請用此 Handle URI 來引用此文件: http://hdl.handle.net/11455/31083
標題: 台灣中南部木瓜細菌性病原之鑑定及檢測
Identification and detection of bacterial pathogens infecting papaya in central and southern Taiwan
作者: 楊雯馨
Yang, Wen-Shin
關鍵字: black rot of papaya
木瓜黑腐病
detection
Pectobacterium cypripedii
Erwinia cyrpipedii
檢測
Pectobacterium cypripedii
Erwinia cypripedii
出版社: 植物病理學系所
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摘要: 近年來在台灣中南部發生嚴重之木瓜細菌病害,其病徵如木瓜黑腐病,罹病組織於切口處可見大量細菌湧出。本研究分別自屏東及彰化地區採回罹病之木瓜組織進行病原細菌分離純化,共獲得 36 個病原細菌菌株,經生理生化特性測試,其中 21 個菌株為 Erwinia sp.、10 個菌株為 Pseudomonas aeruginosa、5 個菌株為 Pectobacterium carotovorum subsp. carotovorum。Erwinia sp. 菌株之生理生化特性與 1980 年由木瓜所分離之 Pectobacterium cypripedii (原Erwinia cypripedii) 相似。利用 16S rDNA 序列分析顯示 Erwinia sp. 菌株與 Erwinia papayae 較為接近。利用 40 組隨機引子進行 RAPD (random amplified polymorphic DNA) 分析,結果顯示隨機引子 OPC5 可對 Erwinia sp. 增幅出具專一性之 DNA 片段 (約950 bp)。將此專一性的 DNA 片段選殖及進行核苷酸序列分析,並依其序列設計出對 Erwinia sp. 菌株具有專一性的引子對 C5-1R/C5-1L。以此引子對應用聚合酵素連鎖反應檢測 Erwinia sp. 菌株皆能增幅出 300 bp 之 DNA 專一性片段,而非標的細菌則無法增幅出此條帶,此引子對測試 Erwinia sp. 之 DNA 時,其靈敏度為 10 pg。此專一性引子對可應用於台灣木瓜黑腐病菌 Erwinia sp. 之鑑定與偵測之用。
A bacterial disease with symptoms as that of black rot on papaya, seriously occurred in central and southern Taiwan in recent years. Based on physiological, biochemical and pathogenicity tests, there were 36 strains of plant pathogenic bacteria isolated from diseased papayas from Pingtung and Changhua, including 21 strains of Erwinia sp., 5 strains of Pectobacterium carotovorum subsp. carotovorum and 10 strains of Pseudomonas aeruginosa. Physiological and biochemical assays revealed that the characteristics of the Erwinia sp. strains were similar to that of Pectobacterium cypripedii (Erwinia cypripedii) strain from papaya reported in 1980. Phylogenetic trees based on the sequences of 16S rDNA indicated that the Erwinia sp. strains isolated from papaya were more closely related to the Erwinia papayae than Pectobacterium cypripedii. Sixty different random primers were tested to amplify specific DNA fragments from strains of Erwinia sp. using random amplified polymorphic DNA (RAPD) analysis. A specific DNA fragment (about 950 bp in size) form Erwinia sp. strains was amplified by the random primer OPC 5. This specific DNA fragment was cloned and sequenced to design specific primer pair C5-1L/C5-1R. This primer pair could amplify a distinct band of 300 bp that was specific to strains of Erwinia sp. isolated from papaya by polymerase chain reaction (PCR). The minimum amount of DNA from Erwinia sp. that could be amplified by PCR was 10 pg. The results indicated that the primer pair C5-1L/C5-1R could be a useful tool for identification and detection of Erwinia sp. strains from black rot of papaya.
URI: http://hdl.handle.net/11455/31083
其他識別: U0005-2708200711291900
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-2708200711291900
顯示於類別:植物病理學系

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