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Optimizing the widely used nuclear protein‐coding gene primers in beetle phylogenies and their application in the genus Sasajiscymnus Vandenberg (Coleoptera: Coccinellidae)

机译:优化甲虫系统中广泛使用的核蛋白编码基因引物及其在Sasajiscymnus Vandenberg(Cocoptera:Coccinellidae)中的应用

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摘要

Advances in genomic biology and the increasing availability of genomic resources allow developing hundreds of nuclear protein‐coding (NPC) markers, which can be used in phylogenetic research. However, for low taxonomic levels, it may be more practical to select a handful of suitable molecular loci for phylogenetic inference. Unfortunately, the presence of degenerate primers of NPC markers can be a major impediment, as the amplification success rate is low and they tend to amplify nontargeted regions. In this study, we optimized five NPC fragments widely used in beetle phylogenetics (i.e., two parts of carbamoyl‐phosphate synthetase: CADXM and CADMC, Topoisomerase, Wingless and Pepck) by reducing the degenerate site of primers and the length of target genes slightly. These five NPC fragments and 6 other molecular loci were amplified to test the monophyly of the coccinellid genus Vandenberg. The analysis of our molecular data set clearly supported the genus may be monophyletic but confirmation with an extended sampling is required. A fossil‐calibrated chronogram was generated by BEAST, indicating an origin of the genus at the end of the Cretaceous (77.87 Myr). Furthermore, a phylogenetic informativeness profile was generated to compare the phylogenetic properties of each gene more explicitly. The results showed that COI provides the strongest phylogenetic signal among all the genes, but Pepck, Topoisomerase, CADXM and CADMC are also relatively informative. Our results provide insight into the evolution of the genus , and also enrich the molecular data resources for further study.
机译:基因组生物学的进展和基因组资源的增加可用性允许发展数百核蛋白质编码(NPC)标志物,其可用于系统发育研究。然而,对于低分类水平,选择用于系统发育推理的少数合适的分子基因座可能更实用。遗憾的是,NPC标记物的退化引物的存在可以是主要的障碍,因为扩增成功率低,并且它们倾向于扩增不确定的区域。在这项研究中,我们通过减少引物的退化位点和略微减少靶基因的简并致靶基因的次数,优化了广泛使用的五种NPC片段(即氨基甲酰基磷酸盐合成酶:CADXM和CADMC,TOPOISOMERASE,无翅和PEPCK)。将这五种NPC片段和6个其他分子基因座进行扩增以测试脉络氏植物的单层。对我们的分子数据集的分析清楚地支持该属可以是单噬细胞的,但需要使用扩展采样的确认。通过野兽产生化石校准的计时器,表示白垩纪末端(77.87 MYR)的原点。此外,产生系统发育信息性曲线以比较每个基因的系统发育性能更明确。结果表明,COI在所有基因中提供最强的系统发育信号,但PEPCK,TOPOISOMERASE,CADXM和CADMC也相对卓越。我们的结果提供了对属的演变的洞察力,并丰富了分子数据资源进行进一步研究。

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