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[摘要]
目的 构建人参PgMYB4基因表达载体,并研究该基因对拟南芥的抗旱功能。方法 采用RT-PCR技术克隆人参PgMYB4基因,构建表达载体,通过农杆菌介导的花序浸蘸法将其转化到拟南芥中,获得转基因拟南芥,以野生型拟南芥作对照,检测植株与抗旱相关的生理指标。结果 获得的人参PgMYB4基因全长为735 bp,编码245个氨基酸,预测其相对分子质量为27 914;在干旱胁迫条件下,转基因拟南芥的生长状态明显优于野生型拟南芥,与野生型相比,转基因拟南芥叶片相对叶绿素量降低幅度小、脯氨酸量显著升高、水丢失率较低。结论 人参PgMYB4基因具有抗干旱胁迫的能力。
[Key word]
[Abstract]
Objective To construct the expression vector of PgMYB4 gene in Panax ginseng and study its function of the drought resisting in Arabidopsis thaliana. Methods A P. ginseng gene PgMYB4 was cloned by RT-PCR analysis, further, recombinant plasmid vector with PgMYB4 was transformed into wild-type plants of A. thaliana by Agrobacterium tumefacies-mediated Floral Dip method. Transgenic A. thaliana with the expression of PgMYB4 was obtained, further compared with wild-type plants of A. thaliana, their determination of physiologic index related to drought stress was detected. Results The cDNA (named a PgMYB4) contained a 735 bp open reading frame, encoded 245 amino acids and the predicted molecular weight was 27.914 KDa; Under the condition of drought stress, the growth of transgenic A. thaliana was obviously better than wild-type A. thaliana. Compared with wild-type A. thaliana, the decreased range of relative chlorophyll content in the leaves of transgenic plants of A. thaliana was smaller and the proline content of transgenic plants of A. thaliana increased significantly. The water loss of transgenic plants of A. thaliana was less than that of Wild-type transgenic plants of A. thaliana. Conclusion Ginseng PgMYB4 gene has the ability of resistance to drought stress.
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[基金项目]
国家自然科学基金青年基金项目(81503212);吉林省省级经济结构战略调整引导资金专项项目(2014N155)