Identification and profiling of miRNAs in overwintering Lissorhoptrus oryzophilus via next-generation sequencing.

ABSTRACT

MicroRNAs (miRNAs) are important regulators of various biological processes in organisms. Insects subjected to abiotic stress can regulate gene expression post-transcriptionally through the use of microRNAs. However, the role of miRNAs in response to cold stress in Lissorhoptrus oryzophilus Kuschel remains unknown. The rice water weevil, L. oryzophilus, is an invasive insect that is able to survive cold winters. To characterize changes in miRNAs in response to overwintering in L. oryzophilus, a comprehensive comparative analysis of microRNAs was performed involving an overwintering and a normal adult. High-throughput Illumina sequencing and bioinformatics analyses revealed 121 conserved and 14 potential novel microRNAs in two small libraries. The novel miRNAs exhibit low expression levels in both libraries. After the expression profiles of the miRNAs in the two libraries were normalized, 36 miRNAs in L. oryzophilus were found to be differentially expressed in response to overwintering. In particular, 14 conserved miRNAs and 6 novel miRNAs were up-regulated, while 15 conserved miRNAs and 1 novel miRNA were down-regulated. In addition, the expression patterns of 11 conserved and potentially novel miRNAs were confirmed by quantitative RT-PCR analysis. Most importantly, this work provides a unique resource of characterized miRNAs for overwintering L. oryzophilus and contributes to studies of the functions of cold-related and other L. oryzophilus miRNAs.