Differentiation of Xanthomonas campestris pv vasculorum by DNA probes
| MSI99P0103 | |
| Saumtally, S. Autrey, L. J. C. Daniels, M. J. | |
| Differentiation of Xanthomonas campestris pv vasculorum by DNA probes | |
| 3rd ISSCT Sugar Cane Pathology Workshop, MSIRI, Reduit, 22-26 July 1991 | |
| Conference paper | |
| 1991 | |
| Abstract in Sug. Cane 1991 (6): 8 | |
| En | |
| En | |
| Two new techniques, Random Amplified Polymorphic DNA (RAPD) and genomic subtraction, were evaluated in the differentiation of the three races of Xanthomonas campestris pv vasculorum present in Mauritius. DNA polymorphisms in the 3 races by RAPD involved the use of random primers of 10 oligomers to amplify DNA fragments of the bacterial genome by polymerase chain reaction (PCR). The amplified fragments were then visualised by a garose gel electrophoresis. Of the 180 primers tested, none gave a reproducible differential banding pattern. However, when certain RAPD products were used as probes and hybridized to Eco RI genomic DNA digests on nylon filters, race 1 could be distinguished from the two other races. Another source of DNA probes was obtained by genomic subtraction. This technique enables the enrichment of sequences present in one race but absent from another by removing the common DNA. Using this procedure Sau 3AI enzyme-digested DNA of race 1 was allowed to reassociate with excess of sheared DNA of race 2 in a high inorganic phosphate buffer. The reassociated DNA was then ligated in the vector pBR322, transformed and selected in E. coli DH5a and the inserts amplified by PCR. The amplified inserts were then used as probes and hybridized to Bam HI, Eco RI, Hind III, and Pst I genomic digests. Eight out of 18 probes tested distinguished race 1 from races 2 and 3. Labelling of probes and detections were performed using a non-radioactive method with AMPPD as substrate for alkaline phosphatase. | |
| XANTHOMONAS CAMPESTRIS DNA STRAINS SUGARCANE GUMMOSIS BACTERIAL DISEASES DISEASES | |
| MAURITIUS | |
| Sugarcane: Diseases and disease management | |
| Bacterial diseases: Gumming | |
| 1992-01-21 | |
| En | |
| Mauritius Sugar Industry Research Institute / John Innes Centre for Plant Research. Sainsbury Laboratory, Reduit / UK | |
| CAT | |
| PATH |