MicroRNAs (miRNAs) are a group of small RNAs involved in various biological processes through negative regulation of mRNAs at the post-transcriptional level. Although miRNA profiles have been documented in over two dozen insect species, few are agricultural pests. In this study, both conserved and novel miRNAs in the diamondback moth, Plutella xylostella L., a devastating insect pest of cruciferous crops worldwide, were documented. High-throughput sequencing of a small RNA library constructed from a mixed life stages of P. xylostella, including eggs, 1st to 4th (last) instar larvae, pupae and adults, identified 384 miRNAs, of which 174 were P. xylostella specific. In addition, temporal expressions of 234 miRNAs at various developmental stages were investigated using a customized microarray analysis. Among the 91 differentially expressed miRNAs, qRT-PCR analysis was used to validate highly expressed miRNAs at each stage. The combined results not only systematically document miRNA profiles in an agriculturally important insect pest, but also provide molecular targets for future functional analysis and, ultimately, genetic-based pest control practice.

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Published on PLOS One, v. 8, issue. 11, e78787.

© 2013 Liang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Figure_S1.tif (1362 kB)
Microarray analysis of expression profiles of predicted P. xylostella miRNAs at different developmental stages. After microarray hybridization and statistical analyses, all detectable P. xylostella miRNAs differentially expressed in eggs, larvae, pupae and adults were subjected to one-way ANOVA at P = 0.01 and the hierarchical clustering using the TIGR MeV (MultiExperiment Viewer) v4.1 software, http://www.tm4.org/mev.html. Two replicates were performed. Color coding: red, up-regulated; black, mean; green, down-regulated.

Figure_S2.tif (953 kB)
Alignment of identified pxy-mir-1c with other insect mir-1 registered in the miRBase. The conserved mature miRNA sequences are highlighted in black.

Figure_S3.tif (735 kB)
Alignment of identified pxy-mir-307 with other insect mir-307 registered in the miRBase. The conserved mature miRNA sequences are highlighted in black.

Table_S1.xls (86 kB)
Predicted P. xylostella miRNAs using Bombyx mori genome data as reference.

Table_S2.xls (148 kB)
Predicted P. xylostella miRNAs using its own genome data as reference.

Table_S3.doc (33 kB)
Primers used for qRT-PCR analysis of 11 miRNAs predicted based on P. xylostella genome data.

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