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转录组测序以及豆荚野螟 Maruca vitrata(鳞翅目:螟蛾科)SNP 标记的快速开发和应用。

Transcriptome sequencing, and rapid development and application of SNP markers for the legume pod borer Maruca vitrata (Lepidoptera: Crambidae).

机构信息

Department of Entomology, Purdue University, West Lafayette, Indiana, United States of America.

出版信息

PLoS One. 2011;6(7):e21388. doi: 10.1371/journal.pone.0021388. Epub 2011 Jul 6.

Abstract

The legume pod borer, Maruca vitrata (Lepidoptera: Crambidae), is an insect pest species of crops grown by subsistence farmers in tropical regions of Africa. We present the de novo assembly of 3729 contigs from 454- and Sanger-derived sequencing reads for midgut, salivary, and whole adult tissues of this non-model species. Functional annotation predicted that 1320 M. vitrata protein coding genes are present, of which 631 have orthologs within the Bombyx mori gene model. A homology-based analysis assigned M. vitrata genes into a group of paralogs, but these were subsequently partitioned into putative orthologs following phylogenetic analyses. Following sequence quality filtering, a total of 1542 putative single nucleotide polymorphisms (SNPs) were predicted within M. vitrata contig assemblies. Seventy one of 1078 designed molecular genetic markers were used to screen M. vitrata samples from five collection sites in West Africa. Population substructure may be present with significant implications in the insect resistance management recommendations pertaining to the release of biological control agents or transgenic cowpea that express Bacillus thuringiensis crystal toxins. Mutation data derived from transcriptome sequencing is an expeditious and economical source for genetic markers that allow evaluation of ecological differentiation.

摘要

豆科植物豆荚螟(鳞翅目:螟蛾科)是一种昆虫害虫,主要危害在热带地区种植的自给农民的作物。我们提供了一种新的方法,即通过 454 测序和 Sanger 测序技术对中肠、唾液和成年昆虫组织的 3729 个连续体进行从头组装,以此研究这个非模式物种。功能注释预测,1320 个 Maruca vitrata 蛋白编码基因存在,其中 631 个在 Bombyx mori 基因模型中有同源物。基于同源性的分析将 M. vitrata 基因分配到一组旁系同源基因中,但随后通过系统发育分析将这些基因分配为假定的直系同源基因。经过序列质量过滤后,在 M. vitrata 连续体组装中总共预测到 1542 个可能的单核苷酸多态性(SNP)。在设计的 1078 个分子遗传标记中,有 71 个被用于筛选来自西非五个采集点的 M. vitrata 样本。种群亚结构可能存在,这对释放生物防治剂或表达苏云金芽孢杆菌晶体毒素的转基因豇豆的昆虫抗性管理建议有重要影响。从转录组测序中获得的突变数据是遗传标记的一种快速、经济的来源,可用于评估生态分化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5fa0/3130784/855b1512015c/pone.0021388.g001.jpg

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