西花蓟马对高低温胁迫响应的转录组分析

Transcriptome analysis of responses of western flower thrips Frankliniella occidentalis to high and low temperature stresses

为揭示西花蓟马Frankliniella occidentalis对温度的适应机制,采用高通量测序技术对低温(-13℃)、常温(26℃)和高温(40℃)胁迫1 h后西花蓟马进行转录组测序分析,筛选差异表达的unigenes,并对其进行功能注释与相关代谢通路富集分析,同时随机选择8个西花蓟马耐热性相关unigenes进行实时荧光定量(real-time fluorescence quantification PCR,q RT-PCR)测定,以验证转录组测序数据的正确性。结果显示,转录组测序、组装后共获得79516个unigenes,在NR和KEGG数据库中分别注释到28675个和47285个unigenes。40℃与26℃温度胁迫后差异表达的unigenes数量最多,为333个;-13℃与26℃温度胁迫后差异表达的unigenes最少,只有134个;40℃与-13℃温度胁迫后差异表达的unigenes为230个。差异表达的unigenes主要参与内质网蛋白合成和代谢通路等途径,其中有许多热激蛋白基因和细胞色素P450基因受到高低温胁迫后上调表达,表明这些unigenes可能参与西花蓟马应对极端温度的耐受性作用。随机筛选的8个差异表达unigenes的qRT-PCR结果与转录组测序结果一致,表明转录组数据可靠。

In order to reveal the adaptation mechanisms of western flower thrips Frankliniella occidentalis to temperature,transcriptome sequencing analysis was performed on F.occidentalis exposure to low temperature(-13℃),normal temperature(26℃)and high temperature(40℃)for one hour with highthroughput sequencing technology,and key unigenes of differential expression were screened.The gene function annotation and related metabolic pathway enrichment analysis were carried out.Meanwhile,eight genes related to heat resistance of F.occidentalis were selected for real-time quantification PCR(qRT-PCR)to verify the results from transcriptome sequencing.Total 79516 unigenes were sequenced and assembled,28675 unigenes were annotated in NR and 47285 unigenes were annotated in KEGG.Under temperatures 40℃ and 26℃,333 unigenes were differentially expressed.Only 134 unigenes were differentially expressed under -13℃ and 26℃,and 230 unigenes were differentially expressed under 40℃ and -13℃.The differentially expressed genes were mainly involved in endoplasmic reticulum protein synthesis and metabolic pathways,among which many heat shock protein genes and cytochrome P450 genes were up-regulated under high and low temperature stresses,suggesting that these genes may play an important role in the tolerance of F.occidentalis to extreme temperature.The qRTPCR results of eight differentially expressed genes randomly screened were consistent with the transcriptome sequencing results,indicating that the transcriptome analysis results were reliable.