Department of Entomology and Nematology, The University of Florida, PO Box 110620, Gainesville, FL 32611-0620, USA.
Exp Appl Acarol. 2013 Mar;59(3):283-96. doi: 10.1007/s10493-012-9603-4. Epub 2012 Aug 25.
Next-generation sequencing was applied to the transcriptome of the phytoseiid Metaseiulus occidentalis to characterize gene expression in all life stages reared under different conditions to optimize the recovery of as many genes as possible. One production and one titration run produced a total of 862,069 reads (average size: 314.87 bp), which generated 255.6 Mbp of sequences on the GS-FLX Titanium sequencing platform. After removal of putative prey sequences 850,543 reads were used in NewBler and PTA assemblies to produce 74,172 non-redundant sequences, including 30,691 contigs and 43,481 singlets with 11,994 contigs consisting of more than 500 bp and 37,278 sequences >300 bp, constituting 48.7 % of all sequences. There were 25,888 hits with the NCBI non-redundant database and 15,376 unique transcripts. There were 26,225 hits with the Ixodes scapularis genome and 6,634 unique transcripts. There were 22,225 hits with the RefSeq of Homo sapiens with 6,465 unique transcripts, and 23,656 hits with the RefSeq of Drosophila melanogaster with 9,216 unique transcripts. Selected ESTs corresponding to genes of interest were analyzed including those related to transposable elements, GPCRs, Sox transcription factors, diapause and foraging behavior, and pesticide resistances. Novel and important genes appear to have been discovered that provide insight into the evolution, biology, and physiology of this important predator of pest mites in agriculture and will be useful in analyzing complete genome sequences of this natural enemy.
下一代测序技术被应用于西方盲走螨的转录组,以描述在不同条件下饲养的所有生活阶段的基因表达情况,从而优化尽可能多的基因回收。一次生产和一次滴定运行共产生了 862069 条reads(平均大小:314.87bp),在 GS-FLX Titanium 测序平台上生成了 255.6 Mbp 的序列。在去除可能的猎物序列后,850543 条reads 用于 NewBler 和 PTA 组装,生成了 74172 条非冗余序列,包括 30691 个 contigs 和 43481 个 singlets,其中 11994 个 contigs 由超过 500bp 的序列组成,37278 个序列大于 300bp,占所有序列的 48.7%。与 NCBI 非冗余数据库有 25888 次匹配,有 15376 个独特的转录本。与斑疹热硬蜱基因组有 26225 次匹配,有 6634 个独特的转录本。与 Homo sapiens 的 RefSeq 有 22225 次匹配,有 6465 个独特的转录本,与 Drosophila melanogaster 的 RefSeq 有 23656 次匹配,有 9216 个独特的转录本。分析了对应于感兴趣基因的 ESTs,包括与转座元件、GPCRs、Sox 转录因子、滞育和觅食行为以及抗药性相关的基因。似乎发现了新的和重要的基因,这些基因为了解这种农业中重要的捕食性螨虫的进化、生物学和生理学提供了线索,并将有助于分析这种天敌的完整基因组序列。