Phytophthora cactorum (Lebert et Cohn) Schroeter is an important plant pathogen that can cause serious damage in agricultural and ornamental crops as well as in a wide range of forest species. The identification of this pathogen, based on morphological and physiological characters, is time consuming, labour-intensive and requires specialised staff to be correctly performed. Recently, PCR-based methods have partially resolved these problems, but the primers used cross react with Phytophthora idaei. To prevent any such reaction the use of a new pair of primers (PC1/PC2) with improved specificity, derived from a specific Random Amplified Polymorphic DNA (RAPD) generated fragment, is proposed. The PC1/PC2 primers, used in a simple PCR protocol, gave a single amplification product of approximately 450 bp; a good degree of specificity, with absence of cross reactions with Phytophthora pseudotsugae and R idaei; sensitivity down to 6 pg of R cactorum DNA extracted from pure mycelium; no reactions with the DNA of the host plants tested (downy oak, pear and walnut trees, potato, strawberry, tomato and pea plants). The detection of R cactorum in infected tissues of pear and walnut trees, potato, strawberry, tomato and pea plants was also confirmed. The specificity, sensitivity and robustness of the PC1/PC2 primers together with the possibility of their use in a rapid, simple and reliable diagnostic method are discussed.
An improved method for the detection of Phytophthora cactorum (l. C.) Schroeter in infected plant tissues using SCAR markers
CAUSIN, ROBERTO;SCOPEL, CRISTINA;MONTECCHIO, LUCIO
2005
Abstract
Phytophthora cactorum (Lebert et Cohn) Schroeter is an important plant pathogen that can cause serious damage in agricultural and ornamental crops as well as in a wide range of forest species. The identification of this pathogen, based on morphological and physiological characters, is time consuming, labour-intensive and requires specialised staff to be correctly performed. Recently, PCR-based methods have partially resolved these problems, but the primers used cross react with Phytophthora idaei. To prevent any such reaction the use of a new pair of primers (PC1/PC2) with improved specificity, derived from a specific Random Amplified Polymorphic DNA (RAPD) generated fragment, is proposed. The PC1/PC2 primers, used in a simple PCR protocol, gave a single amplification product of approximately 450 bp; a good degree of specificity, with absence of cross reactions with Phytophthora pseudotsugae and R idaei; sensitivity down to 6 pg of R cactorum DNA extracted from pure mycelium; no reactions with the DNA of the host plants tested (downy oak, pear and walnut trees, potato, strawberry, tomato and pea plants). The detection of R cactorum in infected tissues of pear and walnut trees, potato, strawberry, tomato and pea plants was also confirmed. The specificity, sensitivity and robustness of the PC1/PC2 primers together with the possibility of their use in a rapid, simple and reliable diagnostic method are discussed.
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