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European Journal of Plant Pathology

, Volume 121, Issue 1, pp 1–8 | Cite as

Differentiation of two powdery mildews of sunflower (Helianthus annuus) by a PCR-mediated method based on ITS sequences

  • Ruey-Shyang Chen
  • Chishih Chu
  • Chi-Wei Cheng
  • Wen-Yu Chen
  • Jwu-Guh Tsay
Full Research Paper

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.

Keywords

Multiplex PCR Polymerase chain reaction Powdery mildew Sunflower 

Notes

Acknowledgements

This study was supported by Grant Nos. 94AS-13.3.1-BQ-B1 (9) and 95AS-13.3.1-BQ-B1 (10) from the Bureau of Animal and Plant Health Inspection and Quarantine, the Council of Agriculture, Taiwan, R.O.C.

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Copyright information

© KNPV 2007

Authors and Affiliations

  • Ruey-Shyang Chen
    • 1
  • Chishih Chu
    • 2
  • Chi-Wei Cheng
    • 3
  • Wen-Yu Chen
    • 1
  • Jwu-Guh Tsay
    • 2
  1. 1.Graduate Institute of BiotechnologyNational Chiayi UniversityChiayiTaiwan
  2. 2.Department of Microbiology and ImmunologyNational Chiayi UniversityChiayiTaiwan
  3. 3.Graduate Institute of Agricultural BiotechnologyNational Chiayi UniversityChiayiTaiwan

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