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。
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 CYP86 A,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 PtCYP86 Aand was driven by the 35 S 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.
出处
《草业学报》
CSCD
北大核心
2017年第6期105-110,共6页
Acta Prataculturae Sinica
基金
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)
甘肃省科技支撑项目(144FKCA058)资助
