運用隨機增幅多型性DNA聚合酶連鎖反應技術區別埃及斑蚊（Diptera: Culicidae）腹部背板白鱗分佈的差異

Use of the Random Amplified Polymorphic DNA Polymerase Chain Reaction (RAPD-PCR) to Distinguish Variations of Aedes aegypti based on Abdominal Tergal Scales

蘇兪丞(Yu-Cheng Su) ； 徐爾烈(Err-Lieh Hsu) ； 何兆美(Chau-Mei Ho)

《台灣昆蟲》 23卷3期 (2003/09) Pp. vi+225-237

https://doi.org/10.6661/TESFE.2003021

摘要

埃及斑蚊（Aedes aegypti L.）成蟲腹部背板鱗片的分佈具有明顯的多樣性，依照白色鱗片在腹部各節的背板從基部往後延伸的情形可分為三型：模式型、深色型及淺色型。以CKM分級系統共可分為8個級別，即CKM0至CKM7。屬於埃及斑蚊CKM0級的個體是深色型的埃及斑蚊，僅在非洲地區和印度洋島嶼之Aedes aegypti fomorsus看到。台灣地區的則屬於模式型的埃及斑蚊（CKM1）和淡色型（CKM2-7）共同混雜在一起的族群。本研究以相同CKM級別之親代在實驗室内進行交配，由產下的後代中挑選出與親代同一性狀的子代繼續交尾繁衍，直至篩選出所有的子代個體皆為親代的CKM級別之品系。再萃取此一純系後代的genomic DNA，以10個具有解析能力的引子（primer）進行隨機擴增多型的核酸－聚合酶連鎖反應（RAPD-PCR）。根據兩兩比對各種CKM級別的埃及斑蚊品系的核酸擴增的出來的多型性的DNA片段之相似度，可以明顯地將CKM1-7級的埃及斑蚊品系區分成二群：一群為由淡色型埃及斑蚊個體所組成，另一群為模式型的埃及斑蚊。本實驗証明此二型的埃及斑蚊在自然界雖有部分重疊的生態席位（niche），但可利用成蟲腹節背板的白色鱗片之分布情形和RAPD-PCR的方法區分出來。

關鍵字

埃及斑蚊 ； 腹部背板鱗片 ； CKM ； 隨機增幅DNA多型性 ； 種内變異

並列摘要

There is much variation in the distribution of white scaling on the abdominal tergites of Aedes aegypti. The C. K. Meek's (CKM) classification system was used to identify and rank eight classes in the abdominal scale patterns, but only seven classes were found in Taiwan, including CKM1 which was generalized as the type form of Aedes aegypti and CKM2 to CKM7 which were concluded to be as the pale form. Pure CKM strains were selected by single-pair matings, and genomic DNA of these mosquitoes was extracted in order to detect the variations in RAPD patterns. The RAPD fragments profiles of seven strains with different scale pattern were analyzed and genetic similarities were estimated between each two strains using two different statistic methods. The UPGMA dedrogram method illustrated the relatedness of these strains, and both statistical methods assorted these strains into two major groups: the type form and the pale form. But the genetic distance between each pair of strains differed using these two methods. These results indicate that the RAPD-PCR technique is sufficiently sensitive for studies of mosquito molecular taxonomy, especially at intra-species level.

並列關鍵字

Aedes aegypti ； abdominal tergal scale ； CKM ； random amplified DNA polymorphism ； intra-specific variation