Development of the rapid detection methods of rice sheath blight based on PCR and quantitative PCR techniques

• Main Authors: Shen, Shih-Mao, 沈世茂
• Other Authors: Lin, Ying-Hong
• Format: Others
• Language: zh-TW
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/6d6yst

碩士 === 國立屏東科技大學 === 植物醫學系所 === 106 === Sheath blight of rice (ShB) caused by Rhizoctonia solani is a major limiting factor to rice production. This soil-borne fungal pathogen can survive in the soil by sclerotia and mycelia. They are the primary inoculum in rice fields. The primary inoculum has ability to infect the rice sheath from the stem base and multiply in the tissues causing lesion symptoms on sheath. In order to prevent the outbreak of rice sheath blight and to reduce the economic loss of rice yield, it’s critically important to develop a rapid detection method with specific, fast, and on-site advantages for disease management. In this study, the rapid extraction reagent which can be used for on-site detection was developed. The specificity and sensitivity of the novel primers set SMS RS1-F/SMS RS1-R and the reference-primer set GMRS-3/ITS1 were tested. In addition, this study developed the four different extraction protocols for rice sheath infected by R. solani. Moreover, the intraday- and interday-assays were performed for the reproducibility evaluation of the detection methods developed in this study. The results of reproducibility evaluation indicated that all developed detection protocols were suitable to be used for diseased rice sheath. In the future, an on-site will be developed for field soil to make the detection system to be more completely.