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.
Similar content being viewed by others
References
American Phytopathological Society Committee on the Standardization of Common Names for Plant Diseases (1994). Common names for plant diseases pp. 147–149. St. Paul, USA: APS Press.
Bardin, M., Carlier, J., & Nicot, P. C. (1999). Genetic differentiation in the French population of Erysiphe cichoracearum, a causal agent of powdery mildew of cucurbits. Plant Pathology, 48, 531–540.
Braun, U. (1995). The powdery mildews (Erysiphales) of Europe (pp 1–337). Jena, Germany: G. Fischer Verlag.
Braun, U., Cook, R. T. A., Inman, A. J., & Shin, H. D. (2002). The taxonomy of the powdery mildew fungi. In R. R. Bélanger, W. R. Bushnell, A. J. Dik, & T. L. W. Carver (Eds.), The powdery mildews: A comprehensive treatise (pp. 13–55). St. Paul, USA: APS Press.
Cheng, C. W., Chen, R. S., Chang, W. H., & Tsay, J. G. (2006). The occurrence of powdery mildew of Physalis angulata caused by Podosphaera xanthii. Plant Protection Bulletin (Taiwan), 48, 41–51.
Epinat, C., Pitrat, M., & Bertrand, F. (1993). Genetic analysis of resistance of 5 melon lines to powdery mildews. Euphytica, 65, 135–144.
Fang, H. C. (1973). Powdery mildew of sunflower in Taiwan. Plant Protection Bulletin (Taiwan), 15, 5–12.
Fraaije, B. A., Lovell, D. J., Coelho, J. M., Baldwin, S., & Hollomon, D. W. (2001). PCR-based assay 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.
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.
Gulya, T. J., Woods, D. M., Bell, R., & Mancl, M. K. (1991). Diseases of sunflower in California. Plant Disease, 75, 572–574.
Heiser, C. B. (1954). Variation and subspeciation in the common sunflower, Helianthus annuus. The American Midland Naturalist, 51, 287–305.
Hirata, T., Cunnington, J. H., Paksiri, U., Limkaisang, S., Shishkoff, N., Grigailiunaite, B., et al. (2000). Evolutionary analysis of subsection Magnicellulatae of Podosphaera section Sphaerotheca (Erysiphales) based on the rDNA ITS sequences with special reference to host plants. Canadian Journal of Botany, 78, 1521–1530.
Hirata, T., & Takamatsu, S. (1996). Nucleotide sequence diversity of rDNA internal transcribed spacers extracted from conidia and cleistothecia of several powdery mildew fungi. Mycoscience, 37, 283–288.
Hirata, T., & Takamatsu, S. (2001). Phylogeny and cross-infectivity of powdery mildew isolates (Podosphaera fuliginea s. lat.) on cosmos and cucumber. Journal of General Plant Pathology, 67, 1–6.
Hsieh, H. J. (1983). Notes on host plants of powdery mildew fungi found in Taiwan (1) Host plants of Sphaerotheca fuliginea. Plant Protection Bulletin (Taiwan), 25, 109–113.
Hsieh, S. Y. (2002). Cultivation of sunflower as landscaping plant and manure. Technical Bulletin 91-1 (Bo. 119), Tainan District Agricultural Improvement Station, Taiwan.
Huang, J. W. (1992). Integrated management of vegetable seedling pests with a formulated plant nutrition. Plant Protection Bulletin (Taiwan), 34, 54–63.
Kashimoto, K., Matsuda, Y., Kakutani, K., Sameshima, T., Kakutani, K., Nonomura, T., et al. (2003). Morphological and molecular characterization for a Japanese isolate of tomato powdery mildew Oidium neolycopersici and its host range. Journal of General Plant Pathology, 69, 176–185.
Khodaparast, S. A., Takamatsu, S., & Hedjaroude, G. (2001). Phylogenetic structure of the genus Leveillula (Erysiphales: Erysiphaceae) inferred from the nucleotide sequences of the rDNA ITS region with special reference to the L. taurica species complex. Mycological Research, 105, 909–918.
Kuo, K. C., Lee, C. Y., & Leu, L. S. (1991). Notes on the Erysiphaceae found in Taiwan (I). Transactions of the Mycological Society of Republic of China, 6, 13–44.
Patzak, J. (2005). PCR detection of Pseudoperonospora humuli and Podosphaera macularis in Humulus lupulus. Plant Protection Science, 41, 141–149.
Reifschneider, F. J. B., Boiteux, L. S., & Occhiena, E. M. (1985). Powdery mildew of melon (Cucumis melo) caused by Sphaerotheca fuliginea in Brazil. Plant Disease, 69, 1069–1070.
Saliman, M., Yang, S. M., & Wilson, L. (1982). Reaction of Helianthus species to Erysiphe cichoracearum. Plant Disease, 66, 572–573.
Shi, A., & Mmbaga, M. T. (2006). Perpetuation of powdery mildew infection and identification of Erysiphe australiana as the crape myrtle pathogen in mid-Tennessee. Plant Disease, 90, 1098–1101.
Takamatsu, S., & Kano, Y. (2001). PCR primers useful for nucleotide sequencing of rDNA of the powdery mildew fungi. Mycoscience, 42, 135–139.
Tsay, J. G., & Tung, B. K. (1992). Ecology and control strategy of powdery mildew of cucurbits. Plant Protection Bulletin (Taiwan), 34, 139–148.
White, T. J., Bruns, T., Lee, S., & Taylor, J. (1990). Amplification and direct sequencing of fungi ribosomal RNA genes for phylogenetics. In M. A. Innis, D. V. Gelfand, J. J. Sninsky, & T. J. White (Eds.), PCR protocols: A guide to methods and applications pp. 315–322. New York: Academic.
Yang, S. M., Wei, S. E., Ge, C. F., Liang, K. Z., & Wang, L. (1988). Diseases of cultivated sunflower in Liaoning Province, People’s Republic of China. Plant Disease, 72, 546.
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.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Chen, RS., Chu, C., Cheng, CW. et al. Differentiation of two powdery mildews of sunflower (Helianthus annuus) by a PCR-mediated method based on ITS sequences. Eur J Plant Pathol 121, 1–8 (2008). https://doi.org/10.1007/s10658-007-9234-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10658-007-9234-5