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Characterization of Xanthomonas oryzae pv. oryzae recX, a gene that is required for high-level expression of recA
Authors:Rojana Sukchawalit  Paiboon Vattanaviboon  Supa Utamapongchai  Gary Vaughn  Skorn Mongkolsuk
Affiliation:Department of Plant Biology, Plant Pathology Section, The Royal Veterinary and Agricultural University, Thorvaldsensvej 40, DK-1871 Frederiksberg C, Denmark. mette.lubeck@plbio.kvl.dk
Abstract:A universally primed (UP)-PCR cross hybridization assay was developed for rapid identification of isolates of Rhizoctonia solani into the correct anastomosis group (AG). Twenty-one AG tester isolates belonging to 11 AGs of R. solani were amplified with a single UP primer which generated multiple PCR fragments for each isolate. The amplified products were spotted onto a filter, immobilized and used for cross hybridization against amplification products from the different isolates. Isolates within AG subgroups cross hybridize strongly, whereas between different AGs little or no cross hybridization occurs. Sixteen Rhizoctonia isolates from diseased sugar beets and potatoes were identified using the assay. The results were supported by restriction fragment length polymorphism analysis of the ITS1-5.8S-ITS2 region of the nuclear encoded ribosomal DNA. Through standardization and use of quick non-radioactive labeling techniques, the UP-PCR cross hybridization assay has potential for routine use by modern DNA chip technology.
Keywords:Universally primed-PCR fingerprinting    Cross hybridization    Anastomosis group    Microarray    Rhizoctonia solani
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