Laboratory of Ecology, Genetics and Environmental Protection, Tomsk State University, Tomsk, Russia.
Department of General Biology and Ecology, Moscow Regional State University, Moscow, Russia.
Parasit Vectors. 2018 Mar 27;11(1):211. doi: 10.1186/s13071-018-2657-3.
Anopheles beklemishevi is a member of the Maculipennis group of malaria mosquitoes that has the most northern distribution among other members of the group. Although a cytogenetic map for the larval salivary gland chromosomes of this species has been developed, a high-quality standard cytogenetic photomap that enables genomics and population genetics studies of this mosquito at the adult stage is still lacking.
In this study, a cytogenetic map for the polytene chromosomes of An. beklemishevi from ovarian nurse cells was developed using high-resolution digital imaging from field collected mosquitoes. PCR-amplified DNA probes for fluorescence in situ hybridization (FISH) were designed based on the genome of An. atroparvus. The DNA probe obtained by microdissection procedures from the breakpoint region was labelled in a DOP-PCR reaction. Population analysis was performed on 371 specimens collected in 18 locations.
We report the development of a high-quality standard photomap for the polytene chromosomes from ovarian nurse cells of An. beklemishevi. To confirm the suitability of the map for physical mapping, several PCR-amplified probes were mapped to the chromosomes of An. beklemishevi using FISH. In addition, we identified and mapped DNA probes to flanking regions of the breakpoints of two inversions on chromosome X of this species. Inversion polymorphism was determined in 13 geographically distant populations of An. beklemishevi. Four polymorphic inversions were detected. The positions of common chromosomal inversions were indicated on the map.
The study constructed a standard photomap for ovarian nurse cell chromosomes of An. beklemishevi and tested its suitability for physical genome mapping and population studies. Cytogenetic analysis determined inversion polymorphism in natural populations of An. beklemishevi related to this species' adaptation.
斑须按蚊是疟蚊属 Maculipennis 组的一个成员，其分布最北。虽然已经开发出该物种幼虫唾液腺染色体的细胞遗传学图谱，但仍缺乏高质量的标准细胞遗传学照相图谱，无法对该蚊的成虫进行基因组学和种群遗传学研究。
本研究利用野外采集的蚊子的高分辨率数字成像，从卵巢滋养细胞中开发了斑须按蚊的多线染色体细胞遗传学图谱。根据斑须按蚊的基因组设计了用于荧光原位杂交（FISH）的 PCR 扩增 DNA 探针。从断裂点区域的显微切割程序获得的 DNA 探针在 DOP-PCR 反应中进行标记。对从 18 个地点收集的 371 个标本进行了种群分析。
我们报告了从卵巢滋养细胞的多线染色体中开发出高质量的标准照相图谱。为了确认图谱适用于物理作图，使用 FISH 将几个 PCR 扩增探针映射到斑须按蚊的染色体上。此外，我们鉴定并绘制了 DNA 探针，以定位该物种 X 染色体上两个倒位断点的侧翼区域。在 13 个地理上遥远的斑须按蚊种群中确定了倒位多态性。检测到 4 个多态性倒位。常见染色体倒位的位置在图谱上进行了标注。
本研究构建了斑须按蚊卵巢滋养细胞染色体的标准照相图谱，并测试了其用于物理基因组作图和种群研究的适用性。细胞遗传学分析确定了与该物种适应相关的自然种群中斑须按蚊的倒位多态性。
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