Open AccessA PCR primer‐specific to Cylindrocarpon heteronema for detection of the pathogen in apple wood
Author(s) -
Brown Averil E.,
Muthumeenakshi S.,
Sreenivasaprasad S.,
Mills Peter R.,
Swinburne Terence R.
Publication year - 1993
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.1574-6968.1993.tb06083.x
Subject(s) - primer (cosmetics) , ribosomal dna , dna , biology , spacer dna , microbiology and biotechnology , polymerase chain reaction , internal transcribed spacer , oligonucleotide , pathogen , ribosomal rna , dna sequencing , chemistry , gene , genetics , phylogenetics , organic chemistry
An oligonucleotide primer (ChInt) was synthesised from the variable internally transcribed spacer (ITS) 1 region of ribosomal DNA (rDNA) of Cylindrocarpon heteronema . PCR with primers ChInt and ITS4 (from a conserved sequence of the rDNA) amplified a 470‐bp fragment from several isolates of C. heteronema but not from various apple wood saprophytes. Amplification of this fragment was achieved from 1–2 pg of fungal DNA. These primers amplified a fragment of the same size from DNA extracted from cankered wood but only after impurities were removed from the DNA on a Qiagen tip‐5 column. Southern hybridization analysis confirmed the 470‐bp fragment from C. heteronema DNA and cankered wood to be identical.