European Journal of Plant Pathology

, Volume 120, Issue 3, pp 241–247

The development of a PCR-based method for detecting Puccinia striiformis latent infections in wheat leaves

  • Xiaojie Wang
  • Wenming Zheng
  • Heinrich Buchenauer
  • Jie Zhao
  • Qingmei Han
  • Lili Huang
  • Zhensheng Kang
Full Research Paper

DOI: 10.1007/s10658-007-9212-y

Cite this article as:
Wang, X., Zheng, W., Buchenauer, H. et al. Eur J Plant Pathol (2008) 120: 241. doi:10.1007/s10658-007-9212-y

Abstract

Stripe rust of wheat caused by Puccinia striiformis f. sp. tritici is one of the most important diseases on wheat worldwide, especially in temperate regions with cool moist weather conditions. A rapid and reliable detection of the pathogen in latent infected wheat leaves during overwintering of the fungus in the dormant stage will contribute to determine the initial inoculum potential and thus to predict early outbreak and to improve effective management of the disease. To achieve this aim, a PCR-based method was developed for specific and sensitive detection of P. striiformis. Specific primers were designed according to a genome-specific sequence of P. striiformis. To evaluate the specificity of the primers, seven different isolates and races of P. striiformis as well as six other pathogens of wheat were tested. All isolates of P. striiformis yielded a distinct band of a fragment of 470 bp, while using DNA of the other wheat pathogens as a template no amplification product was detected. The sensitivity of the primers was tested using serial dilutions of total DNA from P. striiformis; the limit of detection was 10 pg of DNA. Using extracts from P. striiformis-infected wheat leaves, the fungus could be determined in the leaves before symptoms appeared. The stripe rust could also be detected in the dormant stage by the PCR assay in samples of wheat leaves taken during the winter season. The application of the PCR assay may be useful for rapid and reliable detection of P. striiformis in latent infected leaves of overwintering wheat plants.

Keywords

WheatStripe rustPuccinia striiformisMolecular diagnosisPCR

Copyright information

© KNPV 2007

Authors and Affiliations

  • Xiaojie Wang
    • 1
  • Wenming Zheng
    • 2
  • Heinrich Buchenauer
    • 3
  • Jie Zhao
    • 1
  • Qingmei Han
    • 1
  • Lili Huang
    • 1
  • Zhensheng Kang
    • 1
  1. 1.College of Plant Protection and Shaanxi Key Laboratory of Molecular Biology for AgricultureNorthwest A&F UniversityYanglingChina
  2. 2.College of Life ScienceHenan Agricultural UniversityZhengzhouChina
  3. 3.Institute of PhytomedicineUniversity of HohenheimStuttgartGermany