Open AccessUP‐PCR cross blot hybridization as a tool for identification of anastomosis groups in the Rhizoctonia solani complex
Author(s) -
Lübeck Mette,
Poulsen Hanne
Publication year - 2001
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2001.tb10737.x
Subject(s) - rhizoctonia solani , biology , restriction fragment length polymorphism , polymerase chain reaction , southern blot , microbiology and biotechnology , primer (cosmetics) , rhizoctonia , ribosomal dna , dna–dna hybridization , dna , genetics , gene , botany , chemistry , phylogenetics , organic chemistry
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.