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Micro RNAs (mi RNAs) are small noncoding RNAs participating in versatile biological processes via post-transcriptionally gene regulation. However, how mi RNAs are modified or degraded remains unknown, despite years of studies have unravelled much details of mi RNA biogenesis and function. Here, we systematically investigated mi RNA modification using six small RNA sequencing libraries generated from cotton seedling as well as cotton fibre at five developmental stages. Our results show that 1-2-nt truncation and addition on both 5′ and 3′ ends of mi RNAs are the major modification forms. The 5′ and 3′ end mi RNA modification was almost equal in the six development stages. Truncation was more common than addition on both 5′ and 3′ end. Structure analysis of the 5′ and 3′ ends of mi RNAs and isomiRs shows that uridine is the preferential nucleotide at the first position of both 5′ and 3′ ends. According to analysis of nucleotides truncated and tailed from mi RNAs, both mi RNAs and isomiRs share a similar positional structure distribution at their 5′ and 3′ ends, respectively. Furthermore, opposite to previous reports, cytodine is more frequently truncated and tailed from the two ends of isomiRs, implying existence of a complex cytodine balance in isomiRs. Comparison of isomiR expression shows differential mi RNA modification amongst the six developmental stages in terms of selective modification form, development-dependent modification and differential expression abundance. Our results globally uncovered mi RNA modification features in cotton, which could contribute us to understanding mi RNA's postmature modification and its regulatory function. [ABSTRACT FROM AUTHOR]

Micro RNAs (mi RNAs) have been found to be differentially expressed during cotton fibre development. However, which specific mi RNAs and how they are involved in fibre development is unclear. Here, using deep sequencing, 65 conserved mi RNA families were identified and 32 families were differentially expressed between leaf and ovule. At least 40 mi RNAs were either leaf or ovule specific, whereas 62 mi RNAs were shared in both leaf and ovule. q RT- PCR confirmed these mi RNAs were differentially expressed during fibre early development. A total of 820 genes were potentially targeted by the identified mi RNAs, whose functions are involved in a series of biological processes including fibre development, metabolism and signal transduction. Many predicted mi RNA-target pairs were subsequently validated by degradome sequencing analysis. GO and KEGG analyses showed that the identified mi RNAs and their targets were classified to 1027 GO terms including 568 biological processes, 324 molecular functions and 135 cellular components and were enriched to 78 KEGG pathways. At least seven unique mi RNAs participate in trichome regulatory interaction network. Eleven trans-acting si RNA (tasi RNA) candidate genes were also identified in cotton. One has never been found in other plant species and two of them were derived from MYB and ARF, both of which play important roles in cotton fibre development. Sixteen genes were predicted to be tasi RNA targets, including sucrose synthase and MYB2. Together, this study discovered new mi RNAs in cotton and offered evidences that mi RNAs play important roles in cotton ovule/fibre development. The identification of tasi RNA genes and their targets broadens our understanding of the complicated regulatory mechanism of mi RNAs in cotton. [ABSTRACT FROM AUTHOR]

Abstract: MicroRNAs (miRNAs) are an important gene regulator, controlling almost all biological and metabolic processes, in both plants and animals. In this study, we investigated the effect of drought and salinity stress on the expression of miRNAs and their targets in cotton (Gossypium hirsutum L.). Our results show that the expression change of miRNAs and their targets were dose-dependent and tissue-dependent under salinity and drought conditions. The expression of miRNAs in leaf was down-regulated under higher salinity stress while shows variable patterns in other conditions. The highest fold-changes of miRNAs were miR398 in roots with 28.9 fold down-regulation under 0.25% NaCl treatment and miR395 in leaves with 7.6 fold down-regulation under 1% PEG treatment. The highest up-regulation of miRNA targets was AST in roots with 4.7 fold-change under 2.5% PEG and the gene with highest down-regulation was CUC1 in leaves with 25.6 fold-change under 0.25% NaCl treatment. Among seven miRNA-target pairs we studied, five pairs, miR156–SPL2, miR162–DCL1, miR159–TCP3, miR395–APS1 and miR396–GRF1, show significant regulation relationship in roots and leaves under salinity stress concentration. [Copyright &y& Elsevier]

Abstract: No study has been performed on identifying microRNAs (miRNAs) and their targets in cotton although cotton is one of the most important fiber and economic crops around the world. In this study, we found 30 potential cotton miRNAs using a comparative genomic approach based on genomic survey sequence analysis and miRNA secondary structure. These cotton miRNAs belong to 22 miRNA families. Expressed sequence tag (EST) analysis indicated that the predicted miRNAs were expressed in cotton plants. Based on the characteristic that miRNAs exhibit perfect or nearly perfect complementarity with their targeted mRNA sequences, a total of 139 potential miRNA targets were identified in cotton genome. A majority of these targets belong to transcriptional factors which regulate cotton growth and development, including leaf, root, stem, flower, and even fiber development. Those miRNAs may also be involved in other cellular and metabolic processes, such as stress response, signal transduction, and secondary wall synthesis and deposition. Some of the newly identified miRNA targets may be unique to cotton species. In this study, we found that at least 3 miRNA families (miR 396, 414, and 782) target callous synthase, fiber protein Fb23, and fiber quinone-oxidoreductase, suggesting that miRNAs play an important role in cotton fiber differentiation and development. [Copyright &y& Elsevier]