Abstract
Wheat powdery mildew, caused by the fungal pathogen Blumeria graminis f. sp. tritici (Bgt), is a destructive wheat disease worldwide. The key issue for the disease forecast is to timely and accurately estimate and quantify the latent infection levels in volunteer seedlings where the pathogen over-winters or over-summers to serve as sources of initial inoculum of epidemics. To improve the conventional method, a real-time PCR assay had been established in this study to quantify latent infection level of wheat leaves. Artificially and naturally infected leaves in wheat fields at different geographical locations in China were collected and processed to determine the latent infection levels. Linear relationships between the molecular disease index (MDX) and the observed disease index (DX) were obtained from artificial inoculation experiments. Field experiments showed that the spatial distribution patterns of MDX matched well with those of DX in the most cases. This study demonstrated that the real-time PCR assay was a useful tool to rapidly and accurately quantify the latent infection levels of wheat powdery mildew and to efficiently estimate the initial inoculum potentials of epidemics in the fields.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Böhm, 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, 409–416.
Cao, X., Zhou, Y., Duan, X., Song, Y., He, W., Wang, B., et al. (2010). The contribution of chasmothesia of Blumeria graminis f. sp. tritici to its infection cycle. Plant Protection, 36(5), 145–148. in Chinese with English abstract.
Frank, J. A., Cole, H., Jr., & Hatley, O. E. (1988). The effect of planting data on fall infections and epidemics of powdery mildew on winter wheat. Plant Disease, 72, 661–664.
Gayoso, C., De Ilárduya, O. M., Pomar, F., & Merino De Caceres, F. M. (2007). Assessment of real-time PCR as a method for determining the presence of Verticillium dahliae in different Solanaceae cultivars. European Journal of Plant Pathology, 118, 199–209.
Hietala, A. M., Eikenes, M., Kvaalen, H., Solheim, H., & Fossdal, C. G. (2003). Multiplex real-time PCR for monitoring Heterobasidion annosum colonization in Norway spruce clones that differ in disease resistance. Applied and Environmental Microbiology, 69, 4413–4420.
Ippolito, A., Schena, L., Nigro, F., Ligorio, V. S., & Yaseen, T. (2004). Real-time detection of Phytophthora nicotianae and P. citrophthorain citrus roots and soil. European Journal of Plant Pathology, 110, 833–843.
Justesen, A. F., Hansen, H. J., & Pinnschmidt, H. O. (2008). Quantification of Pyrenophora graminea in barley seed using real-time PCR. European Journal of Plant Pathology, 122, 253–263.
Liu, W., & Shao, Z. (1994). Epidemiology, occurrence and analysis of wheat powdery mildew in recent years. Plant Protection Technology and Extension, 6, 17–20. in Chinese with English abstract.
Liu, W., & Shao, Z. (1998). Analysis of weather factors influencing interregional epidemics of wheat powdery mildew in China. Plant Protection Technology and Extension, 18, 3–7. in Chinese with English abstract.
Luo, Y., Ma, Z., & Michailides, T. J. (2007). Quantification of allele E198A in beta-tubulin conferring benzimidazole resistance in Monilinia fructicola using real-time PCR. Pest Management Science, 63, 1178–1184.
Luo, Y., Ma, Z., Reyes, H. C., Morgan, D. P., & Michailides, T. J. (2007a). Quantification of airborne spores of Monilinia fructicola in stone fruit orchards of California using real-time PCR. European Journal of Plant Pathology, 118, 145–154.
Luo, Y., Ma, Z., Reyes, H. C., Morgan, D. P., & Michailides, T. J. (2007b). Using real-time PCR to survey frequency of azoxystrobin-resistant allele G143A in Alternaria populations from almond and pistachio orchards in California. Pesticide Biochemistry and Physiology, 88, 328–336.
Luo, Y., Reyes, H. C., Morgan, D. P., & Michailides, T. J. (2008). Strategies to reduce risk of benzimidazole resistance in Monilinia fructicola populations by using real-time PCR. Phytopathology (Supplement), 98, S95.
Mcneil, M., Roberts, A. M. I., Cockerell, V., & Mulholland, V. (2004). Real-time PCR assay for quantification of Tilletia caries contamination of UK wheat seed. Plant Pathology, 53, 741–750.
Rogers, S. L., Atkins, S. D., & West, J. S. (2009). Detection and quantification of airborne inoculum of Sclerotinia sclerotiorum using quantitative PCR. Plant Pathology, 58, 324–331.
Saghai-Maroof, M. A., Soliman, K. M., Jorgensen, R. A., & Allard, R. W. (1984). Ribosomal DNA spacer-length polymorphisms in barley: Mendelian inheritance, chromosomal location, and population dynamics. Proceedings of the National Academy of Sciences of the United States of America, 81(24), 8014–8018.
Schena, L., Nigro, F., Ippolito, A., & Gallitelli, D. (2004). Real-time quantitative PCR: a new technology to detect and study phytopathogenic and antagonistic fungi. European Journal of Plant Pathology, 110, 893–908.
Vandemark, G. J., & Barker, B. M. (2003). Quantifying Phytophthora medicaginis in susceptible and resistant alfalfa with a real-time fluorescent PCR assay. Journal of Phytopathology, 151, 577–583.
Vandemark, G. J., Barker, B. M., & Gritsenko, M. A. (2002). Quantifying Aphanomyces euteiches in alfalfa with a fluorescent polymerase chain reaction assay. Phytopathology, 92, 265–272.
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, 112–116.
Yan, L., Yang, Q., Zhou, Y., Duan, X., & Ma, Z. (2009). A real-time PCR assay for quantification of the Y136F allele in the CYP51 gene associated with Blumeria graminis f. sp. tritici resistance to sterol demethylase inhibitors. Crop Protection, 28, 376–380.
Zadoks, J. C., & Schein, R. D. (1979). Epidemiology and plant disease management. New York: Oxford University Press.
Zadoks, J. C., Chang, T., & Konzak, C. (1974). A decimal code for the growth stages of cereals. Weed Research, 14, 415–421.
Zeng, X., Luo, Y., Zheng, Y., Duan, X., & Zhou, Y. (2010). Detection of latent infection of wheat leaves caused by Blumeria graminis f. sp. tritici using nested PCR. Journal of Phytopathology, 158, 227–235.
Acknowledgments
We thank Profs. Wanquan Chen, Shichang Xu, Yiping Shi, and Dr. Taiguo Liu, Institute of Plant Protection, CAAS, China, and Prof. Yuanyin Cao, Shenyang Agricultural University, China, for providing isolates of different species of wheat pathogens used in this study. We thank Drs. Xiuquan Li and Xianchun Xia, Institute of Crop Sciences, CAAS, China, and Dr Jinghuan Zhu, Institute of Crop Research and Nuclear Technique Utilization in Agriculture, Zhejiang Academy of Agricultural Sciences, China, for providing barley seeds for primer specificity tests. We also thank Dr. Qiang Li, Northwest A & F University, China, Profs. Wenlan He and Gongqiang Yang, Institute of Plant Protection, Henan Academy of Agricultural Sciences, China, for their help during field experiments. This study was supported by National Natural Science Foundation of China (31171793), National Key Basic Research Program of China (2013CB127700), National Key Technology R&D Programs of China (2012BAD19B04) and Special Fund for Agro-scientific Research in the Public Interest (201303016).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Zheng, Y., Luo, Y., Zhou, Y. et al. Real-time PCR quantification of latent infection of wheat powdery mildew in the field. Eur J Plant Pathol 136, 565–575 (2013). https://doi.org/10.1007/s10658-013-0188-5
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10658-013-0188-5