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Please use this identifier to cite or link to this item: http://ir.hwai.edu.tw:8080/ir/handle/310996100Q/27

Title: 蝴蝶蘭細菌性病原ITS序列分析及其檢測之應用
The analysis of internal transcribed spacer and its applications on the detection of orchid bacterial diseases
Authors: 勾國豪
gou guo hau
Contributors: 生物科技研究所
Keywords: 蝴蝶蘭
pectobacterial ITS orchid
Date: 2006
Issue Date: 2008-12-25 10:48:33 (UTC+8)
Abstract: 造成台灣蝴蝶蘭細菌性病害的主要病原有三種,分別為 Pectobacterium chrysanthemi、 Acidovorax avenae subsp. cattleyae以及 Burkholderia gladioli。本研究利用自行建置之ITS資料庫,進行多序列比對分析。可對A.avenae subsp. cattleyae以及 B.gladioli分別設計出具專一性之AAC-ITS引子對及BG-ITS引子對,所有供試菌株經PCR電泳分析皆可產生專一性條帶。在P.chrysanthemi的鑑定係利用P.chrysanthemi所特有之藍色素相關基因之序列來設計專一性引子對,計有pecS、pecM及idgA等3組引子對,可專一性偵測到P.chrysanthemi。本研究中所設計5組引子對AAC-ITS、BG-ITS、pecS、pecM及idgA,測試之結果顯示100﹪之專一性。在敏感度分析上,idgA引子對偵測P.chrysanthemi之敏感度分析,敏感度為1.2x104 cells。BG-ITS引子對偵測B.gladioli之敏感度為1x102 cells。AAC-ITS引子對偵測A.avenae subsp. cattleyae之敏感度為1x102 cells。在多引子對之分析時以各引子對可精確地檢測出各病原菌的存在。
There are three species of bacterial can infect the local Phalaenopsis, three of them are Pectobacterium chrysanthemi, Acidovorax avenae subsp. cattleyae and Burkholderia gladioli. In this study, we use the in-house bacterial internal transcribed spacer database to analysis the sequence between the bacterial that can infect the Phalaenopsi. We design primers AAC-ITS and BG-ITS based on the results of the analysis of Acidovorax avenae subsp. cattleyae and Burkholderia gladioli ITS sequence, and the primers pecS、pecM and idgA designed by indigoidine related gene, to detect the Pectobacterium chrysanthemi. The primers we use in this study reveals perfect results to their on response bacterial. In the sensitivity test, primer idgA can detect the Pectobacterium chrysanthemi with 1.2x104 cells, primer AAC-ITS can detect Acidovorax avenae subsp. cattleyae with1x102 cells and primer BG-ITS can detect Burkholderia gladioli with 1.2x104 cells. The primers designed in this study can obviously and accurately detect the bacterial that can infect the local Phalaenopsis.
Appears in Collections:[醫學檢驗生物技術系碩士班] 博碩士論文

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