Comparative transcriptome analysis of lufenuron-resistant and susceptible strains of Spodoptera frugiperda (Lepidoptera: Noctuidae).
Comparative transcriptome analysis of lufenuron-resistant and susceptible strains of Spodoptera frugiperda (Lepidoptera: Noctuidae).
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DOI：
10.1186/s12864-015-2183-z
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发表时间：
2015-11-21
期刊：
影响因子：
4.4
通讯作者：
Omoto C
中科院分区：
文献类型：
--
作者：
do Nascimento AR;Fresia P;Cônsoli FL;Omoto C
草地贪夜蛾（Spodoptera frugiperda）抗药性的进化已造成巨大的经济损失和对环境和农业生态系统的干扰。对虱螨脲（一种几丁质生物合成抑制剂杀虫剂）的抗性最近在巴西的S.食虫类。因此，我们利用大规模cDNA测序（RNA-Seq分析）来比较虱螨脲抗性（LUF-R）和敏感（LUF-S）S之间的基因表达模式。幼虫，试图确定S. frugiperda对这种杀虫剂。使用约1，960万个100 bp长的单端读数组装转录组，产生了18，506个转录本，N50为996 bp。对NCBI非冗余数据库的检索产生了51.1%（9，457）的功能注释的转录物。大部分比对与昆虫同源，其中大多数（45%）与家蚕（鳞翅目：家蚕科）的序列相似。此外，10%的序列与夜蛾科（鳞翅目：夜蛾科）不同物种的序列相似，其中3%与S.食虫类。LUF-R和LUF-S S基因表达的比较分析。草地贪夜蛾幼虫鉴定了940个差异表达的转录物（p ≤ 0.05，t检验;倍数变化≥ 4）。其中6个与表皮代谢有关。其中，四个在LUF-R幼虫中过表达。参与解毒过程的机器由35个差异表达的转录本代表;其中24个属于P450单加氧酶，4个谷胱甘肽-S-转移酶，6个羧化酶和1个磺基转移酶。通过使用定量真实的时间PCR（qPCR），对许多选择的候选转录物验证RNA-Seq分析。利用基因芯片技术对LUF-R基因的表达谱进行了分析。frugiperda不同于LUF-S幼虫。在一般情况下，基因表达是高得多的抗性幼虫相比，敏感的，特别是那些基因参与的途径，异生物质解毒，主要代表P450单加氧酶转录。我们的数据表明，参与解毒过程的酶，主要是P450，是LUF-RS采用的抗性机制之一。棉铃虫幼虫对虱螨脲的抗性本文的在线版本（doi：10.1186/s12864 - 015 - 2183-z）包含补充材料，可供授权用户使用。
The evolution of insecticide resistance in Spodoptera frugiperda (Lepidoptera: Noctuidae) has resulted in large economic losses and disturbances to the environment and agroecosystems. Resistance to lufenuron, a chitin biosynthesis inhibitor insecticide, was recently documented in Brazilian populations of S. frugiperda. Thus, we utilized large-scale cDNA sequencing (RNA-Seq analysis) to compare the pattern of gene expression between lufenuron-resistant (LUF-R) and susceptible (LUF-S) S. larvae in an attempt to identify the molecular basis behind the resistance mechanism(s) of S. frugiperda to this insecticide. A transcriptome was assembled using approximately 19.6 million 100 bp-long single-end reads, which generated 18,506 transcripts with a N50 of 996 bp. A search against the NCBI non-redundant database generated 51.1 % (9,457) functionally annotated transcripts. A large portion of the alignments were homologous to insects, with the majority (45 %) being similar to sequences of Bombyx mori (Lepidoptera: Bombycidae). Moreover, 10 % of the alignments were similar to sequences of various species of Spodoptera (Lepidoptera: Noctuidae), with 3 % of them being similar to sequences of S. frugiperda. A comparative analysis of the gene expression between LUF-R and LUF-S S. frugiperda larvae identified 940 differentially expressed transcripts (p ≤ 0.05, t-test; fold change ≥ 4). Six of them were associated with cuticle metabolism. Of those, four were overexpressed in LUF-R larvae. The machinery involved with the detoxification process was represented by 35 differentially expressed transcripts; 24 of them belonging to P450 monooxygenases, four to glutathione-S-transferases, six to carboxylases and one to sulfotransferases. RNA-Seq analysis was validated for a number of selected candidate transcripts by using quantitative real time PCR (qPCR). The gene expression profile of LUF-R larvae of S. frugiperda differs from LUF-S larvae. In general, gene expression is much higher in resistant larvae when compared to the susceptible ones, particularly for those genes involved with pathways for xenobiotic detoxification, mainly represented by P450 monooxygenases transcripts. Our data indicate that enzymes involved with the detoxification process, and mostly the P450, are one of the resistance mechanisms employed by the LUF-R S. frugiperda larvae against lufenuron. The online version of this article (doi:10.1186/s12864-015-2183-z) contains supplementary material, which is available to authorized users.