利用PCR快速鑑定銀葉粉蝨（Bemisia argentifolii）（半翅目：粉蝨科）

Development of Rapid Identification of Bemisia argentifolii (Hemiptera: Aleyrodidae) by PCR

王重雄(Chung-Hsiung Wang) ； 柯俊成(Chiun-Cheng Ko) ； 劉中琪(Chung-Chi Liu) ； 陳秋男(Chiou-Nan Chen)

《台灣昆蟲》 24卷3期 (2004/09) Pp. iv+229-246

https://doi.org/10.6661/TESFE.2004020

摘要

本研究利用逢機增幅核酸多形性－聚合酶連鎖反應（RAPD-PCR）方法鑑定形態上不易鑑別之煙草粉蝨（Bemisia tabaci）A及B生物小種。自27個逢機引子中篩選出對於兩生物小種具有鑑別力之4種引子（OPA03, 91167, 91074, F12）。並依據所獲得之RAPD-PCR圖譜，選殖出具有生物小種間之特異性片段，經定序分析後，再設計出專一性引子組Baf/Bar，此引子組可在B生物小種之DNA中增幅出一段約500bp的片段，而在其他生物小種以及不同種類之粉蝨則不會產生任何產物，顯示此專一性引子組適用於煙草粉蝨B生物小種之鑑定。本研究為首先建立台灣地區煙草粉蝨之分子標示鑑定法，並著手煙草粉蝨之全面採集調查，進而提供後續研究之依據。

關鍵字

逢機增幅核酸多形性 ； 專一性引子組 ； 鑑定技術 ； 煙草粉蝨 ； 銀葉粉蝨

並列摘要

Diagnosis between Bemisia tabaci biotypes A and B was first carried out in this investigation using RAPD-PCR analysis. The four primers, OPA01, F12, 91167, and 91074, from 27 random primers were selected for specific discrimination of biotypes A and B. The specific amplicon of biotype B was cloned and sequenced by RAPD-PCR using primer 91074. A primer set, Baf/Bar, was designed based on the nucleotide sequence of this amplicon. An amplicon (of about 500 bp) offered yields with Baf/Bar only with the genomic DNA template by PCR, while none was found with other genomic DNA biotypes. This result shows that the specific primer set can be used to discriminate biotype B from other biotypes of B. tabaci. This is initial work on a molecular diagnostic approach for B. tabaci in Taiwan. Using results from this investigation, we will study more populations of B. tabaci from Taiwan using additional molecular studies.

並列關鍵字

RAPD ； specific primer set ； diagnostic techniques ； Bemisia tabaci ； B. argentifolii