Identification of Novel and Conserved microRNAs in Homalodisca vitripennis, the Glassy-Winged Sharpshooter by Expression Profiling.

Raja Sekhar Nandety,Asokan Ramasamy,Bryce W Falk,Shizuo G Kamita,Almas Sharif,Ken Mills

doi:10.1371/journal.pone.0139771

Abstract

The glassy-winged sharpshooter (GWSS) Homalodisca vitripennis (Hemiptera: Cicadellidae), is a xylem-feeding leafhopper and an important vector of the bacterium Xylella fastidiosa; the causal agent of Pierce’s disease of grapevines. MicroRNAs are a class of small RNAs that play an important role in the functional development of various organisms including insects. In H. vitripennis, we identified microRNAs using high-throughput deep sequencing of adults followed by computational and manual annotation. A total of 14 novel microRNAs that are not found in the miRBase were identified from adult H. vitripennis. Conserved microRNAs were also found in our datasets. By comparison to our previously determined transcriptome sequence of H. vitripennis, we identified the potential targets of the microRNAs in the transcriptome. This microRNA profile information not only provides a more nuanced understanding of the biological and physiological mechanisms that govern gene expression in H. vitripennis, but may also lead to the identification of novel mechanisms for biorationally designed management strategies through the use of microRNAs.

Highlights

MicroRNAs are a class of non-protein coding small RNAs that are known to play important roles in post-transcriptional level gene regulation and expression
A colony of H. vitripennis was maintained at the University of California-Davis Contained Research Facility (CRF) in cages containing a mixture of host plants as previously described [31]
We were able to investigate the secondary structure of each potential precursor microRNA as well as the coordinates of those precursor microRNAs in the transcriptome of H. vitripennis

Summary

Introduction

MicroRNAs (miRNAs) are a class of non-protein coding small RNAs that are known to play important roles in post-transcriptional level gene regulation and expression MicroRNAs are typically 19–25 nt in length and are the most abundant class of endogenous small RNAs in both plants and animals [3]. MicroRNAs can originate from either noncoding or coding regions of transcripts. The primary transcripts of microRNAs are predominantly cleaved and processed within the cell nucleus by the nuclease Drosha to generate precursor microRNAs (pre-miRNAs) with a characteristic hairpin like secondary structure. The pre-miRNAs are transported into the cytoplasm by Exportin–5, where the terminal loop.

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