameter like survival of rooted and

shooted cutting (48.83%), earliness (21.32 days) for sprouting, root

developement (31.83 roots per cuting), shoot growth (11.03 cm),

number of leaves (7.33 per cutting) and leaves area (610.31 cm2 per

cutting) as compared to other treatment.

Parihar et al. (1999) conducted an experiment to see the

effect of NAA spray on vegetative growth of phalsa. They reported

that 30 ppm NAA spray after pruning increased shoot length

significantly over control. They also noted the maximum shoot

diameter with 20 ppm NAA spray.

Mannan et al. (2006) reported that the highest number

of shoot bud per explant (2.33) and longest shoot (1.45 cm) were

obtained from treatment NAA 0.5 mg/l + BA 1.5 mg/l in jack fruit.

Das et al. (2008) recorded the highest percentage of

multiple shoots (91.23 %) in MS medium augmented with 2.0 mg/l

BAP + 0.2 mg/l NAA. The maximum numbers of multiple shoots

(22.7) per culture were obtained in MS medium enriched with 2.0

mg/l BAP + 0.2 mg/l NAA within fourteen days of inoculation in

wood apple.

Sirchi et al. (2008) studied the plant regeneration as

affected by plant growth regulators in mangosteen (Garcinia

mangostana L.) and reported that shoot tip explants produced the

highest mean number of shoots per explant on media

supplemented with 1.0 mg/l 6- benzylaminopurine (BAP) and 0.05

mg/l kinetin (KIN) (73.3), while the highest mean shoot height was

observed with the treatment combination 0.1 mg/l BAP and 0.05

mg/l Kin.

Zulfiqar et al. (2009) observed good response in axillary

buds of avocado with 1.0 mg/l BAP which leads to the best rate of

shoot multiplication (4.80). Whereas 1.5 mg/l BAP favoured the

good shoot length developement (4.06 cm) from apical buds.

Khan et al. (2010) reported that 1.5 mg/l BAP gave the

best results in terms of maximum number of shoots (4.66) per

proliferated explant. Which was followed by 2.0 mg/l BAP and 1.0

mg/l BAP, which produced 2.66 and 2.11 shoots per proliferated

explant in jack fruit.

Singh et al. (2010) recorded the highest number of

shoots (2.0 ± 0.29) on MS medium augmented with 2.0 mg/l BAP

in Sapindus mukorossi. The medium supplemented with 2.0 mg/l

BAP + 1.0 mg/l NAA responded better than all other media

combinations.

Warrier et al. (2010) inoculated the nodal segments of

bael fruit on three different media combination with varying growth

regulator combinations. Initial treatment with BAP (0.10 to 1.0

ppm) for 5 weeks in MS medium followed by transfer to higher

concentration of BAP 2.5 mg/l in WPM proved most beneficial for

the induction of shoot multiplication.

Al-Saif et al. (2011) conducted an experiment to

evaluate the pineapple regeneration and shoot growth as

affected by 6-benzylaminopurine (BAP) at 2.0 mg/l and

naphthalene acetic acid (NAA) at 0.2 mg/l in vitro. They

observed that BAP at 2.0 mg/l significantly increased the

production of shoots per explant and shoot length.

Hosseini et al. (2011) observed that the media

containing 2 mg/l of BAP lead to the best results with the

average number of shoots 3.19 and shoot length 16.3 mm,

while the media containing 1.5 mg/l IBA and 0.5 mg/l NAA

was the best media for rooting on the base of shoots (3)

and length (3.25 mm) in comparison with others in

mahaleb.

MATERIALS

AND

METHODS

III. MATERIALS AND METHODS

An investigation on “Ex-situ macropropagation

studies in banana cv. Grand Naine under South Gujarat

condition" was conducted at Fruit Research station, Navsari

Agricultural University, Gandevi during 2011- 2012.

3.1 Experimental site

The experiment was conducted in green shade net

house at Fruit Research station, Navsari Agricultural University,

Gandevi, which is situated in Navsari district of Gujarat

state at elevation of 7.6 meter above mean sea level

latitude of 21° N and east 73° E longitude and on the bank of

river Vengania. It is 4 km away from Gandevi town.

3.2 Climate:

Climatically, this region is typically tropical

characterized by fairly hot summer, moderate cold winter and more

humid and warm monsoon with heavy rain. The maximum

temperature of 38 °C in month of April and minimum temperature

of 10 °C in the month of January were recorded. Monsoon mostly

starts from second week of June and lasts up to the second week of

September. The annual mean precipitation of this region is

1500 to 1800 mm. Most of the rainfall is received from South West

monsoon concentrating in the months of July and August and

grouped under south Gujarat heavy rainfall zone (I).

The temperature starts declining in beginning of

November and continues till the middle of February. The

temperature becomes lowest in months of December and

January. Frost occurence is rare in this area.

The summer commences from mid February and

prolongs up to first fortnights of June. The temperature

rising from February onwards. April and May are the

hotest months of the year. The meteorological data in

respect of maximum and minimum temperature, relative

humidity and sunshine hours recorded at meteorological

observatory of Fruit Research Station, Navsari Agricultural

University, Gandevi, during course of investigation from

September 2011 to January 2012 presented in Appendix-

III.

3.3 Growing media:

The paddy husk and vermicompost with a ratio 2:1 was

used for raising the sucker of banana.

3.4 Experimental area:

A shade house of dimension 18 m x 24 m with a height

of 2.4 m was used for experimentation. It was of green shade net

and allowing only 25 per cent sunlight. This structure maintained

relatively a low temperature with high humidity in comparison to

the outside environment.

3.5 Preparation of bed:

Three beds of 9 x 1 m were prepared with bricks and

paddy husk + vermicompost + biopeat were filled in the bed up to

15 cm height.

Plate 1: Preparation of bed.

3.6 Selection of material:

Healthy, robust as well as diseases and pests free

suckers from harvested banana plants were obtained in required

quantity i.e. 540 suckers.

3.7 Preparation of growth substance solution:

3.7.1 Indole Butyric Acid (IBA):

IBA 15 gm was weighed on elecronic balance and

paste was prepared in 0.1 N NAOH and then dissolved in

distilled water. The solution was diluted in distilled water

to make 6000 ml of solution of 2500 ppm.

3.7.2 Benzylaminopurine (BAP):

To prepare 2 and 4 ppm of solution, 1.2 and 2.4

mg of BAP were taken and dissolved in 95 per cent

absolute alcohol and the solution was made to 600 ml by

adding distilled water.

3.7.3 Napthalene Acetic Acid (NAA):

To prepare 2 and 4 ppm of solution, 1.2 and 2.4

mg of NAA were taken and dissolved in 0.1 N NAOH and

the solution was made to 600 ml by adding distilled water.

3.8 Preparation of suckers:

The roots are removed by paring using a sharp knife.

The pseudostem of mother corm or sword sucker is cut

transversely at 2 cm above the collar region and then the

apical meristem is removed, leaving a cavity of 2 cm

diameter and 4 cm depth. The corms are washed and put in

1 2

3 4

1. Healthy and robust suckers with no symptoms of disease

2. The roots are removed by paring using a sharp knife

3. An incision (X) is made at the growth points

4. Arrange in paddy husk and are covered completely

Plate 2: The macropropagation technique.

disinfectant for sometime. Then suckers were arranged in

preapared beds for sprouting and rooting.

3.9 Laying out of experiment:

In each bed within 1 x 1 m2 space 20 suckers were

arranged at 20 x 20 cm distance.

3.10 Application of growth regulator solution:

IBA was sprayed with the knap sack sprayer in

bed of paddy husk and vermicompost mixture before one

week of planting. While, 5 ml of solution of BAP and NAA

taken with the help of pippete and applied in cavity of

sucker at the time of planting according treatment wise.

Then arranged in previously prepared beds in randomized

manner repeating thrice. Then each bed of 1 x 1 m 2 area

covered with 5 kg biocompost.

3.11 Cultural practices:

3.11.1 Weed management and irrigation:

Weed management was done by hand weeding.

The suckers were irrigated three days interval in early

stage and daily in later stage with the help of spraying

cane.

3.11.2 Fertilizer application:

19-19-19 fertilizer 1 kg + Kombi-F 200 gm

dissolved in 200 liter water. Taken 200 ml in 5 liter water

through spraying cane and sprayed uniformally in 1 m 2

plot at every irrigation.

Plate 3: Laying out of experiment.

3.11.3 After care:

The secondary shoots were removed from each

sucker upto uniformity in height of new sprouts of sucker.

3.12 Experimental design and treatment details:

3.12.1 Experimental design:

The experiment was laid out in a Completely

Randomized Design (CRD) with nine treatments of different

concentration of BAP and NAA alone and combintion of both. All

treatments were repeted thrice and each treatment having 20

suckers.

3.12.2 Treatment details:

Sr. No.

TREATMENT

1 2 ppm BAP

2 4 ppm BAP

3 2 ppm NAA

4 4 ppm NAA

5 2 ppm BAP + 2 ppm NAA




9 Control

NAA - α-Naphthalene Acetic Acid

BAP – 6-Banzylaminopurine

3.12.3 Observations recorded:

1. Days to bud initiation

2. No. of shoot /suckers (30 days after treatment)

3. Shoot length (30, 60 and 90 days)

4. Shoot girth (30, 60 and 90 days)

5. No. of leaves (30, 60 and 90 days)

6. Leaf area (30, 60 and 90 days)

7. No. of roots (30, 60 and 90 days)

8. Root length (30, 60 and 90 days)

9. Survival percent (30 and 90 days)

10. Per cent of contamination

11. Per cent of browning

12. Pest and Disease incidence

3.12.4 Methodology adopted for recording

observations:

Six suckers were selected from net plot area in all

treatments and repetations for each observation and the

mean value were presented for meaningful interpretation.

3.12.4.1 Days to bud initiation:

The average days required for bud initiation after

planting was recorded.

3.12.4.2 No. of shoots /sucker (30 days after

treatment):

The number of shoots sprouted from a sucker recorded

randomaly at thirty days of experiment initiation.

3.12.4.3 Shoot length (30, 60 and 90 days):

The length of sprouted shoots from net plot area

recorded at thirty days interval after planting in terms of

centimeters with the help of standard scale. Then the

recorded data was then averaged out.

3.12.4.4 Shoot girth (30, 60 and 90 days):

The girth of shoots on suckers under net plot

area were recorded at thirty days interval after planting in

terms of centimeters with the help of measure tap . Girth of

shoots was measured from one inch above the ground

level. Then the recorded data was then averaged out.

3.12.4.5 No. of leaves (30, 60 and 90 days):

The number of leaves after sprouting, present on

the shoots under net plot area were recorded at thirty days

interval after planting. The total number of leaves was

then averaged out treatment wise and presented as no. of

leaves per shoot.

3.12.4.6 Leaf area (30, 60 and 90 days):

The length of leaf emerged on shoots under net plot area

was measured from the base of leaf petiole to the tip and breadth

was measured at the maximum breadth of leaf blade. The leaf area

was worked out as the multiplication of the length and breadth of

the leaf with leaf area factor (0.8) suggested by Obiefuna and

Ndubizu, (1979).

3.12.4.7 No. of roots (30, 60 and 90 days):

Total number of roots on each sucker under net

plot area were recorded in each bed and mean number of

roots recorded replicat ion wise.

3.12.4.8 Root length (30, 60 and 90 days):

The length of fine root on each sucker under net

plot area were measured out in centimeter with the help of

a measure tap and mean root length was recorded

replication wise.

3.12.4.9 Survival per cent (30 and 90 days):

The survival of shoots was recorded at thirty and

ninety days after planting. The total number of shoots

survived at thirty days and ninety days was counted as

survival shoots per sucker. The total number of shoots

considered as no. of plants per sucker success percentage :

Number of shoots survival

Survival percentage = x 100

Total number of shoots

3.12.4.10 Per cent of contamination:

The number of contaminated plants were counted and

converted into percentage.

3.12.4.11 Per cent of browning:

The number of brown spot in plants were counted and

converted into percentage.

3.12.4.12 Pest and Disease incidence:

The incidence of diseases and pest was observed

at 90 days after planting. The number of affected shoots

per treatment were recorded.

3.13 Economics

The economics of different macropropagation

treatments were computed considering 1000 suckers per

treatment.

3.14 Statistical analysis:

The data recorded during the experiment were

subjected to the statistical analysis in Completely

Randomized Design (CRD) as described by Panse and

Sukhatma (1967). The significance of treatment differences

was tested by 'F' test on the basis of null hypothesis. The

appropriate (S.Em. ±) were calculated in each case and the

critical difference (CD.) at 5 percent level of probability

was worked out to compare the three treatment effects.

The data on percentage were transformed using

arcsine transformation and analyzed.

EXPERIMENTAL

RESULTS

IV. EXPERIMENTAL RESULTS

An experiment on “Ex-situ macropropagation study in

banana (Musa paradisiaca L.) cv.Grand Naine under South

Gujarat condition” was conducted at Fruit Research Station,

Navsari Agricultural University, Gandevi, during September 2011

to December 2011. The effect of different concentration of growth

regulators on bud initiation, growth and survival per cent was

assessed.

The data so collected for different characters are

subjected to statistical analysis and results obtained are presented

under different headings in this chapter.

4.1 Effect on days to bud initiation

The data presented in table 4.1 and depicted in fig.1

revealed that there were significant differences in days required to

bud initiation in suckers due to different concentration of growth

regulators.

Significantly the minimum i.e. 9.21 days required to

bud initiation was recorded when sucker treated with 2 ppm BAP

+ 4 ppm NAA (T7), which was at par with T5 (9.90) i.e., 2 ppm BAP

+ 2 ppm NAA. While the maximum i.e. 14.18 days required in

control treatment T9 i.e. without any tretment.

4.2 Effect on no. of shoot /suckers (30 days after treatment)

The data pertaining to effect of growth regulators

on sprouting of banana suckers are presented in Table 4.2 and

depicted in fig. 2.

Table 4.1: Effect of growth regulators on bud initiation

days of banana cv. Grand Naine.

Treatment

Days

T1

2ppm BAP

12.92

T2

4 ppm BAP

11.11

T3

2 ppm NAA

13.64

T4

4 ppm NAA

12.33

T5

2 ppm BAP + 2 ppm NAA

9.90

T6


11.85

T7


9.21

T8


10.57

T9

Control

14.18

S.Em. ±

0.26

C.D. at 5 %

0.77

C.V. %

3.83

0.00

2.00

4.00

6.00

8.00

10.00

12.00

14.00

16.00

T1 T2 T3 T4 T5 T6 T7 T8 T9

Days to bud initiation (Days)

Fig. 1: Effect of growth regulators on bud initiation days of banana cv. Grand Naine.

Days

Treatment

Table 4.2: Effect of growth regulators on no. of shoots per

sucker of banana cv. Grand Naine. (30 days after

treatment)

Treatment

No. of shoots per sucker

T1

2ppm BAP

2.95

T2

4 ppm BAP

4.88

T3

2 ppm NAA

2.79

T4

4 ppm NAA

3.72

T5


5.81

T6


4.79

T7


5.93

T8


5.64

T9

Control

2.63

S.Em. ±

0.12

C.D. at 5 %

0.35

C.V. %

4.69

0

1

2

3

4

5

6

T1 T2 T3 T4 T5 T6 T7 T8 T9

Fig. 2: Effect of growth regulators on no. of shoots per sucker of banana cv. Grand Naine. (30 days after treatment)

No. of shoots per suker

No. of

shoots

Treatment

There were significant differences in sprouting of

suckers due to use of different plant growth regulators at

different concentaration and their combinations. The

significantly maximum shoots per sucker (5.93) were recorded

in 2 ppm BAP + 4 ppm NAA (T7), which was at par with T5

(5.81) i.e., 2 ppm BAP + 2 ppm NAA and T8 (5.64) i.e., 4 ppm

BAP + 4 ppm NAA. The minimum shoots per sucker (2.63)

were recorded in control treatment T9 i.e. without any

tretment.

4.3 Effect on shoot length

The length of the shoot was measured at 30 days

interval from day after planting (DAP) up to the 90 days after

planting and data are presented in Table- 4.3 and graphically

depicted in Fig. - 3.

The data revealed that the length of the shoot was

significantly influenced by the various treatments of growth

regulators at different growth stages. Consistently the higher

length of the shoots (14.45, 21.27 and 24.68) at 30, 60 and 90

DAP were observed in treatment T7 (2 ppm BAP + 4 ppm NAA)

respectively which was at par with T5 (2 ppm BAP + 2 ppm

NAA) at 30, 60 and 90 DAP. Consistently the minimum length

of the shoots (6.90, 14.90 and 19.15) were recorded under

control (T9) during whole growth stage of banana plant.

4.4 Effect on shoot girth

The data pertaining to the shoot girth at different stages

of plant growth are presented in Table – 4.4 and depicted in

Fig.-4.

Table 4.3: Effect of growth regulators on shoot length (cm)

at different stage of growth of banana cv. Grand Naine.

Treatment

Days after planting

30

60

90

T1

2ppm BAP

8.67

16.24

19.97

T2

4 ppm BAP

9.76

17.73

22.49

T3

2 ppm NAA

8.44

15.57

19.62

T4

4 ppm NAA

8.91

16.80

20.38

T5


14.17

20.58

23.53

T6


9.42

17.18

22.24

T7


14.45

21.27

24.68

T8


10.93

19.64

22.75

T9

Control

6.90

14.90

19.15

S.Em. ±

0.30

0.49

0.51

C.D. at 5 %

0.88

1.44

1.53

C.V. %

5.04

4.73

4.11

0

10

20

30

T1 T2 T3 T4 T5 T6 T7 T8 T9

30 DAP 60 DAP 90 DAP

Sh

oot

len

gth

(cm

)

Treatment

Fig. 3: Effect of growth regulators on shoot length (cm) at different stage of growth of banana cv. Grand Naine.

Table 4.4: Effect of growth regulators on shoot girth (cm)


Treatment

Days after planting

30

60

90

T1

2ppm BAP

6.04

7.49

8.57

T2

4 ppm BAP

6.56

8.46

9.51

T3

2 ppm NAA

5.33

7.26

8.41

T4

4 ppm NAA

6.25

7.75

8.94

T5


7.59

8.87

10.03

T6


6.50

8.10

9.17

T7


8.13

9.28

10.26

T8


6.73

8.58

9.76

T9

Control

4.96

7.09

8.08

S.Em. ±

0.16

0.24

0.26

C.D. at 5 %

0.48

0.70

0.78

C.V. %

4.35

5.03

4.96

0

5

10

15

T1 T2 T3 T4 T5 T6 T7 T8 T9


Fig. 4: Effect of growth regulators on shoot girth (cm) at different stage of growth of banana cv. Grand Naine.

sh

oot

gir

th (cm

)

Treatment

The data on shoot girth indicated significant differences due

to growth regulators at all the stages of shoot growth.

At 30, 60 and 90 DAP the maximum shoot girth

(8.13, 9.28 and 10.26 cm) were measured in treatment T7 (2

ppm BAP + 4 ppm NAA) which was statistically at par with T5

and T8 at 60 DAP and with T5, T8 and T2 at 90 DAP.

The minimum shoot girth (4.96, 7.09 and 8.08 cm)

were recorded under T9 i.e. control at all the stages of banana

shoot growth, respectively.

4.5 Effect on no. of leaves

The data recorded on number of leaves per shoot are

presented in Table- 4.5 and graphically depicted in fig.-5.

The number of leaves were significantly affected due

to growth regulators at all the stages of growth. The highest

number of leaves (4.66, 6.33 and 7.33, respectively) were

found in shoots treated with 2 ppm BAP + 4 ppm NAA from 30

DAP which were at par with T5 at 60 and 90 DAP. The

minimum number of leaves (2.80, 3.50 and 3.83) were

observed in control (T9) during the respective growth stages.

4.6 Effect on leaf area

Leaf area differed significantly due to growth

regulators at all the stages of shoot growth presented in Table-

4.6 and graphically depicted in fig -6.

The maximum leaf area (170.93, 280.37 and 370.35

cm2) were recorded by T7 (2 ppm BAP + 4 ppm NAA) as

compared to other treatments at 30, 60 and 90 DAP which

was statistically at par with T5 at 30, 60 and 90 DAP, T8 at 60

Table 4.5: Effect of growth regulators on no. of leaves at

different stage of growth of banana cv. Grand Naine.

Treatment

Days after planting

30

60

90

T1

2ppm BAP

3.00

3.80

4.20

T2

4 ppm BAP

3.60

5.33

5.50

T3

2 ppm NAA

2.83

3.60

4.00

T4

4 ppm NAA

3.03

4.25

5.00

T5


4.16

6.00

7.00

T6


3.16

4.83

5.33

T7


4.66

6.33

7.33

T8


3.66

5.66

6.66

T9

Control

2.80

3.50

3.83

S.Em. ±

0.09

0.12

0.14

C.D. at 5 %

0.28

0.35

0.42

C.V. %

4.77

4.19

4.46

0

2

4

6

8

T1 T2 T3 T4 T5 T6 T7 T8 T9


Fig. 5: Effect of growth regulators on no. of leaves at different stage of growth of banana cv. Grand Naine.

No.of

leaves

Treatment

Table 4.6: Effect of growth regulators on leaf area (cm2) at


Treatment

Days after planting

30

60

90

T1

2ppm BAP

133.72

239.53

340.82

T2

4 ppm BAP

150.83

261.82

354.57

T3

2 ppm NAA

128.24

230.94

333.66

T4

4 ppm NAA

139.55

246.75

344.74

T5


164.36

274.11

365.15

T6


142.67

252.45

349.60

T7


170.93

280.37

370.35

T8


157.66

268.28

360.23

T9

Control

123.40

223.60

328.40

S.Em. ±

3.38

5.27

6.26

C.D. at 5 %

10.05

15.67

18.61

C.V. %

4.02

3.61

3.10

0

100

200

300

400

T1 T2 T3 T4 T5 T6 T7 T8 T9


Fig. 6: Effect of growth regulators on leaf area (cm2) at different stage of growth of banana cv. Grand Naine.

Leaf

are

a (cm

2)

Treatment

and 90 DAP and also with T2 at 90 DAP. The minimum leaf

area (123.40, 223.60 and 328.40 cm2) recorded at 30, 60 and

90 DAP under treatment T9 which was control.

4.7 Effect on no. of roots


revealed that there were significant differences in no. of roots

in suckers due to different concentration of growth regulators.

The maximum number of roots (6.40, 8.20 and

10.60) were recorded by T7 (2 ppm BAP + 4 ppm NAA) as

compared to other treatments at 30, 60 and 90 DAP

respectively, which was statistically at par with T5 at 60 DAP.

The minimum number of roots (3.60, 5.40 and 6.80) at

respective growth stage were recorded under treatment T9

(control).

4.8 Effect on root length


revealed that there were significant differences in root length

in suckers due to different concentration of growth regulators.

The maximum root length (16.83, 21.40 and 28.19

cm) were recorded by T7 (2 ppm BAP + 4 ppm NAA) as

compared to other treatments at 30, 60 and 90 DAP which

was statistically at par with T2 at 30 and 90 DAP and T5 and

T8 at 30, 60 and 90 DAP. The minimum root length (13.13,

16.46 and 23.71 cm) were recorded at respective growth

stages under treatment T9 (control).

Table 4.7: Effect of growth regulators on no. of roots at


Treatment

Days after planting

30

60

90

T1

2ppm BAP

4.00

6.00

7.80

T2

4 ppm BAP

4.80

7.00

9.00

T3

2 ppm NAA

3.80

5.80

7.40

T4

4 ppm NAA

4.00

6.40

8.00

T5


5.60

7.80

9.80

T6


4.20

6.80

8.20

T7


6.40

8.20

10.60

T8


5.20

7.60

9.40

T9

Control

3.60

5.40

6.80

S.Em. ±

0.13

0.16

0.18

C.D. at 5 %

0.38

0.48

0.53

C.V. %

4.79

4.14

3.63

0

2

4

6

8

10

12

T1 T2 T3 T4 T5 T6 T7 T8 T9


Fig. 7: Effect of growth regulators on no. of roots at different stage of growth of banana cv. Grand Naine.

No. of

roots

Treatment

Table 4.8: Effect of growth regulators on root length (cm)


Treatment

Days after planting

30

60

90

T1

2ppm BAP

14.04

18.05

24.89

T2

4 ppm BAP

15.79

19.70

26.50

T3

2 ppm NAA

13.73

17.50

24.37

T4

4 ppm NAA

14.61

18.67

25.13

T5


16.35

20.93

27.86

T6


15.26

19.12

25.78

T7


16.83

21.40

28.19

T8


16.02

20.26

27.26

T9

Control

13.13

16.46

23.71

S.Em. ±

0.42

0.43

0.62

C.D. at 5 %

1.25

1.28

1.83

C.V. %

4.82

3.91

4.11

0

5

10

15

20

25

30

T1 T2 T3 T4 T5 T6 T7 T8 T9


Fig. 8: Effect of growth regulators on root length (cm) at different stage of growth of banana cv. Grand Naine.

Root

length

(cm

)

Treatment

4.9 Effect on survival per cent

Data on survival percentage of banana shoots per

sucker as affected by different treatents of growth regulators

along with statistical inferences are presented in Table 4.9

and illustrated in Fig. 9.

A persual of data clearly revealed significant effect of

plant growth regulators on survival of banana shoots per

sucker. Significantly maximum survival percentage (73.52 %)

was observed in the treatment T7 (2 ppm BAP + 4 ppm NAA)

which was at par with T5. Survival percentge (40.30 %) was

minimum at respective growth stage under control treatment

(T9).

4.10 Effeect on per cent of contamination

There was no any contamination observed on raising

shoots during the experiments.

4.11 Effeect on per cent of browning

There was no any browning observed on raising

shoots during the experiments.

4.12 Effeect on pest and disease incidence

The data on disease and pest affected shoots per

treatment were recorded and presented in table 4.10. The

result showed that there was a non-significant difference

among the various treatments.

4.13 Economics

The economics of different treatments presented in

table 4.11. The higher BCR 3.27 was computed in treatment

T7 (2 ppm BAP + 4 ppm NAA) followed by T5 i.e. 2 ppm BAP +

2 ppm NAA (3.16).

Table 4.9: Effect of growth regulators on survival per cent


Treatment

No. of shoot at 30

DAP

No. of

shoot survival

at 90 DAP

Survival per cent

(%)

T1

2ppm BAP

2.95

1.48

50.16 (45.09)

T2

4 ppm BAP

4.88

3.31

67.82 (55.44)

T3

2 ppm NAA

2.79

1.29

46.23 (42.84)

T4

4 ppm NAA

3.72

2.12

56.98

(49.02)

T5


5.81

4.21

72.46

(58.35)

T6


4.79

3.06

63.88

(53.06)

T7


5.93

4.36

73.52

(59.03)

T8


5.64

3.97

70.39

(57.03)

T9

Control

2.63

1.06

40.30

(39.40)

S.Em. ±

0.12

0.04

1.02

C.D. at 5 %

0.35

0.12

3.02

C.V. %

4.69

2.46

2.93

(Figures in the parentheses are arcsine transformed value)

0.00

10.00

20.00

30.00

40.00

50.00

60.00

70.00

80.00

T1 T2 T3 T4 T5 T6 T7 T8 T9

Survival …

Surv

ival (%

)

Treatment

Fig. 9: Effect of growth regulators on survival per cent at different stage of growth of banana cv. Grand Naine.

Table 4.10: Effect of growth regulators on pest and

disease incidence of banana cv. Grand Naine.

Treatment

No. of affected shoots

T1

2ppm BAP

4.33

T2

4 ppm BAP

3.67

T3

2 ppm NAA

4.67

T4

4 ppm NAA

4.00

T5


3.67

T6


4.00

T7


3.00

T8


3.33

T9

Control

5.00

S.Em. ±

0.46

C.D. at 5 %

NS

C.V. %

20.02

Table 4.11: Economics of different macropropagation treatments.

Treatment

No. of plants obtained/ sucker

No. of plants obtained/1000

suckers

Treatment cost/1000 suckers (Rs.)

Total cost (Rs.)

Total income (Rs.)

BCR

T1

1.48

1480

1.50

8001.50

8880.00

1.11

T2

3.31

3310

3.00

8003.00

19860.00

2.48

T3

1.29

1290

0.08

8000.08

7740.00

0.97

T4

2.12

2120

0.16

8000.16

12720.00

1.59

T5

4.21

4210

0.79

8000.79

25260.00

3.16

T6

3.06

3060

1.54

8001.54

18360.00

2.29

T7

4.36

4360

0.83

8000.83

26160.00

3.27

T8

3.97

3970

1.58

8001.58

23820.00

2.98

T9

1.06

1060

0.00

8000.00

6360.00

0.79

The cost of sucker considered Rs. 3/- per sucker

Price of BAP and NAA considered Rs. 150/- and Rs. 8/- per gram respectively

The selling price of macropropagated plant considered Rs. 6/- per plant

Common expenditure Rs. 5000/- added in total cost

DISCUSSION

V DISCUSSION

The present study was conducted to study the effect

of different concentration of growth regulators on bud initiation,

growth and survival per cent on banana shoots. The effects of

these treatments were recorded and the results obtained are

discussed.

5.1 Effect on days to bud initiation

The minimum days required to bud initiation was

recorded when suckers were treated with 2 ppm BAP + 4 ppm

NAA (T7). Early bud initiation with BAP treatments might be due

to the rapid multiplication of cells in sprouting corms. Similar

result found by Bhaskar et al. (1993) in banana, Murkute et

al. (2008) in citrus and Sontakke et al. (1996) in fig.

5.2 Effect on no. of shoots /sucker

There were significant differences in sprouting

of suckers due to use of different plant growth

regulators at different concentaration and their

combinations. The maximum shoots per sucker were

recorded in 2 ppm BAP + 4 ppm NAA (T7). It might be due to

BAP in prolifertion and by allowing the apical meristem to

proliferate along with the non dominant lateral meristems for

enhancing the required response. BAP quite more effective in

breaking dormancy of lateral buds, accelerating the sprouting of

lateral buds and resulting more sprouts per sucker. BAP was

found to be as ideal to obtained mass proliferation from shoot

primordial meristems. The effect of cytokinin on axillary bud

brusting and shoot production have been demonstrated by

Murashige (1974). Cytokinin in which should be used to release

apical dominance in the latent buds. The dominant bud of

vegetative apex are stimulated to grow and obligated into

presence of cytokinin and also produce new axes. There is also

one universally accepted concept that morphogenetic response

is regulated by the relative concentration of auxins and

cytokinins as pointed out by Skoog and Miller (1957). When,

BAP level was above 6.0 mg/l it has some inhibitory effect on

shoot elongation. The result agreed with previous findings of

Mukunthakumar et al. (2011), Al-amin et al. (2009), Karim et al.

(2009), Kalimuthu et al. (2007), Rahman et al. (2004) in

banana, Kabir et al. (2007), Naik and Shah (1996) in papaya

and Al-Saif et al. (2011) in pineapple and Das et al. (2008)

also observed similar trend while working in wood apple.

5.3 Effect on shoot length

The data revealed that the length of the shoot was



length of the shoot at 30, 60 and 90 DAP was observed in

treatment T7 (2 ppm BAP + 4 ppm NAA). It might be due to the

NAA which promote linear growth by cell elongation and cell

division along with the longitudinal axis. Cell elongation was

occurred with NAA due to uptake of large quantity of water

which leads to enlargement of vacuole and it has direct effect on

length of plant. Role of BAP i.e. cytokinin has rapid multiplicate

as cell division. Similar result was also reported by Azam et al.

(2010), Al-amin et al. (2009), Karim et al. (2009), Rahman et

al. (2004) in banana, Murkute et al. (2004) in

pomegranate and in mangosteen NAA was only a plant

growth regulator supplement improving the effect of BAP result

found by Sirchi et al. (2008).

5.4 Effect on shoot girth

The data revealed that the girth of the shoot was



girth of the shoot at 30, 60 and 90 DAP was observed in

treatment T7 (2 ppm BAP + 4 ppm NAA). It might be due to the

stimulatory effects of auxin on growth vary considerably

between tissues. In general, the most marked stimulations were

obtained in stem and coleoptile. Because of this reason, shoot

diameter might have increased. Similar result was found by

Gubbuk and Pekmezcl (2004) in banana.

5.5 Effect on no. of leaves

The number of leaves was significantly affected due to

growth regulators at all the stages of growth. The highest

number of leaves was found in plants treated with 2 ppm BAP +

4 ppm NAA. Increased number of leaves per shoot may

probabely be due to the effect of auxin which participates widely

in the regulation of differentiation phenomena, e.g. the polar

differentiation of xylem strands in leaves and cell enlargement of

leaves and retention of chlorophyll in plant and it has direct

effect on number of leaves. Similar result found by Karim et al.

(2009) in banana, Patil et al. (2011) in pomegranate and

Krishan Kumar et al. (2001) in citrus and Sontakke et al.

(1996) in fig.

5.6 Effect on leaf area

The maximum leaf area was recorded by T7 (2 ppm

BAP + 4 ppm NAA). It might be due to NAA and cytokinin

resulted from increased cell division and cell expansion

resulting in enhanced vegetative growth and in the presence of

NAA which increase the carbohydrates and induced

translocation of sugar resulting in supply of food to the leaves

and this may increase leaf area in plant. Result supported by

Karim et al. (2009) in banana, Sontakke et al. (1996) in fig.

5.7 Effect on no. of roots

The data revealed that there were significant

differences in root length in suckers due to different

concentration of growth regulators. The maximum root length

was recorded by T7 (2 ppm BAP + 4 ppm NAA) as compared to

other treatments. It might be due to enhancment of the root

initiation in the presence of auxin in plant. The result was

agreed with previous findings of Viehmannova et al. (2007),

Bekheet and Saker (1999) in banana, Murkute et al. (2004)

in pomegranate, Haripyaree et al. (2011), Begum et al. (2003)

in citrus, Hosseini et al. (2011) in mahaleb and Sontakke et al.

(1996) in fig.

5.8 Effect on root length

The maximum root length was recorded by T7 (2

ppm BAP + 4 ppm NAA). It might be due to cell enlargement of

root which has direct effect on length of root. Similar result was

found by Mukhtar et al. (2005) in citrus and Hosseini et al.

(2011) in mahaleb.

5.9 Effect on survival per cent

A persual of data clearly revealed significant

effect of plant growth regulators on surviva l of banana

suckers. Significantly maximum survival percentage

was observed in the treatment T7 (2 ppm BAP + 4 ppm

NAA). Similar result found by Rahman et al. (2004) in

banana, Rahaman et al. (1992) in papaya, Upadhyay

and Badyal (2007) in pomegranate and Sontakke et al.

(1996) in fig.

5.10 Effect on contamination % and percent of

browning

The contamination % and percent of browning

did not seen on shoot during experiment.

5.11 Effect on pest and disease incidence

There was a non-significant difference in respect on

pest and disease incidences on banana shoot in experiment

period.

SUMMARY AND CONCLUSION

VI SUMMARY AND CONCLUSION

An experiment on “Ex-situ macropropagation

study in banana (Musa paradisiaca L.) cv.Grand Naine

under South Gujarat condition” was conducted at Fruit

Research Station, Navsari Agricultural University, Gandevi,

during 2011- 2012.

The experiment was laid out in a Completely

Randomized Design (CRD) with nine treatments of different

concentration of BAP and NAA alone and combination of both

including control. The treatments were replicated thrice. The

findings pertained during the course of study are summarized

in this chapter.

6.1 Days to bud initiation

Significantly the minimum days (9.21 days) required

to bud initiation was recorded when suckers were treated with 2

ppm BAP + 4 ppm NAA, while the maximum i.e. 14.18 days

required to bud initiation in control treatment i.e. without any

tretment.

6.2 No. of shoots /sucker (30 days after

treatment)

There were significant differences in sprouting

of sucker due to use of different plant growth regulators

at different concentaration and their combinations. The

maximum shoots per sucker (5.93) were recorded in 2

ppm BAP + 4 ppm NAA. The minimum shoots per sucker

(2.63) were recorded in control.

6.3 Shoot length

Consistently the higher length of the shoot at 30, 60

and 90 DAP was observed in treatment 2 ppm BAP + 4 ppm

NAA, while the minimum length of the shoot was recorded

under control during growth stage of banana shoot.

6.4 Shoot girth

At 30, 60 and 90 DAP the maximum shoot girth was

observed with treatment 2 ppm BAP + 4 ppm NAA, while the

minimum shoot girth was recorded under T9 treatment at all the

stages of growth of banana shoot.

6.5 No. of leaves

The number of leaves was significantly affected due

to growth regulators at all the stages of growth. The highest

number of leaves was found in shoots in the suckers treated

with 2 ppm BAP + 4 ppm NAA, while the minimum number of

leaves was observed in control treatment (T9).

6.6 Leaf area

The maximum leaf area was recorded by T7 (2 ppm

BAP + 4 ppm NAA) treatment as compared to other treatments

at 30, 60 and 90 DAP, whereas the minimum leaf area recorded

under treatment T9 (control).

6.7 No. of roots

The maximum number of roots per sucker were

recorded by T7 (2 ppm BAP + 4 ppm NAA) as compared to other

treatments at 30, 60 and 90 DAP, and it was reported the

minimum in control.

6.8 Root length

Root length was recorded the maximum with the

treatment T7 (2 ppm BAP + 4 ppm NAA) as compared to other

treatments at 30, 60 and 90 DAP, while the minimum root

length recorded under treatment T9 (control).

6.9 Survival per cent

A persual of data clearly revealed significant

effect of plant growth regulators on survival of banana

shoots per sucker. The survival percentage was

observed significantly maximum in the treatment T7 (2

ppm BAP + 4 ppm NAA), whereas it was minimum under

control (T9).

6.10 Percent of contamination

There was no any contamination observed on

raising shoots during the experiments.

6.11 Percent of browning

During the experimentation, browning was not

observed in shoots.

6.12 Pest and Disease incidence

There were non significant effect of pests and

diseases on raising shoots during the experimentation.

6.13 Economics

The maximum BCR (3.27) was found in the

treatment T7 (2 ppm BAP + 4 ppm NAA) and minimum BCR

(0.79) in control (T9).

CONCLUSION:

Based on the present investigation on Ex-situ

macropropagation study in banana (Musa paradisiaca

L.) cv.Grand Naine, it can be concluded that, the early

sprouting of buds, maximum number of shoots, shoot length,

shoot girth, number of leaves per shoot, leaf area, number of

roots per shoot and root length were recorded with the

treatment 2 ppm BAP + 4 ppm NAA at all growth stages. The

survival percentage (73.52 %) was recorded the highest after

bagging in polythene bag at 90 days. Other combination of PGR

had also positive effect on growth and survival as compared to

control. After 90 days, four to five healthy plants of banana can

be obtained from each sucker in the said treatment. These

plants can be used as healthy planting material. This method of

propagation was also found economically beneficial.

Thus macropropagation provides cheap, simple and

relatively rapid technique for vegetative multiplication of Musa

species that could be amenable to the resource poor, unskilled,

small and marginal farmers.

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APPENDIX

Appendix-I: State wise area and production

(estimated) of Banana in India in year

2010-2011.

Name of State

Area in 000 HA

Production in 000 MT

Productivity in MT/Ha

Tamilnadu 125.4 8253.0 65.8

Maharashtra 82.0 4303.0 52.5

Gujarat 64.7 3978.0 61.5

Andhra

Pradesh 79.3 2774.8 35.0

Karnataka 111.8 2281.6 20.4

Madhya

Pradesh 38.1 1719.6 45.2

Bihar 31.9 1517.1 47.6

Uttarpradesh 32.4 1346.1 41.5

West Bengal 42.0 1010.1 24.0

Assam 47.6 723.6 15.2

Other 175.3 1873.1 10.7

Total 830.5 29779.9 35.9

Appendix-II District wise area and production of

banana fruit crop in the year 2010-11.

Sr.no Name of District

Area

in 00

HA

Production 00 M.T

Productivity Kg./Ha

1 AHMEDABAD 125 3278 26.22

2 AMRELI 35 1138 32.51

3 BANASKANTHA 00 00 00

4 BHARUCH 15200 990280 65.15

5 DAHOD 0 0 0

6 DANG 20 660 33

7 GANDHINAGAR 58 2291 39.50

8 KHEDA 806 36270 45

9 MEHSANA 0 0 0

10 NARMADA 6000 384000 64

11 NAVSARI 460 25760 56

12 PANCHMAHAL 300 9150 30.50

13 PATAN 0 0 0

14 SABARKANTHA 204 6149 30.14

15 SURAT 11235 719040 64

16 TAPI 1600 88000 55

17 VADODARA 9390 569130 60.61

18 VALSAD 770 43120 56

19 ANAND 14045 877391 62.47

20 BHAVNAGAR 1600 88000 55

21 JAMNAGAR 20 316 15.80

22 JUNAGADH 1800 81000 45

23 KUTCH 1000 52690 52.69

24 PORBANDAR 0 0 0

25 RAJKOT 12 360 30

26 SURENDRANAGAR 0 0 0

Total 64680 3978023 61.50

Appendix-III: Weekly meteorological data recorded at meteorological observatory of FRS, Gandevi.

Month and Year

Standard week

Temperature

(0C)

Relative Humidity

(%)

Rainfall (mm) Max. Min.

Aug

2011

32

30.24

24.38 90.64 8.01

33 29.20 24.90 90.85 9.12

34 30.25 23.75 90.57 9.71

35 28.05 23.22 97.42 31.02

36 30.75 23.78 89.07 20.05

Sep

2011

37 29.75 24.64 88.92 14.38

38 29.72 23.70

88.50

11.47

39 30.64 22.51 81.71 4.74

40 31.82 22.30 81.42 0

Oct

2011

41 33.21 21.78 74.14 3.71

42 36.22 23.58 69.64 0

43 36.27 19.65 61.28 0

44 36.19 16.82 55.28 0

45 35.45 18.05 55.28 0

Nov

2011

46 35.72 15.95 61.35 0

47 34.95 14.01 59.57 0

48 33.72 14.75 53.71 0

49 34.40 17.48 65.07 0

Dec 2011

50 33.13 14.34 66.78 0

51 32.36 12.41 59.64 0

52 32.05 11.51 63.35 0

53 29.85 9.04 63.64 0

Jan 2012

54 29.08 9.38 67.28 0

55 28.9 7.92 56.71 0

56 27.89 8.8 66.64 0

57 29.20 8.9 70.28 0

C E R T I F I C A T E

This is to certify that I have no objection to

supply one copy of any part of this thesis at a time to

any scientist through reprographic process for rendering

reference services in a library or documentation centre.

Place: Navsari

Date: 5th April 2013 (Ravani Dineshkumar G.)

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