中间锦鸡儿参与叶片衰老基因CibHLH027的克隆和功能研究

发布时间：2017-12-28 13:18

本文关键词：中间锦鸡儿参与叶片衰老基因CibHLH027的克隆和功能研究　出处：《内蒙古农业大学》2017年硕士论文　论文类型：学位论文

【摘要】：叶片衰老是植物生长发育的最后阶段,也是必经的主动应答阶段。当植物经历逆境胁迫,比如黑暗、干旱和极端温度等环境因素,会加速叶片衰老过程,严重影响了植物的生长发育和作物产量。运用基因工程对植物进行改良,能够提高植物在逆境胁迫下的耐受性,增强其适应环境的能力,从而有效延缓衰老。锦鸡儿属(Caragana)植物具有抗寒、抗旱及抗热特性,赋予其适应各种环境中不良因素的能力,是改善生态环境的理想植被,因此也成为研究抗逆机制的优良材料。本研究从中间锦鸡儿(Caragana intermedia)转录组数据库筛选到一个受非生物胁迫诱导的中间锦鸡儿基因CibHLH027,并对其功能进行了研究,主要结果如下:1.CibHLH027编码区长738bp,起始密码子为ATG,终止密码子为TGA,编码245个氨基酸。与拟南芥bHLH蛋白系统进化分析显示,CibHLH027与聚类到同一分支上的 AtbHLH027、AtbHLH035 的一致性分别为 58%和 52%。将 CibHLH027与不同植物中一致性较高的bHLH蛋白进行多序列比对和聚类分析显示,N-端碱性区域都包含N-E-R基序,CibHLH027与同一分支上的鹰嘴豆CabHLH035 一致性为90%。2.CibHLH027的表达受ABA、NaCl、高pH、热、黑暗及干旱的诱导。利用拟南芥叶肉原生质体瞬时表达发现CibHLH027-GFP定位于细胞核中。3.过表达CibHLH027基因的拟南芥,离体叶片和整株植株在黑暗诱导下,都表现出叶片衰老加速,叶绿素含量降低、离子渗透率增大和细胞死亡增多的表型:并且干旱处理下,CibHLH027过表达株系与野生型相比,耐旱能力明显降低。
[Abstract]:Leaf senescence is the last stage of plant growth and development, and it is also a required stage of active response. When plants experience environmental stresses, such as darkness, drought and extreme temperature, they will accelerate leaf senescence and seriously affect plant growth and crop yield. The use of genetic engineering to improve plants can improve the tolerance of plants under stress, enhance their ability to adapt to the environment, and effectively delay aging. Caragana (Caragana) has the characteristics of cold resistance, drought resistance and heat resistance, giving it the ability to adapt to various adverse factors in the environment, and is an ideal vegetation for improving the ecological environment. Therefore, it has also become an excellent material for studying the mechanism of stress resistance. This study from Caragana intermedia (Caragana intermedia) transcriptome database to a screening by Caragana intermedia gene CibHLH027 induced by abiotic stress, and its function is studied, the main results are as follows: 1.CibHLH027 encoding region of 738bp, ATG as initiation codon and termination codon was TGA, encoding 245 amino acids. The phylogenetic analysis of bHLH protein in Arabidopsis thaliana showed that the consistency of CibHLH027 and AtbHLH027 and AtbHLH035 on the same branch was 58% and 52% respectively. Multiple sequence alignment and cluster analysis of bHLH protein with high consistency between CibHLH027 and different plants showed that the alkaline region of N- contains N-E-R motifs, and the consistency of CibHLH027 with chickpea CabHLH035 on the same branch was 90%. The expression of 2.CibHLH027 is induced by ABA, NaCl, high pH, heat, darkness and drought. CibHLH027-GFP was located in the nucleus by transient expression of protoplasts of Arabidopsis thaliana. 3. expression of CibHLH027 gene in Arabidopsis leaves and whole plants in the dark induced, showed accelerated leaf senescence, chlorophyll content decreased, ion permeability increases and increased cell death phenotype and under drought treatment, CibHLH027 overexpression lines compared with the wild type, drought resistant ability decreased significantly.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：Q943.2;S793.3

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