过量表达棉花CBF2基因提高转基因拟南芥抗旱耐盐能力

发布时间：2019-03-08 12:38

【摘要】：CBF/DREB是一类植物中特有的转录因子,在植物抵抗逆境胁迫过程中发挥重要功能。本研究从陆地棉(Gossypium hirsutum L.)Coker 312中克隆获得1个棉花CBF/DREB基因,命名为Gh CBF2,该基因编码一个由216个氨基酸组成的CBF蛋白。序列分析结果显示,Gh CBF2与其他植物的CBF蛋白类似,含有AP2转录因子典型的保守结构域。干旱或高盐胁迫处理明显增加了Gh CBF2基因的表达量。亚细胞定位分析结果发现Gh CBF2定位在细胞核中。将Gh CBF2基因构建到由35S启动子调控的植物表达载体p MD上并转化拟南芥(Arabidopsis thaliana L.),结果表明,在干旱和盐胁迫条件下,过量表达Gh CBF2基因拟南芥的成活率显著高于野生型,并且游离脯氨酸和可溶性糖含量也高于野生型,说明转Gh CBF2基因提高了拟南芥的耐盐抗旱能力。采用实时荧光定量PCR方法分析胁迫相关标记基因COR15A、RD29A和ERD6的表达情况,结果显示转基因株系中的表达量显著高于野生型,说明Gh CBF2参与调控拟南芥干旱和盐胁迫相关基因的表达。
[Abstract]:CBF/DREB is a special transcription factor in plants, which plays an important role in the process of plant resistance to adversity stress. In this study, a cotton CBF/DREB gene was cloned from upland cotton (Gossypium hirsutum L.) Coker 312 and named Gh CBF2, which encodes a CBF protein consisting of 216 amino acids. Sequence analysis showed that Gh CBF2 was similar to other plants in CBF protein and contained a typical conserved domain of AP2 transcription factor. Drought or high salt stress significantly increased the expression of Gh CBF2 gene. Subcellular localization analysis showed that Gh CBF2 was localized in the nucleus. The Gh CBF2 gene was constructed into the plant expression vector p MD regulated by 35s promoter and transformed into Arabidopsis thaliana (Arabidopsis thaliana L.),. The results showed that the survival rate of Arabidopsis thaliana overexpressing Gh CBF2 gene under drought and salt stress was significantly higher than that of wild type Arabidopsis thaliana. The contents of free proline and soluble sugar in Arabidopsis thaliana were also higher than those in wild type, which indicated that the transgenic Gh CBF2 gene improved the salt tolerance and drought resistance of Arabidopsis thaliana. Real-time fluorescence quantitative PCR was used to analyze the expression of stress-related marker genes COR15A,RD29A and ERD6. The results showed that the expression of COR15A,RD29A and ERD6 in transgenic lines was significantly higher than that in wild-type lines. These results suggest that Gh CBF2 is involved in regulating the expression of drought and salt stress related genes in Arabidopsis thaliana.
【作者单位】：喀什大学生命与地理科学学院新疆维吾尔自治区教育厅叶尔羌绿洲生态与生物资源研究重点实验室;新疆师范大学生命科学学院;华中师范大学生命科学学院;
【基金】：新疆维吾尔自治区高校科研计划科学研究重点项目(XJEDU2014I037) 喀什大学高层次人才科研启动经费项目(GCC16ZK-002)~~
【分类号】：Q943.2

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