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白邊大葉蟬媒介葡萄皮爾斯病在不同品系葡萄之傳播效率比較

By Fei-Ting Hu and 胡斐婷

Abstract

Pierce’s disease (PD) of grapevines is caused by a xylem-limited bacterium, Xylella fastidiosa, and is transmitted to plants by xylem sap-feeding insects. Symptoms of Pierce’s disease of grapes are marginal leaf necrosis, yellow band between the green and necrotic tissues and dried up raisins. Based on the results of field inspection and molecular detection, Kolla paulula has been considered as a vector for PD. The goals of this study were to: 1). access the acquisition and retention of X. fastidiosa from the honeydew of K. paulula, 2). evaluate the transmission efficiency of X. fastidiosa to different grape cultivars, and 3). determine the genetic relatedness of 4 Pierce’s disease strains. K. paulula acquired the suspension of X. fastidiosa. The acquisition rate of K. paulula was 80%. The positive rate on day 5 and 8 were the highest in the experiment. In a group of three K. paulula transmission test, K. paulula acquired X. fastidiosa from symptomatic Golden Muscat and Black Queen grapevines for 24 hr, the acquisition rate were 49.56 and 54.67%, respectively. Then K. paulula transmitted X. fastidiosa to different grape cultivars for 3-7 days. The transmission rate of K. paulula transmitting from Golden Muscat source cultivar to Golden Muscat and Black Queen recipient cultivar were 17.27 and 15.45%, respectively; from Black Queen source cultivar to Golden Muscat and Black Queen recipient cultivar were 28.15 and 28.52%, respectively. In an individual K. paulula transmission test, K. paulula acquired X. fastidiosa from symptomatic Golden Muscat and transmitted to Golden Muscat: the acquisition rate was 83.33%; the transmittion rate was 13.33%. Phylogenetic analyses were performed by comparing the 16S rRNA and 16S-23S ITS of four PD strains (GMb, BQa, BQ7f and GV148) from Taiwan with the sequences of X. fastidiosa strains from different hosts and different geographical areas. The PD strains of Taiwan were closely related to the American X. fastidiosa grape strains, the similarity index was 99-100%. And the similarity index of four Taiwanese’s PD strains with the strains from citrus, coffee, peach, plum and peach were 98-99% and 99-100% based on 16S rRNA and 16S-23S ITS. The results of all tests showed that K. paulula could acquire X. fastidiosa from symptomatic grapevines, retention for X. fastidiosa in K. paulula, transmit X. fastidiosa to health grapevines. Comparing to Golden Muscat, K. paulula was more effective in transmitting X. fastidiosa to Black Queen.葡萄皮爾斯病 (Pierce’s disease, PD) 之病原係木質部難養菌 Xylella fastidiosa (Wells et al., 1987),可造成葉緣焦枯壞死、提早落葉、植株矮化及果粒乾涸等病徵。其主要的傳播方式為經由葉蟬刺吸罹病植株木質部汁液後,再將病原菌傳至健康植株。依據疫區蟲相調查及病原分子檢測結果,白邊大葉蟬 (Kolla paulula, Walker) 於2011年列入媒介昆蟲之一。故本論文針對白邊大葉蟬對 X. fastidiosa 之傳菌能力、保菌時間,及 X. fastidiosa 對金香 (Vitis labruscana cv. Golden Muscat) 及黑后 (V. labruscana cv. Black Queen) 葡萄植株之致病率進行研究,並以 16S rRNA 及 16S-23S ITS 區段序列之比對,探討本土 PD 之 X. fastidiosa 與其他地區或寄主植物之菌種間的親緣關係。經人工餵予菌液之方式,白邊大葉蟬之成功獲菌率高達 80%,且獲菌試驗第 5 及第 8 天為葉蟬蜜露帶菌之蟲數高峰。在群體傳播試驗中,於金香及黑后葡萄罹病株上經 24 小時獲菌時間處理之白邊大葉蟬獲菌率分別達 49.56 及 54.67%。移至不同品系健康葡萄植株,進行 3~7 天之接種傳菌試驗。結果顯示,於金香病株上獲菌傳至金香和黑后健株之傳菌率為 17.27 和 15.45%;而黑后病株傳至金香和黑后則為 28.15 和 28.52%;單隻試驗中,葉蟬於金香病株之獲菌率 83.33%,將其病原菌傳至金香健株之傳菌率為 13.33%。自田間金香罹病株分離之菌株 GMb、及黑后分離之菌株 BQa 和 BQ7f,以及由藥毒所提供採自巨峰病株之 GV148 等4個菌株經 16S rRNA 及 16S-23S ITS 區段之親緣關係比對,其間有 99~100% 相同度;而與美國之X. fastidiosa間有 99~100% 之相同度。然而與柑桔、咖啡、桃及梅等植物之X. fastidiosa菌株在 16S rRNA 及 16S-23S ITS 區段間各有 98~99% 及 99~100% 之相同度。本研究證實白邊大葉蟬成蟲可自感染 PD 之葡萄病株獲菌後於體內保菌 18 天以上,並達到 15~28% 之傳菌率,在黑后葡萄植株上獲菌且傳播效率高於金香。目 次 中文摘要 I Abstract III 圖 次 VI 表 次 VII 前 言 1 材料與方法 3 一、 供試昆蟲 3 二、 供試植物 3 三、 Xylella fastidiosa 菌種來源、分離及培養 4 四、 不同品系葡萄植株之人工接種 4 五、 PCR 測定檢體中 X. fastidiosa 之靈敏度測試 5 六、 白邊大葉蟬之獲菌處理及蟲體保菌期測試 8 七、 白邊大葉蟬之傳播試驗 9 八、 田間葡萄病株之分子鑑定 10 九、 數據分析 11 結 果 13 一、 X. fastidiosa培養及形態特徵 13 二、 菌株活性檢測 13 三、 蜜露、蟲體及植物檢體中 X. fastidiosa 之靈敏度測試 13 四、 白邊大葉蟬之獲菌及保菌試驗 14 五、 白邊大葉蟬之傳播試驗 15 六、 本土葡萄X. fastidiosa 菌株之親緣關係鑑定 17 討 論 19 參考文獻 24 圖表 3

Topics: 葡萄皮爾斯病, Xylella fastidiosa, 白邊大葉蟬, 親緣分析, Pierce’s disease, Xylella fastidiosa, Kolla paulula, Phylogenetic analyses
Year: 2017
OAI identifier: oai:ir.lib.nchu.edu.tw:123456789/85981
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