Laboratory of Cell Biology, Research Institute of Forestry, Chinese Academy of Forestry, Xiangshan Road, Beijing 100091, People's Republic of China.
Plant Cell Rep. 2013 Sep;32(9):1339-49. doi: 10.1007/s00299-013-1446-x. Epub 2013 May 7.
142 miRNAs were identified and 38 miRNA targets were predicted, 4 of which were validated, in C. intermedia . The expression of 12 miRNAs in salt-stressed leaves was assessed by qRT-PCR. MicroRNAs (miRNAs) are endogenous small RNAs that play important roles in various biological and metabolic processes in plants. Caragana intermedia is an important ecological and economic tree species prominent in the desert environment of west and northwest China. To date, no investigation into C. intermedia miRNAs has been reported. In this study, high-throughput sequencing of small RNAs and analysis of transcriptome data were performed to identify both conserved and novel miRNAs, and also their target mRNA genes in C. intermedia. Based on sequence similarity and hairpin structure prediction, 132 putative conserved miRNAs (12 of which were confirmed to form hairpin precursors) belonging to 31 known miRNA families were identified. Ten novel miRNAs (including the miRNA* sequences of three novel miRNAs) were also discovered. Furthermore, 36 potential target genes of 17 known miRNA families and 2 potential target genes of 1 novel miRNA were predicted; 4 of these were validated by 5' RACE. The expression of 12 miRNAs was validated in different tissues, and these and five target mRNAs were assessed by qRT-PCR after salt treatment. The expression levels of seven miRNAs (cin-miR157a, cin-miR159a, cin-miR165a, cin-miR167b, cin-miR172b, cin-miR390a and cin-miR396a) were upregulated, while cin-miR398a expression was downregulated after salt treatment. The targets of cin-miR157a, cin-miR165a, cin-miR172b and cin-miR396a were downregulated and showed an approximately negative correlation with their corresponding miRNAs under salt treatment. These results would help further understanding of miRNA regulation in response to abiotic stress in C. intermedia.
鉴定了 142 个 miRNA，并预测了 38 个 miRNA 靶标，其中 4 个靶标得到了验证，在中间锦鸡儿中。通过 qRT-PCR 评估了 12 个 miRNA 在盐胁迫叶片中的表达。miRNAs（miRNAs）是内源性小 RNA，在植物的各种生物和代谢过程中发挥重要作用。中间锦鸡儿是一种重要的生态和经济树种，在中国西部和西北部的沙漠环境中突出。迄今为止，尚未报道中间锦鸡儿的 miRNA。在这项研究中，对小 RNA 进行高通量测序，并对转录组数据进行分析，以鉴定中间锦鸡儿中的保守和新型 miRNA 及其靶 mRNA 基因。基于序列相似性和发夹结构预测，鉴定了 132 个推定的保守 miRNA（其中 12 个被证实形成发夹前体），属于 31 个已知 miRNA 家族。还发现了 10 个新的 miRNA（包括 3 个新 miRNA 的 miRNA*序列）。此外，预测了 17 个已知 miRNA 家族的 36 个潜在靶基因和 1 个新 miRNA 的 2 个潜在靶基因；其中 4 个通过 5' RACE 得到验证。在不同组织中验证了 12 个 miRNA 的表达，并在盐处理后通过 qRT-PCR 评估了这些 miRNA 和 5 个靶 mRNA 的表达。7 个 miRNA（cin-miR157a、cin-miR159a、cin-miR165a、cin-miR167b、cin-miR172b、cin-miR390a 和 cin-miR396a）的表达上调，而 cin-miR398a 的表达下调盐处理后。在盐处理下，cin-miR157a、cin-miR165a、cin-miR172b 和 cin-miR396a 的靶标下调，并且与它们对应的 miRNA 呈负相关。这些结果将有助于进一步了解中间锦鸡儿对非生物胁迫的 miRNA 调控。
