department of agronomy sher-e-bangla agricultural

INFLUENCE OF BIOFERTILIZER, NITROGEN AND

PHOSPHOROUS ON NODULATION, GROWTH AND YIELD

OF LENTIL

AFSANA MAHMUD CHOWDHURY

DEPARTMENT OF AGRONOMY

SHER-E-BANGLA AGRICULTURAL UNIVERSITY

DHAKA-1207

JUNE, 2017

INFLUENCE OF BIOFERTILIZER, NITROGEN AND

PHOSPHOROUS ON NODULATION, GROWTH AND YIELD

OF LENTIL

By

AFSANA MAHMUD CHOWDHURY REGISTRATION NO. 15-07013

A Thesis

Submitted to the Faculty of Agriculture,

Sher-e-Bangla Agricultural University, Dhaka,

in partial fulfilment of the requirements for the degree of

MASTER OF SCIENCE

IN

AGRONOMY

SEMESTER: JULY-DECEMBER, 2017

Approved by:

(Prof. Dr. A. K. M. Ruhul Amin)

(Prof. Dr. Md. Fazlul Karim)

Supervisor

Co-Supervisor

(Prof. Dr. Md. Shahidul Islam)

Chairman Examination Committee

CERTIFICATE

This is to certify that the thesis entitled “INFLUENCE OF

BIOFERTILIZER, NITROGEN AND PHOSPHOROUS ON

NODULATION, GROWTH AND YIELD OF LENTIL” submitted

to the Faculty of Agriculture, Sher-e-Bangla Agricultural

University, Dhaka, in partial fulfillment of the requirements for

the degree of MASTER OF SCIENCE (M.S.) in AGRONOMY,

embodies the results of a piece of bona fide research work

carried out by AFSANA MAHMUD CHOWDHURY,

Registration. No. 15-07013 under my supervision and guidance.

No part of this thesis has been submitted for any other degree or

diploma.

I further certify that such help or source of information as has

been availed of during the course of this investigation has duly

been acknowledged.

Dated:

Dhaka, Bangladesh

(Prof. Dr. A. K. M. Ruhul Amin)

Supervisor

i

ACKNOWLEDGEMENTS

All praises to the Almighty Allah, the great, the gracious, merciful and supreme ruler

of the universe who enables me to complete this present piece of work for the degree

of Master of Science (M.S.) in the Department of Agronomy.

The author would like to express her deepest sense of gratitude, respect to her

research supervisor, Prof. Dr. A.K.M. Ruhul Amin, Department of Agronomy, Sher-

e-Bangla Agricultural University, for his kind and scholastic guidance, untiring

effort, valuable suggestions, inspiration, extending generous help and encouragement

during the research work and guidance in preparation of manuscript of the thesis.

The author sincerely expresses her deepest respect and boundless gratitude to her co-

supervisor Prof. Dr. Md. Fazlul Karim, Department of Agronomy, for his helpful

suggestion and valuable advice during the preparation of this manuscript.

The author would like to express her deepest respect and boundless gratitude to all

the respected teachers of Dept of Agronomy, Sher-e-Bangla Agricultural University,

for the valuable teaching, sympathetic co-operation and inspirations throughout the

course of this study and suggestions and encouragement to research work. The author

would like to express her cordial thanks to the departmental and field staff for their

active help during the experimental period.

The author feels proud to express her sincere appreciation and gratitude to Ministry

of Science and Technology, The People’s Republic of Bangladesh for providing her

National Science and Technology (NST) fellowship.

At last but not the least, the author feels indebtedness to her beloved parents and

husband whose sacrifice, inspiration, encouragement and continuous blessing paved

the way to her higher education.

ii

INFLUENCE OF BIOFERTILIZER, NITROGEN AND PHOSPHOROUS

ON NODULATION, GROWTH AND YIELD OF LENTIL

ABSTRACT

To study the influence of Biofertilizer, nitrogen and phosphorous on

nodulation, growth and yield of lentil a field experiment was conducted at the

central farm, Sher-e-Bangla Agricultural University, Dhaka-1207, during rabi

season from November 2016 to March 2017. Treatments consisted of two

biofertilizer levels: (i) control and (ii) Biofertilizer (Rhizobium) with six

combinations of nitrogenous and phosphetic fertilizer: (i) No nitrogen +

phosphorous fertilizer (control), (ii) 50% less of recommended N + P, (iii)

25% less of recommended N + P, (iv) recommended N + P, (v) 25% higher of

recommended N + P and (vi) 50% higher of recommended N + P. The

experiment was conducted in two factor Randomized Complete Block Design

(RCBD) with three replications. Growth and yield parameters like plant

height, no of branch plant-1

, nodule count, dry weight plant-1

, pods plant-1

,thousand seed weight, grain yield, stover yield, biological yield etc. were

collected from this experiment. Data were analyzed by MSTAT-C software.

The significance of difference among the treatment means was estimated by

the Least Significance Difference (LSD) at 5% level of probability. Result

revealed that biofertilizer treated plot B1 (Biofertilizer) was found superior in

producing maximum plant height, branches plant-1

, dry weight plant-1

, nodule

plant-1

, pods plant-1

, seed yield and biological yield of lentil. On the other

hand, N+P fertilizer at recommended dose (F3) gave highest yield, plant

height, branches plant-1


, nodules plant-1

, pods plant-1

, 1000

seed weight (20.73 g), seed yield, stover yield, and biological yield . In case of

interaction, B1F3 was found superior in producing maximum yield and yield

components like pods plant-1

(68.40), 1000 seed weight (20.98 g ), seed yield

(2562.40 kg ha-1

), stover yield (2396.80 kg ha-1

), biological yield (4959.20 kg

ha-1

). From the result of the study, it was revealed that the application of

biofertilizer and recommended N + P combination had a positive impact on

lentil (BARI Mosur-6).

iii

LIST OF CONTENTS

CHAPTER TITLE PAGE NO.

ACKNOWLEDGEMENTS i

ABSTRACT ii

LIST OF CONTENTS iii

LIST OF TABLES viii

LIST OF FIGURES ix

LIST OF APPENDICES xi

LIST OF PLATES xii

LISTS OF ACRONYMS xiii

1 INTRODUCTION 1

2 REVIEW OF LITERATURE 3

2.1 Effect of bio-fertilizer on nodulation, growth and

yield

3

2.2 Nitrogen fixation and Rhizobium inoculation on

lentil

5

2.3 Effect of nitrogen on nodulation, growth and yield 7

2.4 Effect of phosphorus on nodulation, growth and

yield

8

3 MATERIALS AND METHODS 10

3.1 Site description 10

3.1.1 Geographical location 10

3.1.2 Agro-ecological region 10

3.1.3 Climate 10

iv

LIST OF CONTENTS (Contd.)


3.1.4 Soil 11

3.2 Details of the experiment 11

3.2.1 Treatments 11

3.2.2 Experimental design and layout 12

3.3 Crop/Planting Material 12

3.3.1 Description of crop: BARI Mosur-6 12

3.3.2 Description of chemical fertilizer management 12

3.3.3 Description of biofertilizer management 12

3.4 Crop management 12

3.4.1 Seed collection 12

3.4.2 Seed sowing 12

3.4.3 Collection and preparation of initial soil sample 13

3.4.4 Preparation of experimental land 13

3.4.5 Fertilizer application 13

3.4.6 Intercultural operations 13

3.4.6.1 Thinning 13

3.4.6.2 Weeding 14

3.4.6.3 Application of irrigation water 14

3.4.6.4 Drainage 14

3.4.6.5 Plant protection measures 14

3.4.7 Harvesting and post-harvest operation 14

3.4.8 Recording of data 15

3.4.9 Detailed procedures of recording data 15

v



3.4.9.1 Plant height 16

3.4.9.2 Branches plant-1

16

3.4.9.3 Nodules plant-1

16

3.4.9.4 Dry weight of plant 16

3.4.10.1 Pods plant-1

16

3.4.10.2 1000 seed weight 16

3.4.10.3 Seed yield 17

3.4.10.4 Stover yield 17

3.4.10.5 Biological yield 17

3.4.10.6 Harvest index 17

3.4.10.7 Statistical analysis 17

4 CHAPTER IV 18

4.1 Effect of biofertilizer and nitrogen +

phosphorous on growth of lentil

18

4.1.1 Plant height 18

4.1.1.1 Effect of biofertilizer 18

4.1.1.2 Effect of nitrogen + phosphorous 19

4.1.1.3 Interaction effect of biofertilizer and nitrogen +

phosphorous

19

4.1.2 Branches plant-1

20




phosphorous

22

4.1.3 Nodules plant-1

23


vi




4.1.3.3

4.1.4

Interaction effect of biofertilizer and nitrogen +

phosphorous

Dry weight plant-1

25

26

4.1.4.1

4.1.4.2

Effect of biofertilizer

Effect of nitrogen + phosphorous

26

27


phosphorous

27

4.2 Effect of biofertilizer and nitrogen +

phosphorous on yield compponets and yields

29

4.2.1 Pods plant-1

29




phosphorous

30

4.2.2 1000 seed weight 31




phosphorous

33

4.2.3 Seed yield 33




phosphorous

34

4.2.4 Stover yield 35


vii





phosphorous

37

4.2.5 Biological yield 37


4.2.5.2

4.2.5.3

Effect of nitrogen + phosphorous

Interaction effect of biofertilizer and nitrogen +

phosphorous

37

38

4.2.6 Harvest index 38




phosphorous

39

5 SUMMARY AND CONCLUSION 41

REFERENCES 44

APPENDICES

49

viii

LIST OF TABLES

Table No. Title Page No.

1 Interaction effect of biofertilizer and different level of N + P

fertilizer on plant height of lentil at different days after sowing

21


fertilizer on branch plant-1

of lentil at different days after

sowing

23


fertilizer on nodule plant-1

of lentil at different day after sowing

26


fertilizer on dry weight of lentil at different days after sowing

28

5 Interaction effect of biofertilizer and N + P on number of pods

plant-1

at harvest, and thousand seed weight of lentil

31

6 Interaction effect of biofertilizer and N + P on grain yield, stover

yield, biological yield and harvest index of lentil

35

ix

LIST OF FIGURES

Figure No. Title Page No.

1 Effect of biofertilizer on plant height of lentil at different days after

sowing

18

2 Effect of different level (N+P) fertilizers on plant height of lentil at

different days after sowing

20

3 Effect of biofertilizer on branch plant-1


sowing

21

4 Effect of different level (N+P) fertilizers on branch plant-1

of lentil at


22

5 Effect of biofertilizer on nodule plant-1


sowing

24

6

7

8

Effect of different level (N+P) fertilizers on nodule plant-1

of lentil at


Effect of biofertilizer on dry weight of lentil at different days after

sowing

Effect of different level (N+P) fertilizers on dry weight of lentil at


25

27

28

9 Effect of biofertilizer on pods plant-1

of lentil

29

10 Effect of different level of N + P on pods plant-1

of lentil 30

11

12

13

Effect of biofertilizer on thousand seed weight of lentil

Effect of different level of N + P on thousand seed weight of lentil

Effect of biofertilizer on grain yield of lentil

32

32

33

14 Effect of different level of N + P on grain yield of lentil 34

15 Effect of biofertilizer on stover yield of lentil 36

16

Effect of different level of N + P on stover yield of lentil

36

x

Figure No.

LIST OF FIGURES (Cont’d)

Title

Page No.

17 Effect of biofertilizer on biological yield of lentil 37

18 Effect of different level of N + P on biological yield of lentil

38

19 Effect of biofertilizer on harvest index of lentil 39

20 Effect of different level of N + P on harvest index of lentil

40

xi

LIST OF APPENDICES

Appendix No. Title Page No.

I Monthly record of air temperature, relative humidity and rainfall

of the experimental site during the period of November, 2016 to

March 2017

49

xii

LIST OF PLATES

Plates No.

1

2

Title

Field view of seedling stage

Data recording for nodule

Page No.

50

51

xiii

LIST OF ACRONYMS

AEZ Agro-Ecological Zone

BARI Bangladesh Agricultural Research Institute

BAU Bangladesh Agricultural University

BBS Bangladesh Bureau of Statistics

Co Cobalt

CV% Percentage of coefficient of variance

cv. Cultivar

DAE Department of Agricultural Extension

DAS Days after sowing

0C Degree Celsius

et al And others

FAO Food and Agriculture Organization

g gram(s)

ha-1

Per hectare

HI Harvest Index

kg Kilogram

Max Maximum

mg Milligram

Min Minimum

MoP Muriate of Potash

N Nitrogen

No. Number

NS Not significant

% Percent

SAU Sher-e-Bangla Agricultural University

SRDI Soil Resources and Development Institute

TSP Triple Super Phosphate

UPOV Union for the Protection of Plant Varieties

Wt. Weight

1

CHAPTER 1

INTRODUCTION

Lentil (Lens culinaris) belongs to family Fabaceae. It is a nutritious food

legume. It is one of the oldest annual grains legumes more consumed and

cultivated in the world and mostly eaten as dhal. Lentil is originating from

South Western Asia as early as 6000 B.C. Lentil is rich in protein and also

contains high concentration of essential amino acid as isoleucine and lysine, as

well as other nutrients like minerals and fiber, folate, vitamin B1 (Rozan et al.,

2001). Lentil is also known as a „poor man's meat‟ because of its rich protein

content. In South East Asia lentil is also equally liked by all socioeconomic

groups (Bhatty, 1988). Also, it has a positive contribution for increasing the

soil fertility due to the high number of effective nodules in their root that

supply nitrogen into the soil (Omer, 2009).

Omer (2009) reported that lentil is good choice in crop rotations as it produced

130.44 nodules per plant which improve soil health by adding nitrogen and

organic matter for following crops. Lentil crop requires nitrogen for their

growth and development approximately 85% of nitrogen necessity of lentil is

fulfilled with the help of atmospheric nitrogen fixation during symbiotic

relationship of lentil roots with microorganism Rhizobium bacteria in the field

and due to which yield could be increased up to 2 ton ha-1

(Bisen et al., 1980).

Small doses of N fertilizers applied to an annual pulse are beneficial if nodule

initiation is delayed (Mahon and Child, 1979). Phosphorus plays a major role in

many plant processes, including storing and transfer of energy; stimulation of

root growth, flowering, fruiting and seed formation; nodule development and

N2 fixation (Mclaren and Cameron, 1996; Ali et al., 1997). Bremer et al.

(1989) found that P application increased dry matter and grain yield but did not

affect N2 fixation indicating that the legume host was more responsive to P

application than the Rhizobia. Lentils are sensitive to high rates of P fertilizer

placed directly in the seed rows.

2

Gupta and Sharma (1992) reported that yield of lentil was 0.87 - 1.30 t/ha with

0 - 32 kg phosphorus and no inoculation, and 0.89 - 1.68 t/ha with 0 – 32 kg

phosphorus and inoculation. Seeds protein content increased with application

of phosphorus and inoculation. Hossain and Suman (2005) carried out an

experiment to evaluate the effect of Rhizobium and different levels of urea N

on growth, yield and N-uptake of lentil. Among the treatments Rhizobium

inoculation had significant effect on nodule formation, plant height, number of

seeds, seed and stover yields, compared to uninoculated controls. The highest

seed yield was recorded for the treatment Rhizobium treatment that was

statistically similar to that of 100% N and Rhizobium with the corresponding

yields of 1533 and 1458 kg/ha, respectively. The inoculation of Rhizobium

significantly influenced all the crop characters including N contents, N uptake

by seed and shoot as well as protein content of seed. The highest N-uptake by

seed (78.61 kg/ha) was recorded for the treatment Rhizobium and N-uptake by

shoot (53.87 kg/ha) was recorded for the treatment 100% N. Therefore,

inoculation of Rhizobium may be a good practice to achieve higher seed yield

of lentil.

Furthermore, for having maximum nodulation, growth and yield of lentil, it is

necessary to find out the best combination of nitrogen, phosphorous and

biofertilizer. Very little information is available about the influence of nitrogen,

phosphorous and biofertilizer on nodulation, growth and yield of lentil. So,

there is a scope to take research in this aspect. Thus, the present study was

carried out by the following objectives

To select suitable nitrogenous and phosphetic fertilizer management for

maximum yield of lentil,

To study the influence of biofertilizers on growth and yield of lentil, and

To asses interaction of biofertilizer nitrogenous and phosphetic fertilizer

on the yield of lentil.

3

CHAPTER 2

REVIEW OF LITERATURE

An attempt was made in this section to collect and study the relevant

information available in the country and abroad regarding the influence of

Biofertilizer, nitrogen and phosphorous on nodulation, growth and yield of

Lentil to gather knowledge helpful in conducting the present research work and

subsequently writing up the result and discussion.

2.1. Effect of bio-fertilizer on nodulation, growth and yield

Biofertilizers are gaining importance as they are ecofriendly, non-hazardous

and non-toxic. A substantial number of bacterial species, mostly those

associated with the plant rhizosphere, may exert a beneficial effect upon plant

growth. Biofertilizers include mainly the nitrogen fixing, phosphate

solubilizing and plant growth promoting micro-organism. Inoculating pulse

crops with rhizobia to add nitrogen is routine for most growers. The presence

of efficient and specific strains of Rhizobium in the rhizosphere is one of the

most important requirements for proper establishment and growth of grain

legume plant. Phosphate solubilizing bacteria partly solubilizes inorganic and

insoluble phosphate and improves applied phosphorus use efficiency

stimulating plant growth by providing hormone, vitamin and other growth

promoting substances (Gyaneshwar et al.,` 2002).

The application of biofertilizers, micronutrients and RDF enhanced the plant

height appreciably at harvest stages.Increase in plant height might be attributed

to the fact that the better nourishment causes beneficial effects such as

accelerated rate of photosynthesis, assimilation, cell division and vegetative

growth. These results are in agreement with the findings of Singh et al., (2007).

Dhingra et al.(1988) results revealed that the interactions between phosphorus

and Rhizobium inoculation was significantly in 3 out of 5 years, indicating that

the combination of Rhizobium and 20 kg P2O5 /ha gave yield equivalent to 40

4

kg P2O5 /ha without Rhizobium. Gupta and Sharma (1992) reported from the

result of an experiment that yield of lentil 0.87 - 1.30 t/ha with 0 - 32 kg

phosphorus and no inoculation, and 0.89 - 1.68 t/ha with 0 – 32 kg phosphorus

and inoculation. Seeds protein content increased with application of

phosphorus and inoculation.

Rajput and Kushwah (2005) studied that the application of bio-fertilizer on

production of pea. On the basis of three years pooled data, the highest yield

was recorded with the application or recommended doses of fertilizer followed

by soil application of bio-fertilizer mixed 25 kg FYM along with 50%

recommended dose of fertilizer and were at par statistically. So the use of bio-

fertilizer saved 50% N, P (10 kg N, 25 kg P2O5). It also saved the financial

resource as well as FYM.

Sharma and Sharma (2004) determined the effects of P (0, 20 and 40 kg/ha),

potassium (0 or 20 kg/ha) and Rhizobium inoculation on the growth and yield

of lentil cv. L-4147. The mean number of branches, nodules and pods per plant;

100-seed weight and seed yield were highest with the application of 40 kg P/ha,

whereas mean plant height and plant stand row length were highest with the

application of 20 kg P/ha. Application of K resulted in the increase in number

of branches and pods per plant and seed yield, whereas inoculation with

Rhizobium increased the mean plant height; number of branches, nodules and

pods per plant, 100-seed weight and seed yield.

Hossain and Suman (2005) carried out an experiment to evaluate the effect of

Azotobacter, Rhizobium and different levels of urea N on growth, yield and N-

uptake of lentil. Among the treatments Azotobacter plus Rhizobium inoculation

had significant effect on nodule formation, plant height, number of seeds, seed

and stover yields, compared to uninoculated controls. The highest seed yield

was recorded for the treatment Azotobacter+Rhizobium that was statistically

similar to that of 100% N and Rhizobium with the corresponding yields of

1533 and 1458 kg/ha, respectively. The dual inoculation of Azotobacter and

5

Rhizobium significantly influenced all the crop characters including N

contents, N uptake by seed and shoot as well as protein content of seed. The

highest N-uptake by seed (78.61 kg/ha) was recorded for the treatment

Azotobacter+Rhizobium and N-uptake by shoot (53.87 kg/ha) was recorded for

the treatment 100% N. The performances of Azotobacter or Rhizobium alone

were not as good as Azotobacter+Rhizobium in most cases. Therefore,

inoculation of both Azotobacter and Rhizobium together may be a good

practice to achieve higher seed yield of lentil.

Kumar and Uppar (2007) conducted a field experiment to evaluate the effects

of organic manures, biofertilizers, micronutrients and plant growth regulators

on the seed yield and quality of mothbean. RDF + FYM @ 10 t/ha recorded the

highest values for the different seed yield and quality attributes of mothbean.

2.2. Nitrogen fixation and Rhizobium inoculation on lentil

Lentil is a legume and fulfils most of its N requirement through atmospheric

N2fixation with the symbiotic help of rhizobia living in its root nodules.

Generally, the level of N2 fixation in legumes depends on host genotypes,

rhizobial strains, environment and their interactions. Lentil cultivars have

shown genetic variability in their ability to symbiotically fix N2 (Rennie and

Dubetz, 1986), therefore genotypes with high N2 fixation and high seed yield

are desirable for sustainable agriculture. Kurdali et al. (1997) carried out a field

experiment to assess the source of nitrogen (N2 fixation, soil and fertilizer), N

assimilation, partitioning and mobilization in rainfed lentil at various growth

stages using 15N isotopic dilution.

Nitrogen for developing pods can be supplied from soil, atmospheric N2, and

from the mobilization of existing N in plant tissues. The relative importance of

these sources depends on several factors including plant species, genotype,

drought stress, plant and soil N status, and N2 fixation ability (Kurdali et al.,

6

1997). Grain legumes respond most strongly to inoculation when they are

introduced into new areas where soils lack appropriate rhizobia (van Kessel

and Hartley, 2000). There is presumably a yield advantage to crop inoculation

in soils with inadequate inorganic N supply. However, the yield response to

inoculation was highly variable and affected by inherent field variability, and

by differences in environmental and edaphic conditions (van Kessel and

Hartley, 2000).

Effective indigenous strains of Rhizobium leguminosarum biovar viceae are

lacking in most prairie soils, and therefore inoculation is essential to ensure

adequate nodulation and N fixation for maximum yields (Bremer et al., 1988).

When chickpea (Cicer arietinum) and lentil were introduced to North America,

both crops responded strongly to inoculation. In subsequent years, and as the

resident population of effective rhizobia in soils increased, N2 fixation

remained significant but responses to further inoculation diminished (Bremer et

al., 1989). Mengel (1994) concluded that nitrogenase activity is a flexible

process that adjusts to the N demand of the host. The amount of N2 fixed

becomes much more dependent on the demand of N by the host than on the

intrinsic capacity of the rhizobia to fix N.

2.3. Effect of nitrogen on nodulation, growth and yield

Regarding potassium fertilizers, Srinivasarao et al. (2003) concluded that

potassium application increase the pulses pest resistance and improve the seed

yield and quality; the status of potassium in soils depends on soil texture,

nutrient, and agricultural practices. In intensive cropping systems, considerable

amount of potassium is depleted that need to be addressed. Increasing of the

legumes yield and yield components (number of branches, pods and seeds) by

potassium fertilizer has been reported by numerous researchers.

Jahan et al. (2009) stated that the yield of lentil varieties increased when the

rates of potassium fertilization were increased in Bangladesh; the highest seed

7

yield (2.16 t. ha-1

) was found at 35 kg K ha-1

compared to zero, 15, 25, and 45

kgha-1

treatments. While the plant dry weight was not affected; they also stated

that the highest number of nodules plant-1

was obtained when potassium

fertilizer was applied at a rate of 15 kgha-1

compared to control or high levels

of potassium. Also, ElBramawy & Shaban (2010) noticed significant increases

for the most of growth and yield characters of broadbean crop by the

application of potassium fertilizer.

Like most annual legumes, lentil can provide a part of its own N requirement

through symbiotic N2 fixation when the plants are inoculated. Sosulski and

Buchan (1978) reported that rhizobial inoculation alone is not enough for

obtaining high yields of legumes because of poor nodulation and nitrogenase

activity. They concluded that annual legumes may require a high level of plant

N fertility to achieve maximum yield. Indigenous populations of Rhizobia for

legumes may be present in prairie soils, but these indigenous populations may

be ineffective for inducing N2 fixation under semiarid environments (Kucey

and Hynes, 1989). Small doses of N fertilizers applied to an annual pulse are

beneficial if nodule initiation is delayed (Mahon and Child, 1979). In dry pea,

N application at 20 to 60 kg ha–1

increased seed yield by an average of 9% in

one quarter of 58 trials conducted in Alberta (McKenzie et al., 2001). When

spring soil NO3-N (0 to 30 cm depth) was less than 20 kg N ha–1

, the use of

fertilizer N increased pea yield by an average of 11% in one-third of the trials.

Similarly, application of fertilizer N increased dry bean 8 (Phaseolus vulgaris

L.) seed yield proportionally in southern Manitoba (McAndrew and Mills,

2000). Most producers in Western Canada inoculate the seed or the soil with a

rhizobia strain and provide little or no fertilizer N to their lentil crops. Due to

the lag period between rhizobial root colonization infection and the onset of

nodule functioning, the young lentil plants may require a small dose of

additional N (i.e., starter- N) from external sources to achieve vigorous

vegetative growth and establish N2-fixing symbiosis.

8

2.4. Effect of phosphorus on nodulation, growth and yield

Phosphorus plays a major role in many plant processes, including storing and

transfer of energy; stimulation of root growth, flowering, fruiting and seed

formation; nodule development and N2 fixation (Mclaren and Cameron, 1996;

Ali et al., 1997). Phosphorus application on legumes can also increase leaf

area, yield of tops, roots and grain; nitrogen concentration in tops and grain;

number and weight of nodules on roots; and increased acetylene reduction rate

of the nodules (Jessop et al., 1989; Idris et al., 1989; Yahiya et al., 1995).

Research documents evident that the influence of P on nodule development and

N2 fixation by legumes (Israel, 1987). The N2-fixation process in legumes is

sensitive to P deficiency due to reduced nodule mass and decreased ureide

production (Sinclair and Vadez, 2002; Vance, 2001). Nodules are a strong P

sink and nodule P concentration normally exceeds that of roots and shoots (Sa

and Israel, 1991; Drevon and Hartwig, 1997). Therefore, nodule number,

volume, and dry weight can be increased by treating P deficient soils with

fertilizer P (Cassman et al., 1981). However, Bremer et al. (1989) found that P

application increased dry matter and grain yield but did not affect N2 fixation

indicating that the legume host was more responsive to P application than the

Rhizobia.

Saskatchewan soils generally test low to medium in available phosphorus

(Henry, 1980), a nutrient required in relatively large amounts by pulse crops.

Total P in Saskatchewan soils ranges from about 400 to 2200 kg ha-1

in the top

15 cm of soil, but only a very small amount of the total P is available to the

crop during a growing season (Saskatchewan Ministry of Agriculture, 2006).

Although crops can sometimes be grown for a few years without adding P

fertilizer, yields sooner or later begin to decline. Phosphorous is relatively

immobile (moves very little) in the soil. Most crops recover only 10 to 30% of

the P in fertilizer the first year following application (Havlin et al., 2005).

Recovery varies widely depending on soil type and conditions, the crop grown

9

and application method. However, Saskatchewan research has shown 9 that the

newly formed soil P reaction products are more plant available than the native

soil P minerals and crops can continue to recover fertilizer P for several years

after application (Saskatchewan Ministry of Agriculture, 2006).

Granular monoammonium phosphate (MAP) (12-51-0 or 11-55-0) is the most

common P fertilizer used in Saskatchewan (Saskatchewan Ministry of

Agriculture, 2006). Lentils are sensitive to high rates of P fertilizer placed

directly in the seed rows. Research conducted over a three year period

indicated that increasing rates of seed-placed MAP (11-55-0) resulted in

reduced stands of lentil but high yield per plant as compared to side-banded P

application (McVicar et al., 2010). Lentil has a relatively high requirement for

phosphorus to promote development of its extensive root systems and vigorous

seedlings; and may benefit from improved frost, disease, and drought tolerance

because of P application (McVicar et al., 2010). Bremer et al. (1989) reported

that P response is more prevalent in the Black soils, which had the most

favorable growing conditions and lowest available soil P levels, than in Brown

or Dark Brown soils of Saskatchewan.

10

CHAPTER 3

MATERIALS AND METHODS

The experiment was conducted at the Agronomy field, Sher-e-Bangla

Agricultural University, Dhaka-1207 during the period from November 2016

to March 2017. Detailed of the experimental materials and methods followed

in the study are presented in this chapter. The experiment was conducted to

study the influence of biofertilizer, nitrogen and phosphorous on nodulation,

growth and yield of lentil.

3.1 Site description

3.1.1 Geographical location

The experimental area was situated at 2377N latitude and 9033E longitude

at an altitude of 8.6 meter above the sea level (Anon., 2004).

3.1.2 Agro-ecological region

The experimental field belongs to the Agro-ecological zone of “The Modhupur

Tract”, AEZ-28 (Anon., 1988a). This was a region of complex relief and soils

developed over the Modhupur clay, where flood plain sediments buried the

dissected edges of the Modhupur Tract leaving small hillocks of red soils as

„islands‟ surrounded by floodplain (Anon., 1988b).

3.1.3 Climate

This area characterized by high temperature, high relative humidity and heavy

rainfall with occasional gusty winds in Kharif season (April-September) and

scanty rainfall associated with moderately low temperature during the Rabi

season (October-March) as it is sub-tropical climate,. Weather information

regarding temperature, relative humidity and rainfall prevailed at the

experimental site during the study period were presented in Appendix I.

11

3.1.4 Soil

Soil pH ranged from 5.6-6.5 and had organic matter 1.10-1.99%. The soil of

the experimental site belongs to the general soil type, Shallow Red Brown

Terrace Soils under Tejgaon Series. Top soils were clay loam in texture, olive-

gray with common fine to medium distinct dark yellowish brown mottles. The

experimental area was flat having available irrigation and drainage system and

above flood level. Soil samples from 0-15 cm depths were collected from

experimental field. The analyses were done by Soil Resource and Development

Institute (SRDI), Dhaka.

3.2 Details of the experiment

3.2.1 Treatments

The experiment consisted of 2 factors:

Factors A: Biofertilizer (2)

(a) B0 = 0 (Without Biofertilizer)

(b) B1 = Biofertilizer (50 gm per 2.5 kg of seeds)

Factors B: Rates of Nitrogen (N) + Phosphorus (P) (6)

(a) F0 = 0 (N + P)

(b) F1 = 50% Less recommended N + P

(c) F2 = 25% Less recommended N + P

(d) F3 = Recommended N + P (50 kg N + 100 kg P) / ha

(e) F4 = 25% Higher recommended N + P

(f) F5 = 50% Higher recommended N + P

The nitrogenous (N) and phosthatic (P) fertilizers were applied in the form of

urea and triple super phosphate (TSP). The rate of the urea and TSP, and other

fertilizers has been presented in section 3.4.5.

Treatment combination

B0F0, B0F1, B0F2, B0F3, B0F4, B0F5, B1F0, B1F1, B1F2, B1F3, B1F4, B1F5

12

3.2.2 Experimental design and layout

The experiment was laid out in a two factor RCBD (Factorial) with three

replications. There were 12 treatment combinations. The total numbers of unit

plots were 36. The size of unit plot was 3.0 x 2.0 m2. The distances between

plot to plot and replication to replication were 0.50 m and 1.0 m, respectively.

3.3 Crop/Planting Material

BARI Mosur-6 was used as plant material.

3.3.1 Description of crop: BARI Mosur-6

Lentil variety BARI Mosur-6 was used as experimental material. BARI Mosur-

6 was developed by Pulses Research Centre, Ishurdi, Pabna. BARI Mosur-6 is

a semi erect and medium saturated and bushy cultivar. The average plant height

of the variety is 35-40 cm. The leaves are dark green, with broad leaflets

without tendrils. Flowers are light blue, the pods and leaves turn straw color

during maturity stage. Seed color is deep brown and cotyledons are bright

orange. It has a 1000 seed weight of 19.84 g compared to 11.5 g or less for the

local cultivars. Seed size is larger than BARI Mosur-5. The duration of this

crop is 110-115 days. Its yield is 2200-2300 kg/ha. It is resistant to rust/STB

and tolerant to foot rot and moderately resistant to aphid.

3.4 Crop management

3.4.1 Seed collection

Seeds of BARI Mosur-6 were collected from Bangladesh Agricultural

Research Institute (BARI), Joydebpur, Gazipur, Bangladesh.

3.4.2 Seed sowing

Seeds were sowed in the field on 30 November, 2016. The field was labeled

properly and was divided into 36 plots. The seeds of BARI Mosur-6 were

sowed by hand in 30 cm apart from lines with continuous spacing at about 3

cm depth at the rate of 40 g plot-1

on 30 November, 2016.

13

3.4.3 Collection and preparation of initial soil sample

The soil sample of the experimental field was collected before fertilizer

application. The initial soil samples were collected before land preparation

from a 0-15 cm soil depth. The samples were collected by an auger from

different location covering the whole experimental plot and mixed thoroughly

to make a composite sample. After collection of soil samples, the plant roots,

leaves etc. were removed. Then the samples were air-dried and sieved through

a 10-mesh sieve and stored in a clean plastic container for physical and

chemical analysis.

3.4.4 Preparation of experimental land

A pre- sowing irrigation was given on November 22, 2016. After that the land

was open with the help of a tractor drawn disc harrow, then ploughed with

rotary plough twice followed by laddering to achieve a medium tilth required

for the crop under consideration. All weeds and other plant residues of previous

crop were removed from the field. Immediately after final land preparation, the

field layout was made on November 30, 2016 according to experimental

specification. Individual plots were cleaned and finally prepared the plot.

3.4.5 Fertilizer application

The recommended chemical fertilizer dose was 50, 100, 55 and 1 kg ha-1

of

Urea, TSP, MOP and Boric acid respectively. All the fertilizers according to

the treatment with half of urea were applied by broadcasting and was mixed

with soil thoroughly at the time of final land preparation after making plot. The

rest half of urea was applied on later stage as basal dose.

3.4.6 Intercultural operations

3.4.6.1 Thinning

The plots were thinned out on 15 days after sowing to maintain a uniform plant

stand which facilitates proper aeration and light for optimum growth and

development of the crops.

14

3.4.6.2 Weeding

The crop was infested with some weeds during the early stage of crop

establishment. Two hand weedings were done, first weeding was done at 15

days after sowing followed by second weeding at 15 days after first weeding.

3.4.6.3 Application of irrigation water

Irrigation water was added to each plot, first irrigation was done as pre- sowing

and other two irrigation were given 3 days before weeding.

3.4.6.4 Drainage

Drainage channel were properly prepared to easy and quick drained out of

excess water.

3.4.6.5 Plant protection measures

The crop was infested by insects and diseases, those were effectively and

timely controlled by applying recommended insecticides and fungicides.

Malathion 18 ml/L and Ripcord 20ml/L uses as protection measure.

3.4.7 Harvesting and post-harvest operation

Maturity of crop was determined when 80-90% of the pods become straw

color. The harvesting of BARI Mosur-6 were done up to 5 March, 2017. Five

pre-selected plants per plot were harvested from which different yield

attributing data were collected and 1 m2 area from middle portion of each plot

was separately harvested and bundled, properly tagged and then brought to the

threshing floor for recording grain and straw yield data. The grains were

cleaned and sun dried to a moisture content of 12%. Straw was also sun dried

properly. Finally grain and straw yields plot-1

were determined and converted

to kg ha-1

.

15

3.4.8 Recording of data

Emergence of plants were counted from starting to a constant number of plants

m-2

area of each plot. Experimental data were determined from 15 days of

growth duration and continued until harvest. Dry weights of plant were

collected by harvesting respective number of plants at different specific dates

from the inner rows leaving border rows and harvest area for grain. The

following data were recorded during the experimentation.

A. Crop growth characters

i. Plant height (at 30, 50, 70, 90 DAS and at harvest)

ii. Branch plant-1

(at 30, 50, 70 DAS and at harvest)

iii. Nodule count (at 50, 60, 70 and 80 DAS)

iv. Dry weight plant-1

(at 30, 50, 70 and 90 DAS)

B. Yield and other crop characters

i. Pods plant-1

(no.)

ii. Thousand seed weight (g)

iii. Grain yield (kg ha-1

)

iv. Stover yield (kg ha-1

)

v. Biological yield (kg ha-1

)

vi. Harvest index (%)

3.4.9 Detailed procedures of recording data

A brief outline of the data recording procedure followed during the study given

below:

16

A. Crop growth characters

3.4.9.1 Plant height

The height of 10 randomly selected plants from each plots for every treatments

of all three replication was taken carefully at harvest and after 30, 50, 70, 90

days of sowing of the seeds of BARI Mosur-6. Plant height was measured from

the above ground portion of the plants.

3.4.9.2 Branch plant-1

The branches plant-1

were counted carefully from 10 randomly selected plant

from each plot for every treatments of all three replications when it became 30,

50 and 70 days after sowing and at harvest.

3.4.9.3 Nodule count

The nodule plant-1

were counted carefully from 10 randomly selected plant

from each plot for every treatments of all three replications when it became 50,

60, 70, 80 days after sowing. Then it was averaged.

3.4.9.4 Dry weight of plant (kg ha-1

)

10 randomly selected plant from each plot were harvest after 30, 50, 70, 90

days of sowing. Then the plants were dried properly and individual plant

weight was taken to make them average for each treatment.

B. Yield and other crop characters

3.4.10.1 Pods plant-1

The pods of five preselected plants were collected from each plot at the time of

harvest and then counted total number and then averaged them to get pods

plant-1

.

3.4.10.2 Thousands seed weight (g)

Thousand seeds from of each plot were collected and their weight were taken

by digital electric balance in g.

17

3.4.10.3 Grain yield (kg ha-1

)

Grains of 1 m2 area in each plot was weighed and then converted into kg ha

-1.

The grain weight was taken at 12% moisture content of the grains.

3.4.10.4 Stover yield (kg ha-1

)

Stover of central 1 m2 area in each plot was sun dried and weighed. Then the

weight was converted in kg ha-1

.

3.4.10.5 Biological yield (kg ha-1

)

Biological yield was calculated by adding the grain yield and stover yield.

Biological yield = grain yield + Stover yield.

3.4.10.6 Harvest index

Harvest index denotes the ratio of economic yield (seed yield) to biological

yield and was calculated with following formula (Donald, 1963; Gardner et al.,

1985).

Harvest index (%) =𝑆𝑒𝑒𝑑 𝑦𝑖𝑒𝑙𝑑

𝐵𝑖𝑜𝑙𝑜𝑔𝑖𝑐𝑎𝑙 𝑦𝑖𝑒𝑙𝑑× 100

3.4.10.7 Statistical analysis

All the collected data were analyzed following the analysis of variance

(ANOVA) technique using a statistical computer software MSTAT-C program

and the means were adjusted by Statistics 10 at 0.05% level of significance.

18

CHAPTER IV

RESULTS AND DISCUSSION

This chapter represents the result and discussions of the present study.

Summary of mean square values at different parameters are also given in the

appendices from III to IX.

4.1 Effect of biofertilizer and nitrogen + phosphorous on growth of lentil

4.1.1 Plant height

4.1.1.1 Effect of biofertilizer

Plant height of BARI Mosur-6 exerted non-significant variation due to

biofertilizer application treatment (Figure 1). Although the variation due to

biofertilizer treatments was non-significant, numerically the values of plant

height showed an increasing trend with the advance of growth stages upto 90

DAS. It was also inferred from the table that biofertilizer treated plots showed

higher values of plant height than untreated (control) plots for all growth stages

except at 30 DAS.

B0 = 0 (Biofertilizer), B1 = Biofertilizer.

Figure 1. Effect of biofertilizer on plant height of lentil at different days

after sowing (LSD.05 = 0.79, 0.90, 1.41, 2.03, and 1.62 at

30, 50, 70, 90 DAS and at harvest respectively)

0

5

10

15

20

25

30

35

30 50 70 90 Harvest

Pla

nt

hei

ght

(cm

)

Days after sowing

B0 B1

19

4.1.1.2 Effect of nitrogen + phosphorous

Different levels of N+P fertilizer exhibited statistically significant variation on

plant height of lentil at all growth stages except 70, 90 DAS and at harvest and

the results of plant height have been presented in Figure 2 . The figure showed

that at 30 DAS the treatments F3 and F2 showed highest plant height (11.02 cm

and 10.96 cm, respectively). F5, F4 and F1 showed medium plant height (9.60

cm, 9.58 and 9.38 cm, respectively) in comparison to F3 and F2. Where F0

showed the lowest plant height (8.69 cm). This might be due to the different

level of N + P effect on plant height.

At 30 DAS F3 and F2 showed highest plant height (20.40 cm and 20.22 cm,

respectively). F4, F1 and F5 showed medium plant height at 50 DAS (19.05 cm,

18.75 cm and 18.36 cm) in comparison to F3 and F2. Where F0 showed the

lowest plant height (16.77 cm).

Although non-significant variation exhibited at 70 DAS, but numerically F2

and F3 showed highest plant height (26.95 cm and 26.55 cm, respectively). F4

and F1 showed medium plant height at 70 DAS (25.98 cm and 25.22 cm) in

comparison to F3 and F2. Where F0 showed the lowest plant height (23.34 cm).

F3 and F2 showed numerically the highest plant height at 90 DAS (31.39 cm

and 30.35 cm, respectively). F4, F5 and F1 showed medium plant height at 90

DAS (29.95 cm, 29.22 cm and 28.32 cm, respectively) in comparison to F3 and

F2. Where F0 showed the lowest plant height (27.18 cm).

At harvest plant height of F3 and F2 treatments were (30.71cm and 29.15cm,

respectively). F4, F5 and F1 showed medium plant height at harvest (28.05 cm,

27.79 cm and 27.76 cm, respectively) in comparison to F3 and F2. Where F0

showed the lowest plant height (26.60 cm).

4.1.1.3 Interaction effect of biofertilizer and nitrogen + phosphorous

Interaction of biofertilizer and levels of N+P fertilizer showed non-significant

effect on plant height for all sampling dates (30, 70, 90 DAS and at harvest)

except of 50 DAS lentil (Table 1). Numerically, B1F3 (Biofertilizer with

recommended dose of N+P) interaction showed the tallest plant at 50, 90 DAS

20

and at harvest (20.82, 32.06 and 31.45, respectively). The lowest plant height

was observed with B0F0 (without biofertilizer + without N+P) interaction for all

sampling dates 30, 50, 70, 90 DAS and at harvest (8.25, 15.96, 22.74, 26.01

25.58 cm, respectively).

F0= 0 (N+P); F1= 50% less N+P, F2= 25% less N+P; F3= Recommended N+P, F4= 25%

higher N+P, F5= 50% higher N+P.

Figure 2. Effect of different level (N+P) fertilizers on plant height of lentil

at different days after sowing (LSD.05 = 2.06, 2.35, 3.67, 5.28 and

4.16 at 30, 50, 70, 90 DAS and at harvest, respectively)

4.1.2 Branches plant-1


Biofertilizer application showed non-significant variation on number of

branches plant-1

of lentil for all sampling dates (Figure 3). Figure shows that

the values of number of branches plant-1

increased gradually with the advances

of growth stages upto 90 DAS, but it reduced slightly at harvest. However, the

highest branches plant-1

was found at 90 DAS for both biofertilizer treated or

untreated plants.

0

5

10

15

20

25

30

35

30 50 70 90 Harvest

Pla

nt

hei

gh

t (c

m)

F0 F1 F2 F3 F4 F5

Days after sowing

21

Table 1. Interaction effect of biofertilizer and different level of N + P

fertilizer on plant height of lentil at different days after sowing

B0 = 0 (Biofertilizer), B1 = Biofertilizer, F0= 0 (N+P); F1= 50% less N+P, F2= 25% less N+P;

F3= Recommended N+P, F4= 25% higher N+P, F5= 50% higher N+P, NS = non-significant.


Figure 3. Effect of biofertilizer on branch plant-1

of lentil at different days

after sowing (LSD.05 = 0.17, 0.67, 0.37, 0.36 and 0.95 at 30, 50, 70,

90 DAS and at harvest, respectively)

0

2

4

6

8

10

12

30 50 70 90 Harvest

Bra

nch

/pla

nt

B0 B1

Days after sowing

Treatments Plant height (cm)

30 DAS 50 DAS 70 DAS 90 DAS At harvest

B0F0 8.25 15.96 b 22.74 26.01 25.58

B0F1 9.00 17.92 ab 23.90 26.74 26.96

B0F2 11.42 19.92 a 26.31 29.85 28.80

B0F3 11.17 19.98 a 26.01 30.72 29.98

B0F4 9.15 19.09 ab 25.74 29.28 26.41

B0F5 9.72 17.96 ab 23.41 27.92 28.33

B1F0 9.14 17.58 ab 23.94 28.34 27.61

B1F1 9.75 19.58 ab 26.54 29.90 28.56

B1F2 10.51 20.50 a 27.58 30.86 29.50

B1F3 10.87 20.82 a 27.09 32.06 31.45

B1F4 10.01 19.02 ab 26.22 30.61 29.68

B1F5 9.48 18.76 ab 25.35 30.52 27.26

LSD.05 NS 3.88 NS NS NS

CV (%) 11.23 6.51 7.37 7.97 9.15

22


No. of Branch plant-1

showed significant response due to N+P fertilizer level

treatment and the data has been presented in Figure 4. The Figure indicated that

irrespective of sampling dates higher doses of fertilizer increased the higher

trend of number of branches plant-1

in lentil and the highest value was recorded

into F3 fertilizer treatment. A further increase of fertilizer dose reduced the No.

of branches plant-1

marginally. On the other hand branches plant-1

increased

steadily with the increase of growth stages and the highest increase was found

in 90 DAS irrespective fertilizer doses. Branchs plant-1

showed different

response on different nitrogen + phosphorous levels.



Figure 4. Effect of different level (N+P) fertilizers on No. of branch plant-1

of lentil at different days after sowing (LSD.05 = 0.44, 1.75, 0.97,

0.96 and 0.95 at 30, 50, 70, 90 DAS and at harvest,

respectively)


Interaction of biofertilizer and N+P fertilizer level showed significant effect on

branches plant-1

of lentil at 90 DAS and at harvest sampling dates (Table 2). At

90 DAS maximum number and statistically similar branches plant-1

(8.80) was

0

2

4

6

8

10

12

14

30 50 70 90 Harvest

Bra

nch

/pla

nt

F0 F1 F2 F3 F4 F5

Days after sowing

23

found in the treatments of B1F3, B1F4 and B0F3 which was statistically similar

with the interaction of B1F2, B0F2, B1F5 and B0F1. Significantly lowest number

of branches plant-1

was found in the interaction of B0F5 (6.06) which was

statistically similar with the B1F1, B1F0, B0F4, B0F1 and B0F2 interactions. On

the other hand, highest branches plant-1

(13.20) was found with B0F3 interaction

and lowest was recorded with B0F0 interaction.


fertilizer on branch plant-1

of lentil at different days after sowing



4.1.3 Nodule plant-1


Biofertilizer treatment showed significant effect on nodules plant-1

of lentil at

70 DAS but it showed non-significant effect at 50, 60 and 80 DAS(Figure 8).

Irrespective of biofertilizer treatment nodule plant-1

increased stedily with the

advancement of growth stages upto 70 DAS after that the values of nodule

plant-1

reduced slightly. At 70 DAS the highest nodule plant-1

(30.67) was

Treatments No. of Branch plant-1

30 DAS 50 DAS 70 DAS 90 DAS At harvest

B0F0 1.00 4.80 5.46 6.73 b-d 9.00 b

B0F1 1.33 5.80 5.40 7.46 a-d 9.52 ab

B0F2 1.33 5.80 5.60 7.93 ab 9.60 ab

B0F3 2.66 6.83 6.46 8.80 a 13.20 a

B0F4 1.33 5.00 6.06 7.13 b-d 11.80 ab

B0F5 1.00 5.20 5.80 6.06 d 10.06 ab

B1F0 1.00 5.33 5.66 6.33 cd 8.93 b

B1F1 1.33 5.40 5.40 6.73 b-d 9.73 de

B1F2 1.50 4.80 6.53 8.06 ab 10.73 ab

B1F3 1.66 6.20 6.86 8.80 a 12.26 ab

B1F4 1.33 5.73 5.80 8.80 a 10.60 ab

B1F5 1.00 5.60 5.53 7.64 a-c 9.53 ab

LSD.05 NS NS NS 1.57 4.08

CV (%) 17.69 17.23 9.25 12.66 9.52

24

observed with biofertilizer treated plot and the lowest (27.42) was found with

control treatment B0. The figure clearly medicated that biofertilizer appplicatin

increased nodule plant-1

than without biofertilizer (control) irrespective growth

stages.


Figure 5. Effect of biofertilizer on nodule plant-1

of lentil at different days

after sowing (LSD 0.05 = 0.58, 1.52, 1.28, and 1.32

at 50, 60, 70 and 80 DAS, respectively)


Nodules plant-1

had significant effect due to different levels of N+P fertilizer

application in lentil. The values of nodule plant-1

presented in Figure 6.The

figure showed that irrespective sampling dates nodule number increased

gradually upto F3 treatment after that it reduced marginally. On the other had

irrespective of fertilizer treatment, nodules plant-1

increased upto 70 DAS after

that it reduced slightly. The highest increase was found at 70 DAS. Result also

indicated that F3 showed the highest (31.48) nodules plant-1

which was

followed by F2 and F4 treatment.

0

5

10

15

20

25

30

35

50 60 70 80

No

du

les/

pla

nt

B0 B1

Days after sowing

25



Figure 6. Effect of different level (N+P) fertilizers on nodule plant-1

of

lentil at different days after sowing (LSD.05 = 1.52, 3.96, 3.33, and

3.45 at 50, 60, 70 and 80 DAS, respectively)


Interaction effect of biofertilizer and N + P showed significant result on no. of

nodules plant-1

of lentil for all sampling dates i.e. 50, 60, 70 and 80 DAS (Table

3). At 50 DAS the highest value (17.86) was found with B1F3 and the lowest

value (13.65) with B0F0.

At 60 DAS, the interaction of B0F3 showed highest result (23.97) and B1F0

showed lowest result (15.88).

At 70 DAS interaction effect of B1F3 showed highest result (33.14) which was

close to B1F2 interaction (31.44) and B0F0 showed lowest result (25.53).

Interaction effect of biofertilizer B1F3 showed highest result (29.66) and B0F0

showed lowest result (19.66).

0

5

10

15

20

25

30

35

50 60 70 80

No

du

les/

pla

nt

F0 F1 F2 F3 F4 F5

Days after sowing

26


fertilizer on nodule plant-1

of lentil at different days after sowing


F3= Recommended N+P, F4= 25% higher N+P, F5= 50% higher N+P.

4.1.4 Dry weight plant-1


Dry weight plant-1

of lentil due to biofertilizer used treatment have been

presented in Figure 7. The figure shows that irrespective biofertilizer treated

and untreated treatments dry weight plant-1

increased steadily with advance of

growth stages and it continued upto 90 DAS. It was also observed that at 70

and 90 DAS dry weight plant-1

differed significantly among the biofertilizer

treated and untreated treatments. This might be due to the response of

biofertilizer on plant height. Phosphate-solubilizing bacteria (PSB) alone

recorded significantly more number and dry weight of nodules only with seed

inoculation method at 60 and 90 DAS and was at par with the uninoculated

control by producing 19.7 and 6.7% more grain yield and 13.6 and 5.9% more

straw yield with seed and soil inoculation methods (Karmakar et al, 2006)

Treatments Nodule number/plant

50 DAS 60 DAS 70 DAS 80 DAS

B0F0 13.65 c 16.77 b 25.53 ac 19.66 e

B0F1 14.63 c 20.55 ab 27.11 bc 26.00 a-d

B0F2 17.86 a 23.66 a 27.72 a-c 27.66 a-c

B0F3 17.86 a 23.97 a 29.77 a-c 28.66 ab

B0F4 15.20 bc 18.88 ab 27.43 bc 23.33 a-c

B0F5 14.73 bc 15.88 b 26.99 bc 22.66 c-e

B1F0 14.44 c 15.88 b 27.76 a-c 20.66 de

B1F1 15.53 a-c 19.88 ab 29.32 a-c 26.67 cd

B1F2 17.20 ab 22.22 ab 31.44 ab 27.33 a-c

B1F3 17.86 a 23.66 a 33.14 a 29.66 a

B1F4 15.66 a-c 21.87 ab 31.25 ab 27.33 a-c

B1F5 14.76 bc 21.99 ab 30.88 a-c 27.00 a-c

LSD0.05 2.50 6.53 5.05 5.70

CV (%) 5.21 12.38 5.95 7.85

27


Figure 07. Effect of biofertilizer on dry weight of lentil at different days

after sowing (LSD.05 = 0.15, 0.11, 0.58, and 1.17 at 30, 50, 70

and 90 DAS respectively)


Dry weight plant-1

gave significant variation due to N+P level of fertilization

for all the sampling dates except 30 DAS in lentil (Figure 8). The result

persecuted in figure 8 indicated that irrespective fertilizer (N+P) levels dry

weight plant increased progressively with advances of growth stages and the

highest increase was recorded at 90 DAS. Among the fertilizer levels dry

weight showed increasing trend up F3. However, the lowest dry weight plant-1

was observed with F0 fertilization level for all sampling dates.


Interaction effect of biofertilizer and N + P showed significant variation on dry

weight plant-1

for all sampling dates except 30 DAS(Table 4). The interaction

of B1F3 showed highest dry weight at 30, 50, 70 and 90 DAS (1.11, 1.97, 9.07

and 14.48 g, respectively). Interaction of B0F3 showed statistically similar dry

weight with the same sampling dates (50, 70 and 90 DAS). However, the

lowest dry weight value was found with B0F0 interaction treatment for 30, 50,

70 and 90 DAS (0.78, 1.02, 4.33 and 8.74 g, respectively).

0

2

4

6

8

10

12

14

30 50 70 90

Dry

wei

gh

t o

f p

lan

t (g

)

B0 B1

Days after sowing

28



Figure 08. Effect of different level (N+P) fertilizers on dry weight of lentil

at different days after sowing (LSD.05 = 0.41, 0.28, 1.51,

and 3.05 at 30, 50, 70 and 90 DAS, respectively)


fertilizer on dry weight of lentil at different days after sowing



0

2

4

6

8

10

12

14

16

18

30 50 70 90

Dry

wei

gg

ht

of

pla

nt

(g)

F0 F1 F2 F3 F4 F5

Days after sowing

Treatments Dry weight (g)

30 DAS 50 DAS 70 DAS 90 DAS

B0F0 0.78 1.02 d 4.33 bc 8.74 b

B0F1 0.80 1.11 d 4.85 bc 9.63 b

B0F2 0.86 1.36 cd 5.32 bc 10.78 b

B0F3 0.73 1.67 a-c 6.66 ab 13.08 ab

B0F4 1.00 1.42 b-d 5.60 bc 10.64 b

B0F5 0.73 1.01 d 4.10 c 8.99 b

B1F0 0.81 1.06 d 5.25 bc 10.70 b

B1F1 0.90 1.17 d 6.19 bc 11.44 b

B1F2 0.90 1.47 b-d 6.61 ab 12.42 b

B1F3 1.11 1.97 a 9.07 a 17.48 a

B1F4 1.08 1.84 ab 6.69 ab 11.84 b

B1F5 0.71 1.05 d 6.52 bc 10.13 b

LSD.05 NS 0.47 2.50 5.03

CV (%) 25.39 12.41 14.4 14.65

29

4.2 Effect of biofertilizer and nitrogen + phosphorous on yield compponets

and other yield characters

4.2.1 Pods plant-1


BARI Mosur-6 had a significant effect on number of pods plant-1

at harvest for

the application of Biofertilizer. BARI Mosur-6 showed higher number of pods

plant-1

(58.59) with biofertilizer (B1) and B0 (without biofertilizer) showed the

lower number of pods plant-1

(55.84) (Figure 9). This might be due to the

response of biofertilizer on plant height.


Figure 9. Effect of biofertilizer on pods plant-1

of lentil (LSD.05 = 1.60)


Number of pods plant-1

showed different response on different nitrogen +

phosphorous levels. All six combinations of N + P levels are significant where

F3 showed highest number of pods plant-1

(66.16). F2 and F4 showed medium

number of pods plant-1

(59.95 and 59.36, respectively) in comparison to F3.

54

54.5

55

55.5

56

56.5

57

57.5

58

58.5

59

B0 B1

Pod

s/p

lan

t

Treatment

30

Where F0 showed the lowest number of pods plant-1

(45.20). This might be due

to the different level of N + P effect on number of pods plant-1

(Figure 10).



Figure 10. Effect of different levels of N + P on pods plant-1

of lentil (LSD.05

= 4.18)


Interaction effect of biofertilizer and N + P showed significant result. Where

B1F3 showed highest result (68.40) and B0F0 showed lowest result (41.33).

(Table 5). Sharma and Sharma (2004) determined the effects of P (0, 20 and 40

kg/ha), potassium (0 or 20 kg/ha) and Rhizobium inoculation on the growth

and yield of lentil cv. L-4147. The mean number of branches, nodules and pods

per plant; 1000 seed weight and seed yield were highest with the application of

40 kg P/ha, whereas mean plant height and plant stand row length were highest

with the application of 20 kg P/ha.

0

10

20

30

40

50

60

70P

od

s/p

lan

t

F0 F1 F2 F3 F4 F5

Fertilizer level

31

Table 5. Interaction effect of biofertilizer and N + P on number of pods

plant-1

and thousand seed weight of lentil

Treatments Pods plant-1

(no.) Thousand seed weight(g)

B0F0 41.33 e 19.11 d

B0F1 56.06 c 19.52 c

B0F2 58.96 bc 19.54 c

B0F3 63.93 ab 20.49 b

B0F4 58.06 bc 19.57 c

B0F5 56.66 c 19.56 c

B1F0 49.06 d 19.14 d

B1F1 56.40 c 19.53 c

B1F2 60.93 bc 19.57 c

B1F3 68.40 a 20.96 a

B1F4 60.66 bc 19.58 c

B1F5 56.09 c 19.56 c

LSD.05 6.90 0.256

CV (%) 4.60 0.64


F3= Recommended N+P, F4= 25% higher N+P, F5= 50% higher N+P.

4.2.2 1000 seed weight (g)


Biofertilizer treatment showed significant variation on thousand seed of

lentil(Figure 11). Biofertilizer treatment showed higher 1000 seed weight over

control. Biofertilizer treated plot gave higher 1000 seed weight (19.72 g) and

untreated plot gave lower 1000 seed weight (19.63 g).


Thousand seed weight of lentil exerted significant variation due N + P

fertilization at different levels (Figure 12). The figure indicated that the trend of

seed weight increased sharply up to F3 treatment, a further increase of fertilizer

(N+P) dose reduced the 1000 seed weight marginally. Howevr, the lowest

weight was found with F0 treatment and that of highest with F3 treatment.

Phosphorus application on legumes can also increase leaf area, yield of tops,

32

roots and grain; nitrogen concentration in tops and grain; number and weight of

nodules on roots; and increased acetylene reduction rate of the nodules (Jessop

et al., 1989; Idris et al., 1989; Yahiya et al., 1995).

B0 = 0 (Biofertilizer), B1 = Biofertilizer

Figure 11. Effect of biofertilizer on thousand seed weight of lentil (LSD.05 =

0.059)



Figure 12. Effect of different level of N + P on thousand seed weight of

lentil (LSD0.5 = 0.15)

19.58

19.6

19.62

19.64

19.66

19.68

19.7

19.72

19.74

B0 B1

Th

ou

san

d s

eed

wei

gh

t (g

)

Treatmnet

18

18.5

19

19.5

20

20.5

21

Th

ou

san

d s

eed

wei

gh

t (g

)

F0 F1 F2 F3 F4 F5

Fertilizer level

33



B1F3 showed highest result (20.96 gm) and B0F0 showed lowest result (19.11

gm). (Table 5).

4.2.3 Grain yield (kg ha-1

)


Biofertilizer had a non-significant impact on grain yield of BARI Mosur-6.

BARI Mosur-6 showed higher grain yield (1971.10 kg ha-1

) in with of B1 and

B0 showed the lower grain yield (1859.40 kg ha-1

) (Figure 13). Effective

indigenous strains of Rhizobium leguminosarum biovar viceae are lacking in

most prairie soils, and therefore, inoculation is essential to ensure adequate

nodulation and N fixation for maximum yields (Bremer et al., 1988).


Figure 13. Effect of biofertilizer on grain yield of lentil (LSD.05 = 21.35)


Grain yield showed different response on different nitrogen + phosphorous

levels. All six combinations of N + P levels are significant where F3 and F2

34

showed highest grain yield (2534.00 kg ha-1

and 2309.40 kg ha-1

) respectively.

F1 and F4 showed medium Grain yield (1872.90 kg ha-1

and 1751.60 kg ha-1

) in

comparison to F3 and F2. Where F0 showed the lowest grain yield (1442.90 kg

ha-1

). This might be due to the different level of N + P effect Grain yield

(Figure 14). Phosphorus application on legumes can also increase leaf area,

yield of tops, roots and grain; nitrogen concentration in tops and grain; number

and weight of nodules on roots; and increased acetylene reduction rate of the

nodules (Jessop et al., 1989; Idris et al., 1989; Yahiya et al., 1995).



Figure 14. Effect of different level of N + P on grain yield of lentil (LSD.05 =

55.54 )



B1F3 showed highest result (2505.70 kg ha-1

) and B0F0 showed lowest result

(1385.70 kg ha-1

). (Table 6 ).

35

Table 6. Interaction effect of biofertilizer and N + P on grain yield, stover

yield, biological yield and harvest index of lentil

Treatments Grain yield

(kg ha-1

)

Stover yield (kg

ha-1

)

Biological yield

(kg)

Harvest index

(%)

B0F0 1385.70 d 1211.50 ae 2597.00 d 53.29

B0F1 1825.40 a-d 1595.90 b-e 3420.20 a-d 53.23

B0F2 2193.60 a-d 2031.80 a-d 4225.50 a-d 53.51

B0F3 2505.70 ab 2259.40 ab 4765.00 ab 53.57

B0F4 1638.30 b-d 1320.00 de 2958.10 d 53.32

B0F5 1607.80 b-d 1443.10 c-e 3050.10 cd 52.57

B1F0 1500.10 d 1446.00 c-e 2945.80 d 53.31

B1F1 1920.30 a-d 1764.40 a-e 3685.00 a-d 53.43

B1F2 2425.10 a-c 2230.60 a-c 4656.00 a-c 53.59

B1F3 2562.40 a 2396.80 a 4959.20 a 54.00

B1F4 1864.90 a-d 1746.80 a-e 3611.70 a-d 53.00

B1F5 1553.80 cd 1425.20 de 3187.40 b-d 52.65

LSD.05 91.68 79.76 167.30 NS

CV (%) 14.85 14.68 14.11 1.93



4.2.4 Stover yield (kg ha-1

)


Biofertilizer had a significant impact on stover yield of BARI Mosur-6. B1

treatment showed higher stover yield (1835.00 kg ha-1

) B0 showed the lower

stover yield (1643.60 kg ha-1

) (Figure 15). The presence of efficient and

specific strains of Rhizobium in the rhizosphere is one of the most important

requirements for proper establishment and growth of grain legume plant

(Gyaneshwar et al., 2002).


Stover yield showed different response on different nitrogen + phosphorous

levels. Treatment F3 and F2 showed highest stover yield (2328.10 kg ha-1

and

2131.30 kg ha-1

, respectively). F1 and F4 showed medium stover yield (1680.20

36

kg ha-1

and 1533.40 kg ha-1

, respectively) in comparison to F3 and F2. Where F0

showed the lowest stover yield (1328.70 kg ha-1

). This might be due to the

different level of N + P effect grain yield (Figure 16).


Figure 15. Effect of biofertilizer on stover yield of lentil (LSD.05 = 18.58)



Figure 16. Effect of different level of N + P on stover yield of lentil (LSD.05

= 48.31)

37



B1F3 showed highest result (2396.80 kg ha-1

) and B0F0 showed lowest result

(1311.50 kg ha-1

). (Table 6 ).

4.2.5 Biological yield (kg ha-1

)


Biofertilizer had a non-significant effect on biological yield of BARI Mosur-6.

Treatment of B1 showed higher biological yield (3840.80 kg ha-1

) and B0

showed the lower biological yield (3502.70 kg ha-1

) (Figure 17). The dual

inoculation of Azotobacter and Rhizobium significantly influenced all the crop

characters including N contents, N uptake by seed and shoot as well as protein

content of seed (Hossain and Suman, 2005)


Figure 17. Effect of biofertilizer on biological yield of lentil (LSD.05 =

38.97)


Biological yield showed different response on different nitrogen + phosphorous

levels and the values of biological yield have been presented in fiugure18. It

can be inferred from the figure that biological yield showed an increasing trend

38

with the increased fertilizers and it continued up to F3 treatment. After that a

further increase in fertilizer dose reduced the biological yield slightly.

However, F3 showed highest biological yield (4862.10 kg ha-1

). F2 showed

medium biological yield (4440.80 kg ha-1

) in comparison to F3. Where F0

showed the lowest biological yield (2771.40 kg ha-1

). This might be due to the

different level of N + P effect on biological yield.



Figure 18. Effect of different level of N + P on biological yield of lentil

(LSD.05 = 101.35)


Interaction effect of biofertilizer and N + P showed non-significant result.

Where B1F3 showed highest result (4959.20 kg ha-1

) and B0F0 showed lowest

result (2597.00 kg ha-1

) (Table 6).

4.2.6 Harvest index (%)


Harvest index had a non-significant impact on biological yield of lentil(Figure

19). The treatment B1 showed higher harvest index (53.33 %) and B0 showed

39

the lower harvest index (53.25 %). This might be due to the response of

biofertilizer on plant height.


Figure 19. Effect of biofertilizer on harvest index of lentil (LSD.05 = 0.78)


Higher harvest showed different response on different nitrogen + phosphorous

levels. F3 and F2 showed highest harvest index (53.78% and 53.55%,

respectively). F1 and F4 showed medium harvest index (53.33% and 53.16%,

respectively) (Figure 20) in comparison to F3. Where F5 showed the lowest -

harvest index (52.61%).


Interaction effect of biofertilizer and N + P showed non-significant result.

Where B1F3 showed highest result (54.00%) and B0F5 showed lowest result

(52.57%). (Table 6).

53.2

53.22

53.24

53.26

53.28

53.3

53.32

53.34

B0 B1

Ha

rves

t in

dex

(%)

Treatment

40



Figure 20. Effect of different level of N + P on harvest index of lentil

(LSD.05 = 2.03)

52

52.2

52.4

52.6

52.8

53

53.2

53.4

53.6

53.8

54

Ha

rves

t in

dex

(%

)

F0 F1 F2 F3 F4 F5

Fertilizer level

41

CHAPTER V

SUMMARY AND CONCLUSION

The experiment was conducted at the Agronomy field, Sher-e-Bangla

Agricultural University, Dhaka-1207 during the period from November 2016

to March 2017 to examine the influence of Biofertilizer, Nitrogen and

Phosphorous on nodulation, growth and yield of lentil. In this experiment, the

treatment consisted of two biofertilizer levels: (i) control and (ii) Biofertilizer

(Rhizobium) with six combinations of nitrogenous and phosphetic fertilizer: (i)

No nitrogen + phosphorous fertilizer (control), (ii) 50% less of recommended

N + P, (iii) 25% less of recommended N + P, (iv) recommended N + P, (v)

25% higher of recommended N + P and (vi) 50% higher of recommended N +

P. The experiment was conducted in two factor Randomized Complete Block

Design (RCBD) with three replications. Data on different growth and yield

parameters like plant height, no of branch plant-1

, nodule count, dry weight

plant-1

, pods plant-1

,thousand seed weight, grain yield, stover yield, biological

yield etc. were recorded. The collected data were statistically analyzed for

assessment of the treatment effect. A significant variation among the

treatments was found while Biofertilizer application and different levels of

N+P fertilizers were applied in different combinations.

Plant height (cm), branch plant-1

showed the best result in case of biofertilizer

application. Whereas biofertilizer combined with F3 and F2 showed the best

result (36.40 cm and 35.19 cm) for plant height at harvest. Biofertilizer

combined with F3 and F2 showed the best result (14.46 and 12.76) for branch

plant-1

at harvest. In case of plant height and branch plant-1

B0F0 gave the

lowest result.

Nodule number showed always better result for biofertilizer application. In

case of biofertilizer application with nitrogen + phosphorous (B1F3) showed the

highest nodule number (33.14) and B0F0 showed the lowest result (25.53).

42

Pod number was higher values for application of biofertilizer. In case of

biofertilizer application with nitrogen + phosphorous (B1F3) showed highest

result (68.40) and B0F0 showed the lowest result (41.33).

Grain yield and stover yield was higher for biofertilizer application.

Biofertilizer in combination with nitrogen + phosphorous, in interaction of

B1F3 and B1F2 showed the highest grain and stover yield. B0F0 showed lowest

result for both of the parameters.

Dry weight of plant, biological yield and harvest index showed higher result for

biofertilizer application. For these parameters B1F3 showed highest result and

B0F0 showed the lowest result.

From the experimental results it revealed that biofertilizer application was

better for nodulation, growth, yield contributing parameters and yield. For most

of the cases F3 showed highest result. On the other hand F2 and F4 showed

medium result in most of the cases. F0 showed lowest result always.

Furthermore, probably the dry matter produced by the biofertilizer application

with N + P contributed to the vegetative growth and was enough to be

partitioned into yield components. Therefore, it can be concluded that the

application of biofertilizer and N + P combination had a positive impact on

lentil (BARI Mosur-6).

Considering the above mentioned result revealed that biofertilizer treated plot

B1 (Biofertilizer) was found superior in producing maximum plant height,

branches plant-1


, nodule plant-1

, pods plant-1

, seed yield and

biological yield of lentil. On the other hand, N+P fertilizer at recommended

dose (F3) gave highest yield, plant height, branches plant-1


,

nodules plant-1

, pods plant-1

, 1000 seed weight (20.73 g), seed yield, stover

yield, and biological yield . In case of interaction, B1F3 was found superior in

producing maximum yield and yield components like pods plant-1

(68.40),

1000 seed weight (20.98 g ), seed yield (2562.40 kg ha-1

), stover yield

(2396.80 kg ha-1

), biological yield (4959.20 kg ha-1

).Therefore, the present

experimental results suggest that lentil yield is improved with Biofertilizer

under recommended dose of nitrogen and phosphorus.

43

Considering the situation of the present experiment, further studies in the

following areas may be recommended:

1. Such study is needed in different agro-ecological zones (AEZ) of

Bangladesh for analogy the accurateness of the experiment.

2. It is needed to have a specific conclusion this experiment may be under

taken by taking more biofertilizer and different concentration of N+P

treatment which can regulate the yield and seed quality of different

varieties of lentil.

44

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49

APPENDICES

Appendix I. Monthly record of air temperature, relative humidity and rainfall of

the experimental site during the period of November, 2016 to

March 2017

Month Air temperature (0C) Relative

humidity

(%)

Total

rainfall(

mm)

Sunshine

(hr)

Maximum Minimum

November, 2016 29.6 19.2 77 34.4 5.7

December, 2016 26.4 14.1 69 12.8 5.5

January, 2017 25.4 12.7 68 7.7 5.6

February, 2017 28.1 15.5 68 28.9 5.5

March, 2017 32.5 20.4 64 65.8 5.2

Source: SAU mini weather station, Sher-e-Bangla Agricultural University, Dhaka-

1207, Bangladesh

50

Plate 1. Field view of seedling stage

51

Plate 2.Data recording for nodule

department of agronomy sher-e-bangla agricultural

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