Identification and characterization of microRNAs in the leaf of ma bamboo (Dendrocalamus latiflorus) by deep sequencing

Abstract

MicroRNAs (miRNAs), a class of non-coding small endogenous RNAs of approximately 22 nucleotides, regulate gene expression at the post-transcriptional levels by targeting mRNAs for degradation or by inhibiting protein translation. Thousands of miRNAs have been identified in many species. However, there is no information available concerning miRNAs in ma bamboo (Dendrocalamus latiflorus), one of the most important non-timber forest products, which has essential ecological roles in forests. To identify miRNAs in D. latiflorus, a small RNA library was constructed from leaf tissues. Using next generation high-throughput sequencing technology and bioinformatics analysis, we obtained 11,513,607 raw sequence reads and identified 84 conserved miRNAs (54 mature miRNAs and 30 star miRNAs) belonging to 17 families, and 81 novel miRNAs (76 mature miRNAs and five star miRNAs) in D. latiflorus. One hundred and sixty-two potential targets were identified for the 81 novel bamboo miRNAs. Several targets for the novel miRNAs are transcription factors that play important roles in plant development. Among the novel miRNAs, 30 were selected and their expression profiles in response to different light conditions were validated by qRT-PCR. This study provides the first large-scale cloning and characterization of miRNAs in D. latiflorus. Eighty-four conserved and 81 novel miRNAs were identified in D. latiflorus. Our results present a broad survey of bamboo miRNAs based on experimental and bioinformatics analysis. Although it will be necessary to validate the functions of miRNAs by further experimental research, these results represent a starting point for future research on D. latiflorus and related species.

Figure 1. Length, distribution and abundance of small RNAs in D. latiflorus.

Figure 2. Abundance of conserved miRNA families in D. latiflorus.

Figure 3. Conserved miRNAs from D. latiflorus and their homologs in other some species.

Scale 0: miRNA undetected; Scale 1: miRNA identified using computational programs; Scale 2: miRNA sequenced using experimental cloned or high-throught technology; Scale 3: miRNA/miRNA* accumulation detected; Scale 4: miRNA detected by small RNA blot or RT-PCR or Northern; Scale 5: miRNAs with validated targets. Total information in Figure 3 is from miRBase 19.0 and references ,,-. Abbreviation: ath, Arabidopsis thaliana; bna, Brassica napus; rco, Ricinus communis; mtr, Medicago truncatula; csi, Citrus sinensis; vvi, Vitis vinifera; osa, Oryza sativa; sbi, Sorghum bicolor; zma, Zea mays; pab, Picea abies; pta, Pinus taeda; ppt, Physcomitrella patens; smo, Selaginella moellendorffii.

Figure 4. Expression profiles of novel miRNAs in leaves of D. latiflorus response to different light conditions.

CK: 200 µmol·m^-2·s^-1 for 4 h; High light: 1200 µmol·m^-2·s^-1 for 4 h; Dark: dark for 24 h.