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研究生:鄭翊宏
研究生(外文):Yi-Hong Cheng
論文名稱:台灣產小麥赤黴病菌之鑑定與其真菌毒素化學型分析
論文名稱(外文):Identification and Mycotoxin Chemotypes of Fusarium Head Blight Pathogens of Wheat in Taiwan
指導教授:王智立王智立引用關係
口試委員:黃振文謝廷芳
口試日期:2015-07-15
學位類別:碩士
校院名稱:國立中興大學
系所名稱:植物病理學系所
學門:農業科學學門
學類:植物保護學類
論文種類:學術論文
論文出版年:2015
畢業學年度:103
語文別:中文
論文頁數:71
中文關鍵詞:小麥赤黴病真菌毒素化學型脫氧雪腐鐮孢烯醇雪腐鐮孢烯醇
外文關鍵詞:Fusarium head blightchemotypeDONNIVFusarium asiaticumFusarium graminearumFusarium meridionale
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小麥赤黴病 (Fusarium head blight of Wheat﹔FHB) 是造成全球小麥重大經濟損失的病害之一,可分別由17種不同的Fusarium species所造成,並以Fusarium graminearum, Fusarium culmorum, Fusarium avenaceum, Fusarium poae, 及 Microdochium nivale為主要病原菌,其中又以F. graminearum所造成的FHB最普遍。目前經由多基因的分子演化分析,認為過去依據形態所鑑定的F. graminearum是一個複合種 (Fusarium graminearum sepcies complex﹔FGSC),因此進一步區分出16種不同的親緣種 (phylogenetic species)。台灣近年來部分地區小麥赤黴病發生嚴重,但過去對於小麥病害研究甚少,有關小麥赤黴病菌僅紀錄F. graminearum,對於台灣產小麥赤黴病菌所產生的真菌毒素之化學型 (chemotype)相關紀錄甚少。本篇研究主要係利用形態及分子生物學方法,針對195株台灣產小麥赤黴病菌的族群進行菌種鑑定及分析真菌毒素之化學型。研究發現造成台灣產小麥赤黴病之病原菌至少有3個屬於FGSC的親緣種,其中以Fusarium asiaticum (98.5%)占最多數,另外亦包含F. graminearum sensu stricto (0.5%)及Fusarium meridionale (0.5%)及未知親緣種(0.5%),且進一步發現F. asiaticum係由3種不同的族群所組成,分別為SCAR type 2 (76.1%)、SCAR type 3 (1.6%) 及SCAR type 4 (21.4%)。其中真菌毒素之化學型分析發現產生脫氧雪腐鐮孢烯醇 (deoxynivalenol﹔DON) 的菌株占多數 (83.1%),產生雪腐鐮孢烯醇 (nivalenol﹔NIV) 的菌株較少 (17.2% ),產生DON的菌株可依據所產生之DON結構再分成兩種化學型 (3-Ac-DON, 15-Ac-DON),其中產生15-Ac-DON的菌株佔多數 (98.1%),產生3-Ac-DON的菌株佔少數 (1.9%),更進一步利用HPLC化學分析,結果顯示利用分子生物學鑑定小麥赤黴病菌之真菌毒素化學型的方法具有可靠性,此外為了瞭解不同SCAR type與不同真菌毒素化學型是否有毒力差異,因而造成族群數量不同,進一步進行不同族群菌株的毒力測試,結果並未發現菌株族群間有毒力差異。本研究顯示造成台灣產小麥赤黴病之病原菌主要係F. asiaticum SCAR type 2引起,所產生的真菌毒素化學型為15-Ac-DON,未來若要進行小麥赤黴病之抗病育種,應以此病原菌族群作為主要防除對象進行植株選拔。

Fusarium head blight (FHB) is a very important wheat disease that causes great economic loss in the world. Seventeen different Fusarium species have been reported as the causal agents and Fusarium graminearum is the most common pathogen. According to the multiple locus genotyping, F. graminearum had been reported as a species complex. As of today, 16 phylogenetic species had been recongized. Recently, FHB severly occurred in some areas in Taiwan. Due to lack of studies on wheat diseases in Taiwan, F. graminearum was the only recorded pathogen and the information regarding the chemotype of mycotoxin was not well known. This study used morphology and molecular methods to identify the phylogenetic species and mycotoxin chemotypes of 195 FHB pathogens from various areas of Taiwan. The result showed that there were at least 3 FGSC phylogenetic species in Taiwan, including Fusarium asiaticum, F. graminearum sensu stricto, Fusarium meridionale and an unknown species. F. asiaticum was the most common species (98.5%), and could be further classified into 3 SCAR types. The most common SCAR type was SCAR type 2 (76.1%), others were SCAR type 3 (1.6%) and SCAR type 4 (21.4%). Three mycotoxin chemotypes were identified by the Tri gene molecular chemotyping. The most common mycotoxin chemotype was 15-Ac-DON (81.4%), others were NIV (17.2%) and 3- Ac-DON (1.57%). The mycotoxin chemotypes of some isolates were further determined by HPLC, indicating that the Tri gene molecular chemotyping was a reliable method for chemotyping. In additional, the virulence assays were carried out to address if pathogen virulence is associated with the uneven distributions of SCAR types or chemotypes in populations. However, the result showed no correlation between them. This study indicated that the major causal agent of FHB in Taiwan was F. asiaticum SCAR type 2 that produced 15-Ac-DON mycotoxin. The population of this fungus should be the target of FHB control.

中文摘要 I
Abstract III
表次索引 VII
圖次索引 VIII
前言 1
材料與方法 8
一、 小麥赤黴病之發生與調查 8
(一) 台中市大雅區小麥赤黴病的發生調查 8
(二) 小麥種子感染小麥赤黴病之調查 8
二、 供試菌株之來源、分離與保存 9
(一) 小麥赤黴病菌之蒐集 9
(二) 病原菌之分離 9
(三) 供試菌株之更新與保存 9
三、 小麥赤黴病菌之病原性測定 10
(一) 小麥赤黴病菌之產孢與孢子懸浮液製備 10
(二) 小麥胚芽軸接種法 10
四、 小麥赤黴病菌之分子鑑定 10
(一) 菌株之總DNA萃取 10
(二) FGSC複合種及SCAR type之鑑定 11
(三) 真菌毒素化學型之分子鑑定 12
(四) F. graminearum s.s 及 F. asiaticum 親緣種鑑定 13
五、 代表性菌株之形態、毒力測試、分子親緣分析及真菌毒素化學分析 13
(一) 菌株形態 14
(二) 麥穗點接種毒力測試 14
(三) 基因序列之分子親緣分析 14
(四) 真菌毒素化學分析 16
結果 18
一、 小麥赤黴病之發生與調查 18
(一) 台中市大雅區小麥赤黴病害調查 18
(二) 小麥種子感染小麥赤黴病之調查 18
二、 供試菌株之分離 18
三、 菌株之病原性測定 19
四、 SCAR type分析結果 19
五、 F. graminearum s.s 及 F. asiaticum 親緣種鑑定 19
六、 真菌毒素化學型分子鑑定 20
七、 代表性菌株之挑選 20
八、 代表性菌株之演化樹親緣分析 21
九、 代表性菌株之無性態及有性態 22
十、 代表性菌株之毒力分析 22
討論 24
參考文獻 31


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