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利用PCR-RFLP技術進行縊管蚜屬（Rhopalosiphum）（常蚜科）昆蟲之分子鑑定
PCR-RFLP Technique for Molecular Identification of Rhopalosiphum (Aphididae)
摘要
近年來分子標記已被用於區分個體微小、外部形態相似不易辨別之物種，其檢測快速、操作容易的優點，相信對微小昆蟲的鑑定，也能提供莫大的幫助。依照形態特徵對於鑑定蚜蟲標本品質的要求相當高，再加上蚜蟲具有多態型，個體間外部形態變異程度大，造成蚜蟲種類鑑定時的困難。本研究藉由DNA分子標記來鑑定目前分布在台灣的縊管蚜屬（Rhopalosiphum）之四個種類：玉米蚜（R. maidis）、睡蓮蚜（R. nymphaeae）、稻麥蚜（R. padi）及紅腹根蚜（R. rufiabdominalis）。首先增幅蚜蟲核DNA之延伸因子1α（elongation factor 1α, EF 1α）以及蚜蟲體內的原生共生菌（primary symbionts）Buchnera sp.所攜帶之質體的基因片段pLeuC，再配合限制酶片段長度多態型（restriction fragment length polymorphism, RFLP）分析，結果以限制酶NlaIII和Eam1104I進行EF 1α增幅片段酶切，以及限制酶DraI進行pLeuC增幅片段的酶切，在四種蚜蟲間可得不同的限制酶片段多態型圖譜，藉以鑑別種類。另外提供上述二個基因片段之核苷酸序列以為種類鑑定之用。最後再探討利用原生共生菌的DNA分子資料做為鑑別蚜蟲種類之可行性。
並列摘要
Aphids are insects with polymorphism. Species identification mostly depends on the morphological features of the adult. However, it is difficult to distinguish species at immature stages. Molecular markers may be helpful in solving this problem. Therefore, in this study, the elongation factor 1α (EF 1α) in the genome of aphids and the partial pLeuC gene of the endosymbionts (Buchnera sp.) in aphids were amplified by polymerase chain reaction (PCR) using specific primers. Then, the restriction fragment length polymorphism (RFLP) of the PCR products was obtained. These DNA fragments were different from each other in the four species of aphids, Rhopalosiphum maidis, R. nymphaeae, R. padi and R. rufiabdominalis. Moreover, the DNA pattern of PCR-RFLP was also stable in immature stages, as those found in both adults. These results indicate that the DNA markers of PCR-RFLP from both host insects and their endosymbionts may be useful in species identification.