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Detection, Identification and Phylogenetic Analysis of Indian Ralstonia solanacearum Isolates by C omparison of Partial hrpB G ene Sequences
Author(s) -
Chandrasekhar Bhadvelu,
Umesha Sharanaiah
Publication year - 2015
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1111/jph.12287
Subject(s) - ralstonia solanacearum , biology , phylogenetic tree , bacterial wilt , genetics , phylogenetics , polymerase chain reaction , sequence analysis , gene , ralstonia , homology (biology) , microbiology and biotechnology , pathogen
A species‐specific Polymerase Chain Reaction ( sPCR ) method was developed to identify and detect isolates of Ralstonia solanacearum , the cause of bacterial wilt disease in chilli. PCR primers for R. solanacearum were identified by alignment of hrpB gene sequences and selection of sequences specific for R. solanacearum at their 3′ ends. The primers were shown to be specific for R. solanacearum , as no PCR product was obtained when genomic DNA from other bacterial species including closely related Ralstonia species, were used as test species. Lone pair of primers (RshrpBF and RshrpBR) was designed using hrpB gene sequence, unique to R. solanacearum which amplified a predicted PCR product of 810 bp from 20 different isolates. Phylogenetic analysis was also attempted to understand the evolutionary divergence of Indian R. solanacearum isolates. Based on phylogenetic analysis, Indian isolates showed homology with the standard reference isolates from other countries but, interestingly, one new isolate showed complete evolutionary divergence by forming an out‐group.