JcERF1基因RNA干扰载体的构建、转化及沉默效果研究
The construction of RNAi vector of JcERF1 gene, transfom tobaccos and silence result analysis
作者：韩海洋(四川大学生命科学学院)；孙 焕(四川大学生命科学学院)；王雪华(四川大学生命科学学院)；严 君(四川大学生命科学学院)；魏 炜(四川大学生命科学学院)
Author：HAN Hai Yang(College of Life Sciences, Sichuan University)；SUN Huan(College of Life Sciences, Sichuan University)；WANG Xue Hua(College of Life Sciences, Sichuan University)；YAN Jun(College of Life Sciences, Sichuan University)；WEI Wei(College of Life Sciences, Sichuan University)
收稿日期：2014-04-22 年卷（期）页码：2015,52(5):1164-1170
期刊名称：四川大学学报: 自然科学版
Journal Name：Journal of Sichuan University (Natural Science Edition)
关键字：RNA干扰, 载体构建, 转基因, 荧光定量PCR
Key words：RNA interference; Vector construction; Transgenic; Fluorescent quantitation PCR
基金项目：国家自然科学基金（31071448）; 四川省应用基础研究项目（2014JY0051）
中文摘要
构建了麻疯树(Jatropha curcas)一个新的ERF类转录因子基因RNA干扰(RNAi)载体,并将之转入过表达JcERF1基因烟草体内, 得到了RNA干扰型烟草. 利用实时荧光定量PCR(QPCR)检测了JcERF1基因的表达量, 发现该基因在干扰型烟草中表达量明显降低. 通过野生型、过表达型和干扰型烟草的种子萌发实验和高盐胁迫实验发现: 与过表达型烟草相比, 干扰型烟草的抗盐能力明显降低, 且干扰型烟草的游离脯氨酸和可溶性糖含量在高盐处理后上升的幅度明显低于过表达型烟草. 以上研究结果说明JcERF1基因在干扰型烟草体内已被有效沉默, 同时说明该基因具有提高转基因烟草抗盐性的功能.
英文摘要
In order to study the function of the JcERF1 gene, the RNAi vector of JcERF1 was designed and transformed into the overexpression tobaccos, then obtained RNAi tobaccos. The expression level of JcERF1 in RNAi tobaccos was detected by QPCR. The salt resistances of them were tested by the experiment of seeds germination and salty treated experiment. The results showed that the expression levels of JcERF1 gene, the contents of free proline and soluble sugar, and the resistance of salt were significantly reduced in RNAi tobaccos compared with overexpression tobaccos. These results suggested that the JcERF1 gene was silenced successfully in the RNAi tobaccos, and JcERF1 gene confers salt tolerances in transgenic tobaccos.
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