Genetic diversity of biovar 3 and 4 of Ralstonia solanacearum causing bacterial wilt of tomato using BOX- PCR, RAPD and hrp gene sequences
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https://doi.org/10.56093/ijas.v84i3.38590
Keywords:
Biovar, Genetic diversity, Ralstonia solanacearum, TomatoAbstract
Genetic diversity of Ralstonia solanacearum, causal agent of bacterial wilt of tomato was assessed by using three different molecular methods such as random amplified polymorphism DNA (RAPD), BOX-PCR and hrp (hypersensitive response and pathogenicity) gene sequence analysis technique. Twelve isolates of Ralstonia solanacearum belonging to biovar 3 (9 isolates) and 4 (3 isolates) were collected from Northern parts of India including Himachal Pradesh and Uttarakhand states from infected tomato plants. Out of 16 primers used in RAPD fingerprinting, four primers (OPA-2, OPA-11, OPC-5, OPE-7) showed monomorphic bands and remaining 12 primers exhibited polymorphic amplified products of both the biovars of R. solanacearum. The primer OPE-10 showed the highest level of genetic diversity among the isolates. Ten isolates of R. solanacearum were classified into two clusters at 20 per cent similarity coefficient and cluster 1 represented all isolates of biovar 3 (UTT-23, UTT-10, UTT-26, HPT11, UTT-9, UTT-32, HPC-3) and cluster 2 comprised the biovar 4 (UTT-22, HPT-3, UTT-24). BOX-PCR fingerprint of R. solanacearum clearly distinguished biovar 3 and 4 grouped into two distinct clusters at 40% similarity coefficient. Cluster 1 represented all isolates of R. solanacearum biovar 3 and cluster 2 comprised the biovar 4 isolates. The isolates of R. solanacearum have genetic diversity in hrpB gene, but it could not differentiate the isolates of biovar 3 and 4. However, the biovars 3 and 4 of R. solanacearum can be genetically distinguished by using BOX- PCR and specific primer of RAPD.Downloads
References
Fegan M and Prior P. 2005. How complex is the Ralstonia solanacearum species complex? (In) Bacterial wilt disease and the Ralstonia solanacearum species complex, pp 449–61. Allen C, Prior P, Hayward A C (Eds). American Phytopathological Society Press, St. Paul.
Grover A, Azmi W, Gadewar A V, Pattanayak D, Naik P S, Shekhawat G S and Chakrabarti S K. 2006. Genotypic diversity in a localized population of Ralstonia solanacearum as revealed by random amplified polymorphic DNA markers. Journal of Applied Microbiology 101(4): 798–806. DOI: https://doi.org/10.1111/j.1365-2672.2006.02974.x
Hayward A C. 2000. Ralstonia solanacearum. (In) Encyclopedia of Microbiology, pp 32–42. Lederberg J (Ed.) Academic Press. Horita M and Tsuchiya K. 2001. Genetic diversity of Japanese strains of Ralstonia solanacearum. Phytopathology 91: 399–407. DOI: https://doi.org/10.1094/PHYTO.2001.91.4.399
Khakvar R, Sijan K, Wong M Y, Radu S and Thong K L. 2008. Genomic diversity of Ralstonia solanacearum strains isolated from banana in West Malaysia. Plant Pathology Journal 7 (2): 162–7. DOI: https://doi.org/10.3923/ppj.2008.162.167
James D, Girija D, Mathew Sally K, Nazeem P A, Babu T D and Varma S A. 2003. Detection of Ralstonia solanacearum race 3 causing Bacterial wilt of solanaceous vegetables in Kerala, using Random amplified polymorphic DNA (RAPD) analysis. Journal of Tropical Agriculture 41: 33–7.
Murray and Thompson. 1980. Rapid islation of high molecular weight DNA. Nucleic Acids Research 8: 4321–5. DOI: https://doi.org/10.1093/nar/8.19.4321
Poussier S, Trigalet- Demery D, Vandewalle P, Goffiner, B, Luisetti J and Trigalet A. 2000. Genetic diversity of Ralstonia solanacearum as assed by PCR-RFLP of the Hrp region, AFLP and 16S rRNA sequence analysis, and identification of an African subdivision. Microbiology 146: 1 679–92. DOI: https://doi.org/10.1099/00221287-146-7-1679
Schaad N W, Jones J B and Chun W. 2001. Laboratory guide for identification of plant pathogenic bacteria. APS Press 174.
Sharma J P, Jha A K, Singh A K, Pan R S, Rai M and Kumar S. 2005. Screening of parental lines and their F2 crosses of brinjal (Solanum melongena) to Ralstonia wilt. Indian Journal of Agricultural Sciences 75 (4): 197–9.
Singh D, Dhar, S and Yadava D K. 2011. Genetic and pathogenic variability of Indian strains of Xanthomonas campestris pv. campestris causing black rot disease in crucifers. Current Microbiology 63: 551–60. DOI: https://doi.org/10.1007/s00284-011-0024-0
Singh D, Sinha S, Yadav D K, Sharma J P, Srivastava D K, Lal H C, Mondal, K K and Jaiswal R K. 2010. Characterization of biovar/races of Ralstonia solanacearum, the incitant of bacterial wilt in solanaceous crops. Indian Phytopathology 63: 261–5. Stevens P and Elsas J D. 2010. Genetic and phenotypic diversity of Ralstonia solanacearum biovar 2 strains obtained from
Dutch waterways. Antonie van Leeuwenhoek 97: 171–88.
Timothy T P. 2006. Plant Pathogenic Ralstonia sp. (In) Plant- Associated Bacteria, pp 573–644. Gnanamanickam S S (Ed). Springer, Netherlands.
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