林木重要根莖腐病原PCR快速檢測技術之建立 (2/3)The establishment of PCR rapid detection technology for major root and stem rot pathogens of forest trees(2/3)

林木根莖腐病害在發生初期不容易被發現，甚至有潛伏感染的現象，難以在發生初期進行防治工作，甚至在不知已感染的狀況之下，藉由造林或移植將病原傳播出去。本年度計畫目標在建立熱帶靈芝根腐病之快速分子檢測技術，本年度已完成熱帶靈芝子實體收集，樹舌靈芝、狹長孢靈芝、南方靈芝及韋伯靈芝等病原菌樣本純化分離培養及菌種保存，，各菌種分別利用通用性引子對ITS4/ITS5進行聚合酶鏈鎖增幅反應後將核酸產物定序，利用軟體比對找出南方靈芝序列之保守性片段設計三組專一性PCR引子對，經過專一性及靈敏度測試後，有一個組合得到良好增幅反應，可應用於日後熱帶靈芝根腐病的檢測鑑定。
Root and basal stem rot diseases are not easy to be found at the initial infection stage and some latent infection also develop, which cause the difficulty of disease control and may spread out the pathogen unintentionally by afforestation or transplantation. The project purpose
in this year is to establish rapid molecular detection technology for Ganoderma root and butt rot. The pure cultures and preservations of G.
tropicum, G. applanatum, G. boninense, Ganoderma austral and G. weberianum are accomplished. The five fungal DNA are subjected to PCR
reaction with the common primer set, ITS4/ITS5, and the products are sequenced respectively. Then the conservative fragments of G. tropicum are discovered by aid of alignment software to design 3 primer combinations. After specificity and sensitivity tests, 3 combinations
showed good amplification results and can be applied in the examination and determination of Ganoderma root and butt rot in the future.