Simultaneous detection of three wheat pathogenic fungal species by multiplex PCR

详细信息    查看全文

• 作者：Si Chen ; Yuan-yin Cao ; Tian-ya Li ; Xian-xin Wu
• 关键词：Blumeria graminis f. sp. tritici ; Puccinia graminis f. sp. tritici ; Puccinia triticina ; Triticum spp
• 刊名：Phytoparasitica
• 出版年：2015
• 出版时间：September 2015
• 年：2015
• 卷：43
• 期：4
• 页码：449-460
• 全文大小：1,399 KB
• 参考文献：Abebe, T., Woldeab, G., & Dawit, W. (2012). Distribution and physiologic races of wheat stem rust in Tigray, Ethiopia. Journal of Plant Pathology and Microbiology, 3, 142.View Article
  Aldrich, J., & Cullis, C. A. (1993). RAPD analysis in flax: optimization of yield and reproducibility using Klen Taq I DNA polymerase, Chelex 100, and gel purification of genomic DNA. Plant Molecular Biology Reports, 11, 128鈥?41.View Article
  Ballabio, A., Ranier, J. E., Chamberlain, J. S., Zollo, M., & Caskey, C. T. (1990). Screening for steroid sulfatase (STS) gene deletions by multiplex DNA amplification. Human Genetics, 84, 571鈥?73.PubMed View Article
  Bates, J. A., Taylor, E. J. A., Kenyon, D. M., & Thomas, J. E. (2001). The application of real-time PCR to the identification, detection and quantification of Pyrenophora species in barley seed. Molecular Plant Pathology, 2, 49鈥?7.PubMed View Article
  Bohm, J., Hahn, A., Schubert, R., Bahnweg, G., Adler, N., Nechwatal, J., et al. (1999). Real-time quantitative PCR: DNA determination in isolated spores of the mycorrhizal fungus Glomus mosseae and monitoring of Phytophthora infestans and Phytophthora citricola in their respective host plants. Journal of Phytopathology, 147, 409鈥?16.View Article
  Cao, L. H., Xu, S. C., Chen, W. Q., Liu, T. G., & Lin, R. M. (2007). Molecular diagnosis and detection of Puccinia triticina in China. Acta Phytopathologica Sinica, 34, 561-566. (in Chinese, with English abstract)
  Cao, Y. Y., Yu, J. C., Liu, Q., & Wei, S. H. (2005). Studies on preservation method for pure isolates of wheat powdery mildew. Acta Phytopathologica Sinica, 32, 271-272. (in Chinese, with English abstract)
  Capote, N., Pastrana, A. M., Aguado, A., & S谩nchez-Torres, P. (2012). Molecular tools for detection of plant pathogenic fungi and fungicide resistance. In C. J. R. Cumagun (Ed.), Plant pathology (pp. 151鈥?02). Rijeka, Croatia: Intech.
  Chen, H. G., Fang, Z., Chen, H. D., Lin, L., & Wang, Y. Z. (2005a). Comparison of Wheat sharp spot pathogenic isolates based on the internal transcribed spacer sequence. Acta Phytopathologica Sinica, 35, 24-29. (in Chinese, with English abstract)
  Chen, M. J., Fang, T., Ke, T., Xiong, Z. Y., & He, G. Y. (2005b). Multiplex PCR 鈥?a molecular biotechnique of high efficiency and speed. Journal of Wuhan University of Technology, 27, 33鈥?6 (in Chinese with English abstract).
  C么t茅, M.-J., Tardif, M.-C., & Meldrum, A. J. (2004). Identification of Monilinia fructigena, M. fructicola, M. laxa, and Monilia polystroma on inoculated and naturally infected fruit using multiplex PCR. Plant Disease, 88, 1219鈥?225.View Article
  Cullen, D. W., Lees, A. K., Toth, I. K., & Duncan, J. M. (2001). Conventional PCR and real-time PCR detection of Helminthosporium solani in soil and on potato tubers. European Journal of Plant Pathology, 107, 387鈥?98.View Article
  Edwards, M. C., & Gibbs, R. A. (1994). Multiplex PCR: advantages, development, and applications. Genome Research, 3, S65鈥揝75.View Article
  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鈥?17.View Article
  He, J. M., Song, Y. L., Zhang, Z. S., & He, W. L. (1998). Prevention and control of wheat powdery mildew I. The distribution symptoms and harm of wheat powdery mildew. Henan Agricultural Science, 1, 17-18. (in Chinese, with English abstract)
  Huang, Y. H., Hu, X. X., & Xu, W. Z. (2003). The factors affecting the efficiency of multiplex PCR. Hereditas, 25, 65-68. (in Chinese, with English abstract)
  Ito, T., Ieki, H., & Ozaki, K. (2002). Simultaneous detection of six citrus viroids and Apple stem grooving virus from citrus plants by multiplex reverse transcription polymerase chain reaction. Journal of Virological Methods, 106, 235鈥?39.PubMed View Article
  Johnson, J. R. (2000). Development of polymerase chain reaction-based assays for bacterial gene detection. Journal of Microbiological Methods, 41, 201鈥?09.PubMed View Article
  Lefeuvre, P., Hoareau, M., Delatte, H., Reynaud, B., & Lett, J. M. (2007). A multiplex PCR method discriminating between the TYLCV and TYLCV-Mld clades of Tomato yellow leaf curl virus. Journal of Virological Methods, 144, 165鈥?68.PubMed View Article
  Lievens, B., & Thomma, B. P. H. J. (2005). Recent developments in pathogen detection arrays: Implications for fungal plant pathogens and use in practice. Phytopathology, 95, 1374鈥?380.PubMed View Article
  Markoulatos, P., Siafakas, N., & Moncany, M. (2002). Multiplex polymerase chain reaction: A practical approach. Journal of Clinical Laboratory Analysis, 16, 47鈥?1.PubMed View Article
  McCartney, H. A., Foster, S. J., Fraaije, B. A., & Ward, E. (2003). Molecular diagnostics for fungal plant pathogens. Pest Management Science, 59, 129鈥?42.PubMed View Article
  Nakamura, T., Vrinten, P., Saito, M., & Konda, M. (2002). Rapid classification of partial waxy wheat using PCR-based markers. Genome, 45, 1150鈥?156.PubMed View Article
  Periasamy, M., Niazi, F. R., & Malathi, V. G. (2006). Multiplex RT-PCR, a novel technique for the simultaneous detection of the DNA and RNA viruses causing rice tungro disease. Journal of Virological Methods, 134, 230鈥?36.PubMed View Article
  Schoske, R., Vallone, P. M., Ruitberg, C. M., & Butler, J. M. (2003). Multiplex PCR design strategy used for the simultaneous amplification of 10 Y chromosome short tandem repeat (STR) loci. Analytical and Bioanalytical Chemistry, 375, 333鈥?43.PubMed
  Sun, Z. G., Cao, Y. Y., Han, J. D., & Chen, W. S. (2010). Comparison of different methods for extraction of DNA of wheat stem rust urediospores. Hubei Agricultural Sciences, 49, 1281鈥?284.
  Thompson, J. R., Wetzel, S., Klerks, M. M., Va拧kov谩, D., Schoenb, C. D., 艩pak, J., et al. (2003). Multiplex RT-PCR detection of four aphid-borne strawberry viruses in Fragaria spp. in combination with a plant mRNA specific internal control. Journal of Virological Methods, 111, 85鈥?3.
  Uga, H., & Tsuda, S. (2005). A one-step reverse transcription-polymerase chain reaction system for the simultaneous detection and identification of multiple tospovirus infection. Phytopathology, 95, 166鈥?71.PubMed View Article
  Wang, N., Wang, J., Yin D. H., Gao, G. P., & Wang, W. (2010). Triplex PCR detection of Botrytis cinerea, Colletotrichum gloeosporioides and Verticillium dahliae in infected strawberry plant tissues. Scientia Agricultura Sinica, 43, 4392鈥?400.
  Wang, X., Liu, T. G., Xiang, W. S., & Chen, W. Q. (2011). Development of a SSR molecular marker for Puccinia graminis f. sp. tritici. Scientia Agricultura Sinica, 44, 4593鈥?599. (in Chinese, with English abstract)
  Ward, E., Foster, S. J., Fraaije, B. A., & McCartney, H. A. (2004). Plant pathogen diagnostics: immunological and nucleic acid-based approaches. Annals of Applied Biology, 145, 1鈥?6.View Article
  Winton, L. M., & Hansen, E. M. (2001). Molecular diagnosis of Phytophthora lateralis in trees, water, and foliage baits using multiplex polymerase chain reaction. Forest Pathology, 31, 275鈥?83.View Article
  Winton, L. M., Stone, J. K., Watrud, L. S., & Hansen, E. M. (2002). Simultaneous one-tube quantification of host and pathogen DNA with real-time polymerase chain reaction. Phytopathology, 92, 112鈥?16.PubMed View Article
  Wittwer, C. T., Herrmann, M. G., Moss, A. A., & Rasmussen, R. P. (1997). Continuous fluorescence monitoring of rapid cycle DNA amplification. Biotechniques, 22, 130鈥?38.PubMed
  Wu, Y. S., & Huang, Z. T. (1987). Racial identification and dynamics analysis of Puccinia graminis f. sp. tritici for past 20 years in China. Journal of Shenyang Agricultural University, 18, 105-138.
  Xie, Z. X., Fadl, A. A., Girshick, T., & Khan, M. I. (1997). Amplification of avian reovirus RNA using the reverse transcriptase-polymerase chain reaction. Avian Diseases, 41, 654鈥?60.PubMed View Article
  Zeng, X. W., Luo, Y., Zhou, Y. L., & Duan, X. Y. (2008). PCR detection of Blumeria graminis f. sp. tritici based on the sequences of rDNA ITS. Acta Phytopathologica Sinica, 38, 211鈥?14.
  Zhao, Y. G., Gao, C. Y., Wang, J. H., Wang, M., Xing, D. F., & Zhao, Q. S. (2003). How to avoid inhibiting by the inhibitor in PCR. Natural Sciences Journal of Harbin Normal University, 19, 81鈥?4.
  
• 作者单位：Si Chen (1)
  Yuan-yin Cao (1)
  Tian-ya Li (1)
  Xian-xin Wu (1)
  
  1. College of Plant Protection, Shenyang Agricultural University, Shenyang, Liaoning, 110866, People鈥檚 Republic of China
  
• 刊物主题：Plant Pathology; Plant Sciences; Ecology; Agriculture; Life Sciences, general;
• 出版者：Springer Netherlands
• ISSN：1876-7184

文摘

Wheat stem rust, wheat leaf rust, and wheat powdery mildew are three of the most economically damaging fungal diseases of wheat in China, caused by Puccinia graminis f. sp. tritici, Puccinia triticina, and Blumeria graminis f. sp. tritici, respectively. In this study, single-step multiplex polymerase chain reaction (PCR) was developed for the simultaneous detection of the three pathogens. Three sets of specific primers were selected, followed by optimization of the annealing temperature and the dNTPs, Taq DNA polymerase, and primer concentrations for multiplex PCR to establish a method for the simultaneous detection of P. graminis f. sp. tritici, P. triticina, and B. graminis f. sp. tritici at an early stage of infection. The specificity and sensitivity of the multiplex PCR were tested. We observed three specific bands of 395 bp, 151 bp, and 464 bp on amplification with a three-reaction system. The detection sensitivity of the multiplex PCR was 1 ng DNA for P. graminis f. sp. tritici, 10 pg DNA for P. triticina, and 10 pg DNA for B. graminis f. sp. tritici. This study pioneered the use of the triplex PCR system for simultaneous detection of P. graminis f. sp. tritici, P. triticina, and B. graminis f. sp. tritici in infected plant tissues. This novel PCR-based method provides a simple and rapid method for detecting various pathogens in wheat, which will assist in the simultaneous diagnosis and monitoring of multiple plant diseases.