Physiological and Molecular Characterizations of Banana finger-tip Rot and Onion Decay Pathogens in Taiwan

台灣香蕉果指尖腐病原菌與洋蔥腐敗病原菌之生理及分子特性分析

李永安(Yung-An Lee) ； 詹志文(Chih-Wen Chan) ； 趙治平(Chih-Ping Chao)

《植物病理學會刊》 15卷2期 (2006/06) Pp. 117-124

https://doi.org/10.6649/PPB.200606_15(2).0006

摘要

香蕉果指尖腐病原菌具有的生理生化特性與洋蔥腐敗病原菌極為相似，不過洋蔥腐敗病原菌在LA洋菜培養基上呈黃色且無法在42℃下生長。利用recA PCR-RFLP分析結果，發現香蕉果指尖腐病原菌的所有測試菌株均屬Burkholderia cepacta complex中的genomovar Ⅲ，而洋蔥腐敗病原菌的菌株則屬genomovar Ⅰ，由此結果可知香蕉果指尖腐病原菌應為B. cenocepacia，而洋蔥腐敗病原菌為B. Cepacia，因此香蕉果指尖腐病原菌與洋蔥腐敗病原菌為兩種不同的病原菌。此外，本報告亦研發出檢測及鑑定此兩種病原菌的PCR及PCR-RFLP方法。在bcscV-PCR 測試，只有香蕉果指尖腐病原菌可得正反應，而洋蔥腐敗病原菌則為負反應，並且bcscV探針只會與香蕉果指尖腐病原菌產生雜合反應，而不會與洋蔥腐敗病原菌產生反應。另一個uvrB-PCR檢測，則可對香蕉果指尖腐病原菌及洋蔥腐敗病原菌擴增出995-bp長的DNA片段，並且發現洋蔥腐敗病原菌的uvrB DNA片段中具有MluI限制酵素切點，因此可藉以研發出PCR-RFLP方法，以區分香蕉果指尖腐病原菌及洋蔥腐敗病原菌。其他測試的植物病原細菌在bcscv及uvrB-PCR測試中，均無DNA片段被擴增出來。因此本報告所研發出的bcscV及uvrB-PCR與PCR-RFLP的測試方法，將可在生態研究中，用於檢測及鑑定B. cenocepacia及B. cepacia兩種病原。

並列摘要

Banana finger-tip rot pathogen had biochemical and physiological characteristics very similar to those of onion decay pathogen, but the isolates of onion decay pathogen were yellow-pigmented and could not grow at 42℃ in LA agar medium. The recA PCR-RFLP analyses indicated that all tested isolates of banana finger-tip rot pathogen belonged to genomovar Ⅲ, and that isolates of onion decay pathogen belonged to genomovar I of Burkholderia cepacia complex, which were named as B. cenocepacia and B. cepacia, respectively. Accordingly, banana finger-tip rot and onion decay in Taiwan are caused by two separate pathogens. The recA PCR-RFLP tests further showed that genetic variability of the B. cenocepacia isolates, which contained recA ⅢA and ⅢB lineages. In addition, PCR and PCR-RFLP assays were developed in this study for the detection and identification of these two pathogens. In bcscV PCR assay, all tested isolates of banana finger-tip rot pathogen were PCR positive but the isolates of onion decay pathogen were negative for bcscV. The bcscV probe hybridized only to B. cenocepacia but not to B. cepacia. The other uvrB PCR test amplified a 995-bp DNA fragment for all isolates of B. cenocepacia and B. cepacia tested. In addition, discovery of a unique Miul restriction site of uvrB of onion decay pathogen led to the development of a PCR-RFLP assay able to discriminate onion decay pathogen isolates from banana finger-tip rot pathogens. No amplification was observed with other plant pathogenic bacteria in the bcscV and uvrB PCR assays. The bcscV and uvrB PCR and PCR-RFLP assays developed in this study will be highly effective for the detection and identification of B. cenocepacia and B. cepacia in the epidemiological studies.

並列關鍵字

Banana finger-tip rot ； Burkholderia cenocepacia ； B. cepacia ； bcscV ； onion decay ； uvrB.