Construction of DNA markers for diagnosis of turfgrass diseases
2006
Yamaga, A.(Meiji Univ., Kawasaki, Kanagawa (Japan). School of Agriculture) | Tsuchida, T. | Mitsuhori, T. | Shingu, Y. | Arie, T. | Yamashita, Y. | Kudo, K. | Yoneyama, K.
Turfgrass diseases are very hard to diagnose, because many diseases have very similar symptoms. Diagnosis of turfgrass diseases requires skillful techniques to identify the pathogens. Recently, the nucleotide sequences in specific genomic DNA regions, particularly the internal transcribed spacer (ITS) region of rDNA, have been used for identification and classification of filamentous fungi. The ITS regions in fungi are not highly conserved among the genera, or among the species as compared with rDNA-specified regions. When the PCR primers characteristic for each fungal pathogen are constructed, they will be useful for PCR diagnosis of plant diseases because a pathogen can be determined by amplification of the specific ITS region. Experiments were made using the fungal pathogens Rhizoctonia solani, a less-sporulated strain of Curuvularia sp., Sclerotinia homoeocarpa and Pythium aphanidermatum, which cause serious diseases in the turf green of golf courses in Japan. The PCR primers for each pathogen were constructed, and named the primers CD1-CD2 for Curuvularia sp., the primers RSY1-RSY2 and R1S1-R1S2 for R. solani, the primers Pas1-Pas2 for P. aphanidermatum, and the primers Dr1-Dr2 for S. homoeocarpa. The amplified PCR products from these primers gave a predicted DNA fragment in each pathogen. Also, we developed a simple alkaline extraction method to obtain the fungal DNA from infected leaves, and succeeded in detection of the causal pathogens from the diseased leaves that occurred in turfgrass fields. Thus, this PCR diagnostic technique is useful for pathogen assay of infected turfgrasses.
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