Characterization and primary functional analysis of Pinus densata miR171
Characterization and primary functional analysis of Pinus densata miR171
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高松 miR171 的表征和主要功能分析
DOI：
10.1007/s10535-018-0774-7
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发表时间：
2018-06-01
影响因子：
1.5
通讯作者：
Li, Y. F.
中科院分区：
文献类型：
--
作者：
Hai, B. Z.;Qiu, Z. B.;Li, Y. F.
miR171是一个保守的microRNA （miRNA）家族，已被证明参与植物的生长发育。然而，miR171在松木中的确切功能在很大程度上仍不清楚。成熟miR171序列比较表明拟南芥和白沙芥具有高度的相似性，并且pre-miR171可以折叠成典型的茎环发夹结构。利用修饰的RNA连接酶介导的cDNA末端5 '快速扩增（RLM-RACE）技术，确定了编码GRAS （GAI-RGA-SCR）家族转录因子和肌动蛋白结合蛋白的基因为pde-miR171的靶点。此外，通过在拟南芥叶片中短暂共表达pdemir171和拟南芥SCL6（稻草人- like6）基因，进一步验证了两者之间的相互作用。实时荧光定量PCR结果显示，过表达mir171的植株中pde-miR171的表达水平明显高于野生型植株，且与拟南芥SCL6基因的表达呈负相关。此外，与野生型相比，pde-miR171在长日照条件下的过表达诱导了更大的叶片和更早的开花。
The miR171 is a conserved microRNA (miRNA) family and has been shown to participate in plant growth and development. However, the precise function of miR171 in Pinus densata remains largely unclear. Mature miR171 sequence comparison reveals high similarity between Arabidopsis thaliana and P. densata and the pre-miR171 could fold into a characteristic stem-loop hairpin structure. Genes encoding GRAS (GAI-RGA-SCR) family transcription factors and actin binding protein were identified as targets of pde-miR171 using a modified RNA ligase mediated 5’ rapid amplification of cDNA ends (RLM-RACE). Furthermore, the interaction between pde-miR171 and Arabidopsis SCL6 (SCARECROW-LIKE6) was further validated through transient co-expression of both genes in Nicotiana benthamiana leaves. Next, results of real-time quantitative PCR demonstrated that the expression of pde-miR171 was significantly up-regulated in miR171-overexpressing plants than in wild-type plants, which was inversely correlated with the expression of Arabidopsis SCL6 genes. In addition, overexpression of pde-miR171 in Arabidopsis induced larger leaves and earlier flowering under long-day conditions compared with the wild type.