European Journal of Plant Pathology

, Volume 120, Issue 3, pp 241–247 | Cite as

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

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

Wheat Stripe rust Puccinia striiformis Molecular diagnosis PCR 

References

  1. Calderon, C., Ward, E., Freeman, J., Foster, S. J., & McCartney, H. A. (2002). Detection of airborne inoculum of Leptosphaeria maculans and Pyrenopeziza brassicae in oilseed rape crops by polymerase chain reaction (PCR) assays. Plant Pathology, 51(3), 303–310.CrossRefGoogle Scholar
  2. Chen, X. M., Band, R. F., & Leung, H. (1993). Relationship between virulence variation and DNA polymorphism in Puccinia striiformis. Phytopathology, 83, 1489–1497.CrossRefGoogle Scholar
  3. Fraaije, B. A., Lovell, D. J., Coelho, J. M., Baldwin, S., & Hollomon, D. W. (2001). PCR-based assays to assess wheat varietal resistance to blotch (Septoria tritici and Stagonospora nodorum) and rust (Puccinia striiformis and Puccinia recondita) diseases. European Journal of Plant Pathology, 107, 905–917.CrossRefGoogle Scholar
  4. Frederick, R. D., Snyder, C. L., Peterson, G. L., & Bonde, M. R. (2002). Polymerase chain reaction assays for the detection and discrimination of the soybean rust pathogens Phakopsora pachyrhizi and P. meibomiae. Phytopathology, 92, 217–227.CrossRefGoogle Scholar
  5. Goodwin, P. H., English, J. T., Nether, D. A., Duniway, J. M., & Kirkpatrick, B. C. (1990). Detection of Phytophthora parasitica from soil and host tissue with a species-specific DNA probe. Phytopathology, 80, 277–281.CrossRefGoogle Scholar
  6. Grote, D., Olmos, A., Kofoet, A., Tuset, J. J., Bertolini, E., & Cambra, M. (2002). Specific and sensitive detection of Phytophthora nicotianae by simple and nested-PCR. European Journal of Plant Pathology, 108, 197–207.CrossRefGoogle Scholar
  7. Ippolito, A., Schena, L., & Nigro, F. (2002). Detection of Phytophthora nicotianae and P. citrophthora in citrus roots and soils by nested PCR. European Journal of Plant Pathology, 108, 855–868.CrossRefGoogle Scholar
  8. Judelson, H. S., & Tooley, P. W. (2000). Enhanced polymerase chain reaction methods for detecting and quantifying Phytophthora infestans in plants. Phytopathology, 90, 1112–1119.CrossRefGoogle Scholar
  9. Lacourt, I., & Duncan, J. M. (1997). Specific detection of Phytophthora nicotianae using the polymerase chain reaction and primers based on the DNA sequences of its elicitin gene ParaA1. European Journal of Plant Pathology, 103, 73–83.CrossRefGoogle Scholar
  10. Li, S., & Hartman, G. L. (2003). Molecular detection of Fusarium solani f.sp. glycines in soybean roots and soil. Plant Pathology, 52, 74–83.CrossRefGoogle Scholar
  11. Li, Z. Q. (1999). Book of academician. Northwest A&F University Press.Google Scholar
  12. Li, Z. Q., & Shang, H. S. (1989). Wheat rusts and their control. Shanghai, China: Shanghai Science and Technology Press.Google Scholar
  13. Mills, P. R., Sreenivasaprasad, S., & Brown, A. E. (1992). Detection and differentiation of Colletotrichum gloeosporioides isolates using PCR. FEMS Microbiological Letters, 98, 137–144.CrossRefGoogle Scholar
  14. Saari, E. E., & Prescott, J. M. (1985). World distribution in relation to economic losses. In A. P. Roelfs & W. R. Boshell (Eds.), The cereal rusts vol 2. Diseases distribution epidemiology and control (pp. 260–298). Orlando, Florida: Academic.Google Scholar
  15. Shan, W. X, Chen, S. Y., Kang, Z. S., Wu, L. R., & Li, Z. Q. (1998). Genetic diversity in Puccinia striiformis f. sp. tritici revealed by pathogen genome-specific repetitive sequence. Canadian Journal of Botany, 76, 587–595.CrossRefGoogle Scholar
  16. Silvar, C., Duncan, J. M., Cooke, D. E. L., Willians, N. A., Diaz, J., & Merino, F. (2005). Development of specific PCR primers for identification and detection of Phytsphthora capsici Leon. European Journal of Plant Pathology, 112, 43–52.CrossRefGoogle Scholar
  17. Tisserat, N. A., Hulbert, S. H., & Nus, A. (1991). Identification of Leptosphaeria korrae by cloned DNA probes. Phytopathology, 81, 917–921.CrossRefGoogle Scholar
  18. Wan, A., Zhao, E., Chen, X., He, E., Jia, S., Jia, Q., et al. (2004). Wheat stripe rust epidemic and virulence of Puccinia striiformis f. sp. tritici in China in 2002. Plant Disease, 88, 896–904.CrossRefGoogle Scholar
  19. Wang, K. N., Wu, L. R., & Meng, Q. Y. (1986). On the physiological specialization of stripe rust of wheat in China during 1975–1986. Acta Phytopathologica, 16(2), 79–85.Google Scholar
  20. Zadoks, J. C., Chang, T. T., & Konzak, C. F. (1974). A decimal code for growth stages of cereals. Weed Research, 14, 415–421.CrossRefGoogle Scholar
  21. Zheng, W. M., Chen, S. Y., Kang, Z. S., Wang, Y., Li, Z. Q., & Wu, L. R. (2000a). Specificity and stability of PSR (Puccinia striiformis Repeat) sequence. Acta Phytopathologica Sinica, 30(3), 222–225.Google Scholar
  22. Zheng, W. M., Liu, F., Kang, Z. S., Chen, S. Y., Li, Z. Q., & Wu, L. R. (2000b). AFLP analysis of predominant races of Puccinia striiformis in China. Progress in Natural Science, 10(6), 532–537.Google Scholar

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

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