Department of Entomology, China Agricultural University, Beijing, 100193, P. R. China.
Sci Rep. 2017 Jan 18;7:40713. doi: 10.1038/srep40713.
The diamondback moth (DBM), Plutella xylostella (L.), is one of the most serious cruciferous pests and has developed high resistance to most insecticides, including chlorantraniliprole. Previous studies have reported several protein-coding genes that involved in chlorantraniliprole resistance, but research on resistance mechanisms at the post-transcription level is still limited. In this study, a global screen of microRNAs (miRNAs) associated with chlorantraniliprole resistance in P. xylostella was performed. The small RNA libraries for a susceptible (CHS) and two chlorantraniliprole resistant strains (CHR, ZZ) were constructed and sequenced, and a total of 199 known and 30 novel miRNAs were identified. Among them, 23 miRNAs were differentially expressed between CHR and CHS, and 90 miRNAs were differentially expressed between ZZ and CHS, of which 11 differentially expressed miRNAs were identified in both CHR and ZZ. Using miRanda and RNAhybrid, a total of 1,411 target mRNAs from 102 differentially expressed miRNAs were predicted, including mRNAs in several groups of detoxification enzymes. The expression of several differentially expressed miRNAs and their potential targets was validated by qRT-PCR. The results may provide important clues for further study of the mechanisms of miRNA-mediated chlorantraniliprole resistance in DBM and other target insects.
小菜蛾（DBM），Plutella xylostella（L.），是十字花科蔬菜中最严重的害虫之一，对大多数杀虫剂包括氯虫苯甲酰胺已产生高度抗性。先前的研究报道了几种与氯虫苯甲酰胺抗性相关的蛋白质编码基因，但在后转录水平上的抗性机制研究仍有限。在本研究中，对小菜蛾中与氯虫苯甲酰胺抗性相关的 microRNAs（miRNAs）进行了全局筛选。构建并测序了敏感（CHS）和两个氯虫苯甲酰胺抗性品系（CHR，ZZ）的小 RNA 文库，共鉴定出 199 个已知和 30 个新的 miRNAs。其中，CHR 和 CHS 之间有 23 个 miRNAs 表达差异，ZZ 和 CHS 之间有 90 个 miRNAs 表达差异，其中 11 个在 CHR 和 ZZ 中都有表达差异。使用 miRanda 和 RNAhybrid，从 102 个差异表达的 miRNAs 中预测了总共 1411 个靶 mRNAs，包括几种解毒酶组的 mRNAs。通过 qRT-PCR 验证了几个差异表达的 miRNAs 及其潜在靶标的表达。这些结果可能为进一步研究 miRNA 介导的氯虫苯甲酰胺抗性在小菜蛾和其他靶标昆虫中的机制提供重要线索。
