Background: Anopheles subpictus s.l. and Anopheles sundaicus s.l. are closely related species, each comprising of several sibling species. Ambiguities exist in the classification of these two nominal species and the specific status of members of An. subpictus complex. Identifying fixed molecular forms and mapping their spatial distribution will help in resolving the taxonomic ambiguities and understanding their relative epidemiological significance. Methods: DNA sequencing of Internal Transcribed Spacer-2 (ITS2), 28S-rDNA (D1-to-D3 domains) and cytochrome oxidase-II of morphologically identified specimens of two nominal species, An. subpictus s.l. and An. sundaicus s.l. collected from the Indian subcontinent, was performed and subjected to genetic distance and molecular phylogenetic analyses. Results: Molecular characterization of mosquitoes for rDNA revealed the presence of two molecular forms of An. sundaicus s.l. (identified as An. epiroticus s.s. and An. sundaicus D) and three molecular forms of An. subpictus s.l. (provisionally designated as Form A, B and C) in the Indian subcontinent. Phylogenetic analyses revealed two distinct clades: (i) subpictus clade, with a single molecular form of An. subpictus (Form A) prevalent in mainland India and Sri Lanka, and (ii) sundaicus clade, comprising of members of Sundaicus Complex., two molecular forms of An. subpictus s.l., (Form B and C) prevalent in coastal areas or islands, and molecular forms reported from Thailand and Indonesia. Based on the number of float-ridges on eggs, all An. subpictus molecular Form B were classified as Species B whereas majority (80%) of the molecular Form A were classified as sibling species C. Fixed intragenomic sequence variation in ITS2 with the presence of two haplotypes was found in molecular Form A throughout its distribution. Conclusion: A total of three molecular forms of An. subpictus s.l. and two molecular forms of An. sundaicus s.l. were recorded in the Indian subcontinent. Phylogenetically, two forms of An. subpictus s.l., (Form B and C) prevalent in coastal areas or islands in the Indian subcontinent and molecular forms reported from Southeast Asia are members of Sundaicus Complex. Molecular Form A of An. subpictus is distantly related to all other forms and deserve a distinct specific status. Presence of An. epiroticus in Indian territory is recorded for the first time.

中文翻译：

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印度次大陆上的按蚊Anpiceles subpictus sl和Sundaicus sl按蚊的分子形式

背景：亚种按蚊（Anopheles subpictus sl）和大按蚊（Sundaicus sl）是密切相关的物种，每个物种都由几个同胞物种组成。这两个标称物种的分类和An成员的具体地位存在歧义。亚群。鉴定固定分子形式并绘制其空间分布图将有助于解决分类学上的歧义，并有助于理解其相对流行病学意义。方法：对两个标称物种的形态学鉴定标本的内部转录间隔物2（ITS2），28S-rDNA（D1至D3结构域）和细胞色素氧化酶II进行DNA测序。subpictus sl和An。从印度次大陆收集的sundaicus sl进行了遗传距离和分子系统发育分析。结果：蚊子对rDNA的分子表征揭示了An的两种分子形式的存在。sundaicus sl（标识为An。epiroticus ss和An。sundaicus D）和An。sundaicus的三种分子形式。印度次大陆上的subpictus sl（临时指定为Form A，B和C）。系统发育分析揭示了两个不同的进化枝：（i）亚分子进化枝，具有单分子形式的An。亚种（A型）在印度大陆和斯里兰卡盛行，（ii）sundaicus进化枝，由Sundaicus Complex。的两种分子形式组成。subpictus sl（B型和C型）流行于沿海地区或岛屿，泰国和印度尼西亚报道了分子形式。根据鸡蛋上的浮脊数，全为 亚种的分子形式B被归类为物种B，而分子形式A的大部分（80％）被归类为同胞种C。在分子形式A的整个分布中，ITS2中存在两个单倍型的固定的基因组内序列变异被发现。结论：An共有三种分子形式。subpictus sl和An的两种分子形式。sundaicus sl记录在印度次大陆。在系统发育上，An的两种形式。在印度次大陆的沿海地区或岛屿中普遍存在的subpictus sl（B和C型）和东南亚报道的分子形式是Sundaicus Complex的成员。An的分子形式A。亚种与所有其他形式有很远的联系，并应有其独特的地位。An的存在。首次记录了印度领土上的埃皮克提克。