Multiplex PCR for the Molecular Identification of Economically Important Thrips

複合性PCR於經濟重要性薊馬的分子鑑定

10.6661/TESFE.2014017

葉文斌(Wen-Bin Yeh)；吳昕縈(Cindy Sin-Ying Wu)；曾美容(Mei-Jung Tseng)；張念台(Niann-Tai Chang)；陳文彬(Wen-Ping Chen)

薊馬 ； 核醣體第二區間 ； 專一性引子 ； 內控制引子 ； thrips ； ITS2 ； specific primer ； internal positive control

台灣昆蟲

34卷3-4期（2015 / 03 / 01）

211 - 225

英文

薊馬是世界上許多農作的重要害蟲，常因其體形微小及形態上的高度相似而難以鑑定，單一步驟複合性PCR的分子方法遂被應用於薊馬的鑑定。本研究分析14種農作上重要性薊馬的核醣體第二區間（ITS2）序列，種內變異小於3%遠低於種間14%以上的變異，使得此ITS2序列非常適合開發專一性引子用以鑑定這些薊馬。這些專一性引子與具內控制組特性的廣效性引子混合，進行23種薊馬的複合性PCR檢測，均顯示出其專一性及穩定性。總結來說，薊馬物種核醣體第二區間的高變異特性，可開發專一性引子用於複合性PCR檢測，提供了一個可靠、方便、快速的鑑定方法，區辨這些重要薊馬害蟲。

Thrips are major pests of many agricultural crops all over the world. However, their minute size and high degree of morphological similarity often makes their identification difficult. Molecular methods, such as single-step and multiplex PCR, deserve more consideration for their identification. In this study, ITS2 sequences of 14 agronomically important thrips were analyzed. Low intraspecific distances (i.e., ＜3%) in contrast with high interspecific distances (i.e., ＞14%) makes ITS2 sequences an ideal marker for designing species-specific primers for thrips identification. The specificity and stability of these species-specific primers, mixed with universal paired primers as an internal positive control, were tested and verified using multiplex PCR on 23 thrips species. In conclusion, the highly variable features of ITS2 sequences among thrips species are useful in the development of species-specific primers for multiplex PCR. It provides a detection tool that allows for rapid, reliable and convenient species discrimination of the important insect pest.