普通野生稻miR160f的克隆和功能分析

生物技术通报 ›› 2014, Vol. 0 ›› Issue (11): 114-118.

普通野生稻miR160f的克隆和功能分析

杨松楠¹,王姣²,陈宗祥²,田新杰¹,张静文²,龙艳²,裴新梧²,袁潜华¹

1. 海南大学农学院,海口 570228；2. 中国农业科学院生物技术研究所,北京 100081

收稿日期:2014-04-22 出版日期:2014-11-07 发布日期:2014-11-07
作者简介:杨松楠,男,硕士研究生,研究方向：植物基因工程
基金资助:
国家转基因重大专项2014ZX08011-001,公益性行业（农业）科研专项经费项目（201403075）,教育部热带作物新品种选育工程中心联合资助项目

Clone and Function Analysis MiR160f in Common Wild Rice(Oryza rufipogon Griff.)

Yang Songnan¹,Wang Jiao²,Chen Zongxiang²,Tian Xinjie¹,Zhang Jingwen²,Long Yan²,Pei Xinwu²,Yuan Qianhua¹

1. College of Agriculture,Hainan University,Haikou 570228；2. Biotechnology Research Institute,Chinese Academy of Agricultural Sciences,Beijing 100081

Received:2014-04-22 Published:2014-11-07 Online:2014-11-07

摘要/Abstract

摘要： MicroRNAs是一类在调节基因转录后表达中起重要作用的非编码RNA。miR160通过调节生长素响应因子（ARF）参与根细胞的分裂和分化,从而影响根的发育。克隆了普通野生稻miR160f基因,并将其转入拟南芥中鉴定功能。结果表明,过表达miR160f的拟南芥莲座叶的数量减少,抽薹时间缩短,开花的时间提前。qPCR检测显示miR160f的靶基因ARF10、ARF16及ARF17在过表达拟南芥植株中的表达下调,而ARF10和ARF16蛋白的缺失或减少会导致根冠细胞分化受阻、分裂失控,并导致根尖干细胞群的异位扩大,因此可以表明miR160不仅影响根的发育,还可能与水稻的开花时间相关。

关键词: miRNA, 花期, 普通野生稻, 拟南芥, 表达载体

Abstract: MicroRNAs are a class of non-coding RNAs involved in post- transcriptional control of gene expression. Former study on Arabidopsis thaliana reveals that miR160 involves in root cell division and differentiation by regulating auxin response factors（ARF）thus influence the root development. This study cloned miR160f gene from common wild rice and transferred into Arabidopsis thaliana to identify its function. The results showed that over-expression of miR160f decreased the number of rosette leaves, shortened bloting time, leading to early flowering. RT-PCR showed the expressions of gene ARF10,ARF16 and ARF17 are down-regulation caused by miR160f in transgenic Arabidopsis thaliana, while the deficiency of ARF10 and ARF16 protein can inhibit root cap cell differentiation, lose control of cell division and lead to ectopic expansion of apical stem cell populations Therefore, the result demonstrates that miR160 not only influence on root development, but also may influence flowering time of common wild rice.

Key words: MiRNA , Flowering time, Common wild rice, Arabidopsis thaliana , Expression vector

杨松楠,王姣,陈宗祥,田新杰,张静文,龙艳,裴新梧,袁潜华. 普通野生稻miR160f的克隆和功能分析[J]. 生物技术通报, 2014, 0(11): 114-118.

Yang Songnan,Wang Jiao,Chen Zongxiang,Tian Xinjie,Zhang Jingwen,Long Yan,Pei Xinwu,Yuan Qianhua. Clone and Function Analysis MiR160f in Common Wild Rice(Oryza rufipogon Griff.)[J]. Biotechnology Bulletin, 2014, 0(11): 114-118.

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