拟南芥AtSEC14基因反义RNA植株的获得及抗病性检测
发布时间：2018-08-31 18:40
【摘要】：为确定拟南芥抗灰霉病相关基因AtSEC14的功能,本试验构建了AtSEC14基因的反义RNA载体;通过农杆菌介导的遗传转化方法,将其转化拟南芥野生型Col-0中;利用潮霉素抗性筛选和PCR检测,获得了阳性转基因植株。利用半定量RT-PCR技术,在转基因株系中未检测到AtSEC14基因的表达,说明该基因反义RNA载体的转入能特异影响AtSEC14基因的表达,表明试验所获得的转基因植株是AtSEC14基因的反义RNA转基因植株。对所获得的反义RNA转基因植株进行抗病性鉴定,发现反义RNA转基因植株对灰葡萄孢的敏感性增强,表明AtSEC14基因在拟南芥抗灰葡萄孢过程中起正调控的作用。
[Abstract]:In order to determine the function of Arabidopsis cinerea resistance gene AtSEC14, the antisense RNA vector of AtSEC14 gene was constructed, transformed into Arabidopsis wild-type Col-0 by Agrobacterium tumefaciens mediated genetic transformation, hygromycin resistance screening and PCR detection were used. Positive transgenic plants were obtained. The expression of AtSEC14 gene was not detected in transgenic lines by semi-quantitative RT-PCR technique, which indicated that the transfer of antisense RNA vector could specifically affect the expression of AtSEC14 gene. The results showed that the transgenic plants were antisense RNA transgenic plants of AtSEC14 gene. The antisense RNA transgenic plants were identified for disease resistance, and it was found that the antisense RNA transgenic plants were more sensitive to grapevine spores, indicating that the AtSEC14 gene plays a positive role in the process of resistance of Arabidopsis thaliana to grapevine spores.
【作者单位】： 河北农业大学真菌毒素与植物分子病理学实验室;河北北方学院农林科技学院;
【基金】：河北省自然科学基金项目(C2014405010)
【分类号】：S432.4
，
本文编号：2215835
[Abstract]:In order to determine the function of Arabidopsis cinerea resistance gene AtSEC14, the antisense RNA vector of AtSEC14 gene was constructed, transformed into Arabidopsis wild-type Col-0 by Agrobacterium tumefaciens mediated genetic transformation, hygromycin resistance screening and PCR detection were used. Positive transgenic plants were obtained. The expression of AtSEC14 gene was not detected in transgenic lines by semi-quantitative RT-PCR technique, which indicated that the transfer of antisense RNA vector could specifically affect the expression of AtSEC14 gene. The results showed that the transgenic plants were antisense RNA transgenic plants of AtSEC14 gene. The antisense RNA transgenic plants were identified for disease resistance, and it was found that the antisense RNA transgenic plants were more sensitive to grapevine spores, indicating that the AtSEC14 gene plays a positive role in the process of resistance of Arabidopsis thaliana to grapevine spores.
【作者单位】： 河北农业大学真菌毒素与植物分子病理学实验室;河北北方学院农林科技学院;
【基金】：河北省自然科学基金项目(C2014405010)
【分类号】：S432.4
，
本文编号：2215835
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