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首页> 外文期刊>Journal of Medical Entomology >Ribosomal DNA ITS2 sequences differentiate six species in the Anopheles crucians complex (Diptera : Culicidae)
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Ribosomal DNA ITS2 sequences differentiate six species in the Anopheles crucians complex (Diptera : Culicidae)

机译：核糖体DNA ITS2序列区分了按蚊cru（Diptera：Culicidae）中的六个物种

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Anopheles crucians Wiedemann (sensu lato) was investigated for the presence of cryptic species using rDNA ITS2 sequences. This complex of species presently contains the named species An. crucians, An. bradleyi King, and An. georgianus King. Adult female mosquitoes were collected at 28 sites in Alabama, Florida, Georgia, North Carolina, Mississippi, and Louisiana, resulting in 245 progeny broods. Species were identified using preliminary morphological characters, and the internal transcribed spacer two (ITS2) was amplified from all broods. The result was five distinct sizes of amplification product, and based on morphological characters, one of the size classes was suspected to consist of two species. All six putative species were then sequenced: five directly, and the sixth, because of extreme intragenomic (each individual with many variants) size variability, cloned. The ITS2 sequences were markedly distinct for all six species. Species designations and ITS2 sequence lengths (base pairs in parentheses) were A (461), B (1,000+), C (204), D (293), E (195), and An. bradleyi (208). Species B showed both large intraspecific and intragenomic sequence variability and is distinguished by having the longest ITS2 found so far in an Anopheles. Based on these data, we found that all species could be identified with polymerase chain reaction (PCR) using a mixture of four primers in a single reaction. Members of this complex were often found in sympatry, with the adults of five species collected at a single site in central Florida.
机译：使用rDNA ITS2序列研究了cru按蚊Wiedemann（sensu lato）是否存在隐性物种。该物种复合体当前包含命名物种An。 ，安。 bradleyi国王和安。乔治亚王。在阿拉巴马州，佛罗里达州，乔治亚州，北卡罗来纳州，密西西比州和路易斯安那州的28个地点收集了成年雌性蚊子，形成了245个子代亲体。使用初步的形态学特征鉴定物种，并从所有亲代扩增内部转录的间隔区二（ITS2）。结果是扩增产物的五个不同大小，并且根据形态学特征，其中一个大小类别被怀疑由两个物种组成。然后对所有六个推定物种进行测序：直接克隆了五个，第六个因为极高的内部基因组（每个个体都有许多变异）的大小变异而被克隆。在所有六个物种中，ITS2序列均明显不同。种类名称和ITS2序列长度（括号中的碱基对）为A（461），B（1,000 +），C（204），D（293），E（195）和An。 bradleyi（208）。物种B表现出较大的种内和基因组内序列变异性，并以迄今在按蚊中发现的最长的ITS2为特征。根据这些数据，我们发现可以在单个反应中使用四种引物的混合物，通过聚合酶链反应（PCR）识别所有物种。这种复合物的成员经常在交配处发现，在佛罗里达州中部的一个地点收集了五种物种的成虫。

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