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標題: | 建立 SSR 分子標誌及分析臺灣晚疫病菌族群遺傳動態 Development of microsatellite markers and genetic assessment for population dynamics of Phytophthora infestans from Taiwan |
作者: | Shun-Yuan Huang 黃順源 |
指導教授: | 劉瑞芬 |
關鍵字: | 晚疫病菌,基因分型,簡單序列重複,RG57, Phytopthora infestans,genotyping,SSR,RG57, |
出版年 : | 2016 |
學位: | 碩士 |
摘要: | 為了追蹤田間晚疫病菌的分佈與族群遺傳組成,以利其防治與管理,過去已經開發出數種分子標誌,包括RG57 基因型指紋分析。雖然RG57 是目前歐美地區廣為接受的基因分型方法,但由於涉及南方雜合分析,有需要大量基因體DNA、耗時且步驟繁複等缺點。本研究的主旨為藉由簡單序列重複(SSR)分子標誌建立一套快速、高效率的基因分型流程,以便應用於分析臺灣的晚疫病菌族群與區隔海外的菌系。經初步測試,已建立一套包含12 組SSR 分子標誌的複合式PCR 反應流程,能夠高通量地分析晚疫病菌的基因分型。為評估該分子標誌區別晚疫病菌菌系的能力,目前已完成超過200 個來自臺灣與海外菌株的基因分型。RG57 分析結果顯示,臺灣的晚疫病菌以2010 年為界,US 11.2 亞型已經取代典型的US-11系成為田間主要的基因型。SSR 分型資料顯示,近十年來,不同年分的臺灣晚疫病菌株基因型有顯著分化,說明晚疫病菌族群有隨時間演替的現象。但晚疫病菌並沒有在地理上和寄主上有遺傳分化。為區別本土與海外的晚疫病菌,以包含至少8 種菌系的海外菌株作為參考菌株,分析顯示臺灣的菌株在基因型層次能顯著的與US-8、US-22、US-23 區分,但US-7、US-11 與US-16 參考菌株的基因型與臺灣菌株相近。綜合上述結果,相對於傳統的RG57 分型方法,此套SSR 基因分型方法是一套高效率、適合高通量分析的工具,能夠精準的鑑別晚疫病菌的菌系,以快速提供菌相的遺傳資訊,幫助田間晚疫病菌的監控與防治管理。 ABSTRACT Late blight caused by Phytophthora infestans is a devastating threat to tomato and potato production worldwide. To facilitate studies of fungicide resistance, population genetics, as well as pathogen dissemination and migration of P. infestans, a variety of molecular markers have been developed, including the RG57 fingerprinting marker. Despite it is widely used, this method has drawbacks of being labor intensive, time consuming, and in need of a large amount of DNA. This study aims to develop a fast, effective genotyping method based on simple sequence repeat (SSR; also known as microsatellites). A one-step multiplex PCR method with 12 SSR markers is applied for high-throughput genotyping of P. infestans. RG57 fingerprinting showed the variant US-11.2 replace the typical US-11, becoming the dominant genotype since 2010. To evaluate whether the SSR markers are effective for distinguishing RG57-based lineages, SSR genotypes of over 200 isolates from Taiwan and overseas were analyzed. The isolates collected from different years showed significant genotypic differentiation, suggesting populations in Taiwan were turnover for the past decade. To identify overseas lineages form local populations, reference isolates including at least 5 lineages were applied. Analysis showed local isolates can be distinguished from US-8, US-22, and US-16 lineages on genotypic level, while US-7, US-11 and US-16 are close to isolates in Taiwan. In summary, compared to traditional RG57 fingerprinting, the SSR markers are effective for genotyping of P. infestans, which may provide important information for monitoring and management of P. infestans in the fields. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/49410 |
DOI: | 10.6342/NTU201602731 |
全文授權: | 有償授權 |
顯示於系所單位: | 植物病理與微生物學系 |
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