PCR-based detection of Colletotrichum acutatum on strawberry |
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| Authors: | S SREENIVASAPRASAD K SHARADA A E BROWN & P R MILLS |
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| Institution: | Department of Applied Plant Science, The Queen's University of Belfast;;, Applied Plant Science Division, Department of Agriculture for Northern Ireland, Newforge Lane, Belfast BT9 5PX;UK |
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| Abstract: | An oligonucleotide primer ( Ca Int 2) was synthesized from the variable internal transcribed spacer (ITS) 1 region of ribosomal DNA (rDNA) from Colletotrichum acutatum . PCR with primers Ca Int2 and ITS4 (from a conserved sequence of the rDNA) amplified a 490 bp fragment from several isolates of C. acutatum but not from other members of the genus Colletotrichum . Amplification of this fragment was achieved from 100 fg of fungal DNA. These primers amplified a fragment of the same size from DNA extracted from strawberry tissues infected by C. acutatum . Southern hybridization analysis confirmed the 490 bp fragment from C. acutatum DNA and infected strawberry to be identical. The species-specific primer ( Ca Int2) developed in this work could be used for the accurate identification of C. acutatum and its detection on other host plants. |
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