Determination of Ralstonia (Pseudomonas) solanacearum rDNA subgroups by PCR tests |
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| Authors: | Seal Taghavi Fegan Hayward & Fegan |
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| Institution: | ;Natural Resources Institute, University of Greenwich, Central Avenue, Chatham Maritime, Kent ME4 4TB, UK;,;Department of Microbiology, The University of Queensland, Queensland 4072,;The Cooperative Research Centre for Tropical Plant Pathology, The University of Queensland, Queensland 4072,;Division of Food Science &Technology, PO Box 3312, Tingalpa DC, Queensland 4173;Australia |
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| Abstract: | Rapid and sensitive polymerase chain reaction (PCR) methods are described for determination of the two 16 S rDNA subgroups of Ralstonia solanacearum, the causal agent of bacterial wilt. A third subgroup consisting of Indonesian R. solanacearum isolates belonging to Division II, the blood disease bacterium and Pseudomonas syzygii can also be identified. Primers were designed to sequences within R. solanacearum 16 S rDNA (equivalent to Escherichia coli 16 S rDNA positions 74–97, 455–475, 1454–1474), and the internal transcribed spacer region between the 16 S and 23 S rDNA genes. Different combinations of forward and reverse primers allowed selective PCR amplification of (a) R. solanacearum Division I (biovars 3, 4 and 5), (b) Division II (biovars 1, N2, and 2) including the blood disease bacterium and P. syzygii , or (c) amplification of Division II only except for five biovar 1, 2 or N2 isolates of R. solanacearum from Indonesia, P. syzygii and the BDB. A total of 104 R. solanacearum , 14 blood disease bacterium and 10 P. syzygii isolates were tested. Simultaneous detection of species and subdivision was achieved by designing a multiplex PCR test in which a 288-base pair (bp) band is produced by all R. solanacearum isolates, and an additional 409-bp band in Division I strains. |
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| Keywords: | bacterial wilt blood disease bacterium genetic groups PCR Pseudomonas syzygii rDNA |
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