College of Horticulture, Nanjing Agricultural University, No. 1 Weigang Road, Nanjing, 210095 Jiangsu Province, People's Republic of China.
Mol Biol Rep. 2012 Oct;39(10):9463-72. doi: 10.1007/s11033-012-1811-9. Epub 2012 Jun 24.
MicroRNAs (miRNAs) are an extensive class of newly identified small RNAs that regulate gene expression at post-transcription level by mRNA cleavage or translation. In our study, we used qRT-PCR and found that Vv-miR164 is expression in grapevine leaves, stems, tendrils, inflorescences, flowers and fruits. In addition, two potential target genes for Vv-miR164 were also found and verified by PPM-RACE and RLM-RACE. The results not only maps the cleavage site of the target mRNA but allowed for detection the expression pattern of cleaved fragments that can indicate the regulatory function of this miRNA on its target genes. These target genes were explored by qRT-PCR where some exhibited different expression patterns from their corresponding miRNA, indicating the cleavage mode of the miRNA on its target genes. The efficient and powerful approach used in this study can help in further understanding of how miRNAs cleaved their target mRNAs. Results from this study prove the importance of Vv-miR164 in regulating development and growth of grapes, and adds to the existing knowledge of small RNA-mediated regulation in grapes.
微 RNA（miRNA）是一类新发现的小 RNA，通过 mRNA 切割或翻译在转录后水平调控基因表达。在本研究中，我们使用 qRT-PCR 发现 Vv-miR164 在葡萄叶片、茎、卷须、花序、花和果实中表达。此外，还通过 PPM-RACE 和 RLM-RACE 找到了两个 Vv-miR164 的潜在靶基因，并进行了验证。这些结果不仅定位了靶 mRNA 的切割位点，还可以检测到可以指示该 miRNA 对其靶基因的调控功能的切割片段的表达模式。通过 qRT-PCR 对这些靶基因进行了探索，其中一些基因的表达模式与其对应的 miRNA 不同，表明 miRNA 对其靶基因的切割模式。本研究中使用的高效有力的方法有助于进一步了解 miRNA 如何切割其靶 mRNA。本研究的结果证明了 Vv-miR164 在调控葡萄发育和生长中的重要性，并为葡萄中小 RNA 介导的调控提供了新的知识。
