Microsatellite markers developed using a next-generation sequencing technique for Neotrogla spp. (Psocodea: Prionoglarididae), cave dwelling insects with sex-reversed genitalia
Microsatellite markers developed using a next-generation sequencing technique for Neotrogla spp. (Psocodea: Prionoglarididae), cave dwelling insects with sex-reversed genitalia
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DOI：
10.1111/ens.12339
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发表时间：
2019-03-01
影响因子：
0.9
通讯作者：
Yoshizawa, Kazunori
中科院分区：
文献类型：
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作者：
Kamimura, Yoshitaka;Abe, Jun;Yoshizawa, Kazunori
Neotrogla属(Psocodea：Philoglarididae)由四个来自巴西的命名种组成。这种穴居昆虫的雌性特征是有一个明显的阴茎状插入器官，称为雌核，在交配时插入阴道状的雄性生殖器。另一种进化上的新结构，精囊板，使雌性能够同时将两个新沉积的精囊(由精子和可能的营养物质组成)存储在她的精子存储器官(受精囊)中。目前尚不清楚这两个精原是否来自两个不同的雄性。为了研究这些性交反转生殖器昆虫的交配生态和种群遗传结构，从一个未知的Neotrogla种分离到了16个新的高度多态的微卫星基因座，并根据其2,275 Mbp的基因组序列进行了分析。我们的第一次筛查发现了99,888个候选基因座。与到目前为止研究的其他半翅目昆虫类似，AAT基序微卫星明显占优势。我们进一步筛选了99个序列，成功地设计了50对聚合酶链式反应引物。其中16个引物在11个Neotrogla sp.中成功扩增出预期大小的产物。从两个洞穴收集的个体。每个座位的等位基因数从2个到9个不等，两个群体都没有明显偏离Hardy-Weinberg平衡。尽管采样的洞穴相距仅约1公里，但在两个种群之间发现了显著的遗传分化。结果表明，在极光、巴西、曲线虫和树干夜蛾中分别检测到13个、12个、13个和11个微卫星座位，表明这些微卫星座位可用于多个新寄主植物的集合种群遗传学研究。
The genus Neotrogla (Psocodea: Prinoglarididae) comprises four named species from Brazil. Females of this cave-dwelling insect are characterized by a conspicuous penis-like intromittent organ, termed a gynosome, which is inserted into the vagina-like male genitalia during copulation. Another evolutionarily novel structure, the spermathecal plate, enables a female to simultaneously store two freshly deposited spermatophores (consisting of sperm and possibly nutritious substances) in her sperm storage organ (spermatheca). It is unknown whether the two spermatophores are derived from two different males. To investigate the mating ecology and population genetic structures of these insects with sex-reversed genitalia, 16 novel highly polymorphic microsatellite loci were isolated and characterized based on similar to 2,275 Mbp genomic sequences from an undescribed Neotrogla species. Our first screening detected 99,888 candidate loci. Similar to other hemipteroid insects studied thus far, AAT motif microsatellites were conspicuously dominant. We further screened 99 sequences, for which 50 pairs of polymerase chain reaction primers were successfully designed. Sixteen of these primers successfully amplified products of the expected size in the 11 Neotrogla sp. individuals collected from two caves. The number of alleles per loci varied from two to nine, with no significant deviation from the Hardy-Weinberg equilibrium in either population. Although the caves sampled were only approximately 1 km apart, significant genetic differentiation was detected between the two populations. In total, 13, 12, 13 and 11 loci were cross-amplified in N. aurora, N. brasiliensis, N. curvata and N. truncata, respectively, indicating the applicability of these microsatellite loci for metapopulation genetic studies in multiple Neotrogla species.