Department of Entomology and the Fralin Life Sciences Institute, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA.
Laboratory of Evolutionary Genomics of Insects, the Federal Research Center Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia.
Malar J. 2021 Mar 10;20(1):141. doi: 10.1186/s12936-021-03676-4.
The malaria mosquito Anopheles punctipennis, a widely distributed species in North America, is capable of transmitting human malaria and is actively involved in the transmission of the ungulate malaria parasite Plasmodium odocoilei. However, molecular diagnostic tools based on Internal Transcribed Spacer 2 (ITS2) of ribosomal DNA are lacking for this species. Anopheles punctipennis is a former member of the Anopheles maculipennis complex but its systematic position remains unclear.
In this study, ITS2 sequences were obtained from 276 An. punctipennis specimens collected in the eastern and midwestern United States and a simple and robust Restriction Fragment Length Polymorphism approach for species identification was developed. The maximum-likelihood phylogenetic tree was constructed based on ITS2 sequences available through this study and from GenBank for 20 species of Anopheles.
The analysis demonstrated a consistent ITS2 sequence length and showed no indications of intragenomic variation among the samples based on ITS2, suggesting that An. punctipennis represents a single species in the studied geographic locations. In this study, An. punctipennis was found in urban, rural, and forest settings, suggesting its potential broad role in pathogen transmission. Phylogeny based on ITS2 sequence comparison demonstrated the close relationship of this species with other members of the Maculipennis group.
This study developed molecular tools based on ITS2 sequences for the malaria vector An. punctipennis and clarified the phylogenetic position of the species within the Maculipennis group.
疟蚊属的 punctipennis 是一种广泛分布于北美的物种，能够传播人类疟疾，并积极参与有蹄类疟原虫 Plasmodium odocoilei 的传播。然而，目前缺乏基于核糖体 DNA 的内转录间隔区 2（ITS2）的分子诊断工具。疟蚊属 punctipennis 曾是斑蚊复合体的成员，但它的系统位置仍不清楚。
本研究从美国东部和中西部采集的 276 只疟蚊属 punctipennis 标本中获得了 ITS2 序列，并开发了一种简单而可靠的限制性片段长度多态性方法来进行物种鉴定。根据本研究中获得的 ITS2 序列和 GenBank 中 20 种疟蚊属的序列构建了最大似然系统发育树。
分析表明 ITS2 序列长度一致，且根据 ITS2 没有显示出样本之间的种内变异迹象，表明在所研究的地理区域中，疟蚊属 punctipennis 代表了一个单一的物种。在本研究中，疟蚊属 punctipennis 出现在城市、农村和森林环境中，这表明它在病原体传播方面可能具有广泛的作用。基于 ITS2 序列比较的系统发育分析表明，该物种与 Maculipennis 组的其他成员密切相关。
本研究基于 ITS2 序列为疟蚊属 punctipennis 开发了分子工具，并阐明了该物种在 Maculipennis 组中的系统发育位置。
