A PCR primer-specific to Cylindrocarpon heteronema for detection of the pathogen in apple wood.
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.
Brown
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AE
Department of Applied Plant Science, Queen's University of Belfast, UK.
Muthumeenakshi
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Sreenivasaprasad
S
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Mills
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PR
Swinburne
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TR
D001483:Base Sequence; D015342:DNA Probes; D004271:DNA, Fungal / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D003904:Mitosporic Fungi / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D008969:Molecular Sequence Data; D010944:Plants / Q000382:microbiology; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D014934:Wood
D015342:DNA Probes; D004271:DNA, Fungal; D004275:DNA, Ribosomal