Isolation and characterization of halotolerant bacilli from chickpea (Cicer arietinum L.) rhizosphere for plant growth promotion and biocontrol traits
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The present study describes the identification and polyphasic characterization of salt tolerant bacilli displaying plant growth promoting and broad spectrum antifungal activities. A total of 110 strains were isolated, out of which nine were selected on the basis of halotolerance, in vitro plant growth promoting attributes and antagonism against multiple phytopathogens. Preliminary identification of strains was done on the basis of phylogenetic analysis and comparison of 16S RNA sequences with type strains. The test strains were identified as Bacillus pumilus, B. subtilis, B. licheniformis, B. safensis and B. cereus. These strains possess ability to tolerate high salt (8–10% NaCl), form endospore, showed broad spectrum antifungal activity and therefore might cope with adverse environments. Scanning electron microscopy of promising test strains (CG18 and CM25) and pathogen interaction indicated destruction of fungal mycelia by halotolerant antagonists due to cytoplasmic extrusion. Moreover, PCR amplification of bacillomycin (bmyB), chitinase (ChiA), and β-glucanase genes and production of hydrolytic enzymes suggested that test strains could have a role to manage plant pathogens under saline conditions. Finally, the selected strains exhibited some plant growth promotion traits. Overall, this study demonstrated that the evaluated strains could be useful in developing microbial products to enhance tolerance to biotic and abiotic stresses and boost plant growth.
KeywordsAntagonism Bacillus Chickpea Plant growth promotion Salinity
The authors thanks to Director, ICAR-NBAIM, Maunath Bhanjan (U.P.) for providing necessary support for conducting the research work.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
Research does not involve human participants and/or animals.
- Cappuccino, J., & Sherman, N. (1992). Microbiology: A laboratory manual (Vol. 125, p. 179). New York: Benjamin/Cummings Pub. Co.Google Scholar
- Gajbhiye, A., Rai, A. R., Meshram, S. U., & Dongre, A. (2010). Isolation, evaluation and characterization of Bacillus subtilis from cotton rhizospheric soil with biocontrol activity against Fusarium oxysporum. World Journal of Microbiology and Biotechnology, 26, 1187–1194.CrossRefPubMedGoogle Scholar
- Maxton, A., Singh, P., & Masih, S. A. (2017). ACC deaminase-producing bacteria mediated drought and salt tolerance in Capsicum annuum. Journal of Plant Nutrition. https://doi.org/10.1080/01904167.2017.1392574.
- Natarajan, A., Kumar, K., Madhuri, K., & Usharani, K. G. (2016). Isolation and characterization of salt tolerant plant growth promoting rhizobacteria from plants grown in tsunami affected regions of Andaman and Nicobar islands. Geomicrobiology Journal, 33, 942–947. https://doi.org/10.1080/01490451.2015.1128994.CrossRefGoogle Scholar
- Nene, Y., Reddy, M., Haware, M., Ghanekar, A., Amin, K., Pande, S., Sharma, M. (2012). Field diagnosis of chickpea diseases and their control. Information Bulletin No. 28 (revised) International Crops Research Institute for the Semi-Arid Tropics.Google Scholar
- Pikovskaya, R. I. (1948). Mobilization of phosphorus in soil in connection with vital activity of some microbial species. Microbiology, 17, 362–370.Google Scholar
- Saitou, N., & Nei, M. (1987). The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution, 4, 406–425.Google Scholar
- Sharma, A., Singh, P., Kumar, S., Kashyap, P. L., Srivastava, A. K., Chakdar, H., Singh, R. N., Kaushik, R., Saxena, A. K., & Sharma, A. K. (2015). Deciphering diversity of salt-tolerant bacilli from saline soils of eastern indo-gangetic plains of India. Geomicrobiology, 32, 170–180.CrossRefGoogle Scholar
- Siddikee, M. A., Chauhan, P. S., Anandham, R., Han, G. H., & Sa, T. (2010). Isolation, characterization, and use for plant growth promotion under salt stress, of ACC deaminase-producing halotolerant bacteria derived from coastal soil. Journal of Microbiology and Biotechnology, 20(11), 1577–1584.CrossRefPubMedGoogle Scholar
- Singh, R. K., Kumar, D. P., Singh, P., Solanki, M. K., Srivastava, S., Kashyap, P. L., Kumar, S., Srivastava, A. K., Singhal, P. K., & Arora, D. K. (2014). Multifarious plant growth promoting characteristics of chickpea rhizosphere associated Bacilli help to suppress soil-borne pathogens. Plant Growth Regulation, 73, 91–101.CrossRefGoogle Scholar
- Smibert, R., & Krieg, N. (1994). Phenotypic characterization. Methods for general and molecular bacteriology. In Methods for general and molecular microbiology (pp. 607–654). Washington, DC: ASM Press.Google Scholar
- Solanki, M. K., Robert, A. S., Singh, R. K., Kumar, S., Pandey, A. K., Srivastava, A. K., & Arora, D. K. (2012a). Characterization of mycolytic enzymes of Bacillus strains and their bio-protection role against Rhizoctonia solani in tomato. Current Microbiology, 65, 330–336.CrossRefPubMedGoogle Scholar
- Solanki, M. K., Kumar, S., Pandey, A. K., Srivastava, S., Singh, R. K., Kashyap, P. L., Srivastava, A. K., & Arora, D. K. (2012b). Diversity and antagonistic potential of Bacillus spp. associated to the rhizosphere of tomato for the management of Rhizoctonia solani. Biocontrol Science and Technology, 22, 203–217.CrossRefGoogle Scholar
- Solanki, M. K., Singh, R. K., Srivastava, S., Kumar, S., Kashyap, P. L., & Srivastava, A. K. (2015). Characterization of antagonistic-potential of two Bacillus strains and their biocontrol activity against Rhizoctonia solani in tomato. Journal of Basic Microbiology, 55, 82–90.CrossRefPubMedGoogle Scholar
- Srivastava, R., Tripathi, B. M., Singh, R. K., Srivastva, P., Kumari, P., et al. (2013). Profiling of plant growth promoting bacteria associated with Jaunpuri giant raddish rhizosphere. International Journal of Agriculture, Environment & Biotechnology, 6(2), 187–196.Google Scholar
- Versalovic, J., Schneider, M., de Bruijn, F. J., & Lupski, J. R. (1994). Genomic fingerprinting of bacteria using repetitive sequence-based polymerase chain reaction. Methods in Molecular and Cellular Biology, 5, 25–40.Google Scholar
- Zahid, M., Abbasi, M. K., Hameed, S., & Rahim, N. (2015). Isolation and identification of indigenous plant growth promoting rhizobacteria from Himalayan region of Kashmir and their effect on improving growth and nutrient contents of maize (Zea mays L.). Frontiers in Microbiology, 6, 207.CrossRefPubMedPubMedCentralGoogle Scholar