盐生植物小花碱茅CYP86A基因的RNAi载体构建

doi:10.11686/cyxb2017076

草业学报 ›› 2017, Vol. 26 ›› Issue (6): 105-110.DOI: 10.11686/cyxb2017076

盐生植物小花碱茅CYP86A基因的RNAi载体构建

王沛^1,2, 崔彦农¹, 高丽¹, 王锁民^1,*

1.草地农业生态系统国家重点实验室,兰州大学草地农业科技学院,甘肃兰州 730020;
2.西南民族大学青藏高原研究院,四川成都 610041

收稿日期:2017-03-02 修回日期:2017-04-10 出版日期:2017-06-20 发布日期:2017-06-20
通讯作者: E-mail:smwang@lzu.edu.cn
作者简介:王沛(1987-),男,甘肃庆阳人,在读博士。E-mail:wp_10@lzu.edu.cn
基金资助:
国家自然科学基金(31470503),教育部博士点基金优先发展领域项目(20130211130001)和甘肃省科技支撑项目(144FKCA058)资助

Construction of RNAi expression vector of CYP86A gene in halophyte Puccinellia tenuiflora

WANG Pei^1,2, CUI Yan-Nong¹, GAO Li¹, WANG Suo-Min^1,*

1.State Key Laboratory of Grassland Agro-ecosystems, College of Pastoral Agriculture Science and Technology, Lanzhou University, Lanzhou 730020, China;
2.Institution of Qinghai-Tibetan Plateau, Southwest Minzu University, Chengdu 610041, China

Received:2017-03-02 Revised:2017-04-10 Online:2017-06-20 Published:2017-06-20

摘要/Abstract

摘要： 拒盐型盐生植物小花碱茅根系具有高度木栓化的内皮层可能是其抵御盐胁迫的重要策略之一。本研究以盐胁迫下小花碱茅幼苗的根为材料,采用RT-PCR方法扩增到木栓质生物合成关键基因CYP86A的RNAi靶片段,以中间载体pHANNIBAL和植物表达载体pART27为基础,采用酶切连接结合In-Fusion的克隆方法成功构建了以35S启动子驱动,含PtCYP86A基因片段反向重复序列的RNAi植物表达载体pARC,为利用RNAi技术深入探究根系木栓化在小花碱茅拒盐中的功能奠定基础。

Abstract: The salt-excluding halophyte Puccinellia tenuiflora exhibits a highly suberized endodermis in the root, which may contribute to its salt tolerance. In this study, the RNAi target fragment of CYP86A, a key gene in suberin biosynthesis, was amplified by RT-PCR from roots of P. tenuiflora seedlings under salt stress. Then based on the intermediate vector pHANNIBAL and the plant expression vector pART27, the RNAi plant expression vector pARC, which contained inverted repeats of PtCYP86A and was driven by the 35S promoter, was constructed successfully by restriction enzyme digestion, ligation and In-Fusion cloning. The work might be a good basis for further investigating the roles of roots suberization in Na⁺ exclusion in P. tenuiflora by RNAi technique.

王沛, 崔彦农, 高丽, 王锁民. 盐生植物小花碱茅CYP86A基因的RNAi载体构建[J]. 草业学报, 2017, 26(6): 105-110.

WANG Pei, CUI Yan-Nong, GAO Li, WANG Suo-Min. Construction of RNAi expression vector of CYP86A gene in halophyte Puccinellia tenuiflora[J]. Acta Prataculturae Sinica, 2017, 26(6): 105-110.

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盐生植物小花碱茅CYP86A基因的RNAi载体构建

Construction of RNAi expression vector of CYP86A gene in halophyte Puccinellia tenuiflora

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