Differentiation of two powdery mildews of sunflower (<Emphasis Type="Italic">Helianthus annuus</Emphasis>) by a PCR-mediated method based on ITS sequences |
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Authors: | Ruey-Shyang Chen Chishih Chu Chi-Wei Cheng Wen-Yu Chen Jwu-Guh Tsay |
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Institution: | (1) Graduate Institute of Biotechnology, National Chiayi University, Chiayi, 600-04, Taiwan;(2) Department of Microbiology and Immunology, National Chiayi University, Chiayi, 600-04, Taiwan;(3) Graduate Institute of Agricultural Biotechnology, National Chiayi University, Chiayi, 600-04, Taiwan |
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Abstract: | Powdery mildew can be found in most sunflower fields during the winter season in Taiwan and causes severe yellowing on the
blade, petiole, stem, and calyx, as well as serious defoliation. Two types of powdery mildew fungi isolated from sunflower
leaves showed variable status for fibrosin bodies. But only the cleistothecium of Podosphaera xanthii, one of the pathogens causing this disease, was observed on samples from Chungpu County at the beginning of 2005. With a
species-specific primer pair, PN23/PN34, no specific PCR product was amplified from the pathogen’s genomic DNA. Based upon
the ITS sequence of rDNA, three PCR primer sets (S1/S2, G1/G2, and L1/L2) specific to P. xanthii, Golovinomyces cichoracearum and Leveillula taurica, respectively, were designed to detect and differentiate pathogens causing powdery mildews on sunflower. Only the primer
pairs S1/S2 and G1/G2 could amplify PCR products, with product sizes of 454 and 391 bp, respectively. Four samples of fungal
DNA were subjected to a multiplex PCR amplification with primer pairs S1/S2 and G1/G2; P. xanthii and G. cichoracearum were successfully detected. These results suggest that the multiplex PCR method is a rapid, simple, and effective technique
to detect and differentiate powdery mildews, for example P. xanthii and G. cichoracearum, found on sunflower. With morphologic characteristics, ITS sequence analysis and pathogenicity testing, P. xanthii and G. cichoracearum, the first case, are two powdery mildews on sunflower in Taiwan. |
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Keywords: | Multiplex PCR Polymerase chain reaction Powdery mildew Sunflower |
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