RAPD-PCR在六種斑潛蠅（Liriomyza spp.）（雙翅目：潛蠅科）快速鑑定技術之應用

The Application of RAPD-PCR to Develop Rapid Diagnostic Technique for Identification of 6 Species of Liriomyza spp. (Dipter: Agromyzidae)

10.6660/TESFE.2000028

邱一中(Yi-Chung Chiu)；吳文哲(Wen- Jer Wu)；蕭旭蜂(Shiuh-Feng Shiao)；石正人(Cheng-Jen Shih)

斑潛蠅 ； 快速診斷鑑定技術 ； 聚合酶連鎖反應 ； DNA逢機增幅多態型 ； Liriomyza ； rapid diagnostic techniques ； identification ； polymerase chain reaction (PCR) ； random amplified polymorphic DNA (RAPD)

中華昆蟲

20卷4期（2000 / 12 / 01）

IV+293 - 309

繁體中文

斑潛蠅蠅(Liriomyza)昆蟲為國際檢疫上的重要害蟲，但其成蟲及幼體時期(卵、幼蟲和蛹)均不易鑑定，在檢疫鑑定上面臨極大的困難。本研究之目的在嘗試應用分子生物技術，分析基因組DNA(genomic DNA)並建立分子診斷標誌，以解決斑潛蠅種類鑑定所面臨的難題。以DNA作為分子標誌的試驗中，利用100個逢機引子(random primer)分別進行聚合酶連鎖反應－DNA逢機增幅多態型分析(random amplified polymorphic DNA-polymerase chain reaction, RAPD-PCR)，結果共篩選出3個逢機引子UBC-70、UBC-71和UBC-96，所增幅出的核酸片段型態，可以清楚區別出供試的6種斑潛蠅，包括昭和草斑潛蠅(L. asterivora)、蕃茄斑潛蠅(L. bryoniae)、中華蔥斑潛蠅(L. chinensis)、南美斑潛蠅(L. huidobrensis)、蔬菜斑潛蠅(L. sativae)以及非洲菊斑潛蠅(L. trifolii)。同種斑潛蠅的不同個體間、不同生活區，或取食不同寄主作物的小族群(colony)間，均增幅出極相似的DNA片段長度多態型，具有高穩定性。然而，不同種的斑潛蠅間其RAPD-PCR多態型，則具有明顯的差異，可供作6種斑潛蠅種類的鑑定依據。

Leafminer flies (Liriomyza spp.) are important quarantined pests in international trade. However, the Liriomyza spp. are difficult to identify in immature stages (eggs, larvae, and pupae) as well as adult stage, so that many obstacles appeared in the international quarantine. This study attempts to develop the molecular technique of genomic DNA marker to identify 6 species of the genus Liriomyz. In the DNA analysis, we found DNA molecular marker to identify the Liriomyza species with similar morphological characters. In the random amplified polymorphic DNA (RAPD) test, we checked 100 random primers and only 3 primers (UBC-70, UBC71, and UBC-96) are available for distinguishing the 6 species of Liriomyza (L. asterivora, L. bryoniae, L. chinensis, L. huidobrensis, L. sativae, and L. trifolii), and there are no difference in the RAPD patterns among the different stages, sexes, strains of each species, but obvious differences between different species. According to this study, t he RAPD patterns is worth to be suggested for the identification of these species.