蝴蝶蘭（Phalaenopsis amabilis）為相當重要的外銷花卉，卻常因病害而造成產值的損失，如軟腐菌(Erwinia chrysanthemi)造成的疾病即為ㄧ嚴重危害。利用化學方法進行防治固然有效，但卻會對環境帶來相當嚴重的傷害，因此，欲以分子生物的研究探討蝴蝶蘭受軟腐菌感染後所誘發的反應，以期進一步解決相關問題。本文中利用suppression subtractive hybridization (SSH)來尋找受軟腐菌感染後24小時有差異表現的基因，分析了170 個cDNA clones，進行序列比對結果共得到73個基因，將其以功能性作分類的結果顯示，與代謝相關基因佔36.45%，另有29.4%和逆境相關，12.35%與能量代謝有關。挑選其中12個基因做進一步的RT-PCR分析，結果發現此12個基因之表現量在蝴蝶蘭受到軟腐菌感染的48小時期間會上升。蝴蝶蘭PaCDPK1基因會受到低溫，物理性傷害和病原菌感染而誘發，且表現有PaCDPK1啟動子-GUS的阿拉伯芥轉殖株也會受低溫，物理性傷害和病原菌感染而偵測到GUS的表現。上述這些結果提供了對蝴蝶蘭在受到軟腐菌感染後其分子層次表現上的進一步了解。

Erwinia chrysanthemi is the causal agent of soft rot, a disease affecting the orchid, Phalaenopsis amabilis. In order to analyze mechanisms leading to compatible interactions between Erwinia and Phalaenopsis, early plant molecular events were investigated. Many defense mechanisms, including reactive oxygen species (ROS) accumulation and superoxide dismutase (SOD) induction were elicited. To identify the genes involved in the responses, a suppression subtractive hybridization strategy was used to isolate genes that were induced at 24 h after infection. A total of 170 clones was sequenced from the whole SSH library to generate 73 nonredundant ESTs. Among the genes with assigned function, 36.45% were associated with metabolism, 29.4% with defense and stress, and 12.35% with energy. cDNA encoding an enoyl-CoA reductase (ECR )of orchid was the most abundant representatives of plant genes, comprising 14.8% of the total clones. The data from the quantitative RT-PCR analysis of the expression patterns of 12 chosen genes show that all of their transcripts were up-regulated during 48 hr after Erwinia infection. Their sequences showed high homology to the chalcone synthase, short chain alcohol dehydrogenase-like protein, enoyl-CoA reductase, methylenetetrahydrofolate reductase, caffeic acid O-methyltransferase, putative PDR-like ABC transporter, glutathione transferase , calmodulin binding protein, acetyl-CoA carboxylase, pathogenesis-related protein 10, bZIP transcription factor and WRKY transcription factor-b. In addition, the PaCDPK1 gene was
transcriptionally activated in response to low temperature, wounding, and pathogen infection. To identify the regulatory role of the PaCDPK1 promoter, a construct containing the PaCDPK1 promoter fused to a β-glucuronidase (GUS) gene was transferred into Arabidopsis by Agrobacterium-mediated transformation. GUS staining revealed that PaCDPK1/GUS expression was induced by cold, wounding, and pathogen challenge in leaves and stems of transgenic Arabidopsis. Our results reported here provide a valuable information towards the understanding the orchid-Erwinia interaction at molecular level.

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