Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/31918
標題: 台灣木瓜黑腐病菌Erwinia 菌株之多基因序列分析
Multilocus sequence analysis (MLSA) of Erwinia strains from black rot disease of papaya in Taiwan
作者: 吳瓔旂
Wu, Ying-Chi
關鍵字: 木瓜黑腐病
Black rot of papaya
多基因序列分析
Multilocus sequence analysis
出版社: 植物病理學系所
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摘要: Papaya (Carica papaya) is an economically important crop in Taiwan. In recent years, a bacterial disease, black rot of papaya, seriously occurred in central and southern Taiwan. Based on physiological, biochemical and pathogenicity tests, the causal agent was identified as Erwinia sp. The 16S rDNA sequences of these Erwinia sp. (EP) strains were 97.9% similarity to Erwinia papayae. In order to clarify the taxonomic status of these EP isolates from Taiwan, multilocus sequence analysis (MLSA) and biochemical tests were used to identify the EP strains from black rot of papaya in this study. In the results of biochemical tests, the EP strains could utilize D-mannitol and L-arabinose, but did not produce blue pigment on KB or YDC medium. However, Erwinia papayae produces blue pigment on KB madium and utilizes L-arabinose but not D-mannitol. E. mallotivora does not produce blue pigment and it utilizes D-mannitol but not L-arabinose. The results of physiological and biochemical tests can not effectively identify the taxonomic status of EP strains. Phylogenetic trees were constructed based on the sequences of the four housekeeping genes, gyrB, rpoD, atpD and infB. The four gene sequence of EP strains alignment with E. papayae and E. mallotivora up to 99% identity. Neighbour-joining (NJ), maximum-parsimony (MP) and maximum-likelihood (ML) phylogenies of gyrB, rpoD, atpD and infB genes were compared. The EP strains formed a separate cluster from the E. papayae and E. mallotivora with bootstrap value of 100%. While the EP strains groups on the border of E. papayae with bootstrap value of 78-93 %. The results of this study revealed that the caused agent of black rot of papaya in Taiwan were E. papayae. The MLSA data suggested that EP strains could be a novel species, phylogenetically related to E. papayae.
木瓜為台灣具經濟重要性之水果,近年來,在台灣中南部地區發生由細菌引起之木瓜黑腐病。藉由生理生化及病原性接種測試,將病原菌鑑定為 Erwinia sp. (EP菌株)。其16S rDNA序列與E. papayae有97.9% 的相似度 (similarity)。為了鑑定在台灣引起木瓜黑腐病之病原菌,本研究以多基因分析法 (multilocus sequence analysis, MLSA ) 配合生化測試來釐清木瓜黑腐病菌在演化上的分類地位。在生理生化測試之結果,供試之EP菌株可利用D-甘露糖醇 (D-mannitol) 以及L-阿拉伯醣 (L-arabinose),但在KB及YDC培養基上不產生藍色色素。然而,Erwinia papayae在KB培養基上可產生藍色色素,可利用L-阿拉伯醣,但不利用D-甘露糖醇;而E. mallotivora在KB培養基上不產生藍色色素,可利用D-甘露糖醇,但不利用L-阿拉伯醣。因此,藉由生理生化測試無法有效鑑定EP菌株之分類地位。應用多基因序列分析法使用四個管家基因 (housekeeping genes),gyrB、rpoB、atpD 以及infB序列構築系統演化樹。結果顯示EP菌株四個基因在序列比對上與E. mallotivora及E. papayae達99% 相同度 (identity)。以neighbor-joining、minimum evolution以及maximun parsimony演算法所得之演化樹,EP菌株在演化上與E. mallotivora及E. papayae各別形成單一菌群 (monophyletic groups),其分支可信度 (bootstrap value) 為100%。而EP菌株之單一菌群,鄰近於E. papayae的單一菌群,其分支可信度為78-93%,顯示EP菌株在演化上與E. papayae較相近。因此,將引起台灣木瓜黑腐病之EP菌株暫定為E. papayae。而根據MLSA之結果顯示EP菌株可能為一新種,其親緣關係接近E. papayae。
URI: http://hdl.handle.net/11455/31918
其他識別: U0005-2506201316144000
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-2506201316144000
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