運用逢機擴增多形性核酸分析長毛捕植蟎(Amblyseius longispinosus)及溫氏捕植蟎(A. womersleyi)(蟎蜱亞綱；捕植蟎科)的遺傳變異
Other Title
Genetic Diversity of Amblyseius longispinosus and A. womersleyi (Acari: Phytoseiidae) Using RAPD Analysis
Journal
中華昆蟲
Journal Volume
20
Journal Issue
4
Pages
335-345
Date Issued
2000-12
Author(s)
葉文斌
何佳霖
許祖法
何琦琛
Wen-Bin Yeh
Chai-Lien Ho
Cho-Fat Hui
Chyi-Chen Ho
Abstract
長毛捕植蟎(Amblyseius longispinosus, AL)及溫氏捕植蟎(A. womersleyi, AW)被廣用於葉蟎之生物防治研究。由於學名曾被混用，且AL及AW的辨識特徵是L8背毛，但有中間型，因此有學者認為AL與AW是同一物種。本報告運用DNA指紋－逢機擴增多形性核酸(random amplified polymorphic DNA)技術，探討長毛捕植蟎與溫氏捕植蟎遺傳物質的歧異度。包括長毛捕植蟎與溫氏捕植蟎的卵、若蟲及成蟲6種樣本。各項反應條件的測試結果顯示，模板DNA的純度、量及其保存液，對反應結果均會有影響。以60個逢機序列的引子分別進行RAPD-PCR反應，再從電泳圖譜分析兩種捕植蟎的遺傳歧異度的結果顯示，AL或AW的卵、若蟲及成蟲複製的的電泳圖譜彼此類似，表示實驗條件的選擇合適。為求客觀，以兩種不同標準計算電泳圖譜上的核酸條帶數，分別得到8.3%與14.9%之遺傳相似性，可見此兩種捕植蟎的遺傳物質已有很大的歧異。其中，引子OPH-17及OPH-18兩個引子所擴增的核酸條帶簡單，可以簡易用於鑑定AL與AW。
Predatory mites of Amblyseius longispinosus and A. womersleyi are used to control spider mites. It has been considered that A. womerslei is a synonym of A. longispinosus since their identification character (dorsal setae L8) with intermediate length has been found. Random amplified polymorphic DNA (RAPD) was used to analysis the genetic diversity of these two morphologically similar mites. Genomic DNAs were extracted separately from egg, nymph, and adult either from A. longispinosus or A. womersleyi, respectively. The optimal reaction condition including the buffer and DNA template were determined. Sixty random primers were used to perform the amplification in a constant condition. The results of PCR amplification showed that the DNA template from egg, nymph, or adult of the same species gave a similar pattern. Whereas, the genetic similarity between A. longispinosus and A. womersleyi were very low either from rough (14.9%) or serious (8.3%) calculation. It implied that there was a great divergence between these 2 mites. Furthermore, the OPH-17 and OPH-18 primers were selected, they provided a clearly different pattern between A. longispinosus and A. womersleyi.
Subjects
捕植蟎
逢機擴增多形性核酸
遺傳變異
predatory mite
rapd
genetic diversity
phytoseiidae
Publisher
台灣昆蟲學會
Type
journal article