Handbook of biofertilizers and biopesticides


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Handbook of biofertilizers and biopesticides

  1. 1. Handbook ofBiofertilizers and Biopesticides
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  3. 3. Handbook ofBiofertilizers and Biopesticides Edited by A.M. Deshmukh R.M. Khobragade P.P. Dixit Oxford Book Company Jaipur, India
  4. 4. ISBN: 978-81-89473-15-0First Published 2007Oxford Book Company267, 10-B-Scheme, Opp. Narayan Niwas,Gopalpura By Pass Road, Jaipur-302018Phone: 0141-2594705, Fax: 0141-2597527e-mail: oxfordbook@sify.comwebsite: www.abdpublisher.com©ReservedTypesetby:Shivangi Computers267, 10-B-Scheme, Opp. Narayan Niwas,Gopalpura By Pass Road, Jaipur-302018Printed at:Rajdhani Printers, DelhiAll Rights are Reserved. No part of this publication may be reproduced, stored in aretrieval system, or transmitted, m any form or by any means, electronic, mechamcal,photocopying, recording, scannmg or otherwise, without the prior written permissionof the copyright owner. Responsibility for the facts stated, opinions expressed,conclusions reached and plagiarism, If any, in this volume IS entirely that of the Author,according to whom the matter encompassed in this book has been originallycreated/ edited and resemblance with any such publication may be incidental. TheVublisher bears no responsibility for them, whatsoever.
  5. 5. Preface Advance agricultural practices are using more input of water, soil and plant protectionchemicals in the soil. Continuous use of chemical fertilizers and pesticides make the soil lessfertile as beneficial and useful microogranism are destroyed by them. Moreover, they createmany serious problems like health ~azards, air and water pollution, killing many beneficialinsects. These facts indicate that there is an urgent need to develop newer approach to increase thesoil fertility and to manage the pests. Use of biotertilizers and biopesticides is one of the mostsuitable and widely accepted approach to chemical control. The research articles in the, book provide information on different types of biofertilizersand biopesticides and their applications for different crops. We hope that book will be helpful to students, teachers and researchers in the field ofagricultural microbiology. We would like to acknowledge the efforts taken by ABD publishers to publish this book intime. A.M. Deshmukh R.M. Khobragade P.P. Dixit
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  7. 7. Contents Preface v Introduction 01 I. Eflect of Bioferiilizers on Seedling Growth. Physiology. Nodule Production and VAM Colonization in Pungam. Pongamia pinnata (L.) Pierre 06 2. Biofertiliz,ef: A Supplementary Nutrient Source for Sugarcane 13 3. Eflect of Composite Bioinoculations and Fertilizer Levels on Growth of Sorghum (CCH-14) 184. Signiticance of Azospirillum brassilense and Pseudomonas striata on Growth and Yield ofRagi (Elucine crocana) in Altisol 26 5. Application ofYesicular Arbuscular Mycorrhizae (Glomus fasciculatum) and Rhizobium on Biomass Production of Acacia nilotica inSaline and Forest Soils JI6. Effect ofVAM Fungi and PSB on Growth and Chemical Composition of Micropropagated Banana (Musa paradisiaca L.) Plants 377. Mungbean [Vigna radiata (L.) Wilczek] Response to Inoculation with N-tixing and phosphate Solubilizing Bacteria 438. Field Performace of Asymbiotic Biofertilizers on Grain Yield of Rain-fed KharifSorghumCSH-14 519. Interaction Eflect of Azospirillum and Phosphate Solubilising Culture on Yield and Quality ofSug~rcane 5710. Use of Bio-Inoculants for Recycling of Banana Wastes 61II. Appl ication of Pressl11ud as Plant Growth Promoter and Pollution Arrester 6612. Biofertilizer for Multipurpose Tree Species 71
  8. 8. viii Contents13. Response of Tree Legumes Seedlings to Bioinoculation of Endomycorrhizae and Rhizobium in Alfisol 8014. Infectivity and Efficacy of Glomus aggregatum and Growth Response of Cajanus cajan (L.) Millsp. CVPT-22I in Cement Dust Polluted Soils 8615. Saline Soil Tolerance of Sap indus emarginatus (Vahl) Seedlings with Established Glomus fasciculatum Infection 9316. Importance of Vesicular Arbuscular Mycorrhizae in Transplanted Crops 9917. Biochemical and Genetic Characterisation of Mineral Phosphate Solubilizing Enterobacter asburiae 10518. Effect of Phospho bacterium on Growth and Seed Yield of Sword bean (Canavalis ensiformis L.) Under Graded Levels of Phosphorus 11219. Effect oflrioculation of Phospho microbes on Yield and Nutrient Uplake in Groundnut 11520. Production and Evaluation of Phosphocompost from Neem with Rock. Phosphate 12021. Effect of Cyanobacteria on Soi I Characteristics and Productivity of Gram Grown in Salt Affected Soil 12422. Crop Response toAlgalization in Rice Variety.BPT-5204 12823. Biofertilizers in Banana Fields: A Case Study from Anekal Taluka, Bangalore District, Kamataka 13124 .. Isolation of Pesticides and Heavy Metal Tolerant Strains of Azotobacter chroococcum from the Rhizospheric Region of Wheat Crop 13725. CharacteriZation and Identification of Azotobacter strains Isolated from Mulberry (Morus alba L.) Rhizosphere Soil 14126. Effects of Sulphatic Biofertilizeron Pigeonpea [Cajanus cajan (L.) Millsp.] 14727. Effect of Gamma Irradiation on the Biomass Production, Nodulation and Nitrogen Fixation by Stem Nodulating Sesbania rostrata 15428. Activity of Ammonia Assimilating Enzymes in NQdules ofSesbania rostrata Mutants 15929. Development ofSesbania rostrata Mutants by Gamma Irradiation 16230. B1Qfertilizer from Sludge of Distillery Waste Treatment Plant: A LaborlJtory Study 16831. Signifi.cance of Bacillus and Pseudomonas in Decolourization and Degradation of Dye Effluent 17332. Bioutilization and Decolorization of Paper Mill Effluent Waste 18033. Phosphate Solubilizing Soil Actinomycetes as Biofertilizers 18634. Vermicompos~ing of Kitchen Waste: A Case Study 18935. Rio-control of Fusarial Wilt of Chick Pea (Cicer arietinum) Variety, - (,haffa in Wilt Sick Field 193
  9. 9. Contents ix36. Response of Pigeon pea to Rhizobium and Trichodelma viride in Acid Soils 19737. Performance of Seed Pelletization with Biofertilizers, Macro- and Micronutrients and Biocides under DitTerent Water Holding Capacities in Acacia leucophloea (Roxb.) 20 I38. Performance of Seed Pelletization with Biotertilizers, Macro- and Micronutrients an9- Biocides under Different Soil Conditions in Acacia nilotica (Linn.) 20739. Bacillus thuringiensis-An Effective Bioirisecticide 21340. Field evaluation of ditTerent formulations of Azospirillum inoculants on Rice Crop 21741. Eflects of Psuedomonas on Wheat Fusarium Root Rot 22342. EtTects of Trichoderma on Wheat Sharp Eye Spot 23043. Inhibitory Effect ofSiderophores Produced by Pseudomonas sp. on Salmonella typhi & its Future Biotechnological Applications 23644. FVllluation oflPM Module against Major Pests of Cotton (Gossypium sp.) 24245. Bradyrhizobiumjaponicum for Soybean Growth 25046. Phenotypic and Functional Characterization of A. caulinodans Endophytic Symbiont ofS. rostrata 25747. Effect of Different Phytoextracts on Spore Gennination of Alternaria tomato (Cooke) G.F. Weber Causing Fruit Rot of Tomato 26248. Effect of Different Phytoextracts on Development of Tomato Fruit Rot Caused by . Alternaria tomato (Cooke) G. F. Weber 26549. Bioefficacy of DifTe.:ent Antagonists against Fruit Rot of Tomato Caused by Alternaria tomato (Cooke) G. F. Weber under in vitro Condition 26850. Eflect of DitTerenl Antagonists on Development of Tomato Fruit Rot Caused by Alternaria tomato (Cooke) G. F. Weber 27151. Chitosan Treatment for Plant Growth Regulation 27352. Screening of Various Microbial Strains Producing Antifungal Biomolecules 27953. Effect of Fertilizer and Bio-fertilizer on Pearl Millet with and without Intercropping under Rainfed Conditions 28454. Survey and Identification of Different Species of Earthworllls from Marathwada Region ofMaharashtra 28955. Life of Perionyx sansibaricus during Vennicomposting of Different Wastes 29456. Positive Effect ofDuallnocululll ofGPPB and AM Fungi on Growth of Anogeissus Latifolia Wall 29857. Use of Press Mud for the Production ofVernlicompost 304
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  11. 11. List of ContributorsA. Venkatesh AjaySinghForest College and Research Institute Department of Plant Pathology. c.P. CollegeMettupalayam-641 301. T.N.. India of Agriculture Sardarkrushinagar Dantiwada Agricultural University Sardarkrushinagar-A.M. Deshmukh 385506P G Department ~f Mkrobiology Y. C. CollegeofScience Karad-415 12-1. Maharashtra. India Almas Zaidi Department of Agricultural MicrobiologyA,M. Thopatel Institute Muslim University Aligarh-202 002.Department (~f Agriculture Mi<.:rohiology u.P. IndiaA.R. Madhav Rao . Anu KaliaDepartment of Agricultural Microbioyogy Department of Microbiology PunjabDepartment ~f Agricultural Sciences G K. v. K. Agricultural University Ludhiana. Punjab-141Compus Bangal()re-560 065. Karnataka. 004India B.D.ShindeA.S. Ponnuswamy Central Sugarcane Research StationDepartment ~f Seed Technology Tamil Nadu Padegaon. P.o. Nira - 412 102 Dist. Pune.Agricultural University Coimhatore-641 003.Tamil Nadu. India Maharashtra. IndiaA.lJ. Narayana B.N. PrasadDepartment of Plant Pathology Dr. Panjahrao Biotechnology Unit. Central Department ofDeshmukh Krish Vidyopeeth Akola--I44 104. Botany. Tribhuvan Unilersit). Kirtipur.Malwrashtra. India Kathmandu. Nepal
  12. 12. xii List (~r ContrihutorsB. V. Prasada Reddy Dr. Masood AhmadBiotechnoloR} Cenle for Tree Improvement Department oi BiochemislI:l; Faculty <?l LtieTirupati-517507, Andhra Pradesh Sciences, A.MU., Alighrh, u.p, IndiaBharose, A.A. Eshwarlal SedamkarMGM, Department of Bioinformatics & f!epartment of Botany Plant Pathology andBiofiechi1OIo.f!J, Aurangahad (M.S.) Mycorrhiza Laboratory Gulharga University Gulbarga-585 106, Karnataka, IndiaBirhare, S.c.Dept. of Biotechnology, Govt. Institute ofScience, Aurangabad 431 004. Foroutan, Abdolreza Agricultural & Natural Resources ResearchC. Narayana Reddy Center of Mazandaran, Saty, Iran.Department of Botany Plant Pathology andMycorrhiza LaboratolY Gulbarga University Foroutan, AidinGulbarga-585 106, Karnataka, India Faculty 0/ Agricultural Sciences of Ma:andaran Universit); Sary, Iran.C. UdayassorianDepartment (~l Environment Sciences TamilNadu Agricultural Unilersity (oimbatore- GArunachalamM I 003, Tamil Nadu. India. Professor and Head, Dept. of Agricultural Chemistty Kllikulam-625 272. Tamil NaduC.A. AgasimaniUniversity of Agricultural Sciences Dharwal- GMani580 005, Karnataka, India Department (~r Seed Technology Tamil Nadu Agricultural University Coimbatore-641 003,D.Anusuya Tamil Nadu, IndiaDepartment (Jl Botany Bangalore UniversityBangalore-560 056, Karnataka G Naresh.Kumar Department ofBiochemistry Faculty o/Science,D.B. Shinde MS. .University Vadodara-390 002, Gujarat,Asst!. Microhiologist CentraJ Sugarcane IndiaResearch Station Padegaon, po. Nira, Dist.Pune Maharrashtra - 412 102, India GRajannanDeshmukh, L.S. Department of Environment Sciences, TNA U,Notional Agril. Research Station, Aurangahad, Coimbatore-641 003, Tamil Nadu.M4U, Parbhani (MS.) GO. RadderDeshpande, V.D. Unilcrsity (l Agricultural Sciences Dharwal-MGM, Department of Bioinforma/ics & 580 005, Kamataka, IndiaBioflechnology, Aurangahad (MS.) GO. Sa rata IeDharmesh Shah Department of Biotechnology Engineering,Institute of Biotechnology, Chudasama Main Tatyasaheb Kore "J,stitute (Jl Engineering andRoad, Near Amrapali Crossing, Rajkol-360 Technology. Warananagar. Tal: Panhala. Dist:fIfI ) {;II;nrnl knlhlJn""-.J Iii 111 (M s: I India.
  13. 13. List of Contrihutors xiiiGR.Pathade K.G ThakarePG Department of Microbiology yc. College Department (?fPlal11 Pathology Dr. fanjabraoof S,:ience Vidyanagar, Karad-415 124, Deshmukh Krishi Vidyapeeth Akola-444 104,Maharashtra Maharashtra. IndiaH.C. Lakshman K.K. KhodeP.G Depaltmcnt. of Botany (Micmhiology Lab.) Sugarcal1e Specialisl Cenlral SugarcaneKamatak University. Dharwad-580 003, India. Research Station Pad<!gaon, po. Nira. Dist. Pune Maharl.lshtra - 412 102.J. PrabakaranDepartment q( Environmental Sciences Tamil K.T. Parthiban and RavichandranNadu Agriclllturc.1 University Coimbatore-64I Forest College and Research Institute003, Tamil Nadu, India Mettupalayam-641 301. T.N., India,Jaydeep Joshi Kamal PrasadInstitute of BiotechnolofO!. Chudasama Main Department (?( Post-Gra4uate Studies andRood. Near Amrapali Crossing, Rajkot-360 Research in Biological Science Rani Gurgavati002. Gujarat. Unile1sity.Jaba/pur--482 001 (M.P). IndiaJ<. Govindan Kamlesh PansheriaFacul(v o/Agriculture and Animal Hushandry 1m tillite of Biotechnology; Chudasama Mail7Gandhigram Rurallnslill/le Gandhigram-.624 Road. ,Veal Amrapali Crossing. Ra;kot-360302. Dist. Dindigal Tamil Nadu 002, GuianlfK. P. Sreenath Kamlesh S. PansheriaDepartment of Botany Bangalore University Il7stitute (!/Biolechn%g): Raiya /?oad, Rajkot-Banglore-560 056, Karnataka, India 360002. MG Science Institute. Ahmedabad (Glljarat)K. SrinivasanDepartment of Environll1ental Sciences Tamil Kavita SharmaNadu Agricultural Universi(v Coimbatore-641 Department of Plant Path%?J. c.p College003. Tamil Nadu. India <?f Agri<:ulture Sardarkrushinagar Dantiwada Agricultural University Sardarkrushinagar-K. Vanangamudi 385 506Department of Seed Tei.·hnology Tamil NaduAgricullural University Coimhalor<!-6.J I (JU3. KhapreTamil ,Vadu. India Agril. Research Slat ion, Badnapur. MA U, Parhhal1iK.D. ThakurDepartlll<!l1t olPlant fatholo,s), Dr. falliahra(J Kohire,O.DDeshl11l1kh Krish Vh(W!J<!eth Akolil-4.J4 1(J.J. Agril. Research Station, BadnallIr. AlA U.,lfaharaslrtro. India !lIrbhani
  14. 14. xiv List of Contributors Md. Saghir KhanKohirepatil, V.o Department of Agricultural MicrobiologyAgril. Research Station, Badnapur, MA U, Institute Muslim University Aligarh-202 002,Parbhani UP. IndiaKulkarni,A.M Mchra, v.P.Agril. Research Station, BadnapU/; MA U, Dept. of Biotechnology, Govt. Institute o.fParbhani Science, Aurangabad 431 004.I.J.Parekh Mohd.Ami!Department ofBiochemislIJl Faculty o.fScience, Department of Agricultural MicrobiologyMS. University Vadodara-390 002, Gujaral, Institute Muslim University Aligarh-202 002,India Up., IndiaL.S.Deshmukh Mrunalini A. GuptcNational Agril. nesearch Slation, Aurangabad, Depar,menl o(flant Palh%:?y Dr. PaniabraoMA U, Parbhani De.~/7Il/lIkh /,idYlipeelh Akola-44-1 10-1, Afaharashtra, India •Laxman B. KadamP.G Department. of Botany (Microbiology Lab.) MudalagiriyappaKarnatak University, Dharwad-580 003, India. University of Agricultural Sciences Dharwal- 580 005, Karnataka, IndiaM.D. Shukla N. Venugopal1nstitllte ofBiotechnology, Raiya Road Rajkot- Department of Botany Bangalore University360002, M.O Science Institute, Ahmedabad Banglore-560 056, Karnataka, India(Gujarat) N.A. PatclM.G Bodhankar Department of Plant Pathology, c.p CollegeDepartment of Microbiology Bharliya of Agriculture Sardarkrushinagar DantiwadaVidyapeelhs A.S.C. College Sangli--I16 416, Agricultural Universily Sardarkrushinagar-Maharashlra, India . 385 506M.N. Sreenivasa N.A. SahasrabudheDeptl. ofAgricultural Microbiology University Scienli~t. Plant Molecular Biology Unitof Agricultural Sciences Dharllad-580 005. Division of Biochemical Sciences NationalKarnataka Chemicai .Laboratory Pune-411 008, Maharashtra, IndiaM. V. KulkarniMaharashtra Pollution Control Board, N.M. VanithaSangli, Maharashtra, India Departmenl of Botany Bangalore University Banglore-560 056, Karnatka. IndiaMallika VanangamudiForest College and .Research Institute N.P. PatilMettupalayam-641 301, TN., India College of Agricultural Engineering. MA U, Parhhani (MS,)
  15. 15. List q(COl1triblllors xvN.S. Kulkarni- P. B. SandipanDepar/lllent of Microhiology R.A. College Department of Plant Palhology. C. P CollegeIf(IShilll, Maharashtra, India of Ag/,iclilture Sardarkrushillagar [)al1/iH"ada Agricultural Universt/)· Sardarkrusllinagar-Nikun.i Chhag 385 500Instillife (d Biotechl1olo?:}~ Chudasama ".fainRoad. Near Al11rapali Crossing. RlIikol-300 P.GLavanya002. Ciu;araf Kri,hi Vigyan Kendra. Katlupakkam--003 ]()3Nikunj H. Chhag P.L. PatilInstilllfe ()/Biotechnology, Razya Roud. Rajkot- Ex. Associa/e Dean dnd Prim;ipal ot College360 ()02. Ue. Sciellce Insfifllle. Ahmedabad. of Agriculture Dhule--I24 00-1, Ma/7a/os17/ra.fGujaratj P. V. HatwalncNirali J. JoshiInstitute q( Biotechnology. Razva Road. Rajkot- Departmellt a/Plant Pat/wIn?:,} D,: Panj([/Jr(Jo.3M) ()02, M. e. Science Instituie. Ahmedabad. Deshmukh Krishi JldYIJpeeth Akola-.J.:f4 10-1,(Oll;arat) Maharashtra, IndiaOJ). Kohire Palak R. JadejllNanf Patholog.,·, Agril. Research Swtion, Instilule of Bioteci1nolugl: Roi)o Road. Ra;kot-Bac/naplII AfAU, ParMani (iI.S) 36U O()2, AI G Scienci! II1,IilUle. Ahmedahad. (GujaratjP. ArjunnnFaCilIty 6t.Agriculllire and Antmal Husbandry Pande, A.K.; Kshirsagar, A.R; Harke S.N;Gandhigram Rural/nstillll:/{ Gandhigram-62.J Puthan, P.N.302, Dist. Dindigal Tamil Nadu Dept. of" Biotechnology. GoV!, Institute of Sciellce, Aurangahad .J31 O(}.J.P. GyaneshwarDepartment (){Biochemisfi) FaclIll) (J(Science, R. SalavananUs. Universil) , Jadodara-390 002, Glijarat, Department 0( Environmenf Science,, TNA U.India Coimhatorc-641 V03. Tall/il Nadll.P. Jothimani R.SowmyaDepartment of Environmental Sciences 7(II/lil Di!f)(frtmen/ o( BOluny Bangulore Lniversif),,(fdu Agricultural (ni,ersity (oilllha{ore-o.J I Bangalwe·-500 056. KOJ"l/al<lkuO(J3, lilmil Nadl~. India,P.A. Hatwalne R.B. Somanif.G. Dcpartmenl of Plant lat/7olog;.I Punjabrao PG D.:parll1lel1l of P/olIl Pathology {ol1jahr([(/Deshmukh Krishi flidyapeeth Akola-·1-/.J 10-1, Des/7l11uf..1 Krishi Vh(vapee/h Akola-.J-/.J 10-1,Maharashtra, India :l/ahar{f.htra, India
  16. 16. xvi List of Contributors R.L. Patel S.Sundaravarathan Department (~( Plant Pathology, c.P. College Department q/Agricuture Microbiolo&VTamil.q(Agriculture Sardarkrushinagar Dantiwada Nadu Agricultural University Coimbatore-64I Agricultural University Sardarkrushinagar- 003, TN., India, 385506 S.A. Patil Department of Biotechnology Engineering,R.M. Kothari Tatyasaheb Kore Institute q( Engineering andSchool q( Life Sciences North Maharashtra Technology, Warananagar. Tal: Panhala, Dist:Universi(v.falgaon-425 001, Maharasht~a, Kolhapllr-416 113 (M.S.) IndiaIndia S.B. Chincholkar,R.PGupta School of Life Sciences North Maharashtra Department of Microbiology Punjab University.falgaon-425 001, Maharashtra, Agricultural University Ludhiana. Punjab-141 India 004 S.B.Jadhav Department of Agriculture Science andR.R. More Technolog)J Vasantdada Sugar Instilute ManiriDepartment q( Agriculture Microhiology (BK.), Dist. [une, Maharashtra, IndiaR.W.lngle s.c. KaleDepartment ofP/ant Pathology Dr. Panjabrao P. G Department of Microbiology Y. C. CollegeDeshmukh Krisni Vidyapeeth Akola-444 104, o( Science Vidyanagar, Karad-415 124,Maharashtra, India MaharashtraRanaAthar S.D. KhambeInstitute ofAgriculture, A. M. u., Aliga.rh, U. P. Departme"r of Microbiology, Miraj Mahavidyalay, Miraj-416 -110, Maharashtra,India IndiaRoshan Sharma Poudyal S.K. TalegaonkarBiotechnology Unit, Central Department of School of Lite Sciences North MaharashtraBotany, Tribhuvan University, Kirtipur, University .falgaon-425 00 I. Maharashtra,Kathmandu. Nepal IndiaS. Kannaiyan S.(). KolteDepartment ofAgricuture Mi...robiolog.1 Tamil Department (~fP{ant Pathology Dr. PanjahraoNadu Agricultural University Coimbatore-6-11 Deshmukh Krish Vidyapeeth Ako{a-44-1 10-1.003, TN., India. Maha"rashtra, India S.R.S. DangeS. Mariappan Department q( Plant Pathology, c.P. CollegeDepartment q( Environmental Sciences Tamil of Agriculture Sardarkrllshinagar DantiwadaNadtt Agricultural University Coill1hatore-64I Agricultural University Sardarkrushinagar-(){)3. Tamil Nadu, l11dia 385506
  17. 17. Li! (?f Contributors .niiS.W. Kulkarni T.VijayaDepartment of Microbiology S. B. Zadbuke Presently at Department of BiotechnologyCollege Barshi--I13 401, Disl. Salapur S. V. University, Tirupati-Sl7 S02, AndhraMaharashtra, India PradeshSandeep Garg V.S. ShindeDepartment o{ Microbiology Unilersi1.l ofDelhi South Campus Benito Juare= Road New Department o{the Agronoll1.l; MA U. ParhhaniDelhi - 110021, India (M.S.)Satwekar,A.A.Dept. (~{ Biotechnology, Govt. Institute of V. W. TarsekarScience, Aurangabad 431 00-1. Department (){Plant Pafhologv Dr. Paniahruo Deshmukh Krish Vidyapeeth Akola--I44 10-1,Shams Tabrez Khan Maharashtra, IndiaInstitute ojAgricuflure, A. M. u., A/igarh, U. PIndia Vandana GuptaT. Satyanarayana Department of Microbiology University o{Department of Microbiology University of Delhi South Campus Benito Juarez Road NewDelhi South Campus Benito Juarez Road New Delhi - 110021, IndiaDelhi - I/O 021, IndiaT.H. Shankarappa Yasari, EsmaeilDepartment (~{ Agricultural MicrobiologyDepartment ofAgricultural Sciences G K. V. K. Agricultural Organization of MazundarallCompus Bangalore-S60 06S, Karnataka, India Province. Sary, Iran.
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  19. 19. IntroductionThe success of green revolution depends upon the availability offertilizers, high yielding varietyof seeds, improved agronomical practices and timely availability of water. The demand fornitrogenous fertilizers has been increasing out its production has always fallen short. In spiteof unlimited supply ofN2 in the air, manufacturing the fertilizer for todays needs required 544 9x 10 MJ of fossil fuel energy which is equivalent to about 13 million tons of oil-a non-renewable source. We need about 230-240 m.t. offoodgrains by 2000 A.D. On the other handthe demand for the fertilizer nitrogen produced by using non-renewable fossil fuels cannot bemet through domestic production. In such a scenario, the use of microbes who do nut needfossil energy is of immense value for increasing soil productivity in India, where most ofagriculture is low input subsistence farming through biological nitrogen fixation or increasedefficiency of fertilizers applied. The part of increasing deficit of nitrogenous fertilizers can be made up if one can tap thevast reservoir of atmospheric nitrogen in a simpler way-in the nodules of roots legumedplants e.g. soybean, ground nut, chickpea etc. These are called natures minifertilizer factories.Some specific bacteria or micro-organisms in the soil convert this nitrogen into ammonia andamino acids. These amino acids can be used by the plants to build up proteins. This processworld wide is known as "biological nitrogen fixation". The nitrogen fixation is done by twoways: 1. Symbiotically by Rhizobium, Azolla and Frankia 2. Non-symbiotically by Azotohacter, Azospirillum, Acetobacter, Beijerinckia, and Blue green algae. Biofertilizers are the preparations containing cells of microorganisms which may be nitrogenfixers, phosphorus solubilizers. sulphur oxidisers or organic matter decomposers. In short,
  20. 20. 2 Introductionthey are called as bioinoculants which on supply to plants improve their growth and yield. Inrecent years a need has arisen for organic fertilizers including biofertilizers to minimise ourdependence on fertilizer nitrogen. The experiments conducted in India and abroad onbiofertilizers revealed that legumes viz, beans, soybean, chickpea, pigeon pea can fix 50-500 kgatmospheric nitrogen per hectare under ideal conditions. In recent years a use of Rhizobium,culture has been routinely recommended as an input in pulse cultivation. In India about 30million hectares of land is under pulses. Therefore use of Rhizobium is made when India canstop the impo11 of nitrogenous fertilizers. The association between non-legumes and N2 fixing bacteria as shown by increasednitrogenase activity is now well established Azotobacter and Azospirillum have been widelytested to increase yields of cereals. Of late, attention has been shifted from Azotobacter toAzospirillum due its wide distribution in soils and is relatively more efficient in utilisation ofcarbon to supp0l1 N2 fixation. In an evaluation of the reported worldwide success ofAzotobacterand Azospirillum increase inoculation. Statistically significant in yield has been obtaint!d. Ingeneral inoculation the C 4 plants corn, sorghum, Paricum and Setaria shoed greater yieldincreases than C3 plants wheat. Most striking effects of these organisms is their ability to stimulatethe germination of seeds, improve plant stand, augment root biomass and accelerate the initialvigour and plant stand, so also synthesis of chlorophyll. There is a conclusive evidence thatthese organisms secrete IAA, Kinetin, Oibberelins and B-vitamins. These factors are partlyresponsible for enhanced growth of plants and the ability of organisms to fix atmosphericnitrogen. It has also been proved beyond doubt that Azotobacter synthesises antifungal antibioticswhich prevent the seedling mortality caused due to fungal pathogens. Therefore, the organismis considered for production of biopesticides. It is also known that the organism secretes gumor polysaccharides which contribute to the soil improvement particularly soil structure.Acetobacter is a non-symbiotic (micro-symbiont) bacteria which is mostly associated withsugarcane crop. It grows along with one inside the root as well as stems to some extent, andfixes the atmospheric nitrogen and benefits the crop. It is rather difficult to isolate the organismand grow artificially on a large scale. It is for the scientists gathered here to find out suitabletechniques in order to make the organism to grow faster in the laboratory for the purpose ofbiofertilizers. If this is done successfully we may save huge quantity of chemical nitrogenousfertilizers used to grow sugarcane crop. Blue green algae are another potential input for low lying rice erop, which are also calleda cyanobacteria. Cyanobacteria mUltiply through vegetative and sexual methods. They are widelydistributed and responsible to increase rice yield as reported from places including Pune. Thistype of biofertilizers could contribute about 30-50 kg nitrogen per ha. thereby saving input costof chemical fe11ilizers. They are very simple to multiply even farmers can adopt techniques tocarry out production of BOA on their farms. Maharashtra has a large acreage of saline-alkali soils which are unsuitable for growing anycrop. Various methods are appl ied to reclaim them including providing proper drainage. adoptionof suitable cropping systems, and using proper irrigation methods. However, these systems
  21. 21. Introduction 3need serious efforts at various levels. There are scanty reports that BOA could reclaim suchsoils and reduce the salinity to 25·50%, pH, electrical conductivity and exchangeable sodium.It was further reported that it increases soil aggregation, hydraulic conductivity, soil nitrogenand permeability. However, the mechanism is yet to be investigated. The commonly knownspecies of BOA are Anabaena, Nostoc, Calothrx and Scytonema. I therefore, appeal to thisaugust gathering to conduct more research to solve soil problems affecting crop yield mostlysalinity and alkalinity. Azolla is an aquatic fern that assimilates atmospheric nitrogen in association with thenitrogen fixing symbiotic blue green algae. Anabaena azollae. The agronomic potential ofAzolla is quite significant particularly for rice crop and it may be applied as biofertilizerincreasing rice yield. It is reported that Azolla also suppresses the weeds in wetland rice besidesproviding additional nitrogen to rice. Temperature is the most important factor responsible forgrowing Azolla in tropics. This is considered as limiting factor for cultivation of Azolla and Irequest the scientists gathered here to do the research for obtaining the strains tolerance to hightemperature. Secondly, Azolla need to be multiplied and kept on water throughout its growth and freshAzolla is used for inoculation. This is one of the constraints to store the Azolla. Besides, theviability of Azalia forming seeds is very poor and hence it will be prime responsibility ofresearch workers to breed Azolla forming seeds having longer viability. Some non-leguminous shrubs and trees are modulated by nitrogen fixing actinorrhizae,F,·.::mkia and actiomycete. There are no species of Frankia 50 fare created. Nitrogen fixation occursin terminal swellings ofthe actiomycetes hyphae called vesicles. The nodule size can range froma few mm to several cm diameter. They are not pink in colour but a leg haemoglobin like moleculeshave recently been detected in the nodules. The genus Frankia has been studied in detail by astudent of Dr. P.L. Patil who reported several new species from Pune for the first time. This study includes growing of Azolla in pure culture. Frankia is a micro-aerophillic.Nitrogen fixation in pure culture appears to be relatively insensitive to oxygen. The Frankiafixes atmospheric nitrogen up to 150 kg per hectare. However, its commercial utilisation ashiofertilizer has not been studied which needs to be investigated by the scientists activelyinvolved in microbiological research. Amongst the most widespread, yet least recognised, relationship between microorganismsand higher plants are symbiosis known as mycorrhizae (fungus + plant root). In this systemplant roots and certain species of fungi form an intimate association one which is beneficial toeach other. Three general types of mycorrhizae are known such as ecto, endo (also-vasiculararbuscular mycorrihzae) and ectoendomycorrhizae. YAM are obligate parasites and hence cannotbe cultured on artificial media. YAM increases efficiently mineral nutrient from soil resultingin enhanced growth. It was also reported that YAM increases water uptake or alter plantsphysiology to reduce stress response to soil drought. There are several other benefits derivedfrom YAM including withstand to higil temperature, help to reduce toxicity etc. Though YAM are beneficial to shrubs or herbs their use has been limited because they areobligate and hence they cannot be cultured on a large scale. I will therefore appreciate if
  22. 22. 4 Introductionanybody works out a method to culture the VAM which can break through the research ofmycorrhizae. For degradation of any agricultural waste organic material of any source needsmicroorganisms although in some cases chemical degradation is possible but not economical.In nature, organic matter is degraded by different groups of micro-organism viz bateria, fungi,Actinomycetes and Protozoa. Bacteria are beginners who take active part in degradation.Secondary flora offungi are developed which also take active part in degradation of agriculturaland city wastes. Agricultural College, Pune is pioneer in developing bio-inoculant for degradation oforganic wastes and preparation of decomposing cultures. These micro-organisms includeAspergillus, Penicillium, Trichoderama, Trichurus, and Paecelomyces. These organisms formdifferent types of acids resulting into decomposition of organic matter. Use of compostlFYMis essential in order to increase the efficiency of chemical fertilizers. Under this situation thedegradation of organic wastes is essentially a biological process. By using these efficientcultures period of degradation of agricultural wastes can be minimised by 30-40%. A groupof microbiologists headed by Dr. P.L. Patil, at Pune has developed a unique process ofproduction of decomposing cultures which was subsequently followed by others. Now mostof the sugarcane factories use decomposing cultures for the process of spentwash-pressmuddegradation. This has resulted into utilisation of spent was and pressmud thereby increasingthe availability of organic matter for agricultural production. The role of P solubilisingmicro-organisms has been extensively studied by different-workers. Next to nitrogen P isimportant nutrient required by plants for their growth. Phosphorous applied to soil in one orother way is fixed and not easily available to crop plants resulting into depraving of plants fromusing the phosphorus. It is very necessary to solubilise this phosphorus so that plants canobtain as and when required by them. The micro-organisms viz, bacteria and fungi playasignificant role in solubilising the applied as wen as native phosphorus. Some bacteriamobilise insoluble forms of phosphate into soluble forms. Pre-treatment of seeds of cerealswith phosphobacteria has been reported to help in reducing fertilizer phosphate requirementsof the crop and increasing its grain yield. The bacteria or fungi are known to dissolve nativephosphate through enzymatio action and made it available for plat growth. Crop rotation inwhich cereal crops follow legumes are known to derive double benefits by way of addedorganic nitrogen and soluble phosphate due to actions of micro-organisms in and around thelegume root system. Although, an extensive research has been made on uti~isation of phosphorus solubilisingmicro-organisms in order to provide soluble form of phosphate to plants its large scaleimplementation has not been seen. Hence it is necessary to extent the use of micro-organism inenhancing the solubility of native as well as applied phosphate more efficient strains need to beisolated and used for preparation of biofertilizers. In order to reach the estimated target of 240 Mt of foodgrains by 2000 A.D. we mustharvest the benefits from microbes particularly BNF. We have large area under pulses as wellas cereals and hence we need biofertilizers in million tons. Besides we have to provide good
  23. 23. Introduction 5quality biofertilizers. I was told that at present there is no law for defaulters those who dontproduce good quality biofertilizers. I therefore, alert the scientists to press for such law inevery state. Why there is specificity in certain bacteria to selected plants? The genus Rhizobium isselective in choosing roots of particular legumes. Similar is the case of other species of microbes.We should find out reasons why this is so. Secondly search for organisms which will have modulating capacity on cereals not to bedone although false nodules have been reported on cereals. Can we evolve nitrogen-fixingplants? So that question of adding chemical fertilizers will not arise. These are some of theconstraints in biological nitrogen fixation. You may ponder during next two clays over theseareas so also other important aspects of microbiology. There are not may micro-organisms which can be used as biopesticides except Bacillusthuringiensis and Trichoderma. Besides, there are no proper methods to use them effectivelyin control of plant diseases. These are some of the future research in microbiology.AuthorsChief Editor: Dr. A.M. Deshmukh, Reader & Head, Dept. of Microbiology, Dr. Babasaheb Ambedkar Marathwara University, Subcentre Osmanabad-413 501 (MS.)Co-editors: Shri R.M. Khobragade & Shri P.P. Dixit, Lecturers, Dr. Babasaheb Ambedkar Marathwara University, Subcentre Osmanabad-413 501 (MS.)
  24. 24. 1 Effect of Biofertilizers on Seedling Growth, Physiology, Nodule Production and VAM Colonization in Pungam, Pongamia pinnata (L.) PierreIntroductionPungam is extensively used for afforestation of watersheds in the drier parts of the country. Itis drought resistant. moderately frost hardy ad highly tolerant to salinity. It is being propagatedby direct sowing or by transplanting one year old seedlings raised in nursery (Troup, 983). Micro-organisms that are sued as biofertilizers stimulate plant growth by providing necessarynutrients as a result of their colonisation at the rhizosphere (Azotobacter, Azospirillum,Pseudomonas, phosphate-solubilising bacteria, and Cyanobacteria) or by symbiotic association(Rhizobium, mycorrhizae and Frankia). The role of biofertilizers has already been provedextensively in annual crops, but its exproitation in perennial trees in India is scanty. In forestry,few research reports are available to demonstrate that biofertilizers stimulate the growth(Niranjan el at., 1990), biomass (Sekar et at.; 1995), nodulation and YAM-colonization, (Reenand Bagyaraj, 1990). Keeping this in view, the present study was designed to elicit informationon the effect of biofertilizers on the growth and development of pungam (Pongamia pinnata)(L.) Pierre.Material and MethodsThe study was carried out at Forest College and Research Institute (II ° 19 N, 76°56 E, 300above MSL). Six month old uniform sized seedlings were. chosen for biofertilizer inoculation.The seedlings were transplanted in 30 x 45 cm polybags filled with nursery mixtures of redsoil. sand and FYM (2: I : I). The experiment was set up in a completely randomised design andreplicated thrice. The nursery soil mixture of the polybags were inoculated with biofertilizers,vi= .. (i) Rhizobium, (ii) phosphobacteria, (iii) Yesicular-Arbuscular Mycorrhizae (YAM),(iv) Rhizobium + phosphobacteria, (v) phosphobacteria + YAM, (vi) Rhizobium + YAM,
  25. 25. Effect of Biofertilizers on Seedling Growth, Physiology. Nodule Production ... 7(vii) Rhizobium + phosphobacteria + VAM. Un inoculated seedlings were maintained ascontrol. Biofertilizer inocu.1ation was prepared with a base of peat soil. Two hundred grams ofRhizobium and phospobacteria were weighed and mixed with 3 kg of well decomposed andpowdered FYM separately. Fifty grams of this inoculum mixture and twenty grams of VAMinoculum were applied to each polybag at 5 cm depth near the root zone. Six seedlings in each treatment were selected at random and observed initially, 2 and 4months after inoculation for number of leaves, leaf area (LICOR Model LI 3000 Leaf AreaMeter). root volume. total dry matter, DGR, NAR (Williams, 1946), CGR (Watson. 1958),number of nodules and VAM colonisation (Phillips and Hayman, 1970). The results weresubjected to analysis of variance and tested for significant differences (P < 0.05) after Panseand Sukhatme (1967).Results and DiscussionA significant increase in number of leaves (47.7%) (Table 1), total leaf are (44.4%) (Table 1)and root volume (47.3%) (Table 2) was achieved over control by inoculating the seedlings withRhizobium, phosphobacteria and VAM conjointly. The results corroborated the findings ofSekar et al., (1995) in Shola species. The microorganisms that are used as biofertilizers colonisein the rhizosphere and stimulate the plant growth by providing necessary nutrients, thanks totheir symbiotic association (Varma and Schuepp, 1995). The association may also regulate thephysiological process in the ecosystems by involving in the decomposition of organic matter.fixation of atmospheric nitrogen. secretion of growth promoting substances, increasedavailability of mineral nutrients and protection of plants from pathogens besides increasing theavailability of nutrient elements at the root zone (Newman, et al., 1992). An increase of 54.5% in total dry weight over the control was evident in seedlings inoculatedcombined with Rhizobium, phosphobacteria and VAM (Table 1). Similar increase in biomassproduction due to VA-mycorrhizal inoculation was documented in Acacia spp. (Reena andBagyaraj et al., 1990); and Albizia spp. (Jamaluddin et al., 1995). In the present investigation,individual inoculation VAM also increased biomass relative to the control though not to thetune of combined biofertilizer inoculation. The increase in seedling biomass production maybe strongly correlated with increased accumulation ofN due to Rhizobium (Jamaluddin et al.,1995) and P due to VAM and phosphobacteria (Reddy et aI., 1996). Tripartite inoculation of Rhizobium, phosphobacteria and VAM registered higher numberof nodule production (69.9 %) than uninoculated control (Table 2). Dual inoculation withRhizobium and G. Jasciculatum improved the nodulation status in L. leucocephala compared tothe single inoculation (Manjunath et al., 1984). In the present experiment, the combinedinoculation of Rhizobium, phosphobacteria and VAM registered higher VAM-colonisation(65.6%) than the uninoculated seedlings (56.1 %) (Table 2). Such an increase in root colonisationdue to VAM and Rhizobium inoculation was reported in L. leucocephala (Manjunath et al., 1984). The relative growth rate (RGR) of P. pinnata seedlings inoculated with Rhizobium,phosphobacteria and YAM was higher (0.0071 g.g-I.day-I) compated to the control seedlings
  26. 26. oc Table 1 Effect of biofertilizers onnumber of leaves, total leaf area and total dry weight of pungam seedlings. tTl ~ n .-+ 2 l lBiofertilizers (B) Number of leaves Total leaf are (cm planr ) Total dry weight (g. planr ) ..., 0 Months after inoculation (M) Months after inoculation Months after inoculation C::I o· 0 2 4 Mean 0 2 4 Mean 0 2 4 Mean ~ a.Rhizobium (R) 18.7 43.7 48.7 37.0 312.6 431.1 569.0 437.5 9.14 27.56 36.13 24.27 ~. 47.7 . ... tilPhospho bacteria (P) 18.3 45.3 37.1 305.4 437.9 569.0 464.0 9.45 32.35 41.01 24.60 0 :lVAM 19.0 46.3 53.3 39.6 302.0 462.5 648.6 491.6 9.08 27.61 39.80 25.50 (/) (1) (1)R+P 20.0 48.7 54.3 41.0 305.0 498.9 710.2 504.3 9.25 31.15 40.12 26.84 e: :rP+VAM 19.0 47.7 57.3 41.3 286.4 499.6 651.3 479.1 9.44 29.88 40.60 26.64 (JQR+VAM 19.3 52.3 62.0 44.6 297.6 556.1 739.0 531.0 9.11 32.22 42.39 27.91 0 0.01 aR+P+VAM 19.7 58.7 68.3 48.9 303.0 641.2 850.0 589.0 43.18 46.94 33.04 ~Control 19.7 37.3 42.3 33.1 291.8 408.0 524.1 408.0 9.11 23.15 31.88 21.38 .?" -c :rMean 19.2 47.5 54.3 300.4 488.6 675.1 9.20 30.89 39.86 ~ o· 0 SEd CD (P=0.05) SEd CD (P=O.05) SEd CD (P=0.05) ~B 1.04 2.10 24.76 49.76 0.839 1.686 z 0 0- s::M 0.64 1.29 15.15 30.47 0.514 1.033 (i"BxM 86.19 1.453 2.921 -c 1.81 3.64 42.86 a0- s:: n .-+ o· :l
  27. 27. tTl Tablel ~ Effect of biofertilizers on root volume, number of nodules and vesicular-arbuscular mycorrhizae colonisation of ~ pungam seedlings. ...., 0 c::l o·BioJertilizers (B) Root value (mm 3) Number oj nodules VAM colonisation % ~ Months after inoculation (M) I Months after inoculation Months after inoculation 2:. ~ 0 2 4 Mean 0 2 4 Mean 0 2 4 Mean ... en 0 :::IRhizobium (R) 13.0 25.0 29.0 22.3 9.0 19.0 21.3 16.4 51.0 58.3 66.0 58.4 VJ G (45.6) (49.8) (54.3) (50.0) G e:Phosphobacteria (P) 13.3 26.3 29.0 22.9 9.3 12.3 23.3 15.0 51.3 57.7 65.7 58.2 5 OCI (45.8) (49.4) (54.1) (49.8) aVAM 13.5 25.0 30.0 22.8 10.0 11.3 17.3 12.9 51.7 61.7 70.0 61.1 ... 0 (45.9) (51.8) (56.8) (51.5) ~ ?"R+P 12.8 26.0 30.7 23.2 10.0 15.7 18.3 14.7 51.0 59.0 66.0 58.7 "tl (45.6) (50.2) (54.3) (50.0) ~ enP+VAM 13.4 31.3 34.7 26.5 9.0 13.0 17.0 13.0 . 51.3 63.3 71.0 61.9 o· (45.8) (52.7) (57.4) (52.0) 0 62.7 ~R+VAM 12.9 36.7 42.0 30.5 9.0 18.0 20.0 15.7 51.3 64.0 72.7 Z (45.8) (5~.1) (58.5) (62.7) 0 Q.R+P+VAM 13.6 43.3 49.7 35.S 8.3 20.0 29.3 19.2 52.0 66.7 78.0 65.6 s:: cr (46.1) (54.7) (62.0) (54.3) "tlControl 13.3 21.7 27.0 . 20.7 9.0 10.3 14.7 11.3 51.3 55.0 62.0 56.1 a Q. s:: (45.8) (47.9) (52.0) (48.5) ~Mean 13.2 29.4 34.0 9.2 14.9 20.2 51.4 60.7 69.0 o· :::I (45.8) (51.2) (56.2) SEd CD(P=0.05) SEd CD(P=0.05) SEd CD (PO.05)B 1.19 2.40 0.88 1.77 0.44 0.89M 0.7) 1.47 0.54 J.09 0.27 0.55L"M 2.07 4.16 1.53 0.37 0.77 1.55
  28. 28. o Table 3 Effect of biofertilizers on relative growth rate, crop rate and net assimilation rate of pungam seedlings. tTl ~ nBioJertilizers (B) RGR (g. g . day) CGR (g.m 2.day ) NAR (g.m- 2.day-) ..... 0 ....., Months after inoculation (M) Months after inoculation Months after inoculation tIl 0-2 2-4 0-2 2-4 o· 0-2 2-4 Mean Mean Mean ~ 3-.Rhizobium (M) 0.0080 0.0020 0.0050 0.307 0.144 0.225 3.60 1.48 2.54 N .., (1)Phospho bacteria (P) 0.0089 0.0017 0.0053 0.382 0.145 0.263 4.54 1.44 2.99 en 0VAM 0.0080 0.0052 0.0052 0.304 0.203 3.256 3.56 1.47 2.52 ::s en (1)R+P 0.0089 0.0018 0.0053 0.365 0.149 0.257 4.04 1.49 2.77 (1)P+VAM 0.0083 0.0023 0.0053 0.341 0.179 0.260 3.85 1.43 2.64 e: S· (JQR+VAM 0.0091 0.0020 0.0056 0.385 0.169 0.277 4.05 1.53 2.79 a ..,R+P+VAM 0.011 0.0028 0.0071 0.569 0.145 0.357 5.62 1.83 3.73 0Control 0.0067 0.0024 0.0045 0.234 0.062 0.148 2.92 U8 2.15 S ~::r "0Mean 0.0086 0.0022 0.361 0.149 4.02 1.51 ::r < en o· SEd CD(P = 0.05) SEd CD (P = 0.05) SEd CD (P = 0.05) 0- (JQ :;<B 0.00049 0.00100 0.0287 0.0584 2.734 NS ZM 0.00024 0.00050 0.0143 0.0292 1.146 2.341 0 0- cBxM 0.00069 NS 0.0406 0.0826 1.596 3.257 (j) .., "0 0 0- C n ..... o· ::s
  29. 29. Effect of Biofertilizers on Seedling Growth, Physiology, Nodule Production ... 11(0.0045 g.g-I.day-I) (Table 3). Similarly, increase was also evident in crop growth rate (CGR),the quantum of increase being 141.12 per cent over the control. However, the net as simulationrate (NAR) was not significantly influenced due to the treatments. The COR is function of drymatter production (Milthrope, 1967) hence, the increase in CGR and RGR in the presentinvestigation might be due to the increased dry matter production in the seedlings inoculatedwith Rhizobium + phosphobacteria + YAM. The results of the present study are in strong support of combined inoculation of the Ppinnata seedlings with Rhizobium phosphobacteria and YAM. It is hence recommended thatseedlings produced for largescale afforestation should be subjected to such an inoculationtreatment to ensure better plant growth and survival.SummaryAn experiment was c(mducted to study the response of pungam to biofertilizer inoculationviz., Rhizobium, phosphobacteria and YAM individually and conjointly under nursery conditions.The uninoculated seedling formed the control. The results indicated an enhancement in numberof leaves, total leaf area, total dry matter, root volume, number of nodules and YAM colonisationdue to triple inoculation with Rhizobium, phosphobacteria and YAM. This also recordedincreased ROR and COR, relative to the control. ReferencesJamaluddin, Dadwal, V.S. Coudan, J .S. (1995). Efficacy of different Rhizobium strains of forest tress species on Albizia lebbek. Indian For., 121 (7): 647-645.Manjunath, A., Bagyaraj, D.F. and Gopal Gowda, H.S. (1984). Dual inoculation with YAM and Rhizobium is beneficial to Leucaena. PI. Soil., 78: 445-448.Milthrope, F.L. (1967). Some physiological principles determining yield of root crops. Proc. Int. Symp. Trop. Root crops, 1: 1-19. In: Ecophysiology o/Tropical Crops (ed. Aluim, P. T. and T. T. Kozlowski). Academic Press, New York, 187-236.Newman, E.L., Eason, W.R., Eissenstat, D.M. and Ramos, M.I.R.F. (1992). Interaction between plants: the role of mycorrhiza. Mycorrhiza, 1: 47-53.N iranj an, R., Banwarila, S. and Rao, M. (1990). Studies on the effect of Rhi::obium and endomycorrhizal interaction in Dalbergia sissoo. In Proceedings 0/ National Conference qf Mycorrhiza, Hisar Agricultural University, Hisar. India Feb. 14-16, 1990,205-207.Panse, v.G. and Sukhatme, P. V. (1947). Statistical methods/or agricultural workers, Indian Council of Agricultural Research, New Delhi.Phillips J.A. and Hayman, D.S. (1970). Improved procedures for clearing roots and staining parasitic and vesicular arbuscular fungi for rapid assessment of infection. Mycol. Soc., 55: 158-161.Reddy, B., Bagyaraj, OJ. (1990) and Mallesha, B.C. (1996). Selection of efficient CA-mycorrhizal fungi for Acid lime.lndianJ. Microbiol., 36 (3): 13-16.Reena, J. and Bagyaraj, OJ. (1990). Response of Acacia nilotica and Callianadra calothyrsus to different VAM. Arid Soil Research and Rehabilitation, 4(4): 261-268.
  30. 30. 12 Effect of Biofertilizers on Seedling Growth, Physiology, Nodule Production ...Sekar, I., Vanangamudi, K. and Suresh, K.K. (1995). Effect ofbiofertilizers on the seedling biomass, VAM-Colonisation, enzyme activity and phosphorus uptake in the shoal tree species. My forest, 31(4): 21-26.Troup, R.S. (1983). The Silviculture of Indian Tress, Vol IV; The Controller of Publications, Delhi.Varma, A. and Schepp, H. (1995). Mycorrhization of the commercially important micro-propagated plants. Critical Rev.. Biotechnol., 15(3/4): 313-328.Watson, D.G. (1946). The Physiological basis of variation in yield. Adv. in Agron., 4: 101-145.Williams, R.E., (1946). The physiology of plant growth with special reference to the concept of net assimilation rate. Ann. Bot., 41-71.Authors A. Venkatesh, Mallika Vanangamudi, K. Vanangamudi, K.T. Parthiban and Ravichandran Forest College and Research Institute Mettupalayain-641301, T.N., India
  31. 31. 2 Biofertilizer: A Supplementary Nutrient Source for SugarcaneIntroductionBiofertilizer have an important role to play in improving nutrient supplies and their cropavailability in the years to come. They are of environment friendly non-bulky and low costagricultural inputs. A biofertilizer is an organic product containing a specific micro-organismsin concentrated form which is derived either from the plant roots or from the soil of root zone(Rhizozsphere). High productivity in irrigated areas like sugarcane leads to higher nutrientturnover and a larger gap between nutrient supply and removal. Therefore, there is strong needto have complementary use of available source to plant nutrient including biofertilizer alongwith mineral fertilizers fo~ maintenance of soil productivity. The microorganisms responsible for fixing atmospheric nitrogen has been recognised sincetime immemorial, however, commercialisation of their use is a relatively new innovation., Inview of their apparent low-cost and risk-free application leading to yield advantage both forcurrent as well.as for subsequent crop growth or residual fertility. Their large scale applicationin intensive farming call for systematic efforts. Their use especially important in view of thefact that in the country to be self sufficient in the production of nitrogenous and phosphaticfertilizers. Among the biofertilizers Azotobacter, Azospirillum, Acetobacter are the importantfor nitrogen fixation, Bacillus sp. and Aspergillus sp. are important for phosphate solubilisatiO!!and other soil mineral nutrients.Experiment and ResultsAzotobacterThis is a group of bacteria, which are free-living nitrogen fixer. The mechanism by whichthe plants, inoculated with Azotobacter, derive possible benefits in terms of increased grain,
  32. 32. 14 Biofertilizer: A Sllpplementary Nutrient Source for Sugarc~neplant biomass and nitrogen uptake which in turn are attributed to small increase in nitrogeninput from biological nitrogen fixation, development and branching of roots, production ofplant growth hormones, vitamins, enhancement in uptake as N03 , NH 4 , H2 P0 4 K and Fe,.i~proved water status of the plant, increased nitrate reductase activity and antifungalcompounds. Table 1 Effe.ct of Azotobacter culture on sugarcane yield (T/ha). Nitrogen levels Inoculated Uninoculated Mean 250 kglba 146.43 132.97 139.70 300 kglha 152.77 145.70 149.23 350 kglba 156.73 149.07 152.90 400 kglha (control) 154.67 149.98 152.32 Mean 152.65 144.43 SE ± 1.83; C.D. (5%) 5.52 Fixation of nitrogen to the sr-;1 by use of Azotobacter culture is a natural phenomenon and sufficient research work has been carried out on the role of Azotobacter culture in sugarcane cultivation at Sugarcane Research Station, Padegaon. The results, in general, indicate that application of Azotobacter at the rate of 5 kglha helps in reducing nitrogen dose by 50 kglha with increase in cane yield by 5 to 10 per cent (Anonymous, 1992, Patil & Hapase, 1981). However, it can help in reducing nitrogen dose even up to the tune of 100 kglha without loss in. yield. The Azotobacter plays an important role in improving cane germination, establishing of seedlings, survival of tillers, increase the population of millable cane and improve the soil fertility. Azospirilluln It is an associativemicro-aerophilic nitrogen fixer. It colonises the root mass and fixes nitrogen in close association with plant. It fixes nitrogen in an environment of low oxygen tension. These bacteria induce the plant roots to se~rete a mucilage which creates low oxygen environment and helps to fix atmosphet;ic nitrogen. High nitrogen fixation capacity, 16wenergy requirement and abundant estabHsh~ent in the roots of sugarcane and tolerance to high soil temperature makes these most suitable for tropical conditions. Use of Azospirillum inoculum under saline-alkali conditions is also possible because their strains are known to maintain high nitrogen activities under such stress conditions (Hegade & Dwivedi, 1994; Marwaha, 1995). The field experiment revealed that Azospirillum inoculatin increase the cane yield up to 4.47 tonnes/hectare and could save 25% nitrogen dose in medium black soil under Adsali planting (Shinde ef af., 1991).
  33. 33. Biofertilizer: A Supplementary Nutrient Source for Sugarcane 15 Table 2 Effect of Azospirillum culture on sugarcane yieldTreatment Yield (Tlha) Available N in soils (kglha)100% N (control) 122.19 179.5575% N + Azospirillum 126.44 169.9550% N + Azospirillum 117.10 150.60SE ± 1.37; C.D. (5%) 4.10 Table 3 Effect of Azotobacter and Azospirillum inoculation under graded levels of nitrogen on yield and CCS of sugarcane (average of 3 years).Cultures Nitrogen levels kglha 175 200 225 250 . meanControl 103.50 105.73 107.03 II 0.47 106.68 (12.25) (12.71) (13.07) (13.23) (12.81)Azotobacter 106.70 108.20 11.97 112.67 109.88 (12.58) (12.83) (13.42) (13.50) (13.08)Azospirillum 108.23 109.87 112.63 113.43 111.04 (12.59) (12.93) (13.55) (13.69) (13.19)AZT+ AZP 108.53 110.73 II 4.33 II 5.00 112.14 (12.59) (13.11) (13.85) (13.95) (13.37)Mean 106.74 108.63 I 1I.49 112.89 (12.50) (12.89) (13.47) (13.59)AZT-Azotobacter, AZP-Azospirillum*Figures in parentheses denote the CCS in MT/ha. (commercied cane sugar)I. Yield C.D. (P-0.05) CCS C.D. (P-0.5)2. Culture 0.32 0.0323. N levels 1.78 0.1024. Interaction 3.79 0.33Phosphate Solubilisiug MicroogranismsPhosphorous is the most important micro-nutrient required by sugarcane and micro-organismsfor their normal growth and development. However, Indian soil test, generally low to medium
  34. 34. 16 Biofertilizer: A Supplementary Nutrient Source for Sugarcanein available phosphate and not more than 30% of applied phosphate is available to the currentcrop, remaining part gets converted into relatively unavailable forms. Many common heterotrophic bacteria like Bacillus megaterium and B. circulans and fungilike Aspergillus awanari and Penicilium striata posses the ability to bring sparingly soluble/insoluble inorganic and organic phosphorus into soluble forms by secreting organic acids.These organic acids lower soil pH and in turn brings about dissolution of unavailable forms ofsoil phosphorous. Some of the hydroxy acids may chelate Ca, AI, Fe and Mg resulting ineffective availability of soil and hence its higher utilisation by sugarcane plants. The fieldexperiments showed that the inoculation of phosphorus solubilising cultures to sugarcane setsat the time of planting increased cane and sugar yield significantly over uninoculated control(Shinde et al., 1987). It could be reduced the phosphate dose by 50% and could be applied inthe form of rock phosphate which is the cheaper source of phosphorus. Table 4 Effect of phosphate solubilising culture on cane and sugar yieldTreatment Cane yield CCS (Tlha)Control (N -t- K) 145.28 21.71N -t- K20 + P.S. Culture 152.48 22.63Rock phosphate + N + K20 164.07 25.65Rock phosphate + N + K20+ P.S. Culture 166.81 22.49Superphosphate + N + Kp + Culture 194.81 28.67Rock phosphate + 50% + N + K20 149.35 20.95Rock phosphate 50% + N + K20+ P.S. Culture 158.70 23.61Superphosphate 50% + N + K20 171.48 24.29Superphosphate 50 + N + K20 + P.S.c. 174.86 25.68Rock-P Super-Po 50% +N + K20 182.79 26.00Rock-P + Super-P 50% + N 191.87 27.43+ L20 + P.S.c.P.S.c. = Phosphate Solubilising CultureSE=±3.20C.D. 5% = 9.40 The other groups of micro-organisms like Acetabacter dizotrophicus a nitrogen fixer.Thiobacillus thioaxidans sulphur and iron oxidizers and Yasicular Arbuscular Mycorrhiza (YAM)a phosphate solubilisers are found to be effective but needs more experimentations atmultiplications and soil type for confirmation and recommendation.
  35. 35. Biofertilizer: A Supplementary Nutrient Source for Sugarcane 17SummaryBiofertilizers are low COSt agricultural input playing a significant role in improving nutrientavailability to the crop plants. The present paper examines potential of different bio-inoculantsin sugarcane crop for increasing cane and sugar yield as well as saving of chemical fertilizers.the recommendations made to sugarcane growers based on research work done at CentralSugarcane Research Station, Padegaon are highlighted. The field experiments revealed thatAzospirillum and/or Azotobacter inoculation at the rate of 5kg/ha increased the cane yieldranged from 4.57 to 11.50 t/ha with saving of25% nitrogenous fertilizer. However, the beneficialeffect was more pronounced in dual inoculation of Azotobacter plus Azospirillum culture. The phosphate solubilising bacteria Bacillus megaterium could increase the availabilityofphosphatic ferti Iizer and resulted into increased cane andsugar yield upto 15 per cent. Thecostly single super phosphate could be replaced up to 50 per cent by using cheaper rockphosphate with phosphate solubilising biofertilizer. The study on Acetobacter dizotrophicus aendophytic nitrogen fixer in sugarcane has also been reviewed in the paper. ReferencesAnonymous, (1992). AGRESCO Report, Sugarcane research Station Padegaon, pp. 29-45.Hegde, D.M. and Dwinedi, B.S. (1994). Crop response to biofertilizer in irrigated areas. Fert. News., 39 pp. 19-26.Marwaha, B.C. (1995). Biofertilizer-A supplementary source of plant nutrient. Fert. News., 40 (7): 39-50.Patil, P.G. and Hapase, D.G. (1981). Nitrogen economy in sugarcane (Adsali) by use of Azotobacter. Maharashtra Sugar. June, pp. 29-35.Shinde, D.B., Jadhav, S.K. Jadhav. and Jadhav, M.G. (1987). Use of Phosphate solubilising culture in sugarcane cultivation (Co. 7219) DSTA Convention, pp. A 213-218.Shinde, D.B., Navale, A.M. and Hadhav, S.B. (1991). Effect of Azospirillum inoculation on yield of sugarcane crop. DSTA Convention, pp. A-219-223.Authors B.D. Shinde and K.K. Khade Central Sugarcane Research Station Padegaon, Po. Nira- 412102 Dist. Pune, Maharashtra, India
  36. 36. 3 Effect of Composite Bioinoculations and Fertilizer Levels on Growth of Sorghum (CCH-14)IntroductionIndia, due to increase in population, may face the problems offood. So contribution towards theagricultural productivity improvement is necessary. Sorghum is the staple food crop ofMaharashtra State. The main principle of obtaining maximum yield from Sorghum hybrid isjudicious use of chemical fertilizers which at present is proving a set back in increasing the yieldas the chemical fertilizers are not only in short supply but also are expensive. Use of chemicalfertilizers is limited to rich farmers and they show certain drawbacks like their manufacturedepends on energy derived from fossil fuels which are getting depleted at faster rate. Many cropssuffer from potassium d~ficiency because of excessive use of nitrogen based fertilizers, excessivepotassium treatment decrease available nutritive food such as ascorbic acid etc., continuous useof ammonium fertilizers increase soil acidity and radioactivity, lastly chemical fertilizers haveless effect on succeeding crops. On the other hand biofertilizers are advantageous over chemicalfertilizers due to low cost, simple methodology of production, no any hazard to agroecosystemand considerable residual effect on succeeding crop. Tilak et al., 1982 reported that microbialinoculants application gives equivalent output as derived from the application of30 to 40 kglha.N. Biofertilizer makes a significant contribution towards the development of strategies forproductivity improvement and may help for economising the pro"duction cost. Since long effectof nitrogen fixers, phosphate solublisers singly has been studied by many workers on many crops.But little work on combined effect ofinoculant has been reported. Keeping this view in mind an attempt has been made to see the effect of bioinoculants ongrowth parameters of Sorghum using Azotobacter. Azospirillum, YAM and their differentcombinations. The same experiment is compared with recommended dose offertilizertreatmentand with no any treatment as a control.
  37. 37. Effect of Composite Bioinoculations and Fertilizer Levels on Growth ... .19Material and Methods Pure cultures of Azotobacer chrocooccum, Azospirillum brasilense and YAM fungus, Glomousjasiculatum, wher obtained from Sorghum Research Unit, Akola and ICRISAT, Hyderabad. Keeping all the recommended packages of practice the experiment was laid out in split plot design with 3 replication, using 24 treatment combinations. The main plot treatments were recommended full dose offertilizer 80N : 40P : 40K kg/h~, three-forth dose of fertilizer 60N : 30P : 30K and no fertilizer. While the subplot treatment were seed treatment with Azotobacter,Azospirillum, YAM, Nitrogen Fixers + YAM and no seed treatment. Seed treatment was carried out usingjaggery solution as an adhesive which was prepared by boiling 100 gm of jaggery in I litre of water. The seeds were treated in a pan by sprinkling the adhesive on seed cultures 10 gm of bioinoculant was used for treating 250 gms of seed. The seed material was eventually coated with cultures, seeds were treated two hours prior to sowing. Sowing was done by dibbling seeds. Germination studies were carried out in pot cultures. 10 seeds were treated with bioinoculants singly and combination. Germination count was taken 10 days after sowing. Simultaneously germination studies in laboratory was also carried out using Rad dolls paper towel method (lSTA 1976) 100 seeds were plated on two well moistened blotters (48 x 48 cms) and then covered with another well moistened blotters of the same size and then roll was prepared. For each treatment 100 seeds were tested. The rolls were then kept in upright position in a germination chamber at 30°C. To provide continuous moisture to the germinating seed, one end of the roll is kept in small amount of water. Germination count was recorded on lOth day after seeding. Simultaneously field trials was undertaken at various fertilizer levels and bioinoculant seed treatment. Observation regarding growth par.ameters like germination count, root and shoot length, biomass production and seedling vigour index were recorded by selecting 4 plants randomly at an interval of 15, 30, 45, 60 days. The data recorded for the growth parameters was analysed by standard statistical methods and all the treatments were tested for their significance at 5% probability levels.Results and DiscussionEffect of different treatments on germination count in pot cultures (Table I) indicates that withincrease dose of fertilizers germination percentage also increased but the increase was notsignificant, seed germination using single and composite culture inoculation in the pot studiesrevealed that maximum percentage of seed germination was observed when seeds were tre,tedwith Azotobacter + Azospirillum, followed by Azotobacter + Azospirillum + YAM. Germinationpercentage with treatment YAM culture was better than in combined treatment with Azotobacter+ YAM and Azospirillum + YAM and single inoculation of Azotobacter and Azospirillum.Seed germination percentage observed in paper towel test in laboratory conditions also showedsimilar effects those in pot studies. From the literature referred there are few reports on theeffect of the combined culture inoculation especially on the germination in Sorghum. However,present studies revealed that seed inoculation with Azotobacter and Azospirillum along withYAM increased the seed germination of Sorghum compared to single culture inoculation. Seed