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目的利用形态特征与分子特征相结合对我国海南省按蚊进行种类鉴定。方法研究样本为海南省4个不同地理环境所采集的按蚊成虫和幼虫,依据形态特征鉴定成虫后,测定和分析部分成虫和幼虫的rDNA?ITS2和28S?D3序列以进行分子鉴定。结果对采集的336只按蚊成虫进行形态鉴定,结果显示总共分为9种,其中迷走按蚊所占比例最高,为81.55%。测序获得成虫和幼虫的ITS2序列37条,D3序列41条,Blast对比的结果显示,形态鉴定的成虫中,1只乌头按蚊错定为微小按蚊;分子鉴定的幼虫包括中华按蚊、迷走按蚊、腹簇按蚊和未定名种(与圣代克按蚊和浅色按蚊同源性高);序列分析发现在ITS2序列中迷走按蚊和微小按蚊分别存在1处(G/A)和2处(A/T)单碱基双峰,D3序列中迷走按蚊和腹簇按蚊存在3处(G/A)和1处(G/A)单碱基双峰。结论应用形态特征结合分子特征鉴定按蚊的可靠性和实用性强;分子特征提示迷走按蚊、微小按蚊和腹簇按蚊等处于隐种的分化中。
Objective To identify the species of Anopheles sinensis in Hainan Province by combining morphological and molecular characteristics. Methods The samples were collected from 4 adult mosquitoes and larvae of Anopheles sinensis in four different geographical environments of Hainan Province. According to their morphological characteristics, adults and larvae were identified and analyzed for rDNA ITS ITS2 and 28S D3 D3 sequences for molecular identification. Results The morphological identification of 336 Anopheles mosquitoes collected showed that there were 9 species in total, with the highest proportion of Anopheles stephensi (81.55%). 37 ITS2 sequences and 41 D3 sequences were obtained from adults and larvae. The results of Blast comparison showed that one of the morphologically identified adults was falsely identified as Anopheles minimus. Molecularly identified larvae included Anopheles sinensis, Anopheles stephensi, Anopheles stephensi and unknown species (with high homology with Anopheles stephensi and Anopheles stephensi). Sequence analysis showed that there was one G / A) and two (A / T) single base doublets. There were three single base doublets (G / A) and one (G / A) double base in the D3 sequence. Conclusion The morphological characteristics and molecular characteristics of Anopheles sinensis are reliable and practical. The molecular characteristics suggest that Anopheles stephensi, Anopheles minimus and Anopheles guttata are in the hidden differentiation.