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Hindawi Publishing CorporationInternational Journal of AgronomyVolume 2013 Article ID 581627 8 pageshttpdxdoiorg1011552013581627

Research ArticleEffect of Rhizobium and Phosphate Solubilizing BacterialInoculants on Symbiotic Traits Nodule Leghemoglobin andYield of Chickpea Genotypes

G S Tagore1 S L Namdeo2 S K Sharma2 and Narendra Kumar3

1 Department of Soil Science and Agricultural Chemistry JNKVV Jabalpur Madhya Pradesh 482 004 India2 College of Agriculture Indore Madhya Pradesh 452 001 India3 IIPR Kanpur Uttar Pradesh 208 024 India

Correspondence should be addressed to Narendra Kumar nkumaricargmailcom

Received 25 June 2013 Revised 16 September 2013 Accepted 17 September 2013

Academic Editor J K Ladha

Copyright copy 2013 G S Tagore et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

A field experiment was carried out during the rabi season of 2004-05 to find out the effect of Rhizobium and phosphate solubilizingbacterial (PSB) inoculants on symbiotic traits nodule leghemoglobin and yield of five elite genotypes of chickpea Among thechickpea genotypes IG-593 performed better in respect of symbiotic parameters including nodule number nodule fresh weightnodule dryweight shoot dryweight yield attributes and yield Leghemoglobin content (255mg gminus1 of fresh nodule) was also higherunder IG-593 Among microbial inoculants the Rhizobium + PSB was found most effective in terms of nodule number (2766nodules plantminus1) nodule fresh weight (14490mg plantminus1) nodule dry weight (7430mg plantminus1) shoot dry weight (1176 g plantminus1)and leghemoglobin content (229mg gminus1 of fresh nodule) and also showed its positive effect in enhancing all the yield attributingparameters grain and straw yields

1 Introduction

Pulses are the second most important group of crops aftercereals Developing countries contribute about 74 to theglobal pulses production and the remaining comes fromdeveloped countries India China Brazil Canada Myanmarand Australia are the major pulse producing countries withrelative share of 25 10 5 5 and 4 respectively In2009 the global pulses production was 615 million tonnesfrom an area of 706 million hectares with an average yieldof 871 kgha Dry beans contributed about 32 to globalpulses production followed by dry peas (17) chickpea(159) broad bean (75) lentil (57) cowpea (6) andpigeonpea (40) India is the largest producer and consumerof pulses in the world contributing around 25ndash28 of thetotal global production About 75 of the global chickpea(Cicer arietinum L) area falls in India [1] Chickpea is one of

the major post rainy seasonpulse crops in Madhya Pradeshwhich occupies 309m ha with production of 330mt andproductivity of 1071 kg haminus1 [2] The poor productivity ofchickpea in this region ismainly due to imbalance applicationof nutrients and use of traditional varieties Under suchsituations use of Rhizobium and phosphate solubilizingbacteria (PSB) had shown advantage in enhancing chickpeaproductivity [3 4] Microbial inoculants are cost effectiveecofriendly and renewable sources of plant nutrients [5]Rhizobium and PSB assume a great importance on accountof their vital role in N

2

-fixation and P-solubilisation Theintroduction of efficient strains of P-solubilizing species ofBacillus megaterium biovar phosphaticum Bacillus polymyxaPseudomonas striata Aspergillus awamori and Penicilliumdigitatum in the rhizosphere of crops and soils has beenreported to help in increasing phosphorus availability in thesoil [6] Since the information on response of elite genotypes

2 International Journal of Agronomy

of chickpea to inoculation with Rhizobium and phosphatesolubilizing bacterial inoculants is meager under such situ-ation therefore an experiment was designed to assess theproductivity of chickpea genotypes in combinations withmicrobial inoculants inMalwa Region of Madhya Pradesh

2 Material and Methods

21 Experimental Site The field experiment was conductedat the Experimental Farm of College of Agriculture IndoreMadhya Pradesh (22∘431015840 N 75∘561015840 E and 5557m abovemean sea level) The soil of the experimental site belongs tosarol series which is a member of a fine montmorilloniticfamily of Vertic Ustochrept and Vertic Chromusters The soilcharacteristics of the experimental site before start of thestudy were analysed and presented in Table 1

22 Treatments Details and Crop Culture The experimentwas conducted during the winter (rabi) season of 2004-05The experiment was laid out in split-plot design with threereplications The experiment was conducted with twentytreatment combinations comprising five genotypes namelyIG-226 IG-370 IG-379 JG-412 and IG-593 in main plotsand four microbial inoculants (no inoculum RhizobiumPSB and Rhizobium + PSB) in sub-plots The chickpea geno-types were collected from college of Agriculture Indore andmicrobial inoculants from JNKVV Jabalpur The gross plotsize was 5m times 240m2 Chickpea crop was sown on a fineseed bed prepared after presown irrigation with a seed rate of100 kg haminus1 Rhizobium inoculant at 5 g kgminus1 seed was appliedas seed treatment whereas phosphate solubilizing bacterialinoculants were applied in soil at 3 kg haminus1 prior to sowingThe chickpea crop was given irrigation at 40 days after sow-ing

23 Soil Analysis Soil samples from surface soil (0ndash15 cm)were taken for chemical analysis after harvesting of ricecrop Random cores were taken from each plot with a 5 cmdiameter tube auger and bulked The moist soil sampleswere sieved (2mm) after removing plant material and rootsSimilarly initial soil samples were collected from 10 randomplaces of experimental site before start of study All chemicalresults are means of triplicate analyses and are expressed onoven-dry basis Soil was analyzed for pH in 1 25 soil watersuspension [7] SOC by the method of Walkley and Black[8] Kjeldahl N by FOSS Tecator (Model 2200) available Pfollowing the method of Bray and Kurtz [9] available K by1 N NH

4

OAc using a flame photometer [7] and availableS by using 015 CaCl

2

[10] Rhizobium population in thesoil samples was enumerated by plant infection technique ofToomsan et al [11] and phosphate solubilizing bacteria (PSB)by dilution plate count method as described by Sundara Raoand Sinha [12]

24 Nodulation Growth and Yield of Crops Five plants wererandomly selected and removed from each plot and recordedthe nodule fresh weight at 35 55 and 75 days after sowing(DAS) After removal of nodules the plants were first sun

Table 1 Soil properties before the start of the study

Soil parameter ValuepH 78Electrical conductivity dSmminus1 023Soil organic carbon 045Available nitrogen kg haminus1 204Available phosphorous (Olsenrsquo P) kg haminus1 958Available potassium kg haminus1 576Available sulphur kg haminus1 1288

dried for 3 days and then oven dried at 65∘C for 48 hoursto obtain dry weight Leghemoglobin content in nodulartissues collected at 35 55 and 75 DAS was determined by theprocedure outlined by Beau [13] The nodules were dried inoven at 65∘C for 78 hours for dry weight of nodules Plantswere harvested at physiological maturity and plant heightand number of branches per plant number of pods per plantand number of seeds per pod and test weight (1000 seedweight) were recorded from 10 randomly selected plants atthe time of harvest Total drymatter and grain yield were alsorecorded for each plot

25 Statistical Analysis Effect of treatments were evaluatedby split-plot analysis of variance (ANOVA) with chickpeagenotypes as main and microbial inoculants as subfactorsAnalysis of variance was performed using the program SPSS110 for windows The significance of the treatment effect wasdetermined using F-test When ANOVA indicated that therewas a significant value multiple comparisons of mean valuewere performed using the least significant difference method(LSD)

3 Results and Discussion

31 Symbiotic Traits The data on mean nodule numbernodule fresh weight nodule dry weight and shoot dry weightat 35 55 and 75 days after sowing (DAS) are presentedin Figures 1(a) 1(b) 1(c) 1(d) and Table 2 The analysis ofdata revealed that among the genotypes IG-593 exhibitedthe highest nodule number namely 1991 2704 and 2529plantminus1 nodule fresh weight namely 8679 12748 and10016mg plantminus1 and dry weight of nodules namely 46166729 and 6568mg plantminus1 at 35 55 and 75DAS respectively

Genotype IG-593 recorded maximum shoot dry weightnamely 256 1255 and 2553 g plantminus1 and the minimum inIG-370 namely 167 803 and 1498 g plantminus1 at 35 55 and75 DAS respectivelyThe data on symbiotic traits of chickpeagenotypes indicated that shoot dry weight increased progres-sively and nodule number nodule fresh weight nodule dryweight also followed the similar trend at 35 and 55 DAS butthe decline was noted in nodule number fresh weight anddryweight of nodules at 75DASThiswasmainly due to decayof nodular tissues at pod formation which start from 60 to 65DAS

Coinoculation of Rhizobium and PSB recorded signifi-cantly higher nodule number and its fresh as well as dry

International Journal of Agronomy 3

5

10

15

20

25

30

35 55 75Days after sowing

Mean number of nodules plantminus1

Nod

ule n

umbe

r pla

ntminus1

(a)

Days after sowing

40

50

60

70

80

90

100

110

120

130

140

35 55 75

Mean nodule fresh weight (mg plantminus1)

Nod

ule f

resh

wei

ght (

mg

plan

tminus1

)

(b)


20

30

40

50

60

70

80Mean nodule dry weight (mg plantminus1)

Nod

ule d

ry w

eigh

t (m

g pl

antminus

1

)

(c)


1

6

11

16

21

26

31 Mean shoot dry weight (g plantminus1)

Shoo

t dry

wei

ght (

g pl

antminus

1

)

(d)

IG-226IG-370IG-379

JG-412IG-593


1

15

2

25

3

Legh

emog

lobi

n (m

g gminus1

of fr

esh

nodu

le)

Mean nodule leghemoglobin (mg gminus1 of fresh nodule)

(e)

Figure 1 ((a)ndash(d)) Symbiotic parameters and (e) leghemoglobin content in nodular tissues of chickpea genotypes at different intervals


Table2Symbioticparametersa

ndlegh

emoglobincontentinno

dulartissuesa

tdifferentintervals

Chickp

eageno

types

Nod

ulen

umberp

lantminus1

Nod

ulefresh

weight(mgp

lantminus1 )

Nod

uled

ryweight(mgp

lantminus1 )

Shoo

tdry

weight(gp

lantminus1 )

Nod

uleleghemoglobin

(mggminus1of

fresh

nodu

le)

Daysa

ftersow

ing

Daysa

ftersow

ing

Daysa

ftersow

ing

Daysa

ftersow

ing

Daysa

ftersow

ing

3555

7535

5575

3555

7535

5575

3555

75Ch

ickp

eageno

type

IG-226

117

181

158

523

879

708

268

497

481

1894

202

1315

15IG

-370

95146

133

412

667

630

245

408

384

1780

185

1112

12IG

-379

140

217

193

670

948

813

327

543

534

21

104

219

1618

17JG

-412

174

246

233

805

1178

942

417

617

599

23

114

236

1821

20

IG-593

199

270

253

868

1275

1002

462

673

657

26

126

255

21

26

25

SEmplusmn

1212

09

20

31

22

06

1306

004

03

05

01

07

01

LSD(119875=005)

39

36

28

66

101

7319

41

20

01

1017

02

02

03

Microbialinoculants

Con

trol

94156

148

462

569

541

243

390

380

1992

166

1215

15Rh

izobium

189

254

247

671

1209

1021

403

646

631

22

108

246

1720

19PS

B100

168

157

548

646

616

263

421

409

20

96207

1315

15Rh

izobium

+PS

B207

277

267

951

1449

1204

466

743

705

23

118

265

20

23

23

SEmplusmn

06

08

07

31

39

28

08

1313

005

02

03

004

005

01

LSD(119875=005)

1722

1991

112

80

23

37

38

01

04

08

01

01

02

PSB

phosph

ates

olub

ilisin

gbacteria


weight than Rhizobium and PSB alone The increase innodulation might be due to synergistic effect of the two typesof microorganisms for biological nitrogen fixation as againsttheir individual application Results of the similar kind havealso been reported by Rudresh et al [4] It is also due to thefact that phosphate solubilizing bacteria by virtue of theirproperty of producing organic acids solubilize insoluble orfixed form of phosphorus in the rhizosphere and make itavailable to the growing plants which promotes root develop-ment in plants [14] In the present study a significant responseof dual inoculation with Rhizobium and PSB was observedwith respect to shoot dry weight per plant Observations ofthe similar kind have been recorded by Gupta and Namdeo[15] and Barea et al [16]

32 Leghemoglobin Content in Root Nodules The resultsgiven in Figure 1(e) and Table 2 showed that the leghe-moglobin content in chickpea root nodules increased withthe advancement of crop age and was maximum at 55 DASand thereafter decline at 75 DAS Among the genotypes IG-593 possessed the highest nodule leghemoglobin of 210 255and 247mg gminus1 of fresh nodule and lowest 107 123 and145mg gminus1 of fresh nodule in IG-370 at 35 55 and 75 DASrespectively In the present study coinoculation ofRhizobiumand PSB performed better than Rhizobium and PSB alonewith respect to leghemoglobin content in the nodular tissuesof chickpea crop In case of microbial inoculants higherleghemoglobin content in nodular tissues was observed inRhizobium + PSB in comparison to their individual inocu-lation The better nodulation under chickpea genotype IGminus593 might be resulted in higher content of leghemoglobinin nodular tissues Similarly higher leghemoglobin content inRhizobium + PSB was mainly due to better root and nodulesdevelopment [17]

33 YieldAttributes Thesignificant differenceswere found inyield attributing parameters due to genotypes and microbialinoculants while their interaction effect was nonsignificant(Table 3) Among the genotypes IG-593 exhibited the highestmean number of branches and pods per plant and seeds perpod that is 1595 6396 plantminus1 and 159 podminus1 respectivelyand the lowest values were recorded in IG-370 that is 10233471 plantminus1 and 113 podminus1 respectively It was further notedthat among the genotypes IG-593 produced the tallest plant(4207 cm) while the genotype IG-370 had shortest plant(3063 cm) Variation in the above parameters is bound tooccur due to difference in geneticmakeup and inherited char-acters in different genotypesThe results corroborate with thefindings of Singh et al [18] and Tiwari et al [19]

The data further indicated that IG-593 had highesttest weight (33380 g) followed by JG-412 (24913 g) IG-379 (19148 g) IG-370 (15868 g) and IG-226 (15234 g) Thevariation in test weight among the genotypes is likely to occurdue to difference in seed size of the individual genotype Ahigh value of test weight indicated the boldness of seedswhile the lower values indicated small seeds Largesmall seedsize of chickpea is basically a genotypic character [19] Inthe present study seed inoculation with Rhizobium + PSB

significantly increased the plant height number of branchesnumber of pods per plant number of seeds per pod and1000 seed weight (test weight) over no inoculation Thehigher growth and yield attributes under Rhizobium + PSBinoculation were mainly due to more availability of N PK and S in the soil for chickpea plants [20ndash22] More-over growth promoting substances (phytohormones) areproduced by these organisms which further promote plantgrowth [23ndash25] Further inoculation of Rhizobium and PSBalone produced significantly higher number of pods per plantand test weight These results are in close agreement withTakankhar et al [26] and Khoja et al [27]

34 Grain and StrawYield Critical examination of the data inTable 3 revealed that genotype IG-593 produced the highestgrain and straw yields (2 286 and 2 728Kg haminus1) followed byJG-412 (1 995 and 2 291 Kg haminus1) and lowest in IG-370 (1 475and 1 613 Kg haminus1) The observed variation in seed and strawyields in the present investigation seems to be due to geneticdifference in yield potential of different genotypes and alsodue to variable response of different genotypes to microbialinoculants [19 28]

Significant differences in grain and straw yields werealso recorded due to microbial inoculation The grain andstraw yields increased due to microbial inoculation and thehighest seed and straw yields were obtained in inoculation ofRhizobium + PSB that is 2 150 and 2 461 Kg haminus1 and thelowest in the case of control that is 1 587 and 1 901 Kg haminus1respectively The increase in grain and straw yield might beattributed to the increased availability ofN andP in soil whichresulted in higher growth and development and finally yields[20 23 29ndash31]

35 Soil Characteristics Soil pH and EC remain unaffectedunder different chickpea genotypes and microbial inocula-tionHowever highest value of soil organic carbon (SOC)wasobserved under IG-593 (053) and the lowest under IG-370(047) The significant variations in available N P K and Swere also recorded due to chickpea genotypes and microbialinoculationThe highest value of available N (21925 kg haminus1)available P (1218 kg haminus1) available K (5686 kg haminus1) andavailable S (136 kg haminus1) was recorded after harvest of chick-pea genotype 1G-593 and the lowest values of available NP K and S were recorded in IG-370 In case of microbialinoculation the highest values of available N (2203 kg haminus1)available P (141 kg haminus1) available K (5569 kg haminus1) andavailable S (1341 kg haminus1) were recorded under inoculationof both Rhizobium and PSB however the lowest values ofavailable N P K and S were under no-inoculation (control)The variations in available nutrients under different genotypemight be due to variations in compatibility between soilmicroflora and chickpea genotypes HoweverRhizobium andPSB inoculation had resulted in better plant growth nodu-lation and rhizospheric environment which finally resultedin more availability of plant nutrients (NPKS) in the soil[28 32 33]


Table 3 Yield attributes grain and straw yields of chickpea genotypes

Chickpeagenotypes

Plant height(cm)

Number ofbranches plantminus1

Number ofpods plantminus1

Number ofseeds podminus1

Test weight(g)

Mean yield (Kg haminus1)Grain Straw

Chickpea genotypeIG-226 334 124 386 125 1523 1622 1894IG-370 306 102 347 113 1587 1475 1613IG-379 345 133 428 130 1915 1922 2199JG-412 390 144 484 133 2491 1995 2291IG-593 421 160 640 138 3338 2286 2728SEmplusmn 14 03 09 0058 20 88 91LSD (119875 = 005) 45 09 30 0191 65 288 299

Microbial inoculantsControl 341 114 379 11 2131 1587 1901Rhizobium 367 136 489 13 2200 1967 2303PSB 342 121 435 13 2160 1764 2089Rhizobium + PSB 371 160 557 14 2207 2150 2461SEmplusmn 08 03 08 06 14 59 52LSD (119875 = 005) 24 08 22 02 40 170 152PSB phosphate solubilising bacteria

Table 4 Soil fertility as influenced by chickpea genotypes and microbial inoculants

Chickpea genotypes pH ECdSmminus1 SOC () Available nutrients in soil (kg haminus1) Rhizobium population

(times104 gminus1 of soil)

PSB population(times105 gminus1 of soil)N P K S

Chickpea genotypeIG-226 775 036 049 1985 106 5456 125 968 423IG-370 768 035 047 1928 104 5448 124 929 403IG-379 776 036 049 1945 114 5464 130 999 442JG-412 779 037 053 2103 116 5471 134 1060 453IG-593 779 038 053 2193 122 5686 136 1099 509SEmplusmn mdash mdash 001 15 04 16 04 024 013LSD (119875 = 005) NS NS 002 48 14 51 12 078 041

Microbial inoculantsControl 771 036 048 1933 91 5457 125 937 412Rhizobium 776 037 051 2068 92 5527 130 1079 414PSB 773 036 048 1919 125 5467 130 940 477Rhizobium + PSB 781 037 054 2203 141 5569 134 1086 480SEmplusmn mdash mdash 001 26 02 19 03 008 003LSD (119875 = 005) NS NS 002 75 06 55 07 024 010SOC soil organic carbon EC electrical conductivityInitial rhizobial count 956 times 103 gminus1 of soil and phosphate solubilizing bacterial (PSB) count 436 times 104 gminus1 of soil

36 Microbial Population The data on Rhizobium and phos-phate solubilising bacterial counts are given in Table 4 Sig-nificant variations in Rhizobium and phosphate solubilisingbacteria (PSB) was observed due to both chickpea geno-types and microbial inoculation however their interactionswere nonsignificant Among chickpea genotypes signifi-cantly highest Rhizobium population was recorded in IG-593(1099 times 104 gminus1 of soil) followed by JG-412 (1060 times 104 gminus1

of soil) and least in IG-370 (929 times 104 gminus1 of soil) SimilarlyPSB population was significantly higher in 1G-593 (509 times105 gminus1 of soil) over rest of genotypes In case of microbialinoculation the highest values ofRhizobinum (1086times 104 gminus1of soil) and PSB (480 times 105 gminus1 of soil) were recorded in 1G-593 and lowest under control (no-inculcation) The highest

values of microbial counts in 1G-593 might be due to greatercompatibility of this genotype with inoculated microbialstrains However inoculation of both Rhizobinum and PSBmight have given added advantage over native microbialpopulation [34]

4 Conclusion

Based on above results it can be concluded that the chickpeagenotype IG-593 is superior over the remaining genotypeswith respect to nodulation yield attributing parametersnodule leghemoglobin content and yield under limitedirrigation in vertisols of Malwa Region Use of Rhizobiumand PSB inoculation had also shown advantage over no-inoculationThus chickpea genotype IG-593 and inoculation


ofRhizobium and PSBmay be recommended to realize higheryield of chickpea in this region

References

[1] FAOSTAT Food and agriculture organization of the UnitedNations 2010

[2] Govt ofMPAgricultural Statistics Department ofAgricultureGovt of MP Bhopal India 2011

[3] P C Jain P S Kushawaha U S Dhakal H Khan and S MTrivedi ldquoResponse of chickpea (Cicer arietinum L) to phospho-rus and biofertilizerrdquo Legume Research vol 22 pp 241ndash2441999

[4] D L Rudresh M K Shivaprakash and R D Prasad ldquoEffectof combined application of Rhizobium phosphate solubilizingbacterium and Trichoderma spp on growth nutrient uptakeand yield of chickpea (Cicer aritenium L)rdquoApplied Soil Ecologyvol 28 no 2 pp 139ndash146 2005

[5] M S Khan A Zaidi and P A Wani ldquoRole of phosphate-sol-ubilizing microorganisms in sustainable agriculturemdasha reviewrdquoAgronomy for Sustainable Development vol 27 no 1 pp 29ndash432007

[6] A C Gaur Phosphate Solubilizing Microorganisms as Biofertil-izer Omega Scientific Publishers New Delhi India 1990

[7] M L Jackson Soil Chemical Analysis Prentice Hall of IndiaPvt Ltd New Delhi India 1967

[8] A Walkley and I A Black ldquoAn examination of the Degtjareffmethod for determining organic carbon in soils effect of varia-tions in digestion conditions and of inorganic soil constituentsrdquoSoil Science vol 63 pp 251ndash263 1934

[9] R H Bray and L T Kurtz ldquoDetermination of total organic andavailable forms of phosphorous in soilsrdquo Soil Science vol 59 pp39ndash45 1954

[10] L Chesnin and C H Yein ldquoTurbiditymetric determinationof available sulphur in soilrdquo Soil Science Society of AmericaProceedings vol 15 pp 149ndash151 1951

[11] B Toomsan O P Rupela S Mittal P J Dart and K W ClarkldquoCounting Cicer-Rhizobium using a plant infection techniquerdquoSoil Biology and Biochemistry vol 16 no 5 pp 503ndash507 1984

[12] W V B Sundara Rao and M K Sinha ldquoPhosphate dissolvingorganisms in the soil and rhizosphererdquo Indian Journal ofAgricultural Sciences vol 33 pp 272ndash278 1963

[13] A F Beau ldquoA method for hemoglobin in serum and urinerdquoTechnical Bulletin of the Registry of Medical Technologists vol32 pp 111ndash112 1962

[14] N S Subba Rao Soil Microorganisms and Plant Growth Oxfordand IBH Pub Co Pvt Ltd New Delhi India 1986

[15] S C Gupta and S L Namdeo ldquoFertilizer economy throughcomposts and bio-fertilizer in chickpeardquo Annals of Plant andSoil Research vol 2 pp 244ndash246 2000

[16] J-M Barea M J Pozo R Azcon and C Azcon-AguilarldquoMicrobial co-operation in the rhizosphererdquo Journal of Exper-imental Botany vol 56 no 417 pp 1761ndash1778 2005

[17] G S SidhuN Singh andR Singh ldquoSymbiotic nitrogen fixationby some summer legumes in Punjab Role of leghemoglobinin nitrogen fixationrdquo Journal of Research Punjab AgriculturalUniversity vol 4 pp 244ndash248 1967

[18] R C Singh M Singh R Kumar and D P Tomer ldquoResponseof chickpea (Cicer arietinum) genotypes to row spacing andfertility under rainfed conditionsrdquo Indian Journal of Agronomyvol 39 no 4 pp 569ndash572 1994

[19] K P Tiwari L N Yadav and U S Thakur ldquoRelative perfor-mance of gram varieties under different dates of sowingrdquo CropResearch vol 11 no 1 pp 127ndash130 1996

[20] N Togay Y Togay K M Cimrin and M Turan ldquoEffects ofRhizobium inoculation sulfur and phosphorus applications onyield yield components and nutrient uptakes in chickpea (Cicerarietinum L)rdquo African Journal of Biotechnology vol 7 no 6 pp776ndash782 2008

[21] R K Sharma S K Dubey R S Sharma and J P TiwarildquoEffect of irrigation schedules and fertility levels of nodulationyield and water use efficiency in chickpea (Cicer arietinum L)rdquoJNKVV Research Journal vol 28 no 1-2 pp 8ndash10 1998

[22] A Namvar R S Sharifi M Sedghi R A Zakaria T Khandanand B Eskandarpour ldquoStudy on the effects of organic andinorganic nitrogen fertilizer on yield yield components andnodulation state of Chickpea (Cicer arietinum L)rdquo Communi-cations in Soil Science and Plant Analysis vol 42 no 9 pp 1097ndash1109 2011

[23] R S Jat and I P S Ahlawat ldquoEffect of vermicompost biofer-tilizer and phosphorus on growth yield and nutrient uptake bygram (Cicer arietinum) and their residual effect on foddermaize(Zea mays)rdquo Indian Journal of Agricultural Sciences vol 74 no7 pp 359ndash361 2004

[24] M Geneva G Zehirov E Djonova N Kaloyanova GGeorgiev and I Stancheva ldquoThe effect of inoculation of peaplants with mycorrhizal fungi and Rhizobium on nitrogen andphosphorus assimilationrdquo Plant Soil and Environment vol 52no 10 pp 435ndash440 2006

[25] H Ogutcu O F Algur E Elkoca and F Kantar ldquoThe determi-nation of symbiotic effectiveness of Rhizobium strains isolatedfrom wild chickpeas collected from high altitudes in ErzurumrdquoTurkish Journal of Agriculture and Forestry vol 32 no 4 pp241ndash248 2008

[26] V G Takankhar S S Mane B G Kamble and B S IndulkarldquoGrain quality of chickpea as influenced by phosphorus fertil-ization andRhizobium inoculationrdquo Journal of the Indian Societyof Soil Science vol 45 no 2 pp 394ndash396 1997

[27] J R Khoja S S Khangarot A K Gupta and A K KulharildquoEffect of fertilizer and biofertilizers on growth and yield ofchickpeardquo Annals of Plant and Soil Research vol 4 pp 357ndash3582002

[28] N R N Reddy and I P S Ahlawat ldquoResponse of Chickpea(Cicer arietinum) genotypes to irrigation and fertilizers underlate-sown conditionsrdquo Indian Journal of Agronomy vol 43 no1 pp 95ndash101 1998

[29] K N Meena R G Pareek and R S Jat ldquoEffect of phosphorusand biofertilizers on yield and quality of chickpea (Cicerarietinum L)rdquo Annals of Agricultural Research vol 22 pp 388ndash390 2001

[30] S C Gupta ldquoResponse of gram (Cicer arietinum) to types andmethod ofmicrobial inoculationrdquo Indian Journal of AgriculturalScience vol 74 no 2 pp 73ndash75 2004

[31] M Erman S Demir E Ocak S Tufenkci F Oguz and AAkkopru ldquoEffects of Rhizobium arbuscular mycorrhiza andwhey applications on some properties in chickpea (Cicer ariet-inum L) under irrigated and rainfed conditions 1-yield yieldcomponents nodulation and AMF colonizationrdquo Field CropsResearch vol 122 no 1 pp 14ndash24 2011

[32] L K Jain P Singh and P Singh ldquoGrowth and nutrient uptakeof chickpea (Cicer arietinum L) as influenced by biofertilizersand phosphorus nutritionrdquo Crop Research vol 25 pp 410ndash4132003


[33] Reddy B Gopal andM Suryanarayan Reddy ldquoEffect of organicmanures and nitrogen levels on soil available nutrients status inmaize-soybean cropping systemrdquo Journal of the Indian Societyof Soil Science vol 46 pp 474ndash476 1998

[34] J P Singh and J C Tarafdar ldquoRhizospheric microflora as influ-enced by sulphur application herbicide and Rhizobium inocu-lation in summer mung bean (Vigna radiata L)rdquo Journal of theIndian Society of Soil Science vol 50 pp 127ndash130 2002

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of chickpea to inoculation with Rhizobium and phosphatesolubilizing bacterial inoculants is meager under such situ-ation therefore an experiment was designed to assess theproductivity of chickpea genotypes in combinations withmicrobial inoculants inMalwa Region of Madhya Pradesh

2 Material and Methods

21 Experimental Site The field experiment was conductedat the Experimental Farm of College of Agriculture IndoreMadhya Pradesh (22∘431015840 N 75∘561015840 E and 5557m abovemean sea level) The soil of the experimental site belongs tosarol series which is a member of a fine montmorilloniticfamily of Vertic Ustochrept and Vertic Chromusters The soilcharacteristics of the experimental site before start of thestudy were analysed and presented in Table 1

22 Treatments Details and Crop Culture The experimentwas conducted during the winter (rabi) season of 2004-05The experiment was laid out in split-plot design with threereplications The experiment was conducted with twentytreatment combinations comprising five genotypes namelyIG-226 IG-370 IG-379 JG-412 and IG-593 in main plotsand four microbial inoculants (no inoculum RhizobiumPSB and Rhizobium + PSB) in sub-plots The chickpea geno-types were collected from college of Agriculture Indore andmicrobial inoculants from JNKVV Jabalpur The gross plotsize was 5m times 240m2 Chickpea crop was sown on a fineseed bed prepared after presown irrigation with a seed rate of100 kg haminus1 Rhizobium inoculant at 5 g kgminus1 seed was appliedas seed treatment whereas phosphate solubilizing bacterialinoculants were applied in soil at 3 kg haminus1 prior to sowingThe chickpea crop was given irrigation at 40 days after sow-ing

23 Soil Analysis Soil samples from surface soil (0ndash15 cm)were taken for chemical analysis after harvesting of ricecrop Random cores were taken from each plot with a 5 cmdiameter tube auger and bulked The moist soil sampleswere sieved (2mm) after removing plant material and rootsSimilarly initial soil samples were collected from 10 randomplaces of experimental site before start of study All chemicalresults are means of triplicate analyses and are expressed onoven-dry basis Soil was analyzed for pH in 1 25 soil watersuspension [7] SOC by the method of Walkley and Black[8] Kjeldahl N by FOSS Tecator (Model 2200) available Pfollowing the method of Bray and Kurtz [9] available K by1 N NH

4

OAc using a flame photometer [7] and availableS by using 015 CaCl

2

[10] Rhizobium population in thesoil samples was enumerated by plant infection technique ofToomsan et al [11] and phosphate solubilizing bacteria (PSB)by dilution plate count method as described by Sundara Raoand Sinha [12]

24 Nodulation Growth and Yield of Crops Five plants wererandomly selected and removed from each plot and recordedthe nodule fresh weight at 35 55 and 75 days after sowing(DAS) After removal of nodules the plants were first sun

Table 1 Soil properties before the start of the study

Soil parameter ValuepH 78Electrical conductivity dSmminus1 023Soil organic carbon 045Available nitrogen kg haminus1 204Available phosphorous (Olsenrsquo P) kg haminus1 958Available potassium kg haminus1 576Available sulphur kg haminus1 1288

dried for 3 days and then oven dried at 65∘C for 48 hoursto obtain dry weight Leghemoglobin content in nodulartissues collected at 35 55 and 75 DAS was determined by theprocedure outlined by Beau [13] The nodules were dried inoven at 65∘C for 78 hours for dry weight of nodules Plantswere harvested at physiological maturity and plant heightand number of branches per plant number of pods per plantand number of seeds per pod and test weight (1000 seedweight) were recorded from 10 randomly selected plants atthe time of harvest Total drymatter and grain yield were alsorecorded for each plot

25 Statistical Analysis Effect of treatments were evaluatedby split-plot analysis of variance (ANOVA) with chickpeagenotypes as main and microbial inoculants as subfactorsAnalysis of variance was performed using the program SPSS110 for windows The significance of the treatment effect wasdetermined using F-test When ANOVA indicated that therewas a significant value multiple comparisons of mean valuewere performed using the least significant difference method(LSD)

3 Results and Discussion

31 Symbiotic Traits The data on mean nodule numbernodule fresh weight nodule dry weight and shoot dry weightat 35 55 and 75 days after sowing (DAS) are presentedin Figures 1(a) 1(b) 1(c) 1(d) and Table 2 The analysis ofdata revealed that among the genotypes IG-593 exhibitedthe highest nodule number namely 1991 2704 and 2529plantminus1 nodule fresh weight namely 8679 12748 and10016mg plantminus1 and dry weight of nodules namely 46166729 and 6568mg plantminus1 at 35 55 and 75DAS respectively

Genotype IG-593 recorded maximum shoot dry weightnamely 256 1255 and 2553 g plantminus1 and the minimum inIG-370 namely 167 803 and 1498 g plantminus1 at 35 55 and75 DAS respectivelyThe data on symbiotic traits of chickpeagenotypes indicated that shoot dry weight increased progres-sively and nodule number nodule fresh weight nodule dryweight also followed the similar trend at 35 and 55 DAS butthe decline was noted in nodule number fresh weight anddryweight of nodules at 75DASThiswasmainly due to decayof nodular tissues at pod formation which start from 60 to 65DAS

Coinoculation of Rhizobium and PSB recorded signifi-cantly higher nodule number and its fresh as well as dry


5

10

15

20

25

30



Nod

ule n

umbe

r pla

ntminus1

(a)

Days after sowing

40

50

60

70

80

90

100

110

120

130

140

35 55 75


Nod

ule f

resh

wei

ght (

mg

plan

tminus1

)

(b)


20

30

40

50

60

70


Nod

ule d

ry w

eigh

t (m

g pl

antminus

1

)

(c)


1

6

11

16

21

26


Shoo

t dry

wei

ght (

g pl

antminus

1

)

(d)

IG-226IG-370IG-379

JG-412IG-593


1

15

2

25

3

Legh

emog

lobi

n (m

g gminus1

of fr

esh

nodu

le)


(e)




ndlegh


dulartissuesa

tdifferentintervals

Chickp

eageno

types

Nod

ulen

umberp

lantminus1

Nod

ulefresh

weight(mgp

lantminus1 )

Nod

uled

ryweight(mgp

lantminus1 )

Shoo

tdry

weight(gp

lantminus1 )

Nod

uleleghemoglobin

(mggminus1of

fresh

nodu

le)

Daysa

ftersow

ing

Daysa

ftersow

ing

Daysa

ftersow

ing

Daysa

ftersow

ing

Daysa

ftersow

ing

3555

7535

5575

3555

7535

5575

3555

75Ch

ickp

eageno

type

IG-226

117

181

158

523

879

708

268

497

481

1894

202

1315

15IG

-370

95146

133

412

667

630

245

408

384

1780

185

1112

12IG

-379

140

217

193

670

948

813

327

543

534

21

104

219

1618

17JG

-412

174

246

233

805

1178

942

417

617

599

23

114

236

1821

20

IG-593

199

270

253

868

1275

1002

462

673

657

26

126

255

21

26

25

SEmplusmn

1212

09

20

31

22

06

1306

004

03

05

01

07

01

LSD(119875=005)

39

36

28

66

101

7319

41

20

01

1017

02

02

03

Microbialinoculants

Con

trol

94156

148

462

569

541

243

390

380

1992

166

1215

15Rh

izobium

189

254

247

671

1209

1021

403

646

631

22

108

246

1720

19PS

B100

168

157

548

646

616

263

421

409

20

96207

1315

15Rh

izobium

+PS

B207

277

267

951

1449

1204

466

743

705

23

118

265

20

23

23

SEmplusmn

06

08

07

31

39

28

08

1313

005

02

03

004

005

01

LSD(119875=005)

1722

1991

112

80

23

37

38

01

04

08

01

01

02

PSB

phosph

ates

olub

ilisin

gbacteria












Chickpeagenotypes

Plant height(cm)




Test weight(g)













4 Conclusion




References






































Journal of




Agronomy





Microbiology




Volume 2014

























5

10

15

20

25

30



Nod

ule n

umbe

r pla

ntminus1

(a)

Days after sowing

40

50

60

70

80

90

100

110

120

130

140

35 55 75


Nod

ule f

resh

wei

ght (

mg

plan

tminus1

)

(b)


20

30

40

50

60

70


Nod

ule d

ry w

eigh

t (m

g pl

antminus

1

)

(c)


1

6

11

16

21

26


Shoo

t dry

wei

ght (

g pl

antminus

1

)

(d)

IG-226IG-370IG-379

JG-412IG-593


1

15

2

25

3

Legh

emog

lobi

n (m

g gminus1

of fr

esh

nodu

le)


(e)




ndlegh


dulartissuesa

tdifferentintervals

Chickp

eageno

types

Nod

ulen

umberp

lantminus1

Nod

ulefresh

weight(mgp

lantminus1 )

Nod

uled

ryweight(mgp

lantminus1 )

Shoo

tdry

weight(gp

lantminus1 )

Nod

uleleghemoglobin

(mggminus1of

fresh

nodu

le)

Daysa

ftersow

ing

Daysa

ftersow

ing

Daysa

ftersow

ing

Daysa

ftersow

ing

Daysa

ftersow

ing

3555

7535

5575

3555

7535

5575

3555

75Ch

ickp

eageno

type

IG-226

117

181

158

523

879

708

268

497

481

1894

202

1315

15IG

-370

95146

133

412

667

630

245

408

384

1780

185

1112

12IG

-379

140

217

193

670

948

813

327

543

534

21

104

219

1618

17JG

-412

174

246

233

805

1178

942

417

617

599

23

114

236

1821

20

IG-593

199

270

253

868

1275

1002

462

673

657

26

126

255

21

26

25

SEmplusmn

1212

09

20

31

22

06

1306

004

03

05

01

07

01

LSD(119875=005)

39

36

28

66

101

7319

41

20

01

1017

02

02

03

Microbialinoculants

Con

trol

94156

148

462

569

541

243

390

380

1992

166

1215

15Rh

izobium

189

254

247

671

1209

1021

403

646

631

22

108

246

1720

19PS

B100

168

157

548

646

616

263

421

409

20

96207

1315

15Rh

izobium

+PS

B207

277

267

951

1449

1204

466

743

705

23

118

265

20

23

23

SEmplusmn

06

08

07

31

39

28

08

1313

005

02

03

004

005

01

LSD(119875=005)

1722

1991

112

80

23

37

38

01

04

08

01

01

02

PSB

phosph

ates

olub

ilisin

gbacteria












Chickpeagenotypes

Plant height(cm)




Test weight(g)













4 Conclusion




References






































Journal of




Agronomy





Microbiology




Volume 2014


























ndlegh


dulartissuesa

tdifferentintervals

Chickp

eageno

types

Nod

ulen

umberp

lantminus1

Nod

ulefresh

weight(mgp

lantminus1 )

Nod

uled

ryweight(mgp

lantminus1 )

Shoo

tdry

weight(gp

lantminus1 )

Nod

uleleghemoglobin

(mggminus1of

fresh

nodu

le)

Daysa

ftersow

ing

Daysa

ftersow

ing

Daysa

ftersow

ing

Daysa

ftersow

ing

Daysa

ftersow

ing

3555

7535

5575

3555

7535

5575

3555

75Ch

ickp

eageno

type

IG-226

117

181

158

523

879

708

268

497

481

1894

202

1315

15IG

-370

95146

133

412

667

630

245

408

384

1780

185

1112

12IG

-379

140

217

193

670

948

813

327

543

534

21

104

219

1618

17JG

-412

174

246

233

805

1178

942

417

617

599

23

114

236

1821

20

IG-593

199

270

253

868

1275

1002

462

673

657

26

126

255

21

26

25

SEmplusmn

1212

09

20

31

22

06

1306

004

03

05

01

07

01

LSD(119875=005)

39

36

28

66

101

7319

41

20

01

1017

02

02

03

Microbialinoculants

Con

trol

94156

148

462

569

541

243

390

380

1992

166

1215

15Rh

izobium

189

254

247

671

1209

1021

403

646

631

22

108

246

1720

19PS

B100

168

157

548

646

616

263

421

409

20

96207

1315

15Rh

izobium

+PS

B207

277

267

951

1449

1204

466

743

705

23

118

265

20

23

23

SEmplusmn

06

08

07

31

39

28

08

1313

005

02

03

004

005

01

LSD(119875=005)

1722

1991

112

80

23

37

38

01

04

08

01

01

02

PSB

phosph

ates

olub

ilisin

gbacteria












Chickpeagenotypes

Plant height(cm)




Test weight(g)













4 Conclusion




References






































Journal of




Agronomy





Microbiology




Volume 2014



































Chickpeagenotypes

Plant height(cm)




Test weight(g)













4 Conclusion




References






































Journal of




Agronomy





Microbiology




Volume 2014


























References






































Journal of




Agronomy





Microbiology




Volume 2014





























Journal of




Agronomy





Microbiology




Volume 2014
























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