http://www.ahs.ac.cn/images/0513-353X/images/top-banner1.jpg|#|苹果
http://www.ahs.ac.cn/images/0513-353X/images/top-banner2.jpg|#|甘蓝
http://www.ahs.ac.cn/images/0513-353X/images/top-banner3.jpg|#|菊花
http://www.ahs.ac.cn/images/0513-353X/images/top-banner4.jpg|#|灵芝
http://www.ahs.ac.cn/images/0513-353X/images/top-banner5.jpg|#|桃
http://www.ahs.ac.cn/images/0513-353X/images/top-banner6.jpg|#|黄瓜
http://www.ahs.ac.cn/images/0513-353X/images/top-banner7.jpg|#|蝴蝶兰
http://www.ahs.ac.cn/images/0513-353X/images/top-banner8.jpg|#|樱桃
http://www.ahs.ac.cn/images/0513-353X/images/top-banner9.jpg|#|观赏荷花
http://www.ahs.ac.cn/images/0513-353X/images/top-banner10.jpg|#|菊花
http://www.ahs.ac.cn/images/0513-353X/images/top-banner11.jpg|#|月季
http://www.ahs.ac.cn/images/0513-353X/images/top-banner12.jpg|#|菊花

园艺学报 ›› 2019, Vol. 46 ›› Issue (5): 864-876.doi: 10.16420/j.issn.0513-353x.2018-0924

• 研究论文 • 上一篇    下一篇

黄瓜microRNA171的克隆与功能分析

朱早兵,虞夏清*,翟于菲,王盼乔,赵勤政,李 季,娄群峰,陈劲枫*   

  1. 南京农业大学园艺学院,作物遗传与种质创新国家重点实验室,南京 210095
  • 出版日期:2019-05-25 发布日期:2019-05-25
  • 基金资助:

    国家自然科学基金重点项目(31430075);江苏省基础研究计划(自然科学基金)青年基金项目(BK20180536);中国博士后科学基金面上项目(2017M611834)

Cloning and Functional Analysis of microRNA171 in Cucumber

ZHU Zaobing,YU Xiaqing*,ZHAI Yufei,WANG Panqiao,ZHAO Qinzheng,LI Ji,LOU Qunfeng,and CHEN Jinfeng*   

  1. College of Horticulture,Nanjing Agricultural University,National Key Laboratory of Crop Genetics and Germplasm Enhancement,Nanjing 210095,China
  • Online:2019-05-25 Published:2019-05-25

摘要:

为了解microRNA171(miR171)对黄瓜的调控作用,从华北型黄瓜‘北京截头’中克隆了Csa-miR171家族的4个miRNA前体序列Csa-miR171a、Csa-miR171b、Csa-miR171c、Csa-miR171d,对前体序列进行比对与进化分析,预测成熟序列的靶基因并进一步开展了拟南芥遗传转化功能研究。结果表明,4个前体共剪切出两种成熟序列Csa-miR171a和Csa-miR171b,系统进化显示,Csa-miR171a、Csa-miR171b、Csa-miR171c、Csa-miR171d分别与甜瓜Cme-miR171e、Cme-miR171c、Cme-miR171a、Cme-miR171b同源性最近,与拟南芥、水稻、番茄等较远;Csa-miR171a、Csa-miR171b成熟序列主要靶向GRAS转录因子蛋白家族基因,拟南芥中过表达Csa-miR171a前体改变了叶片形态、颜色、抽薹时间、根、果实、花器官形态等多个表型性状。

关键词: 黄瓜, microRNA171, 克隆, 靶基因预测, 转基因

Abstract:

To investigate the function of microRNA171(miR171)in cucumber,four precursors Csa-miR171a,Csa-miR171b,Csa-miR171c,Csa-miR171d of Csa-miR171 family genes were cloned in the northern Chinese cucumber‘Beijingjietou’and followed by sequence alignment and phylogenetic analysis. Target genes of the mature sequence of Csa-miR171 were predicted and functional analysis was performed in Arabidopsis by genetic transformation. The results showed that the two mature sequences,Csa-miR171a and Csa-miR171b were cleaved from the four Csa-miR171 precursors. Phylogenetic analysis showed that Csa-miR171a,Csa-miR171b,Csa-miR171c,Csa-miR171d were closer to Cme-miR171e,Cme-miR171c,Cme-miR171a,Cme-miR171b in melon than those in Arabidopsis,rice and tomato,etc. GRAS transcription factor family was predicted to be the main target gene of Csa-miR171. Overexpression of Csa-miR171a in Arabidopsis significantly affected the phenotypes including leaf morphology and leaf color,blooming time,root,fruit and floral morphology.

Key words: cucumber, microRNA171, clone, target gene prediction, transgenic

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