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. 2016 Mar 4:6:22687.
doi: 10.1038/srep22687.

High throughput sequencing of small RNAs transcriptomes in two Crassostrea oysters identifies microRNAs involved in osmotic stress response

Xuelin Zhao  1 , Hong Yu  1 , Lingfeng Kong  1 , Shikai Liu  1   2 , Qi Li  1
Affiliations

High throughput sequencing of small RNAs transcriptomes in two Crassostrea oysters identifies microRNAs involved in osmotic stress response

Xuelin Zhao et al. Sci Rep. .

Abstract

Increasing evidence suggests that microRNAs post-transcriptionally regulate gene expression and are involved in responses to biotic and abiotic stress. However, the role of miRNAs involved in osmotic plasticity remains largely unknown in marine bivalves. In the present study, we performed low salinity challenge with two Crassostrea species (C. gigas and C. hongkongensis), and conducted high-throughput sequencing of four small RNA libraries constructed from the gill tissues. A total of 202 and 87 miRNAs were identified from C. gigas and C. hongkongensis, respectively. Six miRNAs in C. gigas and two in C. hongkongensis were differentially expressed in response to osmotic stress. The expression profiles of these eight miRNAs were validated by qRT-PCR. Based on GO enrichment and KEGG pathway analysis, genes associated with microtubule-based process and cellular component movement were enriched in both species. In addition, five miRNA-mRNA interaction pairs that showed opposite expression patterns were identified in the C. hongkongensis, Differential expression analysis identified the miRNAs that play important regulatory roles in response to low salinity stress, providing insights into molecular mechanisms that are essential for salinity tolerance in marine bivalves.

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Figures

Figure 1
Figure 1. Length distribution of small RNAs in four groups from C. gigas and C. hongkongensis.
Figure 2
Figure 2. Expression of eight miRNAs determined by qRT-PCR.
The eight miRNAs included scaffold43364_10952 (a), miR-92-3p (b), miR-1984 (c), miR-183 (d), miR-184-3p (e), and miR-2353 (f) in C. gigas and miR-2353 (g) and miR-3205 (h) in C. hongkongensis. 5S gene was used as an internal control to calibrate the cDNA template for all the samples. Each values were shown as mean ± SD (n = 6).
Figure 3
Figure 3
GO distribution of target genes of differentially expressed miRNAs of C. gigas (a) and C. hongkongensis (b). Red: target genes of up-regulated miRNAs; Purple: target genes of down-regulated miRNAs.
Figure 4
Figure 4. Venn diagram of shared enriched GO terms between C. gigas and C. hongkongensis.
See this image and copyright information in PMC

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