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Biological control of root rot fungus Macrophomina phaseolina and growth enhancement of Pinus roxburghii (Sarg.) by rhizosphere competent Bacillus subtilis BN1

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Abstract

Bacterial isolates having antifungal and good plant growth-promoting attributes were isolated from chir-pine (Pinus roxburghii) rhizosphere. An isolate, Bacillus subtilis BN1 exhibited strong antagonistic activity against Macrophomina phaseolina, and other phytopathogens including Fusarium oxysporum and Rhizoctonia solani. It was characterized and selected for the present studies. BN1 resulted in vacuolation, hyphal squeezing, swelling, abnormal branching and lysis of mycelia. The cell-free culture filtrate of BN1 inhibited the growth of M. phaseolina. Pot trial study resulted in statistically significant increase in seedling biomass besides reduction in root rot symptoms in chir-pine seedlings. BN1 treatment resulted in 43.6% and 93.54% increases in root and shoot dry weights respectively, as compared to control. Also, 80–85% seed viability was recorded in treatments receiving BN1 either alone or in the presence of M. phaseolina, compared to 54.5% with M. phaseolina. Bioinoculant formulation study suggested that maximum viability of bacteria was in a sawdust-based carrier. B. subtilis BN1 produced lytic enzymes, chitinase and β-1,3-glucanase, which are known to cause hyphal degradation and digestion of the cell wall component of M. phaseolina. In the presence of M. phaseolina, population of B1 was 1.5 × 104 c.f.u. g−1 root after one month, which increased to 4.5 × 104 c.f.u. g−1 root in three months. Positive root colonization capability of B. subtilis BN1 proved it as a potent biocontrol agent.

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References

  • Altschul SF, Madden TI, Scaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search program. Nucleic Acids 25:3389–3402

    Article  CAS  Google Scholar 

  • Arora NK, Kumar V, Maheshwari DK (2001) Constraints development and future of the bioinoculants with special reference to rhizobial inoculants. In: Maheshwari DK, Dubey RC (eds) Innovative approaches in microbiology. Bishan Singh, Mahender Pal Singh, Dehradun, India, pp 241–254

    Google Scholar 

  • Bai YD′, Aoust F, Smith DL, Driscoll BT (2002) Isolation of plant growth-promoting Bacillus strains from soybean root nodules. Can J Microbiol 48:230–238

    Article  CAS  Google Scholar 

  • Bapat S, Shah AK (2000) Biological control of fusarial wilt of pigeon pea by Bacillus brevis. Can J Microbiol 46:125–131

    Article  CAS  Google Scholar 

  • Barnard EL (1994) The nursery to field carryover and post outplanting impact of M. phaseolina on loblolly pine on a cutover forest site in North Central Florida. Tree Planter’s Notes 45:68–71

    Google Scholar 

  • Bazzicalupo M, Fani R (1995) The use of RAPD for generating specific DNA probes for microorganisms. In: Clap JP (ed) Methods in molecular biology. species diagnostic protocols: PCR and other nucleic acid methods. Humana Press Inc, Totowa NJ

    Google Scholar 

  • Beatty PH, Jensen SE (2002) Paenibacillus polymyxa products fusaricidin-type antifungal antibiotics active against Leptosphaeria maculans, the causative agent of blackleg disease of canola. Can J Microbiol 48:159–169

    Article  CAS  Google Scholar 

  • Bello GMD, Monaco CI, Simon MR (2002) Biological control of seedling blight of wheat caused by Fusarium graminearum with beneficial rhizosphere microorganism. World J Micorbiol Biotechnol 18:627–636

    Article  Google Scholar 

  • Bhagwat AA, Thomas J (1983) Legume–Rhizobium interactions: role of cowpea root exudate in polysaccharide synthesis and infectivity of Rhizobium species. Arch Microbiol 136:102–105

    Article  CAS  Google Scholar 

  • Bric JM, Bustock RM, Silversone SE (1991) Rapid in situ assay for indole acetic acid production by bacteria immobilization on a nitrocellulose membrane. Appl Environ Microbiol 57:535–538

    CAS  Google Scholar 

  • Broglie K, Chet I, Holliday M, Cressman R, Biddle P, Knowlton S, Mauvais CJ, Broglie R (1991) Transgenic plants with enhanced resistance to the fungal pathogens Rhizoctonia solani. Science 254:1194–1197

    Article  CAS  Google Scholar 

  • Cattelan AZ, Hartal PG, Fuhrmann JJ (1990) Screening for plant growth-promoting rhizobacteria to promote early soybean growth. Soil Sc Am J 43:1670–1680

    Google Scholar 

  • Chan YK, Wayne AM, Seifert KA (2003) Characterization of antifungal soil bacterium and its antagonistic activities against Fusarium species. Can J Microb 49:253–262

    Article  CAS  Google Scholar 

  • Chanway CP, Holl FB, Turkington R (1990) Specificity of association between Bacillus isolates and genotypes of Lolium perenne and Trifolium repens from a grass-legume pasture. Can J Bot 68:1126–1130

    Google Scholar 

  • Chanway CP, Radley R, Holl FB (1991) Inoculation of conifer seed with plant growth-promoting Bacillus strains causes increased seedling emergency and biomass. Soil Biol Biochem 23:575–580

    Article  Google Scholar 

  • Claus D, Berkeley RCW (1984) Bergey´ s manual of systematic bacteriology. In: Noel R. Krieg NR (eds) vol 2, Baltimore: Williams & Wilkins, pp 1104–1208, ISBN 0-683-0408-8

  • de Temp J (1963) The blotter method for seed health testing. Proc Int Seed Test Assoc 28:1933–1935

    Google Scholar 

  • Deshwal VK, Dubey RC, Maheshwari DK (2003) Isolation of plant growth- promoting strains of Bradyrhizobium (Arachis) sp. with biocontrol potential against Macrophomina phaseolina causing charcoal rot of peanut. Curr Sci 84:443–448

    Google Scholar 

  • Dubey RC, Dwivedi RS (1988) Effect of heavy metals on growth and survival of Macrophomina phaseolina (Tassi) Goid. Biol Fert Soils 6:311–314

    Article  CAS  Google Scholar 

  • Dubey RC, Maheshwari DK (2002) Practical microbiology. S Chand Co, New Delhi. India

    Google Scholar 

  • Felsenstein J (2004) PHYLIP phylogeny inference package. Version 3.6. Department of Genome Sciences and Department of Biology, University of Washington, Seattle, WA, USA

    Google Scholar 

  • Gilman JC (1956) A manual of soil fungi. Cons and Company, London

    Google Scholar 

  • Glick BR (1995) The enhancement of plant growth by free-living bacteria. Can J Microbiol 41:109–117

    CAS  Google Scholar 

  • Grossman TH, Trukman M, Ellestad S, Osburne MS (1993) Isolation and characterization of Bacillus subtilis genes involved in siderophore biosynthesis; relationship between B. subtilis sfro and E. coli entD genes. J Bacteriol 175:6203–6211

    CAS  Google Scholar 

  • Holl FB, Chanway CP, Turkington R, Radley R (1988) Growth response of crested wheatgrass (Agropyron cristatum L.) white clover (Trifolium repens L.) and perennial ryegrass (Lolium perenne L.) to inoculation with Bacillus polymyxa. Soil Biol Biochem 20:19–24

    Article  CAS  Google Scholar 

  • Jensen CE, Percich JA, Graham PH (2002) Integrated management strategies of bean root rot with Bacillus subtilis and Rhizobium in Minnesota. Field Crop Res 74:107–115

    Article  Google Scholar 

  • Katz E, Demain AL (1977) The peptide antibiotics of Bacillus: chemistry, biogenesis, and possible functions. Bacteriol Rev 41:449–474

    CAS  Google Scholar 

  • McCain AH, Scharpf RF (1989) Effect of inoculum density of Macrophomina phaseolinaon seedling susceptibility of six conifer species. Eur J Forest Pathol 19:119–123

    Article  Google Scholar 

  • McCain AH, Bega RV, Jenkinson JL (1982) Solar heating fails to control Macrophomina phaseolina. Phytopathol 72:985–988

    Google Scholar 

  • Muhammad S, Amusa NA (2003) In-vitro inhibition of growth of some seedling blight including pathogens by compost-inhabiting microbes. Afr J Biotechnol 2:161–164

    Google Scholar 

  • Munnecke DE, van Gundy SD (1979) Movements of fumigants in soil, dosage responses, and differential effects. Ann Rev Phytopathol 17:405–429

    Article  CAS  Google Scholar 

  • Pal KK, Tilak KVB, Saxena AK, Dey R, Singh S (2001) Suppression of maize root disease caused by Macrophomina phaseolina, Fusarium moniliforme and Fusarium graninearum by plant growth-promoting rhizobacteria. Microbiol Res 156:209–223

    Article  CAS  Google Scholar 

  • Pandey P, Kang SC, Maheshwari DK (2005) Isolation of endophytic plant growth-promoting Burkholderia sp. from root nodules of Mimosa pudica. Curr Sci 89:177–180

    CAS  Google Scholar 

  • Pikovaskya RI (1948) Mobilization of phosphorus in soil in connection with the vital activity of micorobial species. Microbiol 17:362–370

    Google Scholar 

  • Rangel-Castro JI, Levenfors JJ, Danell E (2002) Physiological and genetic characterization of fluorescent pseudomonas associates with Cantharellius cibarius. Can J Microbiol 48:739–748

    Article  CAS  Google Scholar 

  • Rennie RJ, Larson RI (1979) Nitrogen fixation associated with disomic chromosome substitution lines of spring wheat. Can J Bot 57:2771–2775

    Article  CAS  Google Scholar 

  • Schisler DA, Slininger PJ, Behle RW, Jackson MA (2004) Formulation of Bacillus spp. for biological control of plant diseases. Phytopatholgia 94:1267–1271

    Article  CAS  Google Scholar 

  • Schreiber LR, Gregory GF, Krause CR, Ichida JM (1988) Production, partial purification and antimicrobial activity of a novel antibiotic produced by a Bacillus subtilis isolate from Ulmus americana. Can J Bot 66:2338–2346

    CAS  Google Scholar 

  • Schwyn B, Neilands JB (1987) Universal chemical assay for the detection and the determination of siderophores. Anal Biochem 160:47–56

    Article  CAS  Google Scholar 

  • Shishido M, Massicott HB, Chanway CP (1996) Effect of plant growth-promoting Bacillus strains on pine and spruce seedling growth and mycorrhizal infection. Ann Bot 77:433–441

    Article  Google Scholar 

  • Shishido M, Loeb BM, Chanway CP (1995) External and internal root colonization of lodgepole pine seedlings by two growth-promoting Bacillus strains originated from different root microsites. Can J Microbiol 41:707–713

    CAS  Google Scholar 

  • Siddiqui ZA, Mahmood I (1993) Biological control of Meloidogyne incognita race 3 and Macrophomina phaseolina by Paecilomyces lilacinus and Bacillus subtilis alone and in combination on chickpea. Fund Appl Nematol 16:215–218

    Google Scholar 

  • Skidmore AM, Dickinson CH (1976) Colony interaction and hyphal interference between Septoria nodorum and phylloplane fungi. Trans Brit Mycol Soc 66:57–60

    Article  Google Scholar 

  • Skujins JJ, Potgieter HJ, Alexander MA (1965) Dissolution of fungal cell wall by a Streptomyces chitinase and β-1, 3-glucanase. Arch Biochem Biophys 111:358–364

    Article  CAS  Google Scholar 

  • Smith RS Jr, Krugman SL (1967) Control of the charcoal root disease of white fir by fall soil fumigation. Plant Dis Rep 51:671–674

    Google Scholar 

  • Somasegaran P, Hoben HJ (1994) Handbook for rhizobia. Springer-Verlag New York Inc, New York, USA

    Google Scholar 

  • Srivastava AK, Singh T, Jana TK, Arora DK (2001) Induced resistance and control of charcoal rot in Cicer arietinuns (Chickpea) by Pseudomonas fluorescens. Can J Bot 79:787–795

    Article  Google Scholar 

  • Stephens JHG, Rask H (2000) Inoculant production and formulation. Field Crops Res 65:249–258

    Article  Google Scholar 

  • Thirumalachar MJ, O’ Brien MJ (1977) Suppression of charcoal rot in potato with a bacterial antagonist. Plant Dis Rep 61:543–546

    Google Scholar 

  • Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DX (1997) The CLUSTALX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucl Acids Res 25:4876–4882

    Article  CAS  Google Scholar 

  • Trivedi P, Pandey A, Palni LMS (2005) Carrier-based preparations of plant growth-promoting bacterial inoculants suitable for use in cooler regions. World J Microbiol Biotechnol 21:941–945

    Article  Google Scholar 

  • Weller DM, Cook RJ (1983) Suppression of take-all of wheat by seed treatments with fluorescent pseudomonads. Phytopatholgia 73:463–469

    Article  Google Scholar 

  • Whipps JH (2001) Microbial interactions and biocontrol in the rhizosphere. J Exp Biol 52:487–511

    CAS  Google Scholar 

  • Yoshida S, Hiradate S, Tsukamoto T, Hatakedo K, Shirata A (2001) Antimicrobial activity of culture filtrate of Bacillus amyloliquefaciens RC-2 isolated from mulbersy leaues. Am Phytopathol Soc 91:181–187

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Botany and Microbiology, Gurukul Kangri University, Hardwar, UA, 249404, India

    Neetu Singh, R. C. Dubey & D. K. Maheshwari

  2. Department of Microbiology, S.B.S.P.G. Institute of Biomedical Sciences and Research Balawala, Dehradun, UA, 248161, India

    Piyush Pandey

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  1. Neetu Singh
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  2. Piyush Pandey
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  3. R. C. Dubey
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  4. D. K. Maheshwari
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Correspondence to D. K. Maheshwari.

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Singh, N., Pandey, P., Dubey, R.C. et al. Biological control of root rot fungus Macrophomina phaseolina and growth enhancement of Pinus roxburghii (Sarg.) by rhizosphere competent Bacillus subtilis BN1. World J Microbiol Biotechnol 24, 1669–1679 (2008). https://doi.org/10.1007/s11274-008-9680-z

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