Identification of Three Aulacaspis Species (Hemiptera: Coccoidea: Diaspididae) by Rapid Molecular Diagnostic Technique for Quarantine Application in Taiwan

用快速分子診斷技術鑑定台灣三種白輪盾介殼蟲（Aulacaspis spp.）（半翅目：介殼蟲總科：盾介殼蟲科）

邱一中(Yi-Chung Chiu) ； 陳淑佩(Shu-Pei Chen) ； 吳文哲(Wen-Jer Wu) ； 石正人(Cheng-Jen Shih)

《植物保護學會會刊》 55卷4期 (2013/12) Pp. 117-128

https://doi.org/10.6715/PPB.201312_55(4).0002

摘要

介殼蟲蟲體微小且外形相似，幼體時期特徵不明顯，在鑑定上不易確定，造成在檢疫工作上困難重重。本研究利用基因組核酸標誌（genomic DNA markers），以分子診斷技術鑑別三種白輪盾介殼蟲屬（Aulacaspis spp.）之介殼蟲，分別為蘇鐵白輪盾介殼蟲（A. yasumatsui Takagi）、月橘白輪盾介殼蟲（A. murrayae Takahashi）和樟白輪盾介殼蟲（A. yabunikkei Kuwana）。以單隻標本DNA進行聚合酶連鎖反應－限制酶片段長度多態型（polymerase chain reaction-restriction fragment length polymorphism, PCR-RFLP）的結果分析，利用PCR引子對（ITS5和ITS4）增幅rDNA之ITS區段，蘇鐵白輪盾介殼蟲增幅出單一片段，大小約0.6 kb；月橘白輪盾介殼蟲和樟白輪盾介殼蟲則增幅出二片段，大小約0.6 kb和0.67 kb。利用TaqI、MspI和CfoI核酸內切酶進行ITS/rDNA PCR產物酶切，所得圖譜可容易區別三種普遍分布且外型進似的白輪盾介殼蟲。

關鍵字

白輪盾介殼蟲 ； 分子鑑定 ； 核醣體去氧核醣核酸 ； 轉錄區內空白段 ； 限制性片段長度多態型分析

並列摘要

Scale insects are small and similar in morphology, and the identification of species especially in the immature stages is difficult. In this study, a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was developed to identify three Aulacaspis species, A. murrayae, A. yabunikkei and A. yasumatsui. Primers ITS5 and ITS4 were used to amplify the internal transcribed spacer (ITS) regions of the ribosomal DNA (rDNA). The genomic DNA isolated from A. yasumatsui yielded a single fragment after PCR amplification. The fragment size was about 0.6 kb. The other species, A. murrayae and A. yabunikkei, gave two PCR products. The sizes of the two fragments were about 0.6 kb and 0.67 kb. These three Aulacaspis species could be easily discriminated according to the species specific patterns by digesting the PCR products with endonucleases TaqI, MspI and CfoI.

並列關鍵字

Aulacaspis spp. ； molecular identification ； nuclear ribosomal DNA ； internal transcribed spacer (ITS) ； PCR-RFLP