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“STUDIES ON EFFECT OF POTASSIUM AND ZINC

SOLUBILIZING MICROORGANISM ON MUNGBEAN’’

By

RAHUL INDAR NAVSARE

B.Sc. (Agri.)

DEPARTMENT OF SOIL SCIENCE AND AGRIL.CHEMISTRY

COLLEGE OF AGRICULTURE, BADNAPUR

VASANTRAO NAIK MARATHWADA KRISHI VIDYAPEETH,

PARBHANI 431 402 (M.S.) INDIA

2017

“STUDIES ON EFFECT OF “POTASSIUM AND ZINC

SOLUBILIZING MICROORGANISM ON MUNGBEAN”

By

MR. RAHUL INDAR NAVSARE

B.Sc. (Agri.)

DISSERTATION

Submitted to the

Vasantrao Naik Marathwada Krishi Vidyapeeth, Parbhani

In partial fulfillment of the

requirement for the Degree of

MASTER OF SCIENCE

(Agriculture)

In

SOIL SCIENCE AND AGRICULTURAL CHEMISTRY

DEPARTMENT OF SOIL SCIENCE AND AGRICULTURAL

CHEMISTRY


VASANTRAO NAIK MARATHWADA KRISHI VIDYAPEETH,

PARBHANI – 431 402 (M.S.), INDIA

2017

Dr.S.S.Mane

Msc.(Agri) Ph.D.

Professor,(CAS)

Section of Soil Science and Agril Chemistry,

College Of Agriculture,

Badnapur-431202(M.S.)

CCEERRTTIIFFIICCAATTEE -- II

This is to certify that the dissertation entitled “STUDIES ON EFFECT OF

POTASSIUM AND ZINC SOLUBILIZING MICROORGANISM ON MUNGBEAN"

submitted by Mr. NAVSARE RAHUL INDAR to the Vasantrao Naik Marathwada

Krishi Vidyapeeth, Parbhani in partial fulfillment of the requirement for the degree of

MASTER OF SCIENCE in the subject of SOIL SCIENCE AND AGRIL. CHEMISTRY

is record of original and bonafide research work carried out by her under my guidance and

supervision. It is of sufficiently high standard to warrant its presentation for the award of the

said degree. I also certify that the dissertation or part thereof has not been previously

submitted for a degree of any university. The assistance and help rendered during the course

of investigation and sources of literature have been duly acknowledged.

BADNAPUR (Dr.S.S.Mane)

Date: / /2017 Research Guide & Chairman

Advisory Committee

CCEERRTTIIFFIICCAATTEE –– IIII

This is to certify that the dissertation entitled “STUDIES ON EFFECT OF

POTASSIUM AND ZINC SOLUBILIZING MICROORGANISM ON MUNGBEAN"

submitted by Mr. Mr. NAVSARE RAHUL INDAR to the Vasantrao Naik Marathwada

Krishi Vidyapeeth, Parbhani in partial fulfillment of the requirement for the degree of

MMAASSTTEERR OOFF SSCCIIEENNCCEE((AAggrriiccuullttuurree))in the subject of SSOOIILL SSCCIIEENNCCEE AANNDD AAGGRRIILL..

CCHHEEMMIISSTTRRYY has been approved by the student's advisory committee after viva-voce

examination in collaboration with the external examiner.

( ) ( S.S.Mane )

External Examiner Research Guide & Chairman

Advisory Committee

Advisory committee: ( V.D Patil)

(G.R.Hanwate)

(Syed Ismail)

(S.J.Supekar)

Associate Dean (P.G.) Associate Dean and Principal College of Agriculture College of Agriculture

VNMKV, Parbhani - 431 402 (M.S.). Badnapur.

CCAANNDDIIDDAATTEE’’SS DDEECCLLAARRAATTIIOONN

I hereby declare that this dissertation or a

part there

of has not been previously

submitted by me for a degree,

diploma or distinction of

any University or

Institution.

Place :Badnapur (Navasare R.I.)

Date: / / 2017

Abbrievation

% per cent

/ per

C.D. Critical difference

cc Cubic centimeter

cm centimeter

Cfu Colony forming unit

CV Coefficient of variation

dSm-1

desi simen per metre

EC Electrical conductivity

et al. and others

etc. etceteras

Fe Iron

Fig. Figure

g Gram(s)

ha Hectare(s)

i.e. that is

K Potassium

kg ha-1

kilogram per hectare

kg kilogram

m meter

mg kg-1

milligram per kilogram

mm millimeter

No. number oC Degree centigrade

P Phosphorus

RBD Randomised Block Design.

SEm± Standard Error

viz., namely

Zn Zinc

ACKNOWLEDGEMENT

I think it is the matter of pleasure to glance back and recall the way one traverse, the days

of hard work and perseverance. It is still great at the juncture to recall the faces and spirits in the

form of teachers, friends, near and dear once. In my opinion, this work is nothing more than

incomplete, without attending to the task acknowledgemending, to overwhelming help I received

during this endeavour of mine.

Everything has its own beauty, but not everyone can see without critical observation and

great vision. Inspiration is the best medicine which can make it possible to run for crippled one and

it is veritable gold of mine to get the talented and inspiring wilful guidance. Today I stand on door

of this vision due to my Guide Dr. S.S.Mane Associate Professor, Department of Soil science and

Agricultural chemistry, College of Agriculture, Badnapur, and Chairman of my advisory committee.

Who in this unique way provided me with constant encouragement inspiring, scholastic guidance,

love and affection offered to me during the course of my study and research works would be a poor

vehicle to communicate him my sense of gratitude. For his constructive criticism, unflagging

enthusiasm and precious guidance during whole tenure of the investigation. It has his most co-

operative and painstaking attitude, which made this thesis a reality.

This memorable occasion provides me with a unique privilege to express my deep sense of

respect and indebtness to members of my advisory committee Dr. V.D.Patil, Head of Department,

Department of Soil science and Agricultural chemistry, VNMKV Parbhani, Dr. Syed Ismail

Associate proffesor, VNMKV, Parbhani, Dr. G.R.Hanwate Assistant Professor, Department of

Soil science and Agricultural chemistry ,College of Agriculture, Badnapur, Shri. S.J.Supekar

Assistant professor, Department of Agricultural Engineering, College of Agricultural,Badnapur

and may be regarded as the light hours for the ocean liners who have been kindly navigate my ship

of academic pursuit and I would like to mention my gratitude to them.

I owe my greatful thanks to authorities of the V.N.M.K.V., Parbhani, Hon’ble Dr. B.

Venkteshwarlu, Vice Chancellor , Dr. G. M. Waghmare, Associate dean and Principal, College of

Agriculture, Badnapur for providing necessary facilities to conduct the research work.

My sincere thanks to Dr.G.R.Hanwate, College of Agriculture, Badnapur, Shri P.L

Sontakke Senior research assistant, A.R.S,Badnapur, Dr. J.E. Jahagirdar, Incharge officer, ARS,

Badnapur, College of Agriculture,Badnapur, Shri. S.J. Supekar, Dept. of Engeeniring, College of

Agriculture, Badnapur, Shri. K.T. Jadhav, Dept of Agronomy, College of Agriculture, Badnapur,

Shri.S.K.Raskar, Dept.of Agronomy, college of Agriculture, Badnapur, Sau.A.A. Chaudhari, Dept.

of Mathematics, Prof. B.V.Patil, Dept.of Entomology, College of Agriculture,

Badnapur…..…..for their positive backing and hands of co-operation during the research work.

I can’t express the depth of feelings to thanks for being my friends forever to

Raju,Gaurav,Balu,Kunal,Prakash,Akshata for their love, moral support and care.

I am extremely thankful from bottom of my heart to all my batchmates

Basvaprasad,Pandu,Ranjeet,Sachin,Sudam, Akshay,Pravin,Antariksh, Vikas, Kailas, Avdhut,

Raju, , Basavraj, Shital,Sarika,Madhushri,Chaitali,Shubhangi and my beloved friends Prasad,

Abhishek, Akash, Sharad, Rahul, Mayur, Ravi, Abhijeet, Ashok, Amol, Sopan, ,Prayag and Zahir

More for their excellent company and valuable help.

The logistical support and advice from my lovable seniors, Mr. Sriram Murme sir, Mr.

Vaibhav Kadam sir, Mr. Swapnil More, Miss. Sonali Dahifale, Miss.Supriya Yadav, Miss.Varsha

Adsure are gratefully acknowledged and I would also like to recognize my all juniors, Laxman,

Sagar, Amol,Dafde.

I take this opportunity to thank my mess owner, Mr. Pravin Huse, for their delicious and

luscious food, tea, snacks and also love, affection, care and brotherly relationship towords me. My

deepest and sincere gratitude goes out to Shri. Jagnnath Gaware.

No words in this mortal world can suffice, to express my feelings, the great sacrifice,

devotion, constant encouragement, inspiration, high appreciation and the deepest sense of

reverence towards my worship, of my respected and adorable my beloved parents Mr. Indar

Navsare and Mrs. Usha Navsare, who helped me on every path of my life and made every step a

great success. who are always with me for any assistance I required. Her moral and emotional

support gave me strength during my study and same will continue for my entire life. I express my

heartfelt gratitude towards my Uncle Mr. Suresh Navsare and Mr. Gajanan Navsare and my

grandfather Shri. Mr. Mahadu Navsare whose obstinate sacrifice, filial affections and blessing

made my path easier.

My special thanks to my brother Anand, Bablu, Bunty, Chinu, and my dearest sisters

Janhavi More and Brother-in-law Dr. Prakash More for their selfless love filial affection constant

encouragement sincere prayers support and obstinate sacrifices without which this dream could not

become a reality.

Last, but far from the least, I would like to place on record my sincere regard, deepest

gratitude, soulful respect and a million thanks to the “God” the almighty, who helped me on every

path of my life and made every step a great success and also my warm acknowledgements to

farmers, soldiers, scientists and great social workers, who have devoted their whole life for the

country.

Place: Badnapur

Date: (Navsare.R.I)

CONTENTS

Chapter Title Pages

I INTRODUCTION 1-5

II REVIEW OF LITERATURE 6-27

III MATERIALS AND METHODS 28-39

IV RESULTS AND DISCUSSION 40-86

V SUMMARY AND CONCLUSIONS 87-89

LITERATURE CITED I-XVII

THESIS ABSTRACT

APPENDIX I-IV

LIST OF TABLES

Table No. Title Page No.

3.1 Initial physico-chemical properties of soil 29

3.2 Weakly Meteorological data prevails in crop growth period 30

3.3 Quantity of nutrient applied in each plot 32

4.1 Soil properties of the experimental soil before sowing and after

harvest of crop 42

4.2 Effect of potassium and zinc solubilizing microorganism on

germination and final plant stand 44


number of pods per plant 45


number of nodules per plant 46

4.5 Effect of potassium and zinc solubilizing microorganism on total

biomass production (g plant-1

) 48

4.6 Effect of potassium and zinc solubilizing microorganism levels

on dry matter (kg ha-1

) production 50


economic yield of green gram 52


protein content 55


nitrogen concentration at critical growth stages 57


phosphorous concentration at critical growth stages 59


potassium concentration at critical growth stage 61

4.12 Effect of potassium and zinc solubilizing microorganism on iron

concentration at critical growth stages 63

4.13 Effect of potassium and zinc solubilizing microorganism on zinc

concentration at critical growth stages 65

4.14 Effect of potassium and zinc solubilizing microorganism on N

uptake 67

4.15 Effect of potassium and zinc solubilizing microorganism on P 70

Table No. Title Page No.

uptake

4.16 Effect of potassium and zinc solubilizing microorganism on K

uptake 72

4.17 Effect of potassium and zinc solubilizing microorganism on Fe

uptake 74

4.18 Effect of potassium and zinc solubilizing microorganism on Zn

uptake 76


microbial population after harvest 78


available N (kg ha-1

) 83


available P (kg ha-1

) 84


available K (kg ha-1

) 85


available Fe and Zn (kg ha-1

) 86

LIST OF FIGURES

Fig

No.

Title Page No.

3.1 Plan of layout and observation unit 33

4.1 Effect of potassium and zinc solubilizing microorganism

levels on dry matter (kg ha-1

) production 51


economic yield of green gram 54


nitrogen concentration at critical growth stages 58


phosphorous concentration at critical growth stages 60


potassium concentration at critical growth stage 62


iron concentration at critical growth stages 64


zinc concentration at critical growth stages 66

4.8 Effect of potassium and zinc solubilizing microorganism on N

uptake 69

4.9 Effect of potassium and zinc solubilizing microorganism on P

uptake 71

4.10 Effect of potassium and zinc solubilizing microorganism on K

uptake 73


Fe uptake 75


Zn uptake 77


bacterial population 80


actinomycetes population 81


fungal population 82

LIST OF PLATES

Plate No. Title In between

pages

I General view of experimental field during kharif 2016 33-34

II Experimental field plots with different treatments 33-34

Introduction

1

CHAPTER -I

INTRODUCTION

Mungbean [Vigna radiata (L.) Wilczek syn. Phaseolus mungo] is cultivated in many

regions of the world because of its considerable nutritional value, particularly for people

encountering malnutrition (Allahmoradi et al., 2011). Mungbean contains bioactive

compounds with antioxidant, antimicrobial and insecticidal properties (Madhujith et al.,

2004; Ahmad et al., 2008). The seeds contain 25-28 per cent protein, 1.0-1.5 per cent fats,

3.5-4.5 per cent fibres, 60- 65 per cent carbohydrates and are rich in lysine, ascorbic acid,

potassium, iron, phosphorus and calcium (Lambrides and Godwin, 2007). Mungbean can

play the major role in national economy of India due to their wider adaptability, easy

digestibility, better palatability and higher market price (Miah et al., 2009; Reddy, 2009).

The growth of the crop is reduced when potassium is not applied sufficiently

(Hermans et al., 2006). When plants are grown under low supply of potassium, drought stress

induced reactive oxygen spacies (ROS) production can be additionally enhanced, at least due

to K deficiency induced disturbances in stomatal opening, water retention and photosynthesis

(Mengel and Kirkby, 2001). Potassium influences the water economy and crop growth

through its effects on water uptake, root growth, maintenance of turgor, transpiration and

stomatal regulation (Nelson, 1980). Although potassium unlike nitrogen and phosphorous,

does not enter into the composition of any product, yet literature on potassium reveals that it

has an important role either direct or indirect, under different environments, in major plant

processes such as photosynthesis, respiration, protein synthesis, enzyme activation, plant

water relations, osmoregulation, growth and yield of plant (Zaidi et al., 1994, Singh et al.,

1997, Garg et al., 2005, Sharma et al., 2008, Arif et al., 2008, Bukhsh et al., 2011 and

Waraich et al., 2011). Potassium helps plant to adjust low water potential under drought

stress (Bukhsh et al., 2012). Under adverse climatic conditions, potassium plays an important

role in different physiological and biochemical processes. Potassium application under water

stress moderates the adverse effects of water shortage on plant growth (Sangakkara et al.,

2000 and 2001; Singh and Kumar, 2009). Yield limiting effects of water deficit could be

overcome by increasing potassium supply (Damon and Rengel, 2007). During stress

conditions, reactive oxygen species (ROS) formation was induced and oxidative damage to

cells occurred and requirement for potassium was increased (Foyer et al., 2002), this shows

the importance of potassium in legume nutrition. Improvement in potassium status of plants

2

seems to be of great importance for sustaining high yields under rainfed conditions. However,

the work done on legumes with potassium application is not well recognized in comparison to

cereals and other crops.

In view of the importance of mungbean and since it is normally cultivated on

relatively poor soil under rainfed conditions, it becomes imperative to study the effect of

potassium on nodulation, water relations and expression of the same in growth, yield and

quality of seeds. Thus an evaluation of the significance of potassium in crop productivity

under soil moisture deficit must analyze the effect of potassium on above mentioned

physiological processes. Moreover, work done on legumes with potassium, less conclusive

and more contradictory depending on environmental conditions and type of legumes.

Biofertilizers is one of the important components of the sustainable agriculture

containing living microorganisms which have the ability to mobilize nutritionally important

elements from non usable to usable form through biological processes. Biofertilizers have

come to stay in Indian agriculture since last three decades in view of their cost effectiveness,

contribution to crop productivity, soil sustainability and ecofriendly characters. Use of

biofertilizers is one of the important components of integrated nutrient management as these

are renewable source of plant nutrients to supplement the chemical fertilizers for sustainable

agriculture. Biofertilizers include: nitrogen fixing bacteria (Rhizobium, BradyRhizobium,

Azospirillum and Azotobacter), phosphate solubilizing bacteria (PSB) (Bacillus,

Pseudomonas, Aspergillus etc.), phosphate mobilizing biofertilizers (Mycorrhiza), plant

growth promoting biofertilizers, potassium solubilizing bacteria and zinc solubilizing

bacteria. Zinc solubilizing potential of few bacterial genera has been studied. Hutchins et al.

(1986) reported that Thiobacillus thioxidans, Thiobacillus ferroxidans and facultative

thermophilic iron oxidizers solubilize zinc from sulphide ore (sphalerite). Zinc is a nutrient at

low concentration but toxic at higher concentrations. The solubilization of zinc might limit

the growth of bacteria at higher levels. Unless the cultures tolerate higher level of zinc, its

solubilization may not continue. A few fungal genera possess immense potential of

solubilizing zinc; Aspergillus niger was found to grow under 1000 mg Zn and this fungus is

used to quantify zinc in soils containing low zinc (2 mg kg-1 available zinc) (Bullen and

Kemila, 1997). Microorganisms inhabiting rhizospheres of various plants are likely to

synthesize and release auxins as secondary metabolites because of rich supplies of substrates

exuded from the roots compared with non rhizospheric soils. Bacteria belonging to the genera

Azospirillum, Pseudomonas, Xanthomonas and Rhizobium as well as Alkaligenes faecalis,

Enterobacter cloacae, Acetobacter diazotrophicus etc. and some fungi and algae are capable

3

of producing auxins which exert pronounced effect on plant growth and establishment (Patten

and Glick, 1996). Indole acetic acid (IAA) is one of the most physiologically active auxins.

IAA is a common product of L-tryptophan metabolism by several microorganisms. Plant

morphogenic effects may also be a result of different ratios of plant hormones produced by

roots as well as by rhizosphere bacteria. Zinc solubilizing bacteria produce IAA products

which may also have effect on plant growth (Rajkumar et al., 2008).

Plants require various nutrients for their growth and metabolism. Bacteria play an

important role in mobilizing nutrients required by the plants. Among these, zinc is one of the

essential micronutrient required for the normal healthy growth and reproduction of crop

plants. It plays a vital role in plant metabolism (Hughes and Poole, 1989). It occurs in soil as

sphalerite, olivine, hornblende, augite and biotite. However, availability of zinc from these

sources is guided by many factors among which biochemical action of rhizomicroorganisms

plays an important role in converting such unavailable sources into available ones (

Bhupinder et al., 2005).

Zinc is present in the enzyme system as co-factor and it is metal activator of many

enzymes (Parisi et al., 1969). Many bacterial enzymes contain zinc in the active center or in a

structurally important site. In soil, it undergoes a complex dynamic equilibrium of

solubilization and precipitation that is greatly influenced by soil pH and microflora and that

ultimately affects its accessability to plant roots for absorption (Goldstein, 1995). Zinc

deficiency displayed as a remarkable reduction in plant height and plants develop whitish

brown patches that turn necrotic subsequently. Zinc deficiency results in reduced membrane

integrity, reduction in synthesis of carbohydrates, cytochromes, nucleotides, auxins,

chlorophyll and increased susceptibility for heat stress. Root cell membrane permeability is

increased under zinc deficiency which might be related to the function of zinc in cell

membranes (Parker et al., 1992). Zinc deficiency in fungi and bacteria is accompanied by

impairment of the formation of pigments such as melanin, chrisogenin, prodigiosin, subtilin

and others (Chernavina, 1970). One of the widest ranging abiotic stresses in the world

agriculture arises from the low zinc (Zn) availability in the calcareous soils. In India, up to

50% of the agricultural land, particularly the whole of the Indo-Gangetic belt, is reeling under

zinc deficiency. This has serious consequences as plants grown on zinc-deficient soils have

reduced grain yield (80%). The major reason for widespread occurrence of zinc deficiency

problem in crop plants is especially its low solubility rather than a total low amount of zinc

(Cakmak, 2008). The solubility of zinc is highly dependent on soil pH and moisture and

hence arid and semi-arid areas of Indian agro-ecosystems are often zinc deficient. It can be

4

corrected through exogenous application of soluble zinc sources but only 20% of applied zinc

is available for plant uptake and rest of the zinc is converted to various unavailable forms.

Zinc thus made unavailable is converted back to available form by inoculating bacterial

strains which can solubilize it by release of organic acids and decrease in pH.

Zinc solubilizing bacteria also exhibit other traits beneficial to plants like phosphate

solubilization. Phosphorus is vital to seed formation and its content is higher in seeds than in

any other part of the plant. It helps plants to survive winter rigors and also contributes to

disease resistance in some plants. It is also known to improve quality of many fruits,

vegetables and grain crops.

Potassium is one of the three main pillars of balanced fertilizer use, along with N and

P. Leaves, straw and strover retain about 70 to 75 % of the K absorbed. The remaining

portion such as grains, fruits, nuts etc. Whenever soil cannot adequately supply the K

required to produce high yields, farmer must supplement soil reserves with fertilizer K.

Pulses growing regions of India consist of different soil types and available K status

varying from low to high (Srinivasarao et al., 2001). Pulses in India are grown mostly on

marginal and sub marginal lands without proper inputs with low inputs such as fertilizer. Patil

et al., (2001) reported that average use of K in Maharashtra was 11.10 kg ha-1

, which is very

low as against its mining 118.20 kg ha-1

causing K depletion in a very high rate. Potassium

(K) is rarely applied to pulse crops despite larger K requirements of pulses and continued

mining of soil potassium (K), resulting in imbalanced nutrient supply and lower crop yield.

Under intensive cropping systems, larger amounts of K are removed leading to serious

depletion of soil potassium (K) reserves. Among production inputs, fertilizer application

plays a key role in enhancing productivity levels.

However, fertilizer recommendation practices for pulse crops have been paid less

attention. There has been a dramatic decrease in the fertilizer consumption of K compared to

fertilizer N and P while K removal from the soil is generally as much as or higher than N;

still its use in fertilizer is negligible. A steep rise in their price accompanied by no such

increase in the price of N greatly made it imbalanced in the favour of N. As a result the N:

P2O₅: K2O ratio which was already imbalanced at 1: 0.41: 0.17 in 1991 deteriorated further to

1: 0.41: 0.15 during 1999-2000 whereas the ideal ratio is 1: 0.5: 0.25 (Pasricha, 2000). This

derived nutrient imbalance in fertilizer consumption shows a distinctive pattern, increasingly

in favour of N and increasing by negative for P and K. Cultivation of high yielding crop and

hybrids and diversification towards P and K demanding crops such as vegetables, potatoes,

5

oilseeds and pulses will place even more strain on K budgets of the soil. This means

diminishing soil fertility and declining fertilizer use efficiency, which increases cost of

production and restricts water use efficiency.

The present investigation was undertaken with the following objectives:

1. To study the growth and productivity of mungbean.

2. To study the nutrient content and their uptake by mungbean.

3. To study the quality of mungbean.

4. To study the fertility status of soil after the harvest of mungbaen.

Review of Literature

6

CHAPTER - II

REVIEW OF LITERATURE

The present study entitled “Studies on effect of potassium and zinc solubilizing

microorganism on growth, yield and quality of mungbean” is aimed to assess the effect of

potassium and zinc solubilizing microbes on yield and yield components, nutrient

availability, nutrient uptake at crop growth stages and quality in terms of protein content of

green gram. The pertinent literature on these aspects is reviewed under following heads.

11.1 Effect of Potassium solubilizing microorganism on plant growth, yield and

quality of crop.

Sheng and Huang (2002) reported that the effect of potassic bacteria on sorghum,

which results in increase biomass and contents of P and K in plants than the control. The

effect of KSB (Bacillus sp.) on grain yield and plant silica content of rice and available silica

in soil (Raj, 2004). The increase K uptake coupled with increased yield in yam and tapioca

while treating the plants with potassium mobilizer in conjunction with biofertilizers and

chemical fertilizers (Clarson, 2004).

Sheng et al. (2003) reported silicate dissolving bacteria could improves soil P, K, Si

reserves and promote plant growth. The effect of potash mobilizer on brinjal has recorded an

increased potash uptake and increased plant biomass in potash mobilizes treated plant as

compared to the control plant (Nayak, 2001).

Sheng et al. (2005) worked on potassium releasing bacterial strain B. edaphicus for

plant growth promoting effect and nutrient uptake on cotton and rape seed in K deficient soil

pot experiments resulted increased root and shoot growth and potassium content was

increased by 30 and 26 per cent respectively and in chilli crop increased biomass and K

uptake due to inoculation of potash solubilizer (Ramarethinam et al., 2005).

Sugumaran and Janarthanam (2007) recorded increase in the dry matter by 25 per cent

and oil content 35.4 per cent in groundnut plant and available P and K is increased from 6.24

and 9.28 mg/kg and 86.57 to 99.60 mg/kg, respectively in soil due to inoculation of B.

mucilaginosus (KSB) compared to uninoculated control.

Archana et al. (2008) conducted an experiment to study the effect of potassium

solubilizing bacteria on growth and yield of maize. Efficient K solubilizing bacteria Bacillus

spp. were used and the result showed that there was a further increase in growth, nutrition and

7

yield of maize. The results indicated that all the inoculated bacterial isolates increased plant

growth, nutrient uptake and yield component of maize plant significantly over absolute

fertilizer control.

Chaveevan et al. (2010) reported that biofertilizers that possess a high capacity for N2

fixation (Azotobacter tropicalis) and consist of phosphate solubilizing bacteria (Burkhoderia

unamae) and potassium solubilizing bacteria (Bacillus subtilis) and produce auxin (KJB9/2

strain), have a high potential for growth and yield enhancement of corn and vegetables

(Chinese kale). For vegetables, the addition of biofertilizer alone enhanced growth 4 times.

Moreover, an enhancement of growth by 7 times was observed due to the addition of rock

phosphate and K feldspar, natural mineral fertilizers, in combination with the biofertilizer.

Prajapati et al. (2012) studied the potassium is vital component of plant nutrition

package limiting crop yield and quality that performs a multitude of important biological

functions to maintain plant growth, isolation of potassium solubillizers was carried out using

feldspar (insoluble potassium) from the soil samples of ceramic industries, on Aleksandrow‟s

agar medium. From the fourteen isolated bacterial isolates five bacterial strains were selected

which exhibiting highest potassium solubilization on solid medium and characterized on the

basis of cultural, morphological and biochemical characteristics. Solubilization of potassium

from the potassium aluminium silicate minerals by the selected bacterial strains resulted to

the action of different organic acids like Citric, Oxalic, Malic, succinic and Tartric acid.

Bagyalakshmi et al. (2012) conducted a field experiment to study the efficacy of

indigenous potassium solubilizing bacteria (KSB) in combination with various dosages of

potash fertilizers along with recommended dose of nitrogen (N) and phosphorus (P) fertilizers

in tea plants. Soil and leaf samples were drawn from the respective plots and they were

subjected for the analysis of various parameters related to nutrients and quality aspects.

Among various treatments, plants treated with N100: P100: K75 + KSB concentration

formulation was found to be the best in terms of high chlorophyll, carotenoid, N, P and K

contents in the crop shoots followed by other treatments. Significantly higher yield of the

green leaf was achieved by the same treatment. K content in soil and also in crop shoots was

greatly improved due to the application of KSB along with possible reduced doses of potash

source. On the other hand, N100: P100: K25 + KSB formulation and untreated control plots

have exhibited least green leaf yield and nutrient status of soil and crop shoots. The

biochemical parameters, total polyphenols, catechins, amino acids and sugars were

significantly at higher level in plants after imposing treatments in combination with KSB.

Biometric parameters such as plucking surface of the tea bush canopy, plucking points per

8

unit area, internodal length, leaf moisture and dry matter contents were analyzed and found to

be high in tea plants treated with N100: P100: K75 + KSB combination. Banji content was

significantly reduced invariably in all the treatments except the untreated control plots.

Evaluation of KSB population in soil revealed that wherever KSB was incorporated, there

was a significant increase in population level and has coincided with dehydrogenase enzyme

activity. The flush shoots of tea comprising of three leaves and a bud were subjected to

manufacture black tea, and it was revealed that almost all the tea quality parameters such as

theaflavin, thearubigin, highly polymerized substances, total liquor colour, caffeine,

briskness, colour and flavour indexes were greatly improved in KSB treated plants, which in

turn improve the quality as well. This finding confirms that the influence of indigenous

potassium solubilizing bacteria upon potassium nutrient exhibited improvement in the

productivity and nutrient uptake in plants and retained in soil including quality parameters in

tea plantations.

Prajapati et al. (2013) conducted pot experiments were a potassium releasing

bacterial Enterobacter hormaechei and fungal Aspergillus terreus strains were examined for

plant growth promoting effects and nutrient uptake on Okra (Abelmoscus esculantus) in K

deficient soil in pot experiments. Inoculation with bacterial strain Enterobacter hormaechei

was found to increase root and shoot growth of Okra and both microorganisms were able to

mobilize potassium efficiently in plant when feldspar was added to the soil. In okra growing

in soils treated with insoluble potassium and inoculated with strain Enterobacter hormaechei

and fungal Aspergillus terreus the potassium content was increased. Among all the three

applications of microbial inoculants, combined application of seed and soil was found to be

more effective on Okra plant growth.

Patil et al. (2014) conducted a field trial during 2014 season at the farm of College of

Agriculture, Nagpur to study the combine effect of Rhizobium and potash solubilizing

microbes in mungbean. The results revealed that the combination effect of Rhizobium and

potash solubilizing microbes significantly improved in plant productivity. Maximum plant

height, root length, nodulation, fresh weight of nodules, dry weight of nodules, fresh weight

of plant were 37.05cm, 11.78 cm, 13 cm, 13.10 g, 5.25 g, 2.10 g, 2.10 g, 0.63 g, respectively

at 30 days after sowing and 45.90 cm, 18.45 cm, 17025 cm, 18.18 g, 7.20 g, 2.83 g, 0.82 g at

45 days after sowing, hundred seeds weights were 6.88 g, 4.28 g and Rhizobial population at

30, 45 and 60 days were 26.50 x 106, 32.50 x 10

6 and 23.25 x 10

6.

Verma et al. (2016) studied that potassium is one of the important component of

plant nutrients to meet the requirement of crop growth, crop yield and quality of product.

9

Isolates of KSB was carried out using mica (insoluble potassium) from the soil sample of

different regions of India basically U.P and M.P on Aleksandrow‟s agar media. Out off 14

isolated bacterial isolates 7 bacterial isolates MPS1C2, MPS2C5, MPS2C4, MPS5C1,

UPS1C1, UPS2C1, UPS3C1 showed highest D/d ratio 3.13, 3.50, 2.0, 3.22, 5.00, 4.13, 3.75.

Optimum pH and temperature was 7 and 28±2° C respectively. Maximum K solubilization

was achieved in KCl and K2SO4. The isolated bacterial strain was identified as Micrococcus

varians and Corynebacterium kutscheri. KSB treated groundnut plant showed maximum

pods of 16 and seeds 72 per plant as compared to control having pods 7 and seeds 21.

Prajapati (2016) reported that soil microorganisms were supportive in the

transformation of soil potassium (K) and are thus an important component of the soil K cycle.

These are effective in releasing K both from inorganic and organic pools of total soil K

through their respective solubilizing and mineralizing abilities. To evaluate this, two

promising organisms (KSB-1 and KSB-7) capable of solubilization of both organic and

inorganic potassium as investigated under in vitro conditions in a pot culture for their

rhizosphere activity and mineralization potential of organic K in soil, plant growth and yield.

In response to inoculation with these selected potassium solubilizing bacteria (KSB),

significant increases in seed germination, root and shoot length and number of leaves, grain

yield were observed which were increase to respectively, over uninoculated control in the

presence of feldspar in Aleksandrov‟s agar medium. The study demonstrated that the use of

KSB having multifaceted beneficial traits would be highly effective for improving growth

and yield of crops.

Prajapati and Modi (2016) conducted an experiment under hydroponics condition

using micronutrient containing nutrient solution to evaluate the effect of potassium

solubilizing bacteria KSB-8 (Enterobacter hormaechei). The results indicated that a

remarkable increase in root length, flowering, fruit setting, fruit maturing, K content and

chlorophyll content. Thus, it might be concluded that KSB-8 (Enterobacter hormoechei)

could be used as crop-enhancer and bio-fertilizer for cucumber (Cucumis sativus) and other K

rich crops under hydroponic condition.

11.2 Effect of Zinc solubilizing microorganism on plant growth, yield and quality of

Mungbean.

Zinc deficiency not only affects crop yield, but also nutritional quality and human

health. Microbial transformation of unavailable forms of soil zinc to plant available zinc is an

important approach contributing to plant zinc nutrition and growth. Increased zinc

concentration by zinc solubilizing bacteria has large implications in terms of overcoming zinc

10

malnutrition. Zinc solubilizing bacteria influenced mobilization of zinc and its concentration

in edible portion, yield and can be utilized as bio-inoculants for biofertilization and

biofortification.

Kumar et al. (2004) conducted a field experiment to study the effect of Zn enriched

organic manures and Zn solubilizers on the yield, curcumin content of turmeric and nutrient

status of the soil. The treatment FYM + zinc solubilizing bacteria showed higher turmeric

rhizome yield increase of 21.6 per cent than the FYM alone treatment (9.1 per cent) than no

manure (control). The dry rhizome yield reflected the promising effect of Zn and Fe enriched

coirpith or FYM, and Zn enriched coirpith or FYM at M1 (no manure) while at M2 (NPK +

FYM) and M3 (NPK + FYM+ Zn solubilizing bacteria), the foliar spray of Zn + Fe + MOP

excelled the remaining treatment.

Saravanan and Raj (2004) made an attempt to isolate zinc solubilizing bacterial (ZSB)

cultures from soil and ore (sphalerite) sources both by direct plating and by enrichment

technique in the modified Bunt and Rovira medium with 0.1% zinc. Three cultures (ZSB-O-

1, ZSB-S-2 and ZSB-S-4) were isolated by direct plating and one (ZSB-S-3) by enrichment

technique. Among these, ZSB-O-1 and ZSB-S-4 were characterized as Bacillus sp. and

ZSBS- 2 as Pseudomonas sp. and their potential to correct the zinc deficiency was assessed

using 10 soybean plants (the plant was selected because of its high Zn requirement). The

results revealed that Pseudomonas sp. (ZSB-S-1) was able to correct the zinc deficiency

when used along with 1% (w/w) zinc oxide.

Subramaniam et al. (2006) in their field experiment found that Thompson seedless

grapes with treatments involving micronutrients as soil application and foliar spray. The

juice content TSS, titrable acidity, specific gravity, total sugar and TSS/acidity ratio were

higher in the treatment of RDF + micronutrients.

Tariq et al. (2007) reported that the activity of plant growth promoting Rhizobacteria

(PGPR) to mobilize indigenous soil zinc (Zn) in rice (Oryza sativa L.) Rhizosphere was

observed in a net house micro plot experiment and compared with available form of chemical

Zn source as Zn-EDTA. The PGPR application alleviated the deficiency symptoms of Zn and

invariably increased the total biomass (23%), grain yield (65%) and harvest index as well as

Zn concentration in the grain. The inoculation had a positive impact on root length (54%),

root weight (74%), root volume (62%), root area (75%), shoot weight (23%), panicle

emergence index (96%) and showed the highest Zn mobilization efficiency as compared with

the un-inoculated control. The PGPR colonized rice plants were more efficient in acquiring

Zn from either added or indigenous source, than non-colonized plants. Zinc mobilization by

11

PGPR was also confirmed in liquid culture medium. It was concluded that, selected PGPR

strains can serve as efficient solubilizer of Zn, allowing farmers to avoid the use of costly

chemical Zn fertilizer in rice crop.

Esitken et al. (2009) studied the effects of plant growth promoting bacteria (PGPB) on

the fruit yield, growth and nutrient element content of strawberry cv. Three PGPB strains 11

(Pseudomonas BA-8, Bacillus OSU-142 and Bacillus M-3) were used alone or in

combination as biofertilizer agent in the experiment. Results for three years showed that the

use of PGPB significantly increased fruit yield, plant growth and leaf P and Zn contents. In

addition, P and Zn contents of strawberry leaves with bacterial inoculation significantly

increased under organic growing condition.

Iqbal et al. (2010) examined the effects of five bacterial isolates (U, 8M, 36, 102,

and 111) on the growth of Vigna radiata. Bacterial isolates were applied alone, or together

with zinc phosphate [Zn3(PO4)2·4H2O]. The maximum increase in all plant growth

parameters was seen when seedlings were inoculated with isolate 102. Isolate 36 with 1 mm

zinc phosphate showed the maximum increase in seedling length (35.1 cm) as compared to

controls. Isolate 111 was the best phosphate solubilizer, releasing 13.29 ppm phosphorous (P)

in soil when used in combination with 1 mm salt, whereas isolate 36 showed maximum

uptake of P, leaving only 4.63 ppm in soil.

Kumar et al. (2012) isolated seven bacterial isolates from rhizosphere of common

bean growing at Uttarakhand Himalaya and screened them for their potential plant growth

promoting (PGP) and antagonistic activities. Strain BPR7 produced IAA, siderophore,

phytase, organic acid, ACC deaminase, cyanogens, lytic enzymes, oxalate oxidase and

solubilized various sources of organic and inorganic phosphates as well as potassium and

zinc. Strain BPR7 strongly inhibited the growth of several phytopathogens such as

Macrophomina phaseolina, Fusarium oxysporum, F. solani, Sclerotinia sclerotiorum,

Rhizoctonia solani and Colletotricum sp. in vitro. Cell free culture filtrate of strain BPR7 also

caused colony growth inhibition of all test pathogens. PGP and antifungal activities of

Bacillus sp. BPR7 suggested that it may be exploited as a potential bioinoculant agent for

Phaseolus vulgaris.

Sharma et al. (2012) recovered one hundred thirty four putative Bacillus isolates from

soybean rhizosphere soils of Nimar region to select effective zinc solubilizers for increased

assimilation of zinc (Zn) in soybean seeds. Evaluation under microcosm conditions showed

that inoculation of isolates significantly increased the Zn concentration in soybean seeds as

compared with uninoculated control (47.14 μg/g).

12

Nyoki and Ndakidemi (2014) conducted field and green house experiments to assess

the effects of B. japonicum inoculation and phosphorus supplementation on the uptake of

micronutrients in the cowpea. The results showed a significant improvement in the uptake of

micronutrients in the B. japonicum inoculated treatments over the control. Phosphorus

supplementation (40 kg P/ha) also showed a significant increase in the uptake of some

micronutrients while decreasing the uptake of Zn in some plant organs. There was also a

significant interaction between B. japonicum inoculation and phosphorus in the root uptake of

Zn for the field experiment.

Joshi et al. (2013) conducted pot experiment on wheat (PBW 373), application of

bacterial isolates and their consortium with ZnSO4.7H2O @ 5mM significantly enhanced

plant height, chlorophyll, grain number per plant over control after 90 days of sowing.

Consortium (B1B2) treated wheat plants showed 31% increase in grain Zn over control.

HPLC analysis of the root extract of bacterized wheat plants showed the dominance of oxalic,

maleic, ketoglutaric and fumaric acid. Presence of higher amount of valine and leucine,

sugar, protein and phenolic compounds was observed in the root extracts of consortium

treated wheat plants than control.

Vaid et al. (2014) reported that comparison study between the isolated indigenous

bacteria and chemical Zn fertilizer (ZnSO4.7H2O) was conducted to evaluate their potential to

augment Zn nutrition of Zn responsive (NDR 359) and Zn non responsive (PD 16) varieties

of rice under green house. Three bacterial strains namely; BC, AX and AB isolated from a Zn

deficient rice-wheat field belonging to the genera Burkholderia and Acinetobacter were

investigated for the growth promotion and Zn uptake in rice plants. The plant growth

promotory properties such as Zn solubilization and IAA production of the isolates was

checked in a previous study. These three isolates when used individually or in combination

were found effective in significantly increasing the mean dry matter yield/pot (12.9%),

productive tillers/plant (15.1%), number of panicles/plant (13.3%), number of grains/panicle

(12.8%), grain yield (17.0%) and straw yield (12.4%) over the control and Zn fertilizer

treatment, respectively. Bacterial inoculations also significantly enhanced the total Zn

uptake/pot (52.5%) as well as grain methionine concentration (38.8%). Effect of bacterial

treatments on the bioavailability of Zn was assessed by estimating the levels of phytic acid in

grains. A reduction of nearly 38.4% in phytate: Zn ratio in grains was observed under

bacterial inoculations.

Nomen et al. (2015) reported the significant effect of Zn bio-fertilizer on growth and

yield attributes (except kernels/pod), pod and haulms yields, shelling percentage and Zn

13

content and uptake. Due to application of Zn bio-fertilizer 4.7, 6.2, 9.2, 5.2 and 7.9% increase

in pod yield, haulms yield, net returns, S uptake in kernel and Zn uptake in kernel was

recorded over control. Based on two years average, response to S and Zn application was

quadratic and economic optimum dose was worked out to be 39.0 and 4.5kg/ha, respectively.

Hence application of 39kg S and 4.5 kg Zn/ha with Zn solubilizer is recommended for

improving the productivity and profitability of groundnut in sandy loam soils low in S and

Zn.

Muhammad et al. (2015) reported that the 198 isolates were collected from native

soils of Pakistan and in vitro testing was done for Zn mobilizing activity. A significant

increase of 54, 68, 57 and 46% in wheat Zn content over chemical Zn fertilizer was observed

under all PGPR treatments. Low increase in grain Zn concentration of 6.5, 7.0, 15.2 and

12.5% was noticed over control by Zn fertilizer treatment with all four wheat genotypes.

Various wheat genotypes showed different response with PGPR applications. Similarly, all

three strains and their consortium increased wheat grain yield by 2.4, 0.7, 2.2 and 8.6% over

chemical Zn fertilizer, respectively. The strains identified by 16S rRNA, gyrB and gyrA gene

analysis were Serratia liquefaciens, S. marcescens and Bacillus thuringiensis. The present

findings show that enhanced rate of PGPR colonization can improve grain yield and Zn

content of wheat as compared to chemical Zn fertilizer. Co-inoculation of PGPR proved to

have more potential of Zn mobilization towards grain. Maintaining suitable density of Zn

mobilizers in the soil through field inoculation might be a promising strategy to enhance

grain yield and Zn content of wheat. Commercial field application of this approach among

farmers is recommended.

Malik et al. (2015) showed that the effect of zinc, molybdenum and urea has been

studied on plant height (cm), number of productive branches, number of leaves, leaf area

(sq.cm.), fresh weight (g), dry weight (g), number of pods per plant, seed yield per plant and

1000 seeds weight (g) (Test weight) of mungbean [Vigna radiata (L.) Wilczek] Var. Pant

Mung-4 and Narendra-1. The experiment was conducted at Meerut College, Meerut (U.P.)

during the years 2011-2012. Randomised Block Design with 4 replications and 11 treatments

was used. The doses of zinc were 5, 10, 15 and 20 ppm. The concentrations of molybdenum

were 1, 2, 3 and 5 ppm and of urea were 1 and 2 per cent along with control. The results were

found significant in both varieties of mungbean.

Muhammad et al. (2015) reported that plant associated rhizobacteria prevailing in

different agro-ecosystems exhibit multiple traits which could be utilized in various aspect of

sustainable agriculture. Two hundred thirty four isolates were obtained from the roots of

14

basmati-385 and basmati super rice varieties growing in clay loam and saline soil at different

locations of Punjab (Pakistan). Out of 234 isolates, 27 were able to solubilize zinc (Zn) from

different Zn ores like zinc phosphate [Zn3 (PO4)2], zinc carbonate (ZnCO3) and zinc oxide

(ZnO). The strain SH-10 with maximum Zn solubilization zone of 24 mm on Zn3 (PO4)2ore

and strain SH-17 with maximum Zn solubilization zone of 14–15 mm on ZnO and ZnCO3

ores were selected for further studies. These two strains solubilized phosphorous (P) and

potassium (K) in vitro with a solubilization zone of 38–46 mm and 47–55 mm respectively.

The strains also suppressed economically important rice pathogens Pyricularia oryzae and

Fusarium moniliforme by 22–29% and produced various biocontrol determinants in vitro.

The strains enhanced Zn translocation toward grains and increased yield of basmati-385 and

super basmati rice varieties by 22–49% and 18–47% respectively. The Zn solubilizing strains

were identified as Bacillus sp. and Bacillus cereus by 16S rRNA gene analysis.

Shivran et al. (2015) conducted a field experiment at S.K.N. College of Agriculture,

Jobner (Rajasthan) during two consecutive rabi seasons. Results indicated that successive

increase in Zinc application up to 5.0 kg ha-1

significantly increased spikes plant , seeds spike

, test weight, seed -1 (13.07 q ha ), straw and biological yields, protein content, husk recovery

(34.82%), total N, P and Zn uptake and net returns (` 29696 ha ). Spike length was improved

only with 2.5 kg zinc application. Contrarily, swelling capacity was 1 -1 -1 significantly

decreased with 5.0 kg ha-1

zinc (10.80 cc g ) as compared to control (10.98 cc g ). The

increase in seed yield and net returns of blond psyllium with 5.0 kg ha-1

zinc were 26.04 and

32.99% over control and 7.57 and 9.03% over 2.5 kg zinc application, respectively. Blond

psyllium should be fertilized with 5.0 kg zinc per hectare to obtain higher productivity and

profitability.

Naz et al. (2016) revealed that wheat treated with Azospirillum, Pseudomonas and

Rhizobium significantly increased zinc contents in different parts of wheat plant at different

growth stages. These microbes also facilitate efficient nutrient uptake which ultimately

produce plants of superior quality making agriculture more productive and lesser harmful to

environment. It may be concluded from the study that beneficial

microorganisms/biofertilizers applied in combination could be a better choice for farmers to

reduce the use of chemical fertilizers for sustainable crop production.

15

11.3 Effect of Potassium solubilizing microorganism on Nutrient content and their

uptake

Laxminarayana (2001) conducted a field experiment during 1991-92 to study the

effect of mixing of black clayey soil and different levels of potassium on a sandy soil, yield

attributed and nutrient composition of groundnut. Number of filled pods per plant, number

of kernels per pod, test weight, shelling percentage, pod yield, haulm yield and crude protein

content were significantly increased with the addition of clay through vertisol and potassium

to a low potassium sandy soil. Protein content was increased from 19.2 to 21.3 % with

respect to increase the application of potassium (i.e. 0 to 75 kg ha-1

).

Singh and Najar (2007) recorded the organic carbon and available K in the soil

after the harvest of soybean increased over the control significantly when FYM was applied

alone or along with bio-inoculants. The highest organic carbon (1.08%) and available K

(222.3 kg/ha-1

) were recorded with combined inoculation of Rhizobium +Azotobacter + PSB

+ FYM (T6), followed by Rhizobium + PSM + FYM (T15). The available soil K status after

soybean harvest was depleted from initial level under all the biofertilizer treatments.

Sheng and Huang (2002) reported that potassium release from minerals was affected

by pH, dissolved oxygen and strains used. The content of potassium in solution inoculated

with bacteria was increased by 84.8-127.9 per cent compared with the control. The extent of

potassium solubilization by B. edaphicus in the liquid media and reported better growth on

illite than feldspar (Sheng and He, 2006).

Kabir et al. (2004) carried out an experiment to observe whether external application

of K mitigates the harmful effect of salinity and concluded that the uptake of nutrients i.e. N,

P, K showed an increasing tendency with the increased level of K (i.e. 14, 40, 60 kg ha-1

) in

mungbean.

Vasanthi et al. (2004) conducted a field experiment during 2000-2001 to evaluate the

effect of organic manures (vermicompost & FYM) and fertilizer on the uptake of nutrients

and crude protein content in black gram. Higher N, P, K concentration and uptake were

recorded in the treatments that received vermicompost @ 2t ha-1

along with 100%

recommended levels of N, P & K.

Han et al. (2006) conducted an experiments to evaluate the potential of phosphate

solubilizing bacteria (PSB) Bacillus megaterium var. phosphaticum and potassium

solubilizing bacteria (KSB) Bacillus mucilaginosus inoculated in nutrient limited soil planted

16

with pepper and cucumber. Results showed that rock P and K applied either alone or in

combination did not significantly enhance soil availability of P and K, indicating their

unsuitability for direct application. PSB was a more potent P solubilizer than KSB and co-

inoculation of PSB and KSB resulted in consistently higher P and K availability than in the

control without bacterial inoculum and without rock material fertilizer. Integrated rock P with

inoculation of PSB increased the availability of P and K in soil, the uptake of N, P and K by

shoot, root and the growth of pepper and cucumber. Similar but less pronounced results were

obtained when rock K and KSB were added concomitantly. Combined together, rock

materials and both bacterial strains consistently increased further mineral availability, uptake

and plant growth of pepper and cucumber, suggesting its potential use as fertilizer.

Gupta et al. (2007) conducted a field experiment at the Fertilizer Research Station of

Chandra Sekhar Azad University of Agriculture and Technology in Pusa, Uttar Pradesh,

during the summer season (March to June) of 2005 and revealed that successive increases in

K application rate enhanced N uptake by 2 to 20%, P uptake by 5 to 22%, and K uptake by 8

to 33% compared to control in black gram (K application rate 0, 20, 40, 60 K2O kg ha-1

).

Patil (2007) conducted a field experiment during kharif 2001-02 to study the effect of

integrated nutrient management on pigeonpea crop with the treatments comprised of

biofertilizers, farmyard manure, recommended dose (25:50:25 kg ha-1

N,P,K ) of fertilizers

and their all possible combinations. The results revealed that pigeon crop fertilized with 75

and 100 % RDF ha-1

were found equally effective and significantly superior over 50 % RDF

ha-1

and control with respect to content and uptake of nitrogen, phosphorus and potassium by

grain as well as stalk. However the highest values of these parameters were recorded with

100 % RDF ha-1

.

A field experiment was conducted by Sathyamoorthi (2007) during kharif and rabi

2002 and summer 2003 at the College of Agriculture Engineering, Triuchirappalli district of

Tamil Nadu to study the effect of green gram to increase plant density and nutrient

management on the nutrient uptake and yield of green gram and revealed that N, K , and S

uptake was higher with 125 per cent or 150 per cent NP along with foliar spraying of DAP

and SOP.

Billore et al. (2009) concluded that application of graded levels of potassium (0, 16.6,

33.2, 49.8 K kg ha-1

) led to an increase in total K uptake (23.45, 28.02, 31.62, 35.16 kg ha-1

)

by soybean.

17

Youssef et al. (2010) conducted a field experiment during summer seasons at Ismailia

Agric. Res. Station to study the effect of some natural minerals combined with potassium

dissolving bacteria inoculation in the presence of different nitrogen forms on chemical

properties of soil, nutritional status and yield of peanut-sesame. Three forms of nitrogen

fertilizer were included along with two natural minerals, in a presence of potassium

dissolving bacteria inoculation, as well as one mineral fertilizer as source potassium fertilizer.

Furthermore, data show high significant increases in available N due to the application of

ammonium nitrate in combination with feldspar and calcium nitrate in combination with

potassium sulphate in presence of inoculation for peanut and sesame, respectively. However,

application of calcium nitrate combined with potassium sulfate and ammonium nitrate in

combination with feldspar in a presence of inoculation led to significant increases in K

available in soil for peanut and sesame respectively. vis-à-vis, the pH values, different to

those of EC, decreased either for inoculation or non-inoculation as compared to control. In

spite of that, the values of EC and pH of soil were higher with application of either bentonite

or bentonite + feldspar in a presence of all nitrogen fertilizer forms. Generally, the highest EC

values in soil, after the two studied seasons were encountered with calcium nitrate fertilizer

as well as bentonite mineral. Moreover, applying feldspar mineral and ammonium nitrate

treatments had recorded the highest values of yield components as well as nutrient (N and K)

uptake by either peanut or sesame crops, particularly in the presence of inoculation as

compared to those given by other treatments.

Patil et al. (2011) studied the effect of potassium humate on nutrient uptake of black

gram and recorded that potassium humate (1.0%) treated crops showed significant increase

on calcium uptake (25 mg /100 g of dry matter) and phosphorous uptake (73.2 mg /100 g of

dry matter) of black gram.

Patil et al. (2011) recorded that significantly higher total uptake of N (173.02 kg ha-1

),

P (12.13 kg ha-1

), K (99.26 kg ha-1

), Ca (22.86 kg ha-1

), Mg (10.64 kg ha-1

) and S (9.68 kg

ha-1

) was recorded with the treatment of N : P2O5 : K2O @ 30 :50: 30 kg ha-1

along with seed

inoculation by cowpea.

11.4 Effect of Zinc solubilizing microorganism on Nutrient content and their uptake

Saravanan et al. (2003) reported that ZSB-O-1 (Bacillus sp.) showed highest

dissolution in the zinc sulphide (Sphalerite ore) 15th

day after inoculation. The ZSB-S-2

(Pseudomonas sp.) showed more solubilizing ability in the zinc oxide, 16.40 mg kg-1

of zinc

18

in the broth assay over the same inoculation period. The isolate ZSB-S-4 (Pseudomonas sp.)

has highest solubilizing potential in zinc carbonate. Thus, the solubilization potential varies

among different cultures. The solubilization might be due to production of acids by the

culture, since the pH of the culture broth has been shifted form 7.0-7.3 to 4.8-6.5 after 15

days of inoculation. The zinc tolerance limit for two cultures (ZSB-O-1 and ZSB-S-2) was

studied and determined to be upto 100 mg kg-1

of zinc in the in vitro broth assay.

A field experiment was conducted by Kumar et al. (2004) to study the effect of Zn

enriched organic manures and Zn solubilizers on the yield, curcumin content of turmeric and

nutrient status of the soil and noted that incorporation of farmyard manure at 12.5 t ha-1

along

with Zn solubilizing bacteria stood superior by registering highest values for available of N, P

and K content in the soil. The Zn solubilizing organism (Bacillus sp.) identified interestingly

proved to have favorable effect on the availability of N, P and K. The effect of micronutrient

treatments comprising of soil application of ZnSO4, FeSO4 and fortified FYM with Zn and

Fe and foliar spray of these two nutrients resulted in synergitic effect on the enhanced

availability of not only micronutrients but also K. The DTPA - Zn content of the soil though

evidenced significant variation for the different treatments of FYM, FYM + ZSB (Zn

solubilising bacteria) and micronutrients on an overall basis did not exceed the deficiency

level. Addition of Fe with Zn either as such or fortified with FYM with coirpith showed

synergitic effect on Zn availability in the soil. The available B content of the soil showed an

upheaval trend for manuring along with Zn and Fe. During different stages of crop growth

and at harvest stage, DTPA-Fe content in none of the treatments exceeded the threshold level.

However, enhancement for treatments with organic manures and micronutrients were

statistically perceptible. The availability of Cu and Mn in the soil, brought out the positive

effect of Zn and Fe added as such or as fortified either alone or along with FYM and FYM +

ZSB. Both content as well as uptake of all the major nutrients in the turmeric plant right from

the early phase of crop growth to harvest were positively altered by FYM, FYM + ZSB and

soil and foliar application of Zn and Fe.

Nweke et al. (2006) assessed toxicity of Zn2+

on four planktonic bacteria isolated

from New Calabar River water via dehydrogenase assay. Pure cultures of the bacterial strains

were exposed to various Zn2+

concentrations (0.2 -2.0 mM) in a nutrient broth amended with

glucose and TTC (Tri phenyl tetrazolium). At 0.2 mM concentration, Zn2+

stimulated

dehydrogenase activity in Proteus sp. PLK2 and Micrococcus sp. PLK4. In all the strains,

dehydrogenase activity was progressively inhibited at concentrations greater than 0.2 mM.

The order of zinc tolerance was : Micrococcus sp. PLK4 > Proteus sp. PLK2 > Pseudomonas

19

sp. PLK5 > Escherichia sp. PLK1. The results of the in vitro study indicated that the bacterial

7 strains are sensitive to Zn2+

stress. Therefore, Zn2+

contamination would pose serious threat

to their metabolism in natural environments.

Rasouli et al. (2008) showed that inoculation with sid+ strain increased dry matter

production in shoots as compared with the control (sterile condition) or with sid - strain.

Likewise, the concentration of chlorophyll a in leaves of sid+ and sid - treatments were 1.27

and 0.41 g mg-1

of fresh weight, respectively and the concentration of chlorophyll b were

measured to be 1.09 and 0.35 g mg-1

of fresh weight, respectively, indicating significantly

more chlorophyll formation due to inoculation with sid+ as compared with sid -. The uptake

of Fe by roots and its rate of translocation to the shoots were greater for the sid+ treated

plants as compared with the sid - treated ones, indicating that siderophores increased the rate

of Fe uptake by wheat. The effect of microbial inoculation on shoot Zn was not significant,

but increased the concentration of Zn on roots compared with control. The results suggested

that the siderophores of Pseudomonads may involve on increasing bioavailability of iron.

Rana et al. (2012) conducted a pot experiment in net house conditions, with three

rhizobacterial strains AW1 (Bacillus sp.), AW5 (Providencia sp.) and AW7 (Brevundimonas

sp.), applied along with 2/3 recommended dose of nitrogen (N) and full dose of phosphorus

(P) and potassium (K) fertilizers (N90P60K60). An enhancement of 14–34% in plant biometric

parameters and 28–60% in micronutrient content was recorded in treatments receiving the

combination of AW1 + AW5 strains, as compared to full dose of fertilizer application. The

treatment involving inoculation with AW5 + AW7 recorded highest values of % P and N,

with a two-fold enhancement in phosphorus and 66.7% increase in N content, over full dose

application of P and K fertilizers. A significant correlation was recorded between plant

biomass, panicle weight, grain weight, N, P and iron (Fe) with acetylene reduction activity,

indicating the significance of N fixation in overall crop productivity. Our study illustrates the

multiple benefits of plant growth promoting rhizobacteria (PGPR) inoculation in integrated

nutrient management and biofortification strategies for wheat crop.

Misra et al. (2012) isolated and characterized mineral phosphate solubilizing (MPS)

bacteria associated with the Achyranthes aspera L. plant (prickly chaff, flower plant). Out of

35 bacterial isolates, 6 isolates with high mineral phosphate solubilizing (MPS) activity were

subjected to the assessment of MPS activity under various stress conditions, viz. ZnSO4

(0.30- 1.5 M), NaCl and temperature. High MPS activity of RP19 at higher ZnSO4

concentrations suggested its potential as efficient biofertilizer for growing plants in metal

ZnSO4 contaminated soil.

http://www.tandfonline.com/author/Rana%2C+Anuj

20

Wang et al. (2013) isolated three bacterial strains from the dripping water in Heshang

cave, central China, identified as Exiguobacterium aurantiacum E11, Pseudomonas

fluorescens P35, and Pseudomonas poae P41, respectively. Microbial capabilities in the

dissolution of phosphorus containing minerals were tested with zinc phosphate (Zn3(PO4)2)

in batch culture at 30° C. P. fluorescens and P. poae, the well-known phosphorus solubilizing

bacteria (PSB) were observed to solubilize Zn3(PO4)2 with an efficiency of 16.7% and

17.6%, respectively. Soluble Zn (II) concentration reached up to 370 mg-1

in the system

inoculated with E. aurantiacum E11, inhibition of microbial growth was not detected by

spectrophotometer.

Kumar et al. (2013) reported that zinc solubilising bacteria are potential alternates for

zinc supplement. Among 10 strains screened for Zn solubilisation, broth amended with

ZnCO3 (17 and 16.8 ppm) and ZnO (18 and 17 ppm), respectively. Short term pot culture

experiment with maize revealed that seed bacterization with P29 @ 10 g·kg−1

significantly

enhanced total dry mass (12.96 g) and uptake of N (2.268%), K (2.0%), Mn (60 ppm), and Zn

(278.8 ppm).

Kumar et al. (2013) conducted short term pot culture experiment with maize and

revealed that seed bacterization with P29 @ 10 g⋅kg−1

significantly enhanced total dry mass

(12.96 g) and uptake of N (2.268%), K (2.0%), Mn (60 ppm), and Zn (278.8 ppm).

A comparitive study between the isolated indigenous bacteria and chemical Zn

fertilizer (ZnSO4.7H2O) was conducted by Vaid et al. (2014) to evaluate their potential to

augment Zn nutrition of Zn responsive (NDR 359) and Zn non responsive (PD 16) varieties

of rice under the green house. Three bacterial strains namely; BC, AX and AB isolated from a

Zn deficient rice-wheat field belonging to the genera Burkholderia and Acinetobacter were

investigated for the growth promotion and Zn uptake in rice plants. The plant growth

promotory properties such as Zn solubilization and IAA production of the isolates was

checked in a previous study. These three isolates when used individually or in combination

were found effective in significantly increasing the mean dry matter yield/pot (12.9%),

productive tillers/plant (15.1%), number of panicles/plant (13.3%), number of grains/panicle

(12.8%), grain yield (17.0%) and straw yield (12.4%) over the control and Zn fertilizer

treatment, respectively. Bacterial inoculations also significantly enhanced the total Zn

uptake/pot (52.5%) as well as grain methionine concentration (38.8%).

Imran et al. (2014) reported that application of bioinoculants to soil can promote Zn

availability to plants.

21

Babich and Stotzky (2014) reported that 10 mm concentration of Zn2+

decreased the

survival of Escherichia coli; enhanced the survival of Bacillus cereus; did not significantly

affect the survival of Pseudomonas aeruginosa and Norcardia coralline completely inhibited

mycelial growth of Rhizoctonia solani and reduced mycelial growth of Fusarium solani,

Cunninghamella echinulata, Aspergillus niger and Trichoderma viride. The toxicity of zinc

to the fungi, bacteria and coliphages was unaffected, lessened, or increased by the addition of

high concentrations of NaCl. The increased toxicity of zinc in the presence of high

concentrations of NaCl was not a result of a synergistic interaction between Zn2+

and elevated

osmotic pressures but of the formation of complex anionic ZnCl species that exerted greater

toxicities than did cationic Zn2+

.

11.5 Effect of potassium solubilizing microorganism on fertility status of soil after the

harvest.

Han and Lee (2005) conducted a study to evaluate the potential of phosphate

solubilizing bacteria (PSB) Bacillus megaterium and potassium solubilizing bacteria (KSB)

Bacillus mucilaginosus inoculated in nutrient limited soil planted with egg plant. Results

showed that rock P and K materials either applied alone or in combination did not

significantly enhance availability of soil P and K. PSB increased higher soil P availability

than KSB, which was recommended as a K-solubilizer. Inoculation of these bacteria in

conjunction with amendment of its respective rock P or K materials increased the availability

of P and K in soil, enhanced N, P and K uptake, and promoted growth of egg plant.

Han et al. (2006) conducted an experiment to evaluate the potential of phosphate

solubilizing bacteria (PSB) Bacillus megaterium var. phosphaticum and potassium

solubilizing bacteria (KSB) Bacillus mucilaginosus inoculated in nutrient limited soil planted

with pepper and cucumber. Results showed that rock P and K applied either alone or in

combination did not significantly enhance soil availability of P and K, indicating their

unsuitability for direct application. PSB was more potent P solubilizer than KSB and co-

inoculation of PSB and KSB resulted in consistently higher P and K availability than in the

control without bacterial inoculum and without rock material fertilizer. Integrated rock P with

inoculation of PSB increased the availability of P and K in soil, the uptake of N, P and K by

shoot, root and the growth of pepper and cucumber. Similar but less pronounced results were

obtained when rock K and KSB were added concomitantly. Combined together, rock

materials and both bacterial strains consistently increased further mineral availability, uptake

and plant growth of pepper and cucumber suggesting its potential use as fertilizer.

22

Sugumaran and Janarthanam (2007) isolated K solubilizing bacteria from soil, rocks

and minerals samples viz., orthoclase, muscovite and mica. Among the isolates

B.mucilaginosus solubilized more potassium by producing lime in muscovite and mica.

Meena et al. (2013) studied the potassium solubilizing microorganisms (KSMs) are a

rhizospheric microorganism which solubilizes the insoluble potassium (K) to soluble forms

of K for plant growth and yield. K-solubilization is carried out by a large number of

saprophytic bacteria (Bacillus mucilaginosus, Bacillus edaphicus, Bacillus circulans,

Acidothiobacillus ferrooxidans, Paenibacillus spp.) and fungal strains (Aspergillus spp. and

Aspergillus terreus). Major amounts of K containing minerals (muscovite, orthoclase, biotite,

feldspar, illite, mica) are present in the soil as a fixed form which is not directly taken up by

the plant. Now a days most of the farmers use application of chemical fertilizers injudicious

for achieving maximum productivity. However, the KSMs are most important

microorganisms for solubilizing of fixed form of K in soil system. The main mechanism of

KSMs is acidolysis, chelation, exchange reactions, complexolysis and production of organic

acid. According to literature, currently negligible use of potassium fertilizer as a chemical

form has been recorded in agriculture for enhancing crop yield. Most of the farmers use only

nitrogen and phosphorus and not use the K fertilizer due to unawareness so that the problem

of K deficiency occurs in rhizospheric soils. Therefore, the efficient KSMs should be applied

for solubilization of a fixed form of K to an available form of K in the soils. Our aim of this

review is to elaborate on the studies of indigenous K solubilizing microbes to develop

efficient microbial consortia for solubilization of K in soil which enhances the plant growth

and yield of crops. This review highlights the future need for research on potassium (K) in

agriculture.

Prajapati et al. (2013) showed that potassium-releasing bacterial Enterobacter

hormaechei and fungal Aspergillus terreus strains were examined for plant growth promoting

effects and nutrient uptake on Okra (Abelmoscus esculantus) in K deficient soil in pot

experiments. Inoculation with bacterial strain Enterobacter hormaechei was found to increase

root and shoot growth of Okra and both microorganisms were able to mobilize potassium

efficiently in plant when feldspar was added to the soil. In okra growing in soils treated with

insoluble potassium and inoculated with strain Enterobacter hormaechei and fungal

Aspergillus terreus the potassium content was increased. Among all the three applications of

microbial inoculants, combined application of seed and soil was found to be more effective

on Okra plant growth.

23

Parmar and Sindhu (2016) reported that the concentrations of soluble potassium in

the soil are very low and more than 90% of potassium in the soil exists in the form of

insoluble rocks and silicate minerals. Rhizosphere bacteria have been found to dissolve

potassium from insoluble K bearing minerals. In this study, bacterial isolates were obtained

from wheat rhizosphere on modified Aleksandrov medium containing mica powder as

potassium source. Twenty bacterial strains, among 137 cultures tested, showed significant

potassium solubilization on mica powder supplemented plates and the amount of K released

by different strains varied from 15 to 48 mg-1

. In glucose amended medium broth, bacterial

strains WPS73 and NNY43 caused 41.0 and 48.0 mg-1

of K solubilization. Bacterial strain

WPS73 caused maximm solubilization (49.0 mg L-1

) at 25°C whereas bacterial strain NNY43

caused maximum solubilization at 30° C. K solubilization was found more when bacterial

strains were grown in medium broth with pH 7.0. Maximum K solubilization occurred when

KCl was used as a potassium source followed by K2SO4. These results suggested that the

environmental conditions could be optimized for growth of potassium solubilizing bacteria

and these bacterial cultures could be exploited for plant growth improvement under field

conditions.

Parmar et al. (2016) noted that from 25 isolates, 5 bacterial isolates were selected

exhibiting highest potassium solubilization and characterized on the basis of colony,

morphological and biochemical characteristics. The highest solubilization (46.52 μg/ml) was

observed in isolate KSB-1 and was identified as Bacillus licheniformis by Biolog system and

was followed by KSB-3 (42.37 μg/ml) which was identified as Bacillus subtilis.

11.6 Effect of Zinc solubilizing microorganism on fertility status of soil after the harvest.

Fasim et al. (2002) isolated airborne bacteria from the tannery air environment and

screened them for the solubilization of insoluble zinc sources like zinc oxide and zinc

phosphate. Out of 10 strains tested, Pseudomonas aeruginosa (CMG 823) strain showed the

best solubilization. Colonies of the bacterium produced clear haloes on solid medium which

contained these insoluble metal compounds but only when glucose was provided as a carbon

source and, no haloes produced around the colony when the plates contained gluconic acid as

a carbon source.

Saravanan et al. (2003) assessed zinc solubilizing ability of Bacillus sp. and

Pseudomonas sp. using zinc oxide, zinc sulphide and zinc carbonate in both plate and broth

assays. ZSB-O-1 (Bacillus sp.) showed highest dissolution in the zinc sulphide (Sphalerite

24

ore), with 2.80 cm of dissolution zone in the plate assay and 13.60 mg kg-1

of zinc in the

broth assay on the 15th

day after inoculation. The ZSB-S-2 (Pseudomonas sp.) showed more

solubilizing ability in the zinc oxide, with 3.30 cm clearing zone in the plate assay and 16.40

mg kg-1

of zinc in the broth assay over the same inoculation period and isolate ZSB-S-4

(Pseudomonas sp.) had highest solubilizing potential in zinc carbonate with 6.20 cm of

dissolution zone in the plate assay and 13.40 mg kg-1

of zinc in the broth assay. Thus, the

solubilization potential varies among different cultures.

Shahab and Ahmed (2008) studied the zinc phosphate solubilization efficiency of ten

soil bacteria for various parameters like carbon sources, temperature, pH, variable

concentrations of sodium chloride and glucose and found that for majority of the isolates, 200

C was found to be the optimum temperature for solubilization of zinc phosphate and glucose

was the most favorable carbon source for solubilization while lactose was the least favorable

carbon source and pH 7 was the most favorable pH for solubilization. Isolates CMG851

(Acinetobacter lwoffi) and CMG852 (Pseudomonas sp.) showed enhanced solubilization in

presence of 1% sodium chloride.

Bapiri et al. (2012) reported that zinc solubilizing ability of Pseudomonas fluorescent

was evaluated using zinc oxide, zinc carbonate and zinc sulphide in both plate and broth

media assays. Forty bacterial strains and 0.1% of each chemical source in six replications

were used. There were no halos observed in zinc sulphide. The concentration of soluble Zn

for ZnO was 28-625 mg-1

and pH was shifted from 7.0-7.2 to 3.90-6.50 and for ZnCO3 was

247-753 mg-1

and pH was shifted from 7.0-7.2 to 3.5-6.3 after 5 days of inoculation in 28º C.

Desai et al. (2012) isolated Azotobacter, Azospirillum, Bacillus and Pseudomonas

strains under in vitro conditions from diverse crop production systems and evaluated them for

solubilization of „Zn‟ and „P‟ in vitro from insoluble zinc (ZnO, ZnCO3) and phosphorus

[tricalcium phosphate (TCP)], respectively. After 15 days of incubation, 15 strains solubilized

zinc and produced >50 cm2 solubilization zone on solid media.

Sharma et al. (2012) recovered one hundred thirty four putative Bacillus isolates from

soybean rhizosphere soils of Nimar region. These isolates were screened in vitro form zinc

solubilization ability and isolates KHBD-6, KHBAR-1, BDSD-2-2C and KHTH-4-1 and the

reference strain ATCC 13061 had higher soluble zinc concentration in liquid medium

supplemented with zinc phosphate and zinc carbonate compounds as compared with the other

isolates and uninoculated control.

Joshi et al. (2013) recovered two bacterial isolates (B1 and B2) from the rice

rhizosphere which produced siderophores, indole acetic acid and solubilized

25

insolublenphosphate, ZnO and ZnSO4.7H2O. Based on routine biochemical tests, both the

isolates were 6 characterized as species of Pseudomonas. A shift in pH from 7 to 5 was

observed in minimal medium supplemented with Zn compounds after 15 days of bacterial

growth. Isolate B1 solubilized maximum amount of ZnO (1.348 ppm) after 15 days of

incubation.

Ramesh et al. (2014) isolated Bacillus aryabhattai related bacterial isolates with zinc

solubilizing abilities and assured that the strains MDSR7 and MDSR14 produced

substantially higher soluble zinc content with significant decline in pH and increase in total

organic acid production in Tris-minimal broth supplemented with insoluble zinc compounds.

They concluded that these strains substantially influenced mobilization of zinc and its

concentration in edible portion, yield of soybean and wheat and can be utilized as

bioinoculants for biofertilization and biofortification. This assumes significance as the

increased zinc concentration found in this study has large implications in terms of

overcoming zinc malnutrition.

Sharma et al. (2014) isolated forty eight endophytic bacterial isolates from soybean

(43) and summer mungbean (5) rhizosphere and screened them for zinc solubilizing ability in

Tris- minimal medium separately amended with inorganic zinc compounds viz. zinc oxide

(ZnO) and zinc phosphate {Zn3(PO4)2} by plate assay method. These zinc solubilizing

isolates were assessed for morphological, biochemical and plant growth promoting (PGP)

traits in vitro. Out of 48 endophytic bacterial isolates, isolate no. 3 and 17 were able to

solubilize ZnO whereas 2 and 12 were able to solubilize Zn3(PO4)2 on Tris-minimal medium.

Endophytes 1J (Klebsiella spp.) and 19D (Pseudomonas spp.) were found to be promising

bacterial isolates as they solubilized both inorganic sources of zinc separately supplemented

in Tris- minimal medium along with good PGP traits (P solubilization & indole acetic acid)

and can be exploited as potential biofertilizers.

Suganya and Saravanan (2014) reported that the application of graded levels of zinc

fertilizer and zinc solubilizing bacteria under simulated moisture conditions in texturally

varied soils, an incubation experiment was conducted during Jan 2014 with graded levels of

zinc fertilizers, with and without zinc solubilizing bacteria inoculation as treatments. Soil

available zinc content analysed at 30, 60 and 90 days after incubation was found to be more

in the clay loam soil as compared to sandy loam soil irrespective of moisture and the doses of

ZnSO4 application. Irrespective of soil and moisture regime the zinc solubilizing bacteria

enhanced the available zinc content of the soils.

26

Kumar et al. (2014) reported that zinc is an essential micronutrient which plays a

major role in the growth and productivity of the plants. Zinc (Zn) deficiency hinders

metabolic and physiological activity in plants due to its inevitable role as an enzyme cofactor.

Many Indian soil exhibit Zn deficiency with the content much below the critical level of 1.5

ppm. The conditions that make unavailability of zinc to plants are high pH, low organic

matter content, high usage of P fertilizer, less textured soil and utilization of synthetic

fertilizer to correct Zn deficiency which results in unavailability of zinc after seven days of

application. An alternative eco-friendly approach to overcome Zn deficiency constraint in

plants is by the application of microbial inoculants as a biofertilizer. Rhizospheric

microorganisms play a vital role in the conversion of unavailable form of metal to available

form through solubilization mechanism.

Pawar et al. (2015) conducted laboratory experiment to evaluate the zinc solubilizing

ability of different microorganisms using zinc oxide, zinc carbonate and zinc phosphate in

both plate and broth media assays. Results indicated that by plate assay Trichoderma viridae

formed significantly highest colony diameter (2.33 cm) and halozone diameter (4.10 cm) with

zinc carbonate amended media. Pseudomonas striata form highest clearing zone (2.03 cm) by

zinc carbonate amended media. Solubilization efficiency (237.77%, 216.07% ) and

solubilization index (3.36, 3.16) of Burkholderia cenocepacia and Pseudomonas striata were

indicated maximum solubility in zinc oxide amended media. In Broth culture assay,

Maximum zinc solubilization (458 mg lit-1

) was observed with the Trichoderma viridae in

zinc carbonate amended media compared to control (140.0 mg lit-1

). Maximum reduction in

pH was recorded in Pseudomonas fluorescence ( 3.95) with zinc carbonate amended media

compared to control.

Suganya and Saravan (2015) reported that the available zinc content of the soil as

influenced by the application of graded levels of zinc fertilizer and zinc solubilizing bacteria

under simulated moisture conditions in neutral and sodic soils, an incubation experiment was

conducted during Jan 2014 and the DTPA-Zn was analysed at monthly interval starting from

30th

day after incubation. The results indicated that available zinc content was found to be

more in the neutral soils as compared to sodic soils at all the stages of incubation irrespective

of moisture. In both the soils and moisture regimes, the zinc solubilizing bacteria inoculation

enhanced the available zinc content of the soils.

Solanki et al. (2016) reported that an animal performance is expected to depend directly

on fodder quality which relies on various factors including soil fertility status, where zinc is

very important nutrient. It is needed in small quantity, but plays indispensable role in the

27

metabolism of dairy cows. Zinc plays a number of important functions. Among many others,

its role in strengthening the immune system and the reproductive system are notable.

Increasing fodder Zn concentrations through fertilization is an effective method of supplying

Zn to cows. In recent years, the crucial importance of optimal Zn supplies for the health and

sustained productivity of high producing dairy cows has been very well documented. For this

reason, Zn uptake by major dairy forage crops such as the ryegrasses, barseem is of particular

concern. Because of the obvious drawbacks of chemical fertilizes, biofertilizers have gaining

importance in agriculture.The present review focuses the importance of Zn fertilization in

fodder crops through inoculation with zinc solubilizing bacteria.

Sunithakumari et al. (2016) showed that the zinc solubilizing bacteria were isolated from

eight different agricultural fields (banana, chilli, field bean, ground nut, maize, sugarcane,

sorghum and tomato) in and around Coimbatore district of Tamil Nadu. Five isolates were

selected as best strains based on their solubilization efficacy in the qualitative estimation. The

selected five isolates were identified using 16S rRNA as Stenotrophomonas maltophilia

(ZSB-1), Mycobacterium brisbanense (ZSB-10), Enterobacter aerogenes (ZSB-13),

Pseudomonas aeruginosa (ZSB- 22) and Xanthomonas retroflexus (ZSB-23). These strains

were subjected to further studies such as quantitative estimation, influence of the isolates on

the pH of the medium and production of gluconic acid as well as IAA. Of the five bacterial

isolates, Pseudomonas aeruginosa showed maximum solubilization of zinc in the broth and

also maximum decrease in the pH from 7 to 3.3 and recorded highest IAA production. HPLC

analysis of gluconic acid production by the selected isolates indicated their potential to

solubilize zinc.

Krithika et al. (2016) reported that the zinc solubilizing bacteria (ZSB) is an inoculant to

the crop plants could be a sustainable input for Zn fertilization as well as for Zn

biofortification. In order to explore these bacteria, we have conducted a microcosm study to

assess its potential in terms of Zn availability in three different soils of semi arid tropics and

its interaction effects with different insoluble Zn amendments. The ZSB strain, Enterobacter

cloacae ZSB14 was inoculated to semiarid tropical red lateritic, wetland and calcareous soils

with or without zinc amendments viz, zinc oxide (ZnO), zinc carbonate (ZnCO3) and zinc

phosphate (Zn3(PO4)2) under controlled condition

Material and

Methods

28

CHAPTER-III

MATERIALS AND METHODS

A field experiment was conducted during kharif season 2016-2017 entitled “Studies

on effect of potassium and zinc solubilizing microorganism on Mungbean” at Departmental

farm of Soil Science and Agril. Chemistry, College of Agriculture, Badnapur. The details of

materials used and methods adopted during the course of present investigation are explained

in this chapter with appropriate heads.

2.1 Experimental site:

The present experiment was conducted at research farm of Department of Soil

Science and Agril. Chemistry, College of Agriculture, Badnapur. Geographically Badnapur is

situated at 190

52‟00‟‟ North latitude and 75

0 44

‟00‟‟ East longitudes 75.733 and at 498 m

altitude above sea level. Topographically experimental plot was fairly levelled. The soil was

well drained, developed over weathered basaltic materials.

2.2 Experimental soil:

The experimental soil had clay texture, moderately calcareous in nature and slightly

alkaline in reaction, normal in salt content. Before sowing, initial soil sample were collected

randomly from 0-15 cm depth covering experimental area and analysed for various physico-

chemical characteristics and data are presented in Table 1.

2.3 Climate and weather condition:

The data revealed that total rainfall received during crop growth period was 716.5

mm with 38 rainy days.

The meteorological data i.e. variation in rainfall distribution, which was recorded at

Agricultural Meteorological Observatory, College of Agriculture, Badnapur presented in

Table 2. The data revealed that total rainfall received during crop growth period was 716.5

mm.

29

Table 1: Chemical composition of experimental soil based on composite sample.

A) Chemical composition Unit 2015-16

1. pH 7.78

2. EC dSm-1

0.29

3. Organic carbon g kg-1

4.86

4. Calcium carbonate g kg-1

43.12

5. Available nitrogen kg ha-1

143.22

6. Available phosphorus kg ha-1

14.00

7. Available potassium kg ha-1

585

9. Available zinc mg kg-1

0.50

10 Available iron mg kg-1

4.46

B) Biological properties

1. Bacteria CFU x 10-7

g-1

soil 35

2. Actinomycetes CFU x 10-5

g-1

soil 16.50

3. Fungi CFU x 10-4

g-1

soil 4.85

30

Season and Crop Condition during 2016

Table 2: Meteorological data recorded during the course of investigation at Badnapur.

M.W. Date

Total

rain

fall

(mm)

Rainy

days

(No.)

Temperature

( 0C )

Relative

Humidity

(%)

Min Max. Mor.

23 31st May-6

th June 1 22.97 33.48 56.42

24 7th

-13th

June 4.5 1 25.94 36.85 67.85

25 14th

-20th

June 12 1 24.51 36 58.14

26 21st-27

thJune 27.5 4 21.18 32.08 57

27 28th

June-4th

July 26 3 21.2 31 79.71

28 5th

-11th

July 241 4 20.42 26.74 91.28

29 12th

-18th

July 16 2 17.92 28.85 97.71

30 19th

-25th

July 21.5 3 21.64 28.28 94.28

31 26th

July-1st Aug 11.5 4 22 28.64 93.71

32 2nd

-8th

Aug 94 4 21.5 26.52 95.14

33 9th

-15th

Aug 7 1 22.21 29.6 94.28

34 16th

-22nd

Aug - 21.64 30.35 87

35 23rd

-29th

Aug 8 1 22.17 31.07 85.28

36 30th

Aug -5th

Sept 27 2 21.65 28.71 87.42

37 6th

-12th

Sept 7 1 21.14 29.04 92.28

38 13th

-19th

Sept 76.5 4 18.74 29.07 90.57

39 22nd

-26th

Sept 13.5 1 22.14 29.07 94.28

40 27th

Sept-3rd

oct 23.5 2 22.28 30.28 92.28

Total 716.5 38 391.5 545.3 1514.63

Average 21.73 30.29 84.14

The total rainfall received during the year 2016 (31st May to 3

rd October) was716.5

mm in 38 rainy days. Monsoon commenced with 4.5 mm precipitation in 1 rainy day of 24th

MW (7th

-13th

June). Likewise 12 mm rainfall in 1 rainy days received in 25th

MW (14th

-20th

June) and 27.5 mm rainfall in 4 rainy days received in 26nd

MW (21nd

-27th

Jun). During 27th

MW (28th

Jun-4th

July), 26 mm rainfall was received in 3 rainy days. Likewise in 28th

MW

(5rd

-11th

July) 241 mm rainfall was received in 4 rainy days and 16 mm rainfall in 2 rainy

days received in 29th

MW (12th

Aug -18th

July).During 30th

MW (19th

-25th

July), 21.5 mm

rainfall was received in 3 rainy days. Likewise 11.5 mm rainfall in 4 rainy days received in

31th

MW (26th

July-1th

Aug) and 94 mm rainfall in 4 rainy days received in 32nd

MW (2nd

-8th

Aug). During 33th

MW (9th

-15th

Aug), 7 mm rainfall was received in 1 rainy days. Likewise

in 35th

MW (23rd

-29th

Aug) 8 mm rainfall was received in 1 rainy days and 27 mm rainfall in

2 rainy days received in 36th

MW (30th

Aug -5th

Sept).During 37th

MW (6th

-12th

Sept), 7 mm

31

rainfall was received in 1 rainy days.During38th

MW (13th

-19th

Sept) 76.5mm rainfall in 4

rainy days. 13.5mm rainfall received in 1 rain days in 39th

MW (22th

-26th

Sept)13.5mm

rainfall received in 1 rainy days. 23mm rainfall received in 2 rainy days in 40th

MW (27th

Sept-3th

Sept).

The sowing of Mungbean Kharif crops was started in 25th

MW i.e. from 20th

June

2016 and harvesting completed in 40th

MW i.e. 227th

Sept. 2016 at Agriculture Research

Station, Badnapur. The total rainfall 716.5 mm was received in 38 rainy days during 23rd

to

40th

MW (31st May – 3

rd Oct).

2.4 Experimental details:

The experiment was laid out in Randomised Block Design (RBD) with 5 treatments

and replicated four times. The green gram was sown on 26th

July 2016 by adopting 30 cm X

10 cm spacing and was harvested on 30th

september 2016 at maturity. The green gram variety

BM 2002-1 was used. The treatment details are given below

2.4.1 Treatment details:

T1= Absolute control ( No fertilizer )

T2= RDF (25:50:00 N, P2O5, K2O kg ha-1

) + Rhizobium

T3= RDF + Rhizobium + PSB + Potassuim solubilizing microorganism

T4 = RDF + Rhizobium + PSB + Zinc solubilizing microorganism

T5 = RDF + Rhizobium + PSB + Potassium solubilizing microorganism + Zinc solubilizing

microorganism

32

Where,

Basal dose of RDF i.e. 25: 50: 00 N, P2O5, kg ha-1

and K2O as per treatment was

applied at the treatment at the time of sowing (before sowing). The quantity of fertilizers

applied per plot as per the treatments are given in Table 3.

Table 3: Quantity of nutrient applied in each plot

The calculated amount of K as Muriate of Potash (MOP) and recommended dose of N

and P was applied as Urea, and DAP was applied at the time of sowing.

Treatment Urea (g) DAP

(g)

MOP

(g)

T1: Absolute control (No fertilizer) - - -

T2: RDF Only (25:50:00 N, P2O5, K2O kg ha-

1) + Rhizobium

173 156 00

T3: RDF + Rhizobium + PSB + KSB

173

156 00

T4: RDF + Rhizobium + PSB + ZnSB 173

156 00

T5: RDF + Rhizobium + PSB + KSB + ZnSB 173

156

00

33

Plan of layout

……..

Net plotGros s plot

15 Rows (4.5 m)

40

pla

nts

(4

m)

Gros s plot : 4 .50 m x 4 .00 m

Net plot s ize : 3 .60 m x 3 .80 m

28

T2

T3

T4

T3

T4

T5

T4

T5

T1

T5

T1

T2

T1

T2

T3

T1

1 m

4 m

1 m

N

T2

T3

T4

T5

34

2.4.2 Seed and sowing:

Green gram variety BM 2002-1 was selected for kharif season for present study. The

germination test was carried out before sowing. The sowing was done by line showing

maintained the spacing of 30 cm from row to row and the plant to plant distance was 10 cm.

Gap filling was done wherever it is necessary to maintain the plant population in each plot,

periodical intercultural operations like thinning, weeding were carried out and treatment plots

were maintained for good crop growth.

2.5 Other details

2.5.1 Soil and plant sample collection:

Green gram plant samples (5 plants from each observation unit of treatment plot)

were uprooted at crop growth stage i.e. Flowering and harvesting for chemical analysis. At

the same time, soil samples from each plot were also collected to study the soil nutrient

content.

2.5.2 Biometric observations:

Observations on the crop characteristics indicating growth of the crop i.e. number of

nodules per plant, pods per plant, total biomass production per plant, dry matter per plant

were recorded at the crop growth stages of the crop flowering and at harvesting stage from

the plants, uprooted from observation unit.

2.5.3 Germination percentage:

The germination percentage was recorded from net plot of experimental unit.

Germination (%) = Total no. of seeds germinated / Total no. of seeds sown × 100

2.5.4 Nodulation:

Five plants from observation plot were randomly removed with the help of the fork

without damaging the roots at flowering and harvesting stage of green gram. The roots were

washed carefully to remove the soil stucking to them and nodules were counted.

2.5.5 Dry matter per plant:

Five plants uprooted from the observation unit for recording the dry matter weight.

After removing the roots, plant samples were kept in well labelled brown paper bag. First the

samples are dried in shade and after that kept in oven at 650C±2

0C, and then weight of dry

matter was taken and expressed on per plant basis.

35

2.5.6 Total number of pods per plant:

Number of pods from five selected plants were counted and average number of pods

per plant was worked out.

2.5.7 Seed yield:

The plants from each net plot were harvested and seeds were separated by threshing,

after sun drying the pods seed yields obtained in each net plot were weighted (kg) and further

it was calculated on the hectare basis (kg ha-1

).

2.6 Soil analysis:

Surface soil samples (0-30 cm) were collected from different plots of the layout and

were thoroughly mixed, air dried and ground with wooden mortar and pestle and passed

through 2 mm sieve. The sieved sample was stored in bag with proper labelling. The methods

given below adopted for analysis of physico-chemical properties of soil.

2.6.1 Soil reaction (pH)

It was determined in (1: 2.5) Soil: Water Suspension using digital pH meter (Jackson,

1973).

2.6.2 Electrical conductivity (EC)

It was estimated in (1: 2.5) Soil: Water suspension using direct read type conductivity

meter (Jackson, 1973).

2.6.3 Organic carbon:

Walkley and Black wet digestion method was used for the determination of organic

carbon from soil (Jackson, 1973).

2.6.4 Free calcium carbonate:

It was determined by rapid titration method as suggested by Piper (1966).

2.6.5 Available soil nitrogen:

It was determined by using alkaline potassium permanganate method as described by

Subbiah and Asija (1956).

2.6.6 Available soil phosphorous:

Phosphorous from soil was extracted by 0.5 M sodium bicarbonate at a constant pH

8.5 and measured colorimetrically at 420 nm as described by Olsen (Olsen et al., 1954).

2.6.7 Available soil potassium:

It was determined by using neutral normal ammonium acetate as an extractant using

flame photometer (Jackson, 1973).

2.7 Plant analysis

36

2.7.1 Preparation of Plant samples:

For the determination of nutrient contents in plant samples, the samples were

collected at different growth stages of the crop. First of all the fresh plants were washed with

tap water and roots were discarded. Preparation of plants samples are carried out first by sun

drying and then oven drying. The dried samples were grind in electrically operated stainless

steel grinder to maximum fineness. All the precautions were taken to avoid the contamination

from other plant materials. The grind plant materials were stored in the paper bags and used

for further chemical analysis.

2.7.2 Digestion:

0.5 g of fine powdered plant sample was taken in 100 ml conical flask.5 ml

concentrated nitric acid added to it and kept for over night. On the next day, 10 ml of diacid

mixture (HNO3 and HClO4 in 9:4) was added and digested in hot plate as described by Piper

(1966). After digestion, known volume was prepared with glass distilled water and filtered.

The same extract was used for the estimation of P, K.

2.7.3 Nitrogen concentration:

Total Nitrogen in plant and seeds were determined by micro-kjeldhal,s method as

described in A.O.A.C (1975). One gram of plant/defatted seed sample was digested with 1 g

K2SO4, 0.5 g CuSO4.5H2O and 25 ml H2SO4 and then it was distilled with 40 % NaOH. The

distillate was collected in a beaker containing 4 % boric acid. The methyl red and

bromocresol green indicator was used. The contents were back titrated with 0.1 N H2S04 until

pink colour was obtained.

2.7.4 Phosphorous concentration:

The digest prepared with diacid mixture was used for determination of phosphorous

by using vanadomolybdate solution. The phosphorous was estimated by

vanadomolybdophosphoric acid yellow colour method prepared using spectrophotometer,

Jackson (1973). The intensity of yellow colour was measured on at 420 nm wavelength.

2.7.5 Potassium concentration:

The diacid extract was used for potassium determination. It was determined on flame

photometer as suggested by Jackson (1973).

2.8 Analysis for quality parameters.

2.8.1 Protein content:

The nitrogen content from the grain samples was estimated by Microkjeldhals method

(A.O.A.C. 1975) and N content was multiplied by 6.25 to get percent crude protein.

37

2.9 Uptake of nutrients:

Nutrient uptake i.e. uptake of N, P, K, was calculated by considering grain and dry

matter yield at harvest in particular treatment plot in relation concentration of the particular

nutrient in respective treatment plot using the formula.

Nutrient concentration % X (dry matter yield (kg ha-1

)

Uptake (kg ha-1

) = -------------------------------------------------------------------------

100

2.10 Economics in terms of green gram:

Economics of cultivation was worked out as per the following formulae.

Gross Monetary Returns (GMR) = Yield X selling price of Green gram.

Net Monetary Returns (NMR) = GMR – Cost of cultivation

NMR

Benefit Cost Ratio (B: C Ratio) = ------------ X 100

COC

Where,

GMR - Gross monetary return.

NMR - Net monetary return.

B : C - Benefit cost ratio.

2.11 Biological properties:

For isolation of bacteria, fungi and actinomycetes from soil. Three different media

were used for specific group of flora. The composition of these media is given below.

2.11.1 Nutrient Agar medium (For Bacteria)

1. Beef extract : 3 g

2. Peptone : 5 g

3. NaCl : 1 g

4. Agar Agar : 15 g

5. Distilled water : 1000 ml

38

6. pH to be adjusted : 7

2.11.2 Ken knight medium (For actinomycetes)

1. Dextrose : 1 g

2. KH2SO4 : 0.1 g

3. NaNO3 : 0.1 g

4. KCl : 0.1 g

5. MgSO4 : 0.1 ml

6. Agar- Agar : 15 g

7. Distilled water : 1000 ml

8. pH to be adjusted : 7

2.11.3 Potato Dextrose Agar media (for fungi)

1. Potato : 200gm

2. Dextrose : 20gm

3. Agar : 20gm

4. Distilled Water : 1 Litre

Preparation of medium:

Agar-Agar was boiled in 500 ml of distilled water in a pan. In another pan

about 500 ml distilled water and all chemical ingredients were added and mixed properly.

Both these ingredients were mixed together properly, filtered and made up the volume to

1000 ml with distilled water. The respective media were distributed in 500 ml conical flasks,

plugged with non absorbent cotton, tied these plugs with paper by thread and sterilized at

6.82 kg (15 lb) pressure for 15 min in an autoclave.

Method:

Dilution plate technique is one of the most popular method for isolation and

enumeration of soil fungi, actinomycetes and bacteria as follows.

Procedure (Dhingra and Sinclair, 1993)

Transfered of 1 g of soil sample in 10 ml of sterile distilled water in test tube

(1:10), shake properly. Then Transfered 1 ml suspension from this test tube to another tube

containing 9 ml of sterile distilled water (1:100) again transfered 1 ml of suspension from this

39

test tube to tube containing 9 ml sterile distilled water (1:1000). Similarly, the dilution

process was continued as per requirement for fungal isolation 1:104. For bacteria 1:10

7 and

for actinomycetes 1:105dilution were preferred. The concerned diluted samples were poured

at rate of 1 ml/plate. The respective melted medium (cool to 450) was poured at rate of 20

ml/plate. Spread the medium by an inclined rotary motion of the plate. After solidification of

medium, these plates were incubated at 30_+ 20 in an inverted position in incubator.

2.12 Statistical Analysis:

The experiment was laid out in Randomised Block Design(RBD) and data obtained

from growth parameters, yield attributing characters, quality parameters, soil analysis and

plant analysis was complied and statistically analysed as per the method given in “Statistical

Methods for Agricultural Workers” by Panse and Sukhatme (1985) using computer

programme. Appropriate Standard Errors (SE) were worked out. Critical differences (C.D.) at

5 % and CV were calculated and presented in chapter results and discussion.

Results and Discussion

40

RESULTS AND DISCUSSION

The present investigation was undertaken to study the effect of potassium and

zinc solubilizing micro-organism on growth, yield and uptake of nutrients in mungbean .The

results obtained in respect of various growth and quality parameters and nutrient uptake

under various treatments are presented, interpreted and discussed in this chapter under

following subheads.

4.1 Effect of potassium and zinc solubilizing microbes on chemical properties of

soil.

4.2 Effect of potassium and zinc solubilizing microbes on germination and final

plant stand.

4.3 Effect of potassium and zinc solubilizing microbes on growth parameters and

grain yield of mungbean.

4.3.1 Effect of potassium and zinc solubilizing microbes on number of pods per plant.

4.3.2 Effect of potassium and zinc solubilizing microbes on number of nodules per plant.

4.3.3 Effect of potassium and zinc solubilizing microbes on total biomass production (g

plant-1

).

4.3.4 Effect of potassium and zinc solubilizing microbes on dry matter (kg ha-1

).

4.3.5 Effect of potassium and zinc solubilizing microbes on economic yield of mungbean

4.4 Effect of potassium and zinc solubilizing microbes on quality parameters on

green gram.

4.4.1 Effects of potassium and zinc solubilizing microbes on protein content.

4.5 Nutrient status of the experimental soil at various crop growth stages.

4.6 Effect of potassium and zinc solubilizing microbes on plant nutrient

concentration at critical growth stages.

4.6.1 Effects of potassium and zinc solubilizing microbes on nitrogen concentration at

critical growth stages.

4.6.2 Effects of potassium and zinc solubilizing microbes on phosphorus concentration at


4.6.3 Effects of potassium and zinc solubilizing microbes on potassium concentration at


41

4.6.4 Effects of potassium and zinc solubilizing microbes on micronutrient (Fe, Zn,)


4.7 Effect of potassium and zinc solubilizing microbes on plant nutrient

concentration and uptake (at harvesting).

4.7.1 Effects of potassium and zinc solubilizing microbes on nitrogen concentration and

uptake at harvesting.

4.7.2 Effects of potassium and zinc solubilizing microbes on phosphorus concentration

and uptake at harvesting.

4.7.3 Effects of potassium and zinc solubilizing microbes on potassium concentration

and uptake at harvesting.

4.7.4 Effect of potassium and zinc solubilizing microbes on micronutrient (Fe, Zn,)

concentration and uptake at harvesting.

4.8 Effect of potassium and zinc solubilizing microbes on microbial population of

Mungbean.

42

4.1 Effect of potassium and zinc solubilizing micro-organism on chemical

properties of soil before sowing and after harvesting of Mungbean.

The soil analysis of the treatment plots were carried out before the establishment of

field experiment and at harvest of the crop. The data there of are presented in Table 4.1.

Table 4.1 Effect of potassium and zinc solubilizing microorganism on chemical

properties of soil before sowing and after harvesting of Mungbean

Treatments

pH EC

( dSm-1

)

Organic

Carbon

(g kg-1

)

CaCO3

(g kg-1

)

Initial

Before Sowing 7.83 0.285 7.72 48.30

After harvest

T1 Absolute control (No fertilizer) 7.60 0.261 7.88 53.37

T2 RDF (20:50 N and P2O5 kg ha-1

) 7.78 0.263 8.01 52.85

T3 RDF + Rhizobium+ PSB+ KSB 7.59 0.268 8.10 54.05

T4 RDF + Rhizobium+ PSB+ ZnSB 7.81 0.266 8.33 55.60

T5 RDF + Rhizobium+ PSB+ KSB+ ZnSB 7.64 0.272 8.54 57.39

SEm± 0.06 0.003 0.033 2.17

CD at 5% NS NS 0.100 NS

The experiment was conducted on research farm of Department of Soil

Science and Agricultural Chemistry, College of Agriculture, Badnapur. The experimental soil

was fine, smectitic calcareous, iso-hyperthermic Typic Haplusterts.

The soil was alkaline in reaction (pH-7.83) where mean pH after harvest was

7.68 which was slightly decreased. This results are similar to Sunitakumari et al. (2016) who

revealed that, among the ZSB five isolates P. aeruginosa (ZSB-22) caused maximum decline

in pH. The FTIR spectral analysis by Fasim et al. (2002) concluded that gluconic acid was

attributed to the release of Zn ions formed from the Zn metal due to decrease in the pH

caused by bacterial growth.

The soil was safe in soluble salt concentration (EC - 0.285 dSm-1

) before

sowing where mean EC after harvest was 0.266 dSm-1

which was slightly decreased.

43

Raghuwanshi et al. (1988) reported that change in EC values were very close margin due to

combined application of bio-inoculants with chemical fertilizers. Further, Bharadwaj and

Narayanasamy (1994) did not found any change in EC with long term effect of manuring and

fertilization.

The soil was high in organic carbon content (7.72 g kg-1

). However, there was

significant increase in organic carbon due to application of the ZnSB. While, Meshram et al.

(2004) concluded that Biofertilizers integrated with chemical fertilizers showed significant

improvement in soil organic carbon. However, Singh and Najar (2007) recorded the increase

in soil organic carbon in the soil after the harvest of soybean over control significantly when

FYM was applied alone or along with bio-inoculants.

The free CaCO3 content was 48.30 g per kg before sowing where mean

calcium carbonate after harvest of crop was 54.65 g kg-1

which was found to be increased by

0.06 % that seems to be very negligible. At harvest pH, EC, and CaCO3 were not influenced

significantly due to application of various treatment. The organic carbon content was varied

inconsistently due to the application of the fertilizers. It was obvious that the primary soil

properties like pH, EC and CaCO3 content could not change significantly but OC change

significantly due to one crop season (26 Jun-28Aug).

4.2 Effect of potassium and zinc solubilizing micro-organism on germination and final

plant stand.

The data presented in Table 4.2 indicates significant impact of potassium and

zinc solubilizing micro-organism on germination percentage of Mungbean crop.

The results revealed that maximum germination count 582 observed in the plot

which is inoculated with RDF + Rhizobium + PSB + KSB + ZnSB (T5 ) which was at par

with the plot which is inoculated with RDF + Rhizobium + PSB + ZnSB (T4 ) (577.25) and

RDF + Rhizobium + PSB + KSB (T3 ) (576). While, minimum germination count 552 was

observed in the control plot.

Similarly, maximum final plant stand 522 observed in the plot which was

inoculated by RDF + Rhizobium + PSB + KSB + ZnSB (T5 ) which was at par with the plot

which was inoculated by RDF + Rhizobium + PSB + ZnSB (T4 ) (519.75) and RDF +

Rhizobium + PSB + KSB (T3 ) (517.25). While, minimum final plant stand 498 was observed

in the control plot.

44

Table 4.2 Effect of potassium and zinc solubilizing microorganism on germination

and final plant stand.

Treatments

Plant Population

Germination

Count

Final plant

Stand

T1 Absolute control (No fertilizer) 552.00

(92%)

498.00

(83%)


) 554.25

(92.37%)

499.50

(83.25%)

T3 RDF + Rhizobium + PSB + KSB 576.00

(96%)

517.25

(86.20%)

T4 RDF + Rhizobium + PSB + ZnSB 577.25

(96.20%)

519.75

(86.62%)

T5 RDF + Rhizobium + PSB + KSB + ZnSB 582.00

(97%)

522.00

(87%)

SEm± 6.70 3.35

CD at 5% 20.67 10.34

Sivasakthivelan and Stella (2012) found that the seed treatment with

bioinoculants consortium significantly increased the germinaton percentage of sunflower

compared with dual, single inoculation and control. The increased germination percentage of

sunflower might be due to increased survivability exhibited by the microbial consortium on

the spermophere and spermoplane which had increased hormones such as auxins, gibberlins

and cytokinins. The highest germination percentage was observed in the liquid microbial

consortium than carrier based inoculums of microbial consortium.

The combined application of the microbes enhanced the seed germination and

plant growth better than individual application. (Noumavo and Kochoni 2013). Prajapati

(2016) reported that inoculation with selected potassium solubilizing bacteria (KSB),

significantly increased seed germination, root and shoot length and number of leaves grain

yield over uninoculated control in the presence of feldspar in Aleksandrov‟s agar medium.

Verma et al. (2016) reported that incubating KSB with Groundnut the number of un-

germinated seeds significantly decreased in the inoculated treatment. Prajapati and Modi

(2016) The potassium solubilizing microorganisms inoculated seeds showed slightly earlier

seed germination compared to the uninoculated controls. This effect could be due to higher

IAA and GA production shown by KSB-8.

Vijya and Ponnuswamy (1998) reported that ZnSO4 significantly increased

green gram and black gram seed germination. Ashrafuzzaman et al., (2009) reported

45

enhancement in seed germination by plant growth-promoting rhizobacterial (PGPR)

treatments.

4.3 Effect of potassium and zinc solubilizing microorganism on growth parameters and

grain yield.

4.3.1 Effect of potassium and zinc solubilizing microorganism on number of pods of per

plant.


zinc solubilizing micro-organism on number of pods per plant.

The results revealed that at flowering maximum number of pods per plant

(11.8) observed in the plot which was inoculated by RDF + Rhizobium + PSB + KSB + ZnSB

(T5 ) followed by the plot which was inoculated by RDF + Rhizobium + PSB + ZnSB (T4 )

(10.1) and RDF + Rhizobium + PSB + KSB (T3 ) (9.6). While, minimum number of pods per

plant (7.8) was observed in the control plot.

Similarly, at harvesting maximum number of pods per plant (13.2) observed

in the plot which was inoculated by RDF + Rhizobium + PSB + KSB + ZnSB (T5 ) followed

by the plot which was inoculated by RDF + Rhizobium + PSB + ZnSB (T4 ) (12.27) and RDF

+ Rhizobium+ PSB + KSB (T3 ) (11.2). While, minimum number of pods per plant (8.8) was


Table 4.3 Effect of potassium and zinc solubilizing microorganism on number of pods

per plant.

Treatments

Number of pods per plant

Flowering

Harvesting

T1 Absolute control (No fertilizer) 7.8 8.8


) 8.9 10.17

T3 RDF + Rhizobium + PSB + KSB 9.6 11.20

T4 RDF + Rhizobium + PSB + ZnSB 10.1 12.27

T5 RDF + Rhizobium + PSB + KSB + ZnSB 11.8 13.20

SEm± 0.29 0.20

CD at 5% 0.92 0.63

46

Improvement in pod bearing capacity of crop could be possibly because of

improved N and P fertilization efficiency in the presence of K and Zn. Increased rate of

photosynthetic and symbiotic activity following balanced application of NPK stimulated

better vegetative and reproductive growth of the crop resulting in higher green pod yield.

Thenua et al., (2010) reported that application of 40 kg K2O ha-1

recorded higher pod yield in

soybean. Similar observation was reported by Thesiya et al., (2013) in black gram, Kushwala

(2001) in field pea and Asghar et al., (1994) in blackgram.

Nomen et al., (2015) reported that effect of zinc solubilizer was significant on

pod and haulms yields and found to improve the pod yield by 4.7% and haulm yield by 6.2%.

Effect of zinc solubilizer was not significant on harvest index. Increase in kernel and pod

yield due to zinc solubilizer may be ascribed to its favourable effect on growth and yield

attributes. Similar effect of bio-fertilizer on yield of pigeonpea was also reported by Pandey

et al. (2013).

4.3.2 Effect of potassium and zinc solubilizing microorganism on number of

nodules per plant.


zinc solubilizing micro-organism on number of nodules per plant.

Table 4.4 Effect of potassium and zinc solubilizing microorganism on number of

nodules per plant

Treatments

Number of nodules per plant

Flowering

Harvesting



) 29.07 19.24




SEm± 0.46 0.39

CD at 5% 1.44 1.20

47

The results revealed that at flowering maximum number of nodules per plant

(45.36) observed in the plot which was inoculated by RDF + Rhizobium + PSB + KSB +

ZnSB (T5 ) followed by the plot which was inoculated by RDF + Rhizobium + PSB + ZnSB

(T4 ) (42.06) and RDF + Rhizobium + PSB + KSB (T3 ) (34.94). While, minimum number of

nodules per plant (22.07) was observed in the control plot.

Similarly, at harvesting maximum number of nodules per plant (22.72)

observed in the plot which was inoculated by RDF + Rhizobium + PSB + KSB + ZnSB (T5 )

followed by the plot which was inoculated by RDF + Rhizobium + PSB + ZnSB (T4 ) (21.47)

and RDF + Rhizobium + PSB + KSB (T3 ) (21.01). While, minimum number of nodules per

plant (17.23) was observed in the control plot.

The data on nodulation of green gram revealed that there was significant

variation in nodule number at all the stages. Potassium is required by adenosine S-

triphosphate phosphohydrolase (ATP ase) for the movement of sugars from the apoplast

between the cells of the phloem. In depth scrutiny of data influenced by growth stages

showed that there was continuous decrease in number of nodules per plant from flowering

(34.70) to harvest (20.33). The study by Collins and Duke (1981) on composition of the

effects of K and N2 fixation and photosynthesis in a legume found that potassium was found

to have large effect on nodulation and N2 fixation. Highest number of nodules was observed

at 45 kg K2O ha-1

in groundnut by Salve and Gunjal (2011). Das et al.,( 2012) reported that

Rhizobium inoculation in combination with different micronutrients recorded higher

nodulation. Similar findings reported by Patra et al., (1999) in groundnut.

48

4.3.3 Effect of potassium and zinc solubilizing microorganism on total biomass

production (g plant -1

)


zinc solubilizing micro-organism on total biomass production.

Table 4.5 Effect of potassium and zinc solubilizing microorganism on total biomass

production (g plant-1

)

Treatments

Biomass production (g plant -1

)

Flowering

Harvesting



) 3.20 6.62




SEm± 0.13 0.12

CD at 5% 0.41 0.39

The results revealed that at flowering maximum total biomass production

(4.54 gm/plant) was recorded in the plot which was inoculated by RDF + Rhizobium + PSB +

KSB + ZnSB (T5 ) which is at par with the plot which is inoculated by RDF + Rhizobium +

PSB + ZnSB (T4 ) (4.40 gm/plant) and followed by RDF + Rhizobium + PSB + KSB (T3 )

(3.50 gm/plant). While, minimum total biomass production (2.24 gm/plant) was observed in

the control plot.

Similarly, at harvesting maximum total biomass production (9.14 gm/plant)


followed by the plot which was inoculated by RDF + Rhizobium + PSB + ZnSB (T4 ) (8.51

gm/plant) and RDF + Rhizobium + PSB + KSB (T3 ) (7.80 gm/plant). While, minimum total

biomass production (3.53 gm/plant) was observed in the control plot.

The data indicated the periodical increase in total biomass of green gram.

The average increase in biomass recovery was from 3.50 g plant-1

to 7.12 g plant-1

from

flowering to harvesting stages of the crop. The accumulation of biomass was relatively more

49

at the later part of the crop. This may be due to the effect of both potassium and zinc

solubilizing microorganism. Zhang et al. (2004) reported that the effect of potassic bacteria

on sorghum, which results in increased biomass and contents of P and K in plants than the

control. Potassium plays a major role in growth as it is involved in assimilation, transport and

storage tissue development (Cakmak 2005). Balai et al., (2005) found that highest dry matter

accumulation g plant-1

was obtained by 40 kg K20 ha-1

in Cowpea and Salve and Gunjal

(2011) found the similar result in groundnut. Similarly, Singh and Ram (2001) studied the

effect of Zn on mungbean biomass, grain yield and quality.

4.3.4 Effect of potassium and zinc solubilizing microorganism on dry matter (kg ha-1

)

production.

The data presented in Table 4.6 and depicted in fig.1 indicates significant

impact of potassium and zinc solubilizing micro-organism on dry matter production (kg ha-1

).

The results revealed that at flowering maximum dry matter production (kg

ha-1

) (1965.50) observed in the plot which was inoculated by RDF + Rhizobium + PSB +

KSB + ZnSB (T5 ) followed by the plot which is inoculated by RDF + Rhizobium + PSB +

ZnSB (T4 ) (1857.26 kg ha-1

) and RDF + Rhizobium + PSB + KSB (T3 ) (1612.80 kg ha-1

).

While, minimum dry matter production (kg ha-1

) (865.61) was observed in the control plot.

Similarly, at harvesting maximum dry matter production (kg ha-1

) (3951.73)


followed by the plot which was inoculated by RDF + Rhizobium + PSB + ZnSB (T4 )

(3593.88 kg ha-1


). While,

minimum dry matter production (kg ha-1


50

Table 4.6 Effect of potassium and zinc solubilizing microorganism on dry matter (kg

ha-1

) production.

Treatments

Dry matter (kg ha-1

)

Flowering

Harvesting



) 1223.72 3084.32




SEm± 19.85 21.74

CD at 5% 61.17 67.01

The results revealed that various treatments resulted in increase in mean dry

matter yield with advancement in crop growth stages i.e. from flowering (1504.98 kg ha-1

) to

harvest (3190.78 kg ha-1

.This was due to the effect of K and Zn nutrition on cell elongation,

turger potential in leaves. Such results were also observed in soybean plants as by Mengal

and Arneke (1982). Patil and Dhonde (2009) observed that highest dry matter kg ha-1

with the

application of 50 kg K2O ha-1

in green gram.

Salam et al. (2005) studied the effect of micronutrients (Zn, Mo, Fe) on

fertilization and productivity potential of mungbean and urdbean gave the highest dry matter

accumulation, pods per plant, seeds per pod, 100 seed weight, seed yield per plant, pod and

seed weight per plant, harvest index and production efficiency.

Fig. 1 Effect of potassium and zinc solubilizing microorganism on dry matter (kg ha-1

)

0

500

1000

1500

2000

2500

3000

3500

4000

T1 T2 T3 T4 T5

Dry

ma

tter

kg

ha

-1

Treatments

Flowering

Harvesting

51

4.3.5 Effect of potassium and zinc solubilizing microorganism on Economic yield of

green gram

The data presented in Table 4.7 and fig.2 indicates significant impact of

potassium and zinc solubilizing micro-organism on economic yield of mungbean.

Table 4.7 Effect of potassium and zinc solubilizing microorganism on economic yield of

Mungbean.

Treatments

Economic yield

Kg/ha-1

Gm/Plant



) 932.75 2.75




SEm± 11.88 0.04

CD at 5% 36.61 0.13

The results revealed that maximum yield of green gram (kg ha-1

) (1229.0)


followed by the plot which is inoculated by RDF + Rhizobium + PSB + ZnSB (T4 ) (1091.75

kg ha-1


). While, minimum

economic yield (kg ha-1


Similarly, maximum yield of green gram (gm/plant) (3.65) observed in the

plot which was inoculated by RDF + Rhizobium + PSB + KSB + ZnSB (T5 ) followed by the

plot which was inoculated by RDF + Rhizobium + PSB + ZnSB (T4 ) (3.23 gm/plant) and

RDF + Rhizobium + PSB + KSB (T3 ) (3.05 gm/plant). While, minimum yield of green gram

(gm/plant) (2.28) was observed in the control plot.

The positive effect of K and Zn on crop yield might also be due to its

requirement in carbohydrate synthesis and translocation of photosynthesis and also may be

due to improved yield attributing characters, shoot growth and nodulation. Billore et al.,

(2009) observed seed yield of soybean increase

52

35.6 % over control with the application of 49.8 kg K ha-1

.Similar findings

were observed by Thesiya et al., (2013) in lentil, Patil and Dhonde (2009) in green gram,

Salve and Gunjal ( 2011) in groundnut, Balai et al., (2005).

Malewar et al., (1990) showed that the grain yield of Mungbean increased

with increasing levels of Zn upto 15 kg ha-1

and then decreased al 20 kg ha-1

. The increases in

grain yield of legumes and its attributes due to Zn application were reported by Kushwaha

(1993). Wankhade et al., (1996) and Vairavan et al.,(1997). Singh and Nayyer (1997)

indicated that insufficient supply of Zn to soybean and Mungbean plants from soil

appreciably decreased the growth of crops despite the presence of adequate quantities of

NPK.

Khurana et al. (1996) reported that zinc application increased the grain and

straw yield of rice. Kumar and Singh (1996) reported significant increase in grain and straw

yield with increasing level of zinc over control.

Fig. 2 Effect of potassium and zinc solubilizing microorganism on Yield (kg ha-1

)

0

200

400

600

800

1000

1200

1400

T1 T2 T3 T4 T5

Yie

ld k

g h

a-1

Treatments

Yield kg/ha

53

4.4 Effect of potassium and zinc solubilizing microorganism on quality

parameters of green gram

4.4.1 Effect of potassium and zinc solubilizing microorganism on protein content.


zinc solubilizing micro-organism on protein content.

Table.4.8 Effect of potassium and zinc solubilizing microorganism on Protien

content.

Treatments Protien content

T1 Absolute control (No fertilizer) 17.27


) 18.19

T3 RDF + Rhizobium + PSB + KSB 18.29

T4 RDF + Rhizobium + PSB + ZnSB 18.68

T5 RDF + Rhizobium + PSB + KSB + ZnSB 19.19

SEm± 0.03

CD at 5% 0.11

The results revealed that highest protein content (19.19 %) recorded in seed

inoculated by RDF + Rhizobium + PSB + KSB + ZnSB (T5 ) which was at par with the plot

which was inoculated by RDF + Rhizobium + PSB + ZnSB (T4 ) (18.68 %) and RDF +

Rhizobium + PSB + KSB (T3 ) (18.29 %0). While, lowest protein content (17.27 %) was


Potassium involved in physiological and biochemical functions of plant

growth i.e. enzyme activation and protein synthesis and its application in legumes might have

improved the nitrogen use efficiency which leads to increase the protein content of the crop.

Similar findings were obtained by Farhad et al., (2010), and Salve and Gunjal (2011).

Kapur et al., (1977) reported that the application of 10 ppm of Zn to soybean

grown on Zn deficient soil gave significant increases in protein concentration in plant parts and its

total uptake. Singh and Manohar (1987) indicated that foliar application of zinc sulphate was most

effective in improving protein content of Mungbean. Ram and Murthy (1983) revealed that

protein content of green gram increased significantly with increasing levels of Zn. Singh and

Badhoria (1986) obtained highest content of crude protein at 10 mg Zn kg-1

of soil in green gram

and lentil. Keshwa and Jat (1992) while studying the effect of Zn on pear millet found that the

54

concentration and total yield of protein increased due to application of zinc. Application of 5 kg

Zn ha-1

recorded highest increase in protein content which was significantly superior over the

control (Singh and Yadav, 1997).

It is also reported that application of potassium and zinc increase the

absorption of nitrogen and the N use efficiency. This results in to improvement in growth,

yield and quality of crop.

4.6 Nutrient status of the experimental soil at various crop growth stages.

The available nutrient status of the experimental soil was presented in Appendix I-

IV.

4.6 Effect of potassium and zinc solubilizing microorganism on plant nutrient

concentration at critical growth stages

4.6.1 Effect of potassium and zinc solubilizing microorganism on nitrogen concentration at


From the data presented in Table 4.9 and fig.3 it was revealed that application

of RDF + Rhizobium + PSB + KSB + ZnSB (T5) significantly increased the N concentration

in plant and seed which was significantly superior over control (T1). Significant differences in

N concentration in plants were noticed which was inoculated with K and Zn at all the critical

growth stages. N concentration was found to be highest in (T6) RDF + Rhizobium + PSB +

KSB + ZnSB at flowering, , harvest and in seed (2.96, 2.54, 3.81% respectively) followed by

(T4) RDF + Rhizobium + PSB + ZnSB (2.78, 2.42, 3.71% N) and (T3) RDF + Rhizobium +

PSB + KSB (2.39, 1.78, 3.46% N).

55

Table.4.9 Effect of potassium and zinc solubilizing microorganism on nitrogen


Treatments

N concentration (%)

Flowering Harvesting Seed

T1 Absolute control (No fertilizer) 1.68 1.10 2.73


) 2.07 1.51 3.34

T3 RDF + Rhizobium + PSB + KSB 2.39 1.78 3.46

T4 RDF + Rhizobium + PSB + ZnSB 2.78 2.42 3.71

T5 RDF + Rhizobium + PSB + KSB +

ZnSB 2.96 2.54 3.81

SEm ± 0.09 0.02 0.02

CD at 5% 0.27 0.08 0.06

The nitrogen concentration in green gram plant was highest at flowering stage

and decline at harvest of the crop but the concentration was higher in grain. This trend may

be due to high mobility of the nitrogen from vegetative tissues to reproductive organs after

flowering stages. Similar results were observed by Kherawat et al., (2013) in clusterbean,

Meena, (2013) in green gram and Laxminarayana (2001) in groundnut.

Fig.3 Effect of potassium and zinc solubilizing microorganism on N Concentration (%)

0

0.5

1

1.5

2

2.5

3

3.5

4

T1 T2 T3 T4 T5

N c

on

cen

tra

tio

n (

%)

Treatments

Flowering

Harvesting

Seed

56

4.6.2 Effect of potassium and zinc solubilizing microorganism on phosphorus


The data on phosphorous in plant concentration are presented in Table 4.10

and fig.4 There was increase in P content in different plant parts with increasing inoculation

with potassium and zinc. Maximum phosphorous content was found in RDF + Rhizobium +

PSB + KSB + ZnSB (T5) at flowering (0.42%) in plant and at harvesting stages in plant

(0.32%) and in seed (0.55%). However, treatment differences could not reach to the level of

significance. Similar results were observed by Kherawat et al., (2013) in clusterbean, Meena,

(2013) in green gram.

Table 4.10 Effect of potassium and zinc solubilizing microorganism on phosphorus


Treatments

P concentration (%)




) 0.36 0.27 0.51




ZnSB 0.42 0.32 0.55

SEm ± 0.014 0.007 0.017

CD at 5% 0.045 0.02 0.054

Fig. 4 Effect of potassium and zinc solubilising microorganism on P concentration (kg

ha-1

).

0

0.1

0.2

0.3

0.4

0.5

0.6

T1 T2 T3 T4 T5

P c

on

cen

tra

tio

n (

%)

Treatments

Flowering

Harvesting

Seed

57

4.6.3 Effect of potassium and zinc solubilizing microorganism on potassium


The data on potassium concentration in plant as a result of inoculation with K

and Zn are presented in Table 4.11 and fig.5 The maximum potassium concentration was

recorded with the application of RDF + Rhizobium + PSB + KSB + ZnSB (T5) at flowering

(1.61%) in plant and at harvesting stages in plant (0.83%) and in seed (0.99%) which was

significantly higher than control (T1). This may be due to the synergistic effects of potassium

with other nutrients. Similar results were observed by Meena, (2013) in green gram and

Laxminarayana (2001) in groundnut.

Table 4.11 Effect of potassium and zinc solubilizing microorganism on potassium


Treatments

K concentration (%)




) 1.43 0.71 0.86




ZnSB 1.61 0.83 0.99

SEm ± 0.01 0.01 0.01

CD at 5% 0.04 0.03 0.05

Fig. 5 Effect of potassium and zinc solubilizing microorganism on K concentration(%).

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

T1 T2 T3 T4 T5

K C

on

cen

trat

ion

(%)

Treatments

Flowering

Harvest

Seed

58

4.6.4 Effect potassium and zinc solubilizing microorganism on micronutrients (Fe, Zn,)


The data in respect of iron and zinc in plant and seed are presented in Table

4.12 to 4.13 and fig.6 to 7. The concentration of these micronutrient in plant and seed

influence significantly due to application of potassium and zinc. However, there was

significant increase in these concentration due to inoculation of potassium and zinc. The

results also showed that concentration of Fe and Zn was more in seed as compared to plant.

The concentration was reduced with advancement of crop growth. Further it was very clear

that there was no significant variation in nutrient concentration due to potassium and zinc

application either through soil and foliar. It was also noted that there was no antagonistic

effect of potassium and zinc application on micronutrient concentration in plant during varied

growth stages.

Table 4.12 Effect of potassium and zinc solubilizing microorganism on iron


Treatments

Fe concentration (mg kg-1

)




) 325.09 172.67 375.99




ZnSB 377.40 192.38 405.69

SEm ± 4.21 1.37 2.36

CD at 5% 12.99 4.25 7.11

Fig.6 Effect of potassium and zinc solubilizing microorganism on Fe Concentration

(mg kg-1)

0

50

100

150

200

250

300

350

400

450

T1 T2 T3 T4 T5

Fe c

on

cen

tra

tio

n (

%)

Treatments

Flowering

Harvesting

Seed

59

Table 4.13 Effect of potassium and zinc solubilizing microorganism on zinc


Treatments

Zn concentration (mg kg-1

)




) 45.70 36.32 53.88




ZnSB 55.49 52.96 71.16

SEm ± 1.43 1.27 0.97

CD at 5% 4.42 3.92 2.99

Tariq et al., (2007) also reported that PGPR inoculation significantly increased the

concentration of Zn (23.6 mg kg-1

) in the rice grain over the control without Zn (9.2 mg kg-1

).

Ekin et al., (2011) found phosphorus fertilizer and PSB applications significantly increased

the concentration of micronutrient Zn in the seed of sunflower.

Fig.7 Effect of potassium and zinc solubilizing microorganism on Zn Concentration

(mg kg-1

).

0

10

20

30

40

50

60

70

80

T1 T2 T3 T4 T5

Zn

co

nce

ntr

ati

on

(%

)

Treatments

Flowering

Harvesting

Seed

60

4.7 Effect of potassium and zinc solubilizing microorganism on nutrient uptake

by Mungbean.

4.7.1 Effect of potassium and zinc solubilizing microorganism on N uptake.

The data regarding the nitrogen uptake in green gram with respect to different

effect of potassium and zinc are presented in Table 4.14 and fig.8.

Table 4.14 Effect of potassium and zinc solubilizing microorganism on N uptake.

Treatments Uptake N ( kg ha

-1)

Flowering Harvesting Seed Total


T2 RDF (20:50 N and P2O5 kg ha-

1)

18.88 46.64 103.08 149.72

T3 RDF + Rhizobium + PSB + KSB 38.64 62.60 141.32 203.92

T4 RDF + Rhizobium + PSB +

ZnSB 51.96 87.05 133.14 220.19

T5 RDF + Rhizobium + PSB + KSB

+ ZnSB 57.93 106.83 151.83 258.66

SEm ± 1.07 0.77 1.54 0.77

CD at 5% 3.22 2.39 4.76 2.39

Among the treatments, application of RDF + Rhizobium + PSB + KSB +

ZnSB (T5) recorded significantly higher uptake in flowering (57.93 kg ha-1

), plant at harvest

(106.83 kg ha-1

) and seed (151.83 kg ha-1

) followed by RDF + Rhizobium + PSB + ZnSB

(T4) and RDF + Rhizobium + PSB + KSB (T3). In presence of potassium, the increase in N

uptake could be attributed to enhanced vigour of crop growth with increased utilization and

translocation of N in to plant and synergy between N and K in soil system resulting in the

enhancement of yield. Similar trend was also reported by Kurhade et al., (2014) in black

gram and Sahay et al., (2013).

Govindrajan et al.,(1964) reported a considerable increase in nitrogen content

in pod of horse gram with the application of ZnSO4. Reedy and Rao (1979) observed

increases in nitrogen content at preflowering stage, but decreases at post flowering stage of

gram due to application of zinc. Pathak (1983) reported that the concentration and total

uptake of nitrogen increase in oil seed plants due to zinc fertilization. Yadav et al,.

(1984,1986) studied that the effects of phosphorus and Zn on cowpea and found that the

concentration and total uptake of nitrogen in roots, nodules,shoots and grains increased with

61

application of 80 ppm P and 5 ppm Zn. Singh and Antil (1996) noticed that the nitrogen

uptake in wheat crop increased with Zn application even when no N was applied.

Fig.8 Effect of potassium and zinc solubilizing microorganism on N uptake (kg ha-1

)

0

20

40

60

80

100

120

140

160

T1 T2 T3 T4 T5

N u

pta

ke

(kg

/ha

)

Treatments

Flowering

Harvesting

Seed

62

4.7.2 Effect of potassium and zinc solubilizing microorganism on P uptake

Table.4.15 and fig.9 represent the data on uptake of P under various

treatments administered. The significantly maximum enhancement in phosphorous uptake by

green gram crop was recorded with RDF + Rhizobium + PSB + KSB + ZnSB (T5) recorded

significantly higher uptake in flowering (8.12 kg ha-1

), plant at harvest (12.63 kg ha-1

) and

seed (22.27 kg ha-1

). Followed by RDF + Rhizobium + PSB + ZnSB (T4) and Rhizobium +

PSB + KSB (T3). Similar, results was observed by Kurhade et al., (2014) in black gram.

Table 4.15 Effect of potassium and zinc solubilizing microorganism on P uptake

Treatments Uptake P (kg ha

-1)



T2 RDF (20:50 N and P2O5 kg ha-

1)

4.35 8.40 4.20 12.60


KSB 6.16 9.77 7.54 17.31


ZnSB 7.44 10.42 9.34 19.76


KSB+ ZnSB 8.12 12.63 12.21 24.84

SEm ± 0.22 0.31 0.13 -

CD at 5% 0.68 0.97 0.42 -

Devarajan et al,.(1980) revealed that P content decreased in seed and straw

with application of Zn in pulse crops. Singh and Manohar (1982) on the other hand, found the

foliar application of ZnSO4 was most effective in improving the protein content of grain and

increasing uptake of nitrogen and phosphorus by Mungbean. Yadav et al,.(1985) reported

that while addition of 10 ppm Zn decreased up to 2.5 ppm increased P concentration in

different plant parts of cowpea. Gupta et al,. (1985) reported that application of zinc 5 ppm

while decreased P concentration in plants, its effect was at deleterious n. It indicated an

antagonism between Zn and P at 5 ppm Zn level.

Fig.9 Effect of potassium and zinc solubilizing microorganism on P uptake (kg ha-1

)

0

5

10

15

20

25

T1 T2 T3 T4 T5

P u

pta

ke

(kg

/ha

)

Treatments

Flowering

Harvesting

Seed

63

4.7.3 Effect of potassium and zinc solubilizing microorganism on K uptake.

The data regarding the K uptake in green gram with respect to treatment of

potassium and zinc are presented in Table 4.16 and fig.10.

Table 4.16 Effect of potassium and zinc solubilizing microorganism on K uptake

Treatments Uptake K (kg ha

-1)


T1 Absolute control (No

fertilizer) 11.64 6.50 10.66 17.16

T2 RDF (20:50 N and P2O5 kg

ha-1

) 16.77 16.27 21.04 37.30


KSB 22.41 20.77 25.64 50.04


ZnSB 26.12 26.86 29.29 57.07


KSB + ZnSB 27.83 28.94 32.99 62.19

SEm ± 0.43 0.38 0.44 -

CD at 5% 1.34 1.18 1.38 -

The significantly maximum enhancement in potassium uptake by green gram

crop was recorded in RDF + Rhizobium + PSB + KSB + ZnSB (T5) which was significantly

higher uptake in flowering (27.83 kg ha-1

), plant at harvest (28.94 kg ha-1

) and seed (32.99

kg ha-1

). followed by RDF + Rhizobium + PSB + ZnSB (T4) and Rhizobium + PSB + KSB

(T3). This might be due to application of higher doses of mineral K with micronutrients

favored higher root and shoot development which might have also increased the K uptake.

Similar, result was observed by Kurhade et al., (2014) in black gram and Kherawat et al.,

(2013) in clusterbean.

Singh et al.(1986) reported that the concentration of potassium increased

significantly due to application of Zn in wheat grain and straw. Singh et al,.(1987) observed

that the uptake of potassium increased with application of zinc upto 10 ppm in wheat and its

utilization also increased with increasing levels of applied zinc. Malewar et al. (1990) found

an increase in concentration of N, K and Zn in Mungbean with the application of Zn upto 15

Kg ha-1

. When the dose of Zn was raised to 20 Kg ha-1

level the concentration of K showed

decreasing trend.

Fig.10 Effect of potassium and zinc solubilizing microorganism on K uptake (kg ha-1

)

0

5

10

15

20

25

30

35

T1 T2 T3 T4 T5

K u

pta

ke

(kg

/ha

)

Treaments

Flowering

Harvesting

Seed

65

4.7.4 Effect of potassium and zinc solubilizing microorganism on Fe uptake.

The data as presented in Table 4.17 and Fig.11 indicated that inoculation of K

and Zn produced significant effect on Fe uptake by plant and grain in all the growth stages.

Table 4.17 Effect of potassium and zinc solubilizing microorganism on Fe uptake

Treatments Uptake Fe (g ha

-1)



fertilizer) 273.75 340.50 93.48 433.98


ha-1

) 396.50 530.75 150.78 681.53


KSB 541.75 626.50 208.45 834.95


ZnSB 666.50 667.25 242.21 909.46


KSB + ZnSB 740.25 758.00 280.38 1038.38

SEm ± 7.01 4.41 3.40 -

CD at 5% 21.62 13.61 10.50 -

The significantly maximum enhancement in potassium uptake by green

gram crop was recorded at of RDF + Rhizobium + PSB + KSB + ZnSB (T5) recorded

significantly higher uptake in flowering (740.25 g ha-1

), plant at harvest (758 g ha-1

) and seed

(280.38 g ha-1

). followed by RDF + Rhizobium + PSB + ZnSB (T4) and Rhizobium + PSB +

KSB (T3).

The increasing Fe uptake may also be attributed due to concentration of Fe in

plant and seed. Significant increase in biomass production is also one of the reasons for better

nutrient uptake. Similar trend was observed by Chaturvedi et al., (2010) in soybean.

Fig.11 Effect of potassium and zinc solubilizing microorganism on Fe uptake (gm ha-1

).

0

100

200

300

400

500

600

700

800

T1 T2 T3 T4 T5

Fe

up

tak

e (g

m/h

a)

Treatments

Flowering

Harvesting

Seed

66

4.7.5. Effect of potassium and zinc solubilizing microorganism on Zn uptake.

The data as presented in Table 4.18 and Fig.12 indicated that inoculation of K

and Zn produced significant effect on Zn uptake by plant and grain in all the growth stages.

The increased grain yield and higher nutrient content in the crop by K and Zn

application, thereby activating more absorption of nutrients from the soil, resulted in higher

uptake of nutrients.

Table 4.18 Effect of potassium and zinc solubilizing microorganism on Zn uptake

Treatments Uptake Zn (g ha

-1)



fertilizer) 42.34 36.68 30.34 67.02


ha-1

) 56.96 51.50 42.18 93.68


KSB 75.77 64.89 54.27 119.16


ZnSB 95.53 85.55 69.94 155.49


KSB + ZnSB 109.31 100.42 85.97 186.39

SEm ± 2.20 1.26 1.60 -

CD at 5% 6.78 3.9 4.95 -

The significantly maximum enhancement in Zn uptake by green gram was

recorded at of RDF + Rhizobium + PSB + KSB + ZnSB (T5) recorded significantly higher

uptake at flowering (109.31 g ha-1

), plant at harvest (100.42 g ha-1

) and seed (85.97 g ha-1

).

followed by RDF + Rhizobium + PSB + ZnSB (T4) and Rhizobium + PSB + KSB (T3).

Fig.12 Effect of potassium and zinc solubilizing microorganism on Zn uptake (gm ha-1

).

0

20

40

60

80

100

120

T1 T2 T3 T4 T5

Zn

up

tak

e (g

m/h

a)

Treatments

Flowering

Harvesting

Seed

67

Table 4.19 Effect of potassium and zinc solubilizing microorganism on microbial

population after harvest of Mungbean.

Treatments Bacteria

(CFU X10-7

)

Actinomycetes

(CFU X10-5

)

Fungi

( CFU X 10-4

)


fertilizer)

30.81 16.71 4.75


ha-1

)

33.99 20.09 6.11


KSB

36.58 21.64 7.12


ZnSB

37.83 22.14 7.16


KSB + ZnSB

39.78 23.29 7.51

SEm ± 0.49 0.24 0.10

CD at 5% 1.51 0.75 0.33

The scrutiny of the data given in Table 4.19 shows significant increase in microbial

population in soil after harvest of mungbean crop was also noted with single as well as dual

inoculation with Rhizobium, KSB and ZnSB along with recommended dose of fertilizers over

control. Significantly highest value of bacteria population (39.78) were noted in T5) RDF +

Rhizobium + PSB + KSB + ZnSB, Significantly highest value of fungi population (7.51) were

noted in (T5) RDF + Rhizobium + PSB + KSB + ZnSB, Significantly highest values of

actinomycetes population (23.29 CFU × 10-5

) were noted in (T5) RDF + Rhizobium + PSB +

KSB + ZnSB.

Karande and Khot (2007) noted that the Rhizobium and potassium

recorded significantly more total microbial count at harvest than that of 100% RDF.

However, Ingle et al. (2003) concluded that the rhizobial population was increased from 30

days onwards and highest was recorded at 60 days in combined application of Rhizobium

japonicum + Azotobacter brasilence in soybean. Kumar et al. (1998) found that seed

inoculation with Rhizobium significantly increased nodulation and root colonization over

uninoculated control treatment in chickpea. Further, Sreenivasa (1994) reported that VAM

fungi and Rhizobium both showed similar root colonization and spore count, spore number

and root colonization increased upto 50% RDF but decreased slightly at 100% RDF in

68

groundnut. Earlier, Balachandran and Nagarjun (1999) also noted that in green gram crop.

Combination of Rhizobium inoculation with levels of zinc increased VAM fungi colonization.

Similar findings were reported by Mandhre et al. (1995).

Fig 13. Bacteria population

0

5

10

15

20

25

30

35

T1 T2 T3 T4 T5

Bact

eria

(C

FU

×10

-7)

TREATMENTS

Fig 14. Actinomycetes population

0

2

4

6

8

10

12

14

16

18

20

T1 T2 T3 T4 T5

(Act

inom

yce

tes

CF

U×

10

-5)

TREATMENTS

Fig 15. Fungal population

0

1

2

3

4

5

6

7

8

9

T1 T2 T3 T4 T5

Fu

ngi

( C

FU

× 1

0-4

)

TREATMENTS

Summary and Conclusion

69

SUMMARY AND CONCLUSIONS

To investigate the effect of “Potassium and zinc solubilizing microorganism on mungbean”.

A field experiment was conducted on research farm, Department of Soil Science and

Agricultural Chemistry, College of Agriculture, Badnapur during 2016-2017. The experiment

comprised of 5 treatments and 4 replications. The salient findings interpreted and discussed in

the previous chapter are summerised below.

Significant impact of bioinoculants in the form of Rhizobium , KSB and ZnSB

in combination was found on growth in mungbean. Number of pods per plant and

germination percentage was found significantly maximum in treatment receiving inoculation

of RDF + Rhizobium + PSB + KSB + ZnSB over the control treatments.

Application of bioinoculants increased the seed yield of mungbean.

Significantly highest seed yield of mungbean was noted in inoculated plots with RDF +

Rhizobium + PSB + KSB + ZnSB over other control.

The concentration in plant and uptake of nitrogen by mungbean was also

found enhanced significantly with seed inoculation of RDF + Rhizobium + PSB + KSB +

ZnSB. The data shows increase in N concentration and its uptake by seed, and total was

maximum in inoculated with RDF + Rhizobium + PSB + KSB + ZnSB over other control.

The concentration in plant and uptake of phosphorus by mungbean was also

influenced significantly with the seed inoculation of RDF + Rhizobium + PSB + KSB +

ZnSB as compared to control. Significantly higher values of concentration and uptake of

phosphorus by seed and total were recorded in inoculated plots (T5) which is followed with

(T4).

The concentration in plant and uptake of potassium by mungbean crop was

increased with inoculation of RDF + Rhizobium + PSB + KSB + ZnSB over the control.

Significantly highest values of potassium concentration at flowering seed (1.61 %) and seed

(0.99 %) were noted in (T5). Simillarly, the uptake of potassium by Mungbean crop was also

increase with inoculation of RDF + Rhizobium + PSB + KSB + ZnSB over the control.

The concentration in plant and uptake of Fe by mungbean crop was also influenced

significantly with the seed inoculation of RDF + Rhizobium + PSB + KSB + ZnSB.

70

Significantly higher values of Fe concentration at flowering and in seed was noted in (T5)

RDF + Rhizobium + PSB + KSB + ZnSB.

The concentration in plant and uptake of Zn by mungbean crop was also influenced

significantly with the seed inoculation of RDF + Rhizobium + PSB + KSB + ZnSB.

Significantly higher values of Fe concentration at flowering and in seed was noted in (T5)

RDF + Rhizobium + PSB + KSB + ZnSB.

The significant increase in nutrient availability in soil after harvest of mungbean crop

was also recorded with bioinoculants. Significantly higher values of available potassium was

noted in (T5) RDF + Rhizobium + PSB + KSB + ZnSB. However, in case of available

nitrogen and available phosphorus highest in (T5) RDF + Rhizobium + PSB + KSB + ZnSB

which are followed by each other.

The available micronutrients (Fe and Zn) in soil after harvest of mungbean crop were

also found significantly highest. Available Fe and Zn was noted maximum in (T5) RDF +

Rhizobium + PSB + KSB + ZnSB.

Soil pH, EC and CaCO3 after harvest of mungbean indicated non-significant results.

The soil organic carbon in mungbean crop was also influenced significantly with the seed

inoculation of Rhizobium and PSB + KSB + ZnSB along with RDF as compared to control.

The significant increase in microbial population in soil after harvest of mungbean

crop was also noted with inoculation of KSB and ZnSB and PSB along with recommended

dose of fertilizers over control. Significantly higher values of actinomycetes, bacteria, fungi

population were noted in seed inoculation of Rhizobium and PSB + KSB + ZnSB along with

RDF as compared to control.

Conclusions :

From the results summarized above following conclusion can be drawn.

1. In mungbean plant germination percentage were significantly increased with seed

inoculants (RDF + Rhizobium + PSB + KSB + ZnSB).

2. Application of RDF + Rhizobium +PSB + KSB + ZnSB significantly improved seed yield

of Mungbean.

71

1. Concentration of nutrients and their uptake also increased with increasing levels of

seed inoculantion (RDF + Rhizobium + PSB + KSB + ZnSB).

2. The seed quality parameters viz., protein content were also improved with seed

inoculantion (RDF + Rhizobium + PSB + KSB + ZnSB).

3. Available N, P, K, Zn and Fe in soil were found more in seed inoculants treatment

(RDF + Rhizobium + PSB + KSB + ZnSB) than control.

4. The organic carbon in mungbean crop was also improved in soil with the seed

inoculantion (RDF + Rhizobium + PSB + KSB + ZnSB).

5. The soil microbial population after harvest of sunflower crop was also increased with

seed inoculantion of Rhizobium and PSB + KSB + ZnSB along with recommended dose of

fertilizers.

In general, it can be noted that in mungbean crop liquid inoculants (RDF +

Rhizobium + PSB + KSB + ZnSB) noted significant effects on yield, chemical and biological

health of soil, nutrient availability, nutrient uptake and quality attributes of Mungbean.

The results inferrered from present investigation suggest further confirmation for

final recommendations.

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Thesis Abstract

ABSTRACT

‘‘Studies on effect of potassium and zinc solubilizing microorganism on mungbean’’

By

NAVSARE RAHUL INDAR

A Candidate for the Degree

of

Master of Science (Agriculture)

In

DEPARTMENT OF SOIL SCIENCE AND AGRICULTURAL CHEMISTRY


VASANTRAO NAIK MARATHWADA KRISHI VIDYAPEETH

PARBHANI 431 402 (M.S.), INDIA.

2017

Research Guide : Dr. S. S.Mane

Department : Soil Science and Agricultural Chemistry

A Field experiment was conducted during kharif season 2016-17 at

experimental farm of Department of Soil Science and Agril. Chemistry, College of

Agriculture, Badnapur using green gram as a test crop to studies on effect of potassium and

zinc solubilizing microorganism on mungbean. The experiment was laid out on Vertisols

with five treatment combination, replicated four times in randomized block design. The

treatment consists of T1 Absolute control (No fertilizer application), T2 RDF (25:50:00 N,

P2O5 and K2O kg ha-1

), T3 (RDF + Rhizobium + PSB + KSB), T4(RDF + Rhizobium + PSB +

ZSB), T5 (RDF + Rhizobium + PSB + KSB + ZSB). The results emerged out clearly indicated

that various growth parameters like number of pods, number of nodules, total biomass

production, dry matter and seed yield was increased due to application of potassium and zinc

solubilizing microorganism. It was inferred from the results that application of RDF and zinc

and potassium solublizing microorganism found superior over only N and P application i.e.

RDF (25:50:00 N, P2O5 and K2O kg ha-1

). The KSB and ZSB application showed synergistic

effects on other nutrients (N, P, K) uptake. Soil fertility was also found to be improved due to

application of ZSB and PSB to green gram.

(R.I.Navsare) (S.S.Mane)

Student Research Guide Head, SSAC

APPENDIX – I

Table 4.20 Effect of potassium and zinc solubilizing microorganism on available N (kg

ha-1

)

Treatments

Available N (kg ha-1

)

Flowering

Harvesting



) 176.01 184.47




SEm± 1.14 2.10

CD at 5% 3.53 6.49

Initial available N 152.24 kg ha-1

APPENDIX – II

Table 4.21 Effect of potassium and zinc solubilizing microorganism on available P (kg

ha-1

)

Treatments

Available P (kg ha-1

)

Flowering

Harvesting



) 14.52 16.32




SEm± 0.25 0.16

CD at 5% 0.80 0.51

Initial available P 13.42 kg ha-1

APPENDIX – III

Table 4.22 Effect of potassium and zinc solubilizing microorganism on available K (kg

ha-1

)

Treatments

Available K (kg ha-1

)

Flowering

Harvesting

T1 Absolute control (No fertilizer) 581 541.50


) 629 577.50

T3 RDF + Rhizobium + PSB + KSB 656 611.50

T4 RDF + Rhizobium + PSB + ZnSB 674 620.00

T5 RDF + Rhizobium + PSB + KSB + ZnSB 712 646.00

SEm± 7.08 7.71

CD at 5% 21.84 23.77

Initial available K 602.46 kg ha-1

APPENDIX – IV

Table 4.23 Effect of potassium and zinc solubilizing microorganism on available Zn

and Fe after harvest (kg ha-1

).

Treatments Available Zn (Kg/ha-1

) Available Fe (Kg/ha-1

)



) 0.55 4.59




ZnSB

0.67 5.47

SEm± 0.005 0.035

CD at 5% 0.016 0.10

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