大豆维生素E合成酶基因的克隆与功能分析

发布时间：2018-05-08 10:39

本文选题：大豆 + 维生素E　；参考：《华中农业大学》2016年硕士论文

【摘要】：大豆是一种重要的粮油作物,是天然维生素E的主要来源。维生素E作为一种重要的脂溶性抗氧化剂,不仅具参与动物体的中枢神经系统、心血管系统和肌肉系统的正常代谢而且还与植物体细胞膜的构建和维持关系密切。由于具有很好美容保健效果,天然维生素E的市场需求很大。目前维生素E的研究集中在模式植物上,对大豆维生素E知道较少。基于以上背景,本研究利用代谢工程的手段,通过过量表达维生素E合成途径中的基因,比较每个基因超量表达对维生素E含量的具体影响;通过对大豆种子不同发育时期的分析,掌握在发育时期大豆种子维生素E变化规律;通过对大豆植株进行逆境胁迫处理,得到具体逆境下维生素E的变化规律,进一步找到对应胁迫的应答基因。取得具体结果如下:基因转入野生拟南芥进行超量表达:GmHPT-1、GmHPT-2、GmHPT-3的超表达株系相对于野生型维生素E含量分别提高了20.3%、11.2%、38.5%;GmMPBQ-1和GmMPBQ-2的超表达株系相对于野生型维生素E含量分别提高了7.3%和11.2%,其中,α-生育酚和野生型相比分别提高了6倍和7.3倍;GmTC-1和GmTC-2的超表达株系相对于野生型维生素E含量分别提高了10.9%和7.8%;GmTMT-2和GmTMT-3的超表达株系相对于野生型维生素E总含量多数株系有所降低,α-生育酚占总生育酚的比例分别提高到了42%和33%。种子发育时维生素E的变化规律:在黄豆种子的发育过程中,维生素E总含量是上升的,其中γ-生育酚和δ-生育酚含量一直升高,α-生育酚含量先降低后升高;在黑豆种子的发育过程中,维生素E的含量是先升高后降低的,其中γ-生育酚含量先升高后不变,δ-生育酚和α-生育酚含量先升高后降低。大豆植株在非生物逆境下维生素E含量及其基因的变化规律:在低温胁迫下,随着处理时间的增加,总维生素E的含量逐渐增加,第24小时维生素E总含量是对照的3.3倍,胁迫下GmHPT-3基因、GmMPBQ-1基因、GmTC-1基因和GmTMT-2基因转录本很高,揭示了这些基因在低温胁迫中起重要作用。
[Abstract]:Soybean is an important crop of grain and oil, and is the main source of natural vitamin E. As an important lipid soluble antioxidant, vitamin E is not only involved in the normal metabolism of central nervous system, cardiovascular system and muscle system, but also closely related to the construction and maintenance of plant cell membrane. As a result of very good beauty care effect, the market demand of natural vitamin E is very big. At present, the research of vitamin E is focused on the model plant, and little is known about soybean vitamin E. Based on the above background, this study used metabolic engineering to overexpression the genes in the vitamin E synthesis pathway, and compared the specific effects of each gene overexpression on the vitamin E content. Through the analysis of the different development stages of soybean seeds, the changes of vitamin E in soybean seeds were grasped during the development period, and the changes of vitamin E in soybean seeds under specific stress were obtained through the stress treatment of soybean plants. Furthermore, the corresponding stress response gene was found. The specific results are as follows: the overexpression of GmHPT-1 and GmHPT-2 + GmHPT-3 of transgenic wild Arabidopsis thaliana increased the content of vitamin E of wild type by 20.33.11.2GmMPBQ-1 and GmMPBQ-2, respectively, relative to the content of vitamin E of wild type of Arabidopsis thaliana, the overexpression lines of GmHPT-1 and GmHPT-2, GmHPT-2 and GmHPT-3 increased respectively the content of vitamin E in wild type. The over-expression of 伪 -tocopherol and wild-type GmTC-1 and GmTC-2 increased by 10.9% and 7.8%, respectively, compared with wild type, and the over-expression of GmTC-1 and GmTC-2 were 10.9% and 7.8% higher than that of wild type, respectively. The percentage of 伪 -tocopherol to total tocopherol increased to 42% and 33% respectively. The changes of vitamin E during seed development: during the development of soybean seeds, the total content of vitamin E increased, and the contents of 纬 -tocopherol and 未 -tocopherol increased, and the content of 伪 -tocopherol decreased first and then increased. During the development of black bean seeds, the content of vitamin E increased first and then decreased. The content of 纬 -tocopherol increased first and then remained unchanged, and the contents of 未 -tocopherol and 伪 -tocopherol increased first and then decreased. Changes of Vitamin E content and its Gene in Soybean plants under abiotic stress: under low temperature stress, the total vitamin E content increased gradually with the increase of treatment time, and the total vitamin E content at 24 hours was 3.3 times of that of the control. The high transcripts of GmTC-1 and GmTMT-2 genes of GmHPT-3 gene GmMPBQ-1 under stress suggest that these genes play an important role in low temperature stress.
【学位授予单位】：华中农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S565.1

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