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題名 | 柑橘潰瘍病菌pthA與pel基因對病徵之影響=Effect of PthA and Pel Genes in Symptom of Xanthomonas axonopodis pv. citri |
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作者 | 林信成; 曾國欽; 張翔; 張佑安; | 書刊名 | 健康與照顧科學學刊 |
卷期 | 1:2 2012.12[民101.12] |
頁次 | 頁165-178 |
分類號 | 435.33 |
關鍵詞 | 單點突變; 果膠分解酵素基因; 病徵; Xanthomonas axonopodis pv citri; Xac; Point mutagenesis; Pectolytic genes; Symptoms; |
語文 | 中文(Chinese) |
中文摘要 | 台灣柑橘潰傷病係由Xanthomonas axonopodis pv. Citri所引起,可將其分成A、Af、Ar及Ap等四種類型。為瞭解柑橘潰傷病菌pthA基因與果膠分解酵素基因,在柑橘潰傷病徵呈現所扮演之角色,將引起典型病徵之A類型柑橘潰傷病菌XW19菌株的pthA基因選殖出,轉入Af類型XW47基因之胺基酸序列,顯示WX47菌株之pthA蛋白,在第49、286、742及767位置上的胺基酸與XW19菌株之氨基酸有差異。利用單點突變技術,在此4個位置上構築帶有單一氨基酸突變之pthA蛋白菌株,結果顯示柑橘潰傷病菌pthA蛋白上第286位置由脯胺酸變為絲胺酸或由絲胺酸變為脯胺酸之單一胺基酸的改變,可影響潰傷病斑之形成。進行XW19及Ar類型菌株XW121果膠分解酵素基因之選殖與分析,結果顯示只有來自XW19菌株之pel1基因具有果膠分解能力,經序列分析pel1基因為一個含377個胺基酸所組成的pel1蛋白;而XW121菌株之pel1基因在3’端第350個胺基酸位置為一個終止碼,將XW19菌株之pel1基因轉入XW121菌株,則獲得之轉殖菌株在葡萄柚葉片能恢復引起具有水浸狀邊緣之潰傷病斑,顯示pel1基因與柑橘潰傷病斑水浸狀邊緣之形成有關。 |
英文摘要 | Xanthomonas axonopodis pv. citri causes citrus bacterial canker and is divided into A, Af, Ar and Ap types in Taiwan. In order to study the role of pthA and pel genes, a pthA homolog was cloned from A type strain XW19 and transferred into Af type strain XW47. The transforment of XW47 induced typical canker lesions on grapefruit leaves. Sequence analysis of pthA homolog from XW47, the amion acid residues located at positions 49, 286, 742 and 767 of PthA were different between XW47 and XW19. The PthA mutants with a single amino acid substitution at each of these four positions were constructed by site-directed mutagenesis. The results indicated that a single amino acid substitution from either serine to proline or proline to serine at position 286 of PthA can alter canker formation. The pel1 gene from XW19 and XW121 encoded a Pel1 protein of 377 amino acids. However, there was a stop codon located at the 350th amino acid residue of pel1 gene from XW121. When the pel1 gene from XW19 was transferred into XW121, the transformant exhibited pectate lyase activity and also induced water-soaked margins surrounding the lesions on grapefruit leaves. The results suggest that expression of the pel1 gene of Xac is associated with formation of the water-soaked margin of the bacterial canker lesion. |
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