Zoological Survey of India, Prani Vigyan Bhawan, New Alipore, Kolkata, India.
Department of Animal Ecology and Conservation Biology, Wildlife Institute of India, Chandrabani, Dehradun, 248001, India.
Mol Biol Rep. 2024 Oct 1;51(1):1033. doi: 10.1007/s11033-024-09916-5.
The butterfly assemblage of Ladakh Trans-Himalaya demands a thorough analysis of their population genetic structure owing to their typical biogeographic affinity and their adaptability to extreme cold-desert climates. No such effort has been taken till date, and in this backdrop, we created a COI barcode reference library of 60 specimens representing 23 species.
Barcodes were generated from freshly collected leg samples using the Sanger sequencing method, followed by phylogenetic clade analyses and divergence calculation. Our data represents 22% of Ladakh's Rhopaloceran fauna with the novel barcode submission for six species, including one Schedule II species, Paralasa mani. Contrary to the 3% threshold rule, the interspecific divergence between two species pairs of typical mountain genus Hyponephele and Karanasa was found to be 2.3% and 2.2%, respectively. The addition of conspecific global barcodes revealed that most species showed little increase in divergence value, while a two-fold increase was noted in a few species. Bayesian clade clustering outcomes largely aligned with current morphological classifications, forming monophyletic clades of conspecific barcodes, with only minor exceptions observed for the taxonomically complicated genus Polyommatus and misidentified records of Aulocera in the database. We also observed variations within the same phylogenetic clades forming nested lineages, which may be attributed to the taxonomic intricacies present at the subspecies level globally, mostly among Eurasian species.
Overall, our effort not only substantiated the effectiveness of DNA Barcoding for the identification and conservation of this climatically vulnerable assemblage but also highlighted the significance of deciphering the unique genetic composition among this geographically isolated population of Ladakh butterflies.
由于拉达克横断喜马拉雅山脉的蝴蝶类群具有典型的生物地理亲缘关系和对极端寒冷沙漠气候的适应性，因此需要对其种群遗传结构进行深入分析。迄今为止，尚未对此进行任何研究。在此背景下，我们创建了一个包含 60 个代表 23 个物种的 COI 条码参考库。
使用 Sanger 测序法从新采集的腿部样本中生成条码，然后进行系统发育分支分析和分歧计算。我们的数据代表了拉达克鳞翅目动物区系的 22%，其中包括一种二级保护物种 Paralasa mani，有 6 个物种的新条码提交，其中包括一种二级保护物种 Paralasa mani。与 3%的阈值规则相反，两个典型高山属 Hyponephele 和 Karanasa 的两个物种对之间的种间分歧分别为 2.3%和 2.2%。添加同种全球条码后，发现大多数物种的分歧值增加不大，而少数物种的分歧值增加了一倍。贝叶斯聚类结果与当前形态分类学基本一致，形成了同种条码的单系聚类，只有少数例外，如分类复杂的 Polyommatus 属和数据库中 Aulocera 的错误鉴定记录。我们还观察到同一系统发育分支内的变异形成嵌套谱系，这可能归因于全球亚种水平上存在的分类复杂性，主要存在于欧亚物种中。
总的来说，我们的努力不仅证明了 DNA 条码在识别和保护这个气候脆弱的类群方面的有效性，还强调了在拉达克蝴蝶这个地理隔离种群中，解析其独特遗传组成的重要性。
