Multiplex-PCR及PCR-RFLP於檢疫害蟲西方花薊馬的鑑定

Molecular Identification of Multiplex-PCR and PCR-RFLP for the Quarantine Pest, Frankliniella occidentalis (Pergande)

林峻賢(Jgun Sian Lin) ； 葉文斌(Wen-Bin Yeh) ； 王清玲(Chin-Ling Wang)

《台灣昆蟲》 23卷4期 (2003/12) Pp. viii+353-366

https://doi.org/10.6661/TESFE.2003031

摘要

西方花薊馬（Frankliniella occidentalis（Pergande），簡稱Fo）為世界性危害嚴重的農業害蟲，尚未在台灣的田野發現，被列為檢疫害蟲；然而在進口的農作產品上，卻常檢測出此害蟲。因此本研究即針對其無法明確鑑定的若蟲階段，應用分子形質，以期達到鑑定的目標；除西方花薊馬之外，採用進口作物上最常檢測到的蔥薊馬（Thrips tabaci Lindeman，簡稱Tt）為比對物種。分析的分子形質包括28S rDNA及16S rDNA；其中28S rDNA主要的複製長度約689個鹼基，Fo及Tt分別有8及14隻個體，未發現種內變異，兩種間差異為12.3%；16S rDNA複製長度約300 bp，Fo及Tt分別有7及11隻個體，蔥薊馬種內最大變異為3個鹼基位，兩種間差異為17.5%。應用已知之序列採用專一性引子及酵素切位，進行multiplex-PCR及PCR-RFLP兩分析方法，分別針對單一及混合之害蟲樣品，進行鑑定之分析運用；結果顯示兩種方法均可如期達到鑑定的目的。

關鍵字

西方花薊馬 ； 蔥薊馬 ； multiplex-PCR ； PCR-RFLP ； 16S rDNA ； 28S rDNA

並列摘要

The polyphagous insect, Frankliniella occidentalis (Pergande) (Fo), causes serious damage to economic plants in the world, whereas it is not found on the list of quarantine pests in Taiwan. However, the pest is occasionally found in imported agriculture products. In this study, molecular characters were used to identify the nymph stage of this quarantine pest; the onion thrips, Thrips tabaci Lindeman, found mostly in imported agriculture products was used as a reference sample. Molecular markers including both 28S rDNA and 16S rDNA sequences were analyzed. For 28S rDNA, 22 individuals of these two species were used, and intraspecific sequence variation were not found; interspecific variation was 12.3% in 689 bp. For 16S rDNA, 18 individuals of these two species were used, and intraspecific sequence variations were nearly zero, while interspecific variation was 17.5% between these two species. Specific primers designed from 28S rDNA and restriction enzyme from 16S rDNA were respectively applied for multiplex- PCR and PCR-RFLP to identify F. occidntalis. Results showed that application of the two molecular markers was useful in identifying this quarantine pest.

並列關鍵字

Frankliniella occidentalis ； Thrips tabaci ； multiplex-PCR ； PCR-RFLP ； 16S rDNA ； 28S rDNA