ABB  Vol.3 No.8 , December 2012
Genetic characterization of Burkholderia cepacia strain from Northeast India: A potential bio-control agent
Abstract: This paper reports the characterization of naturally occurring rhizobacteria from NE India for the control of wilt and damping-off diseases in French beans. French bean (Phaseolus vulgaris L.) is one of the most important food legumes for direct human consumption which was originated from South and Central America. In NE India wilt and damping-off in beans are serious diseases which are mainly caused by Fusarium spp and Rhizoctonia solani. Out of 625 bacterial isolates obtained from French bean growing fields of Manipur, 15 potential isolates were selected, whose inhibition zone width were more than 15 mm. Strain IBSD-LKBI exhibited strong antagonistic activity and can inhibit both the pathogens. Besides, this strain also produced extracellular hydrolytic enzymes (protease, chitinase, β-1, 3-glucanase), secondary metabolites (Siderophore, HCN). It also produces plant growth promoting hormone (IAA) and can solubilize organic phosphate. Burkholderia cepacia IBSD-LKBI exhibited the highest plant growth promotion and BCA. After 60 days of treatment, plant height, root length, fresh weight and % of disease suppression were significantly (P < 0.05) higher than in control plants. Productions of secondary metabolites were detected using gene specific primer. This bacterium was phenotypically characterized and sequence data of 16S rDNA revealed the closest identity with Burkholderia cepacia (98%). Multiple sequence alignment & phylogram with closely related sequence accessions using MEGA Software revealed this naturally occurring biocontrol bacteria belongs to Burkholderia cepacia complex genomovar I based on the Biolog identification system with 16S rDNA gene sequence analysis. This organism is one of the new reports from NE India for the control of wilt and damping-off diseases in French beans.
Cite this paper: Devi, S. , Somkuwar, B. , Potshangbam, M. and Talukdar, N. (2012) Genetic characterization of Burkholderia cepacia strain from Northeast India: A potential bio-control agent. Advances in Bioscience and Biotechnology, 3, 1179-1188. doi: 10.4236/abb.2012.38144.

[1]   Copping, G.L.. and Menn, J.J. (2000) Biopesticides: A review of their action, applications and efficacy. Pest Management Sciences, 65, 651-676. doi:10.1002/1526-4998(200008)56:8<651::AID-PS201>3.0.CO;2-U

[2]   Parmeter, J.R. (1970) R. solani. Biology of Pathology. University of California Press, Berkeley Los Angles & London.

[3]   Govindarajan, M., Balandreau, J., Kwon, S.W., Weon, H.Y. and Lakshminarasimhan, C. (2008) Effects of the inoculation of Burkholderia Vietnamensis and related endophytic diazotrophic bacteria on grain yield of rice. Microbial Ecology, 55, 21-37. doi:10.1007/s00248-007-9247-9

[4]   Payne, G.W., Vandamme, P., Morgan, S.H., LiPuma, J.J., Coenye, T., Weightman, A.J., Jones, T.H. and Mahenthiralingam, E. (2005) Development of a recA gene-based identification approach for the entire Burkholderia genus. Applied and Environmental Microbiology, 71, 3917-3927. doi:10.1128/AEM.71.7.3917-3927.2005

[5]   Indira, S., Talukdar, N.C., Chandradev Sharma, K., Jeyaram, K and Rohinikumar. M. (2011) Screening of rhizobacteria for their plant growth promotion ability and antagonism against damping off and root rot diseases of broad bean (Vicia faba L.). Indian Journal of Microbiology, 51, 14-21. doi:10.1007/s12088-011-0069-6

[6]   Samad, A., Ajaykumar, P,V., Gupta, M.K., Shukla, A.K., Darokar, M.P., Somkuwar, B.G and Alam, M. (2008) Natural infection of periwrinkle (Catharanthus roseus) with Cucumber mosaic virus, Subgroup IB. Australian. Plant Disease Notes, 3, 30-34. doi:10.1071/DN08013

[7]   Berg, G., Roskot, N., Steidle, A., Ebert, L., Zock, A. and Smalla, K. (2002) Plant-dependent genotypic and phenotypic diversity of antagonistic rhizobacteria isolated from different Verticillium host plants. Applied and Environmental Microbiology, 68, 3328-3338. doi:10.1128/AEM.68.7.3328-3338.2002

[8]   Chernin, L.S., D.E. Fuente, L, Sovolev, V., Haran, S., Vorgias, C.E., Oppenheim, A.B. and Chet. (1997) Molecular cloning, structural analysis, and expression in Escherichia coli of a chitinase gene from Enterobacter agglomerans. Applied and Environmental Microbiology, 63, 834-839.

[9]   Katatny, El., M.H., Gudelj, M., Robra, KH., Elnaghy, M.A. and Gubitz, G.M. (2001) Characterization of a chitinase and an endo-?-1,3-glucanase from Trichoderma harzianum Rifai T24 involved in control of the phytopathogen Sclerotium rolfsii. Applied and Microbial Biotechnology, 56, 137-143.

[10]   Millar, R.L. and Higgins, V.J. (1970) Association of cyanide with infection of birds foot trefoil by Stemphyllium loti. Phytopathology, 60, 104-110. doi:10.1094/Phyto-60-104

[11]   Raajmakers, J.M., Weller, D.M. and Thomashow, L.S. (1997) Frequency of antibiotic-production Pseudomonas spps. in natural environments. Applied and Environmental Microbiology, 63, 881-887.

[12]   Zhang, Y.C., Ronimus, R.S., Turner, N. Zhang, Y. and Morgan, H.W. (2002) Enumeration of thermophillic Bacillus species in composts and identification with a Random Amplified Polymorphic DNA (RAPD) protocol system. Applied Microbiology, 25, 618-626. doi:10.1078/07232020260517760

[13]   Zhang, Z., Schwartz, S., Wagner, L. and Miller, W. (2000) A greedy algorithm for aligning DNA sequences. Journal of Computational Biology, 7, 203-214. doi:10.1089/10665270050081478

[14]   Poirot, O., O’Toole, E. and Notredame, C. (2003) Tcoffee@igs; A web server for computing, evaluating and combining multiple sequence alignments. Nucleic Acid Research, 13, 3503-3506. doi:10.1093/nar/gkg522

[15]   Kumar, S., Tamura, K., Jakobsen, I.B. and Nei, M. (2001) MEGA2: Molecular evolutionary genetics analysis software. Arizona State University, Tempe.

[16]   Vandamme, P., Holmes. B., Vancanneyt, M., Coenye, T., Hoste, B., Coopman, R., Revets. H., Lauwers, S., Gillis, M., Kersters, K. and Govan, J.R. (1997) Occurrence of multiple genomovars of Burkholderia cepacia in cystic fibrosis patients and proposal of Burkholderai multivorans sp. nov. International Journal of Systemic Bacteriolog, 47, 1188-1200. doi:10.1099/00207713-47-4-1188

[17]   Coenye, T., Vandamme, P., Govan, J.R. and Lipuma, J.J. (2001c) Taxonomy and identification of the Burkholderia cepacia complex. Journal of Clinical Microbiology, 39, 3427-3436. doi:10.1128/JCM.39.10.3427-3436.2001

[18]   Perin, L., Martinez-Aguilar, L., Castro-Gonzalez, P., Estrada-de, LSP., Cabellos-Avelar, T., Guedes, H.V., Reis V.M. and Caballero-Mellado, J. (2006) Diazotrophic Burkholderia species associated with field grown maize and sugercane. Applied and Environmental Microbiology, 72, 3103-3110. doi:10.1128/AEM.72.5.3103-3110.2006

[19]   Mahenthiralingam, E. and Vandamme, P. (2005) Taxonomy and pathogenesis of the Burkholderia cepacia complex. Chronic Respiratory Disease, 2, 209-217. doi:10.1191/1479972305cd053ra

[20]   Spreet, D.P. (2001) Understanding Burkholderia cepacia: Epidemiology, genomovars, and virulent. Infections in Medicine, 18, 49-56.

[21]   Weisburg, W.G., Barns, S.M., Pelletier, D.A. and Lane, D.J. (1992) 16S ribosomal DNA amplification for phylogenetic study. Journal of Bacteriology, 173, 697-703.

[22]   Allan, N.D., Kooi, C., Sokol, P.A. and Beveridge, T.J. (2003) Putative virulence factors are released in association with membrane vesicles from Burkholderia cepacia. Canadian Journal of Microbiology, 49, 613-624. doi:10.1139/w03-078

[23]   Cardona, S.T., Wopperer, J., Eberl L. and Valvano, M.A. (2005) Diverse pathogenecity of Burkholderia cepacia complex strains in the Caenorhabditis elegans host model. FEMS Microbiology, 14, 277-286.

[24]   Chiarini, L., Bevivino, A., Dalmastri, C., Tabacchioni, S. and Visca, P. (2006) Burkholderia cepacia complex species: Health hazards and biotechnological potential. Trends in Microbiology, 14, 277-286. doi:10.1016/j.tim.2006.04.006

[25]   Milus, E.A. and Rothrock, C.S. (1997) Efficacy of bacterial seed treatments for controlling Pythium root rot of winter wheat. Plant Disiese, 81, 180-184. doi:10.1094/PDIS.1997.81.2.180

[26]   Cartwright, D.K. and Benson, O.M. (1995) Comparison of Pseudomonas species and application techniques for biocontrol of Rhizoctonia stem rot of poinsettia. Plant Disease, 79, 309-313. doi:10.1094/PD-79-0309

[27]   de los Santos-Villalobos, S., Barrera-Galicia, G.C., Miranda-Salcedo, M.A. and Pe?a-Cabriales J.J. (2012) Burkholderia cepacia XXVI siderophore with biocontrol capacity against Colletotrichumgloeosporioides. World Journal of Microbiology and Biotechnology, 28, 2615-2623. doi:10.1007/s11274-012-1071-9

[28]   Zaiton, S., Sariah, M. and Zainal Abidin, M.A. (2008) Effect of endophytic bacteria on growth and suppression of Ganoderma infection in oil palm. International Journal of Agriculture and Biology, 10, 127-132.

[29]   Bevivino, A., Sarrocco, S., Dalmastri, C., Tabacchion, I.S., Cantale, C. and Chiarini, L. (1998) Characterization of a free-living maize-rhizosphere population of Burkholderia cepacia: Effect of seed treatment on disease suppression and growth promotion of maize. FEMS Microbiology Ecology, 27, 225-237. doi:10.1111/j.1574-6941.1998.tb00539.x

[30]   McLoughlin, T.I., Quinn, J.P., Bettermann, A. and Bookland, R. (1992) Pseudomonas cepacia suppression of sunflower wilt fungus and role of antifungal compounds in controlling the disease. Applied and Environmental Microbiology, 58, 1760-1763.

[31]   Larralde, C.P., Santiago-Mena, M.R., Sifuentes-Rincon A.M., Rodrigue-Luna, I.C., Rodriguez-Perez, M.A., Shiraj, K. and Narvaez-Zapata, J.A. (2008) Biological potential and polyphasic characterization of novel native Trichoderma strains against Macrophomina phaseolina from Sorgum and common bean. Applied Microbiology and Biotechnology, 80, 167-177. doi:10.1007/s00253-008-1532-0

[32]   Raaijmakers, J.M., Vlami, M. and de Souza, J.T. (2002) Antibiotics production by bacterial biocontrol agents. Antonie Van Leeuvenhoek International Journal, 81, 537- 547. doi:10.1023/A:1020501420831

[33]   Kim, P.I. and Chung, K.C. (2004) Production of an antifungl protein for control of Colletotrichum lagenarium by Bacillus amyloliquefaciens MET0908. FEMS Microbiology Leters, 234, 177-183.

[34]   Ogawa, K., Yoshida, N., Kariya, K., Ohnishi, C. and Ikeda, R. (2002) Purification and characterization of a novel chitinase from Burkholderia cepacia strain KH2 isolated from the bed log of Lentinus edodes, Shiitake mushroom. The Journal of General and Applied Microbiology, 48, 25-92. doi:10.2323/jgam.48.25

[35]   Fridlender, M., Inbar, J. and Chet, I. (1993) Biological control of soilborne plant pathogens by a ?-1,3 glucanase-producing Pseudomonas cepacia. Soil Biology and Biochemistry, 25, 1121-1221. doi:10.1016/0038-0717(93)90217-Y

[36]   Bal, A.S. and Chanway, C.P. (2000) Isolation and identication of endophytic bacteria from lodgepole pine and western red cedar. Auburn University website.