Randeep Singh Aditi Sharma A. K. Gupta


Native Rhizobium rhizogenes strain UHFBA-212 [141/1A (NCBI: KC488174)]was isolated from rhizosphere soil of peach nursery plant of wild peach collected from Himachal Pradesh. In addition to this,159 isolates were also collected and were screened in vitro for their biocontrol potential against Agrobacterium tumefaciens. Out of these strain, UHFBA-212 showed maximum zone of inhibition i.e. 4.16 and 3.57cm without and after exposure to chloroform against C58.Sequence analysis (16SrDNA) of the strain showed nucleotide homology similar to Rhizobium sp. Amplification of total genomic DNA of the strain with Vir D2 andipt primers didn’t showed amplification with these virulence genes suggesting the absence of tumorigenic factors. In the field conditions, maximum population (329.33x106 cfu/g of soil) was observed in antibiotic resistant mutant of R. rhizogenes strain K84 applied on cherry rootstock Colt followed by 285.33 (x 106 ) cfu/g of soil in UHFBA-212 after 9 months at the time of uprooting of plants when applied alone as root dip. Minimum incidence of crown gall (2.00%) was observed in strain UHFBA- 212 co inoculated with strain C58 as seed treatment on behmi seeds. The data on population indices in rhizosphere and incidence of crown gall further suggested that for better management of disease R. Rhizogenes isolates should be either equal or more in population than that of A. tumefaciens isolates. Strain UHFBA-212 controls crown gall as effectively as strain K84 and can be exploited against tumorigenic isolates under field conditions.




Colonization, Disease incidence, Resistant, Rhizobium rhizogenes

Bouzar, H., Daouzli, N., Krimi, Z., Alim, A. and Khemici, E. (1991). Crown gall incidence in plant nurseries of Algeria, characteristics of Agrobacterium tumefaciens strains, and biological control of strains sensitive and resistant to agrocine 84. Agronomie., 11: 901-908
Bruce, K.D., Hiorns, W.D., Hobman, J.L., Osborn, A.M., Strike, P. and Ritchie, D.A. (1992). Amplification of DNA from native populations of soil bacteria by using the polymerase chain reaction. Applied and Environmental Microbiology, 58:3413-3416
Bull, C.T., Weller, D.M. and Thomashow, L. S. (1991). Relationship between root colonization and suppression of Gaeumannomyces graminis var. tritici by Pseudomonas fluorescens strain 2-79. Phytopathology, 81:954-959
Canfield, M. L. and Moore, L. W. (1991). Isolation and characterization of opine utilizing strains of Agrobacterium tumefaciens and fluorescent strains of Pseudomonas spp. from rootstocks of Malus. Phytopathology, 81(4):440-443
Conn, H. J. (1942). Validity of the genus Alcaligenes. Journal of Bacteriology, 44: 353-360
Costechareyre, D., Rhouma, A., Lavire, C., Portier, P., Chapulliot, D., Bertolla, F., Boubaker, A., Dessaux, Y. and Nesme, X. 2010. Rapid and efficient identification of Agrobacterium species by recA allele analysis. Microbial Ecology, 60: 862-872
Escobar, M. A. and Dandekar, A. M. (2003). Agrobacterium tumefaciensas an agent of disease. Trends Plant Science., 8: 380-386
Farrand, S.K., van Berkum, P.B. and Oger, P. (2003). Agrobacterium is a definable genus of the family Rhizobiaceae. Int. J. Syst. Evol. Microbiol., 53, 1681–1687
Felsenstein, J. (1985). Confidence limits on phylogenies: an approach using the bootstrap. Evolution. 39: 783–791. doi:10.2307/2408678
Gross, D. C. (1988). Maximizing rhizosphere populations of fluorescent pseudomonads on potatoes and their effects on Erwiniacarotovora. Potato Association of America,65: 697-710
Gupta, A. K., Kamal, B. and Khosla, K. (2005). Effect of fertilizers, soil amendments and antibacterial compounds on the incidence of crown gall on cherry rootstock colt. Journal of Mycology and Plant Pathology, 35(2): 286-288
Gupta, A. K., Khosla, K., Bhardwaj, S. S., Thakur, A., Devi, S., Jarial, R. S., Sharma, C., Singh, K. P., Srivastava, D. K. and Lal, R. (2010). Biological control of crown gall on peach and cherry rootstock Colt by native Agrobacterium radiobacter isolates. Open Horticulture Journal, 3: 1-10
Haas, J. H., Moore, L. W., Ream, W. and Manulis, S. (1995). Universal PCR primers for detection of phytopathogenic Agrobacterium strains. Applied Environmental Microbiology,61: 2879- 2884
Jindal, K. K. and Sharma, R. C. (1988). Occurrence of biovar 1 of Agrobacterium tumefaciens on almond in India. Indian Phytopathology, 41(4): 614-615
Kapshuk, A. A. (1933). Bacteriological study of crown gall of fruit trees. In: Russian English Summary, Bull. N Caucasian Inst. for Plant Protection, Rostoff on Do., 2: 69-78
Kawaguchi, A., Inoue, K. and Ichinose, Y. (2008). Biological control of crown gall of grapevine, rose, and tomato by non-pathogenic Agrobacterium vitisstrain VAR03-1. Phytopathology, 98:1218–1225
Kawaguchi, A. (2013). Biological control of crown gall on grapevine and root colonization by nonpathogenicRhizobium vitisstrain ARK-1. Microbes and Environments, 28:306–311
Kawaguchi, A. (2014). Reduction of pathogen populations at grapevine wound sites is associated with the mechanism of biological control of crown gall by Rhizobium vitis strain ARK-1. Microbes and Environments, 29: 296–302
Kawaguchi, A. (2015). Biological control agent Agrobacterium vitis strain ARK-1 suppresses expression of the virD2 and virE2 genes in tumorigenic A. vitis. European Journal of Plant Pathology. DOI 10.1007/s10658-015-0730-8
Kerr, A. and Htay, K. (1974). Biological control of crown gall through bacteriocin production. Physiological Plant Pathology, 4: 37-44
Kerr, A. (1969). Transfer of virulence between isolates of Agrobacterium. Nature, 223: 1175–1176
Kerr, A. (1972). Biological control of crown gall: seed inoculation. Journal of Applied Bacteriology, 35: 493-497
Lindstrom, K. and Young, J.P.W. (2011). International Committee on Systematics of Prokaryotes Subcommittee on the taxonomy of Agrobacterium and Rhizobium. International Journal of Systematic and Evolutionary Microbiology, 61: 3089-3093
Loper, J. E., Haack, C. and Schroth, M. N. (1985). Population dynamics of soil pseudomonads in the rhizosphere of potato (Solanum tuberosum L.). Applied and Environmental Microbiology, 49: 416-422
Lippincott, B.B., Whatley, M.H. and Lippincott, J.A. (1977). Tumour induction by Agrobacterium involves attachment of the bacterium to a site on the host plant cell wall. Plant Physiology, 59: 388-90
Moore, L.W. and Allen, J. (1977). Comparison of selective and differential media for Agrobacterium species. Proceedings of American Phytopathological Society, 4: 209
Moore L W and Canfield M. 1996. Biology of Agrobacterium and management of crown gall disease. In: Hall R (Ed.), Principles and practices of managing soil borne plant pathogens. Ontario, USA: APS Press, pp. 151-191
Moore, L. W., Kado, C. I. and Bouzar, H. (1988). Gram negative bacteria –Agrobacterium .In: Laboratory guide for identification for pathogenic bacteria 2ndedn, Schaad N W (ed.), The American Phytopathological Society, St. Paul, Minnesota, pp. 16-36
Mousavi, S. A., Osterman, J., Wahlberg, N., Nesme, X., Lavire, C., Vial, L., Paulin, L., Lajudie, P. De. and Lindstrom, K. (2014). Phylogeny of the Rhizobium-Allorhizobium-Agrobacterium clade supports the delineation of Neorhizobium gen. nov.Systematic and Applied Microbiolgy, 37: 208-215
Murugesan, S., Manoharan, C., Vijayakumar, R. and Panneerselvam, A. 2010. Isolation and characterization of Agrobacterium rhizogenesfrom the root nodules of some leguminous plants. International Journal of Microbiological Research, 1(3): 92-96
New, P. B. and Kerr. (1972). Biological control of crown gall: field measurements and glasshouse experiments. Journal of Applied Bacteriology, 35(2): 279-287
Penalver, R. and Lopez, M. M. (1999). Co-colonization of the rhizosphere by pathogenic strains K-84 and K-1026, used for crown gall bio-control. Applied and Environmental Microbiology, 65: 1936-1940
Pulawska, J. (2010). Crown gall of stone fruits and nuts, economic significance and diversity of its causal agents: tumorigenic Agrobacterium spp. Journal of Plant Pathology, 92(1): 87-98
Raio, A., Zoina, A. and Moore, L. W. (1997). The effect of solar heating of soil on natural and inoculated agrobacteria. Plant Pathology, 46: 320–328
Raio, A., Peluso, R., Puopolo, G. and Zoina, A. (2009). Evidence of pAgK84 transfer from Agrobacterium rhizogenes K84 to natural pathogenic Agrobacterium spp. in an Italian peach nursery. Plant Pathology, 58:745–753
Sawada, H., Ieki, H., Oyaizu, H. and Matsumoto, S. (1993). Proposal for rejection of Agrobacterium tumefaciens and revised descriptions for the genus Agrobacterium and for Agrobacterium radiobacter and Agrobacterium rhizogenes. Int J SystBacteriol., 43: 694-702
Smith, E. F. and Townsend, C. O. (1907). A plant tumour of bacterial origin. Science, 25:671- 673
Stockwell, V.O., Kawalek, M.D., Moore, L.W. and Lopper, J.E. (1996). Transfer of pAgK84 from the biocontrol agent Agrobacterium radiobacter K84 to A. tumefaciens under field conditions. Phytopathology, 86:31-7.
Thakur, D., Yadav A., Gogoi B. K. and Bora, T.C. (2007). Isolation and screening of Streptomyces in soil of protected forest areas from the states of Assam and Tripura, India, for antimicrobial metabolites. Journal of Medical Mycology, 17: 242—249
Trembley, G., Lambert, R., Lebeuf, H. and Dion, P. (1987). Isolation of bacteria from soil and crown-gall tumors on the basis of their capacity for opine utilization. Phytoprotection, 68:35-42.
Velazquez, E., Palomo, J. L., Rivas, R., Guerra, H., Peix, A., Trujillo, M. E., Garc?´a-Benavides, P., Mateos, P. F., Wabiko, H. and Mart?nez- Molina, E. (2010). Analysis of core genes supports the reclassification of strains Agrobacterium radiobacter K84 and Agrobacterium tumefaciens AKE10 into the species Rhizobium rhizogenes. Syst. Appl. Microbiol., 33: 247–251
Vicedo, B., Penlaver, R., Asins, M. J. and Lopez, M. M. (1993). Biological control of Agrobacterium tumefaciens, colonization, and pAGK84 transfer of the Ti plasmid of Agrobacterium radiobacter K84 and the Tra-mutant strain K1026. Applied and Environmental Microbiology, 59: 309-315
Wang, H.M., H.X. Wang, T.B. Ng, and J.Y. Li. (2003). Purification and characterization of an antibacterial compound produced byAgrobacterium vitis strain E26 with activity against A. tumefaciens. Plant Pathology, 52:134–143
Young, J. M., Kuykendall, L. D., Martinez-Romero, E., Kerr, A. and Sawada, H. (2001). A revision of Rhizobium Frank 1889, with and emended description of the genus, and the inclusion of all species of Agrobacterium Conn. 1942 and Allorhizobium undicola de Lajudie et al. 1989 as new combinations: Rhizobium radiobacter, R. rhizogene, R. Rubi and R. vitis. International journal of Systematic and Evolutionary Microbiology, 51: 89-103
Zambryski, P.C. (1998). Basic processes underlying Agrobacterium-mediated DNA transfer to plant cells. Ann. Rev. Genet., 22: 1-33
Research Articles

How to Cite

Rhizosphere competence of native Rhizobium rhizogenes strain and its use in management of crown gall. (2017). Journal of Applied and Natural Science, 9(3), 1772-1781. https://doi.org/10.31018/jans.v9i3.1437