中间锦鸡儿CiCHIL的克隆与功能分析

发布时间：2019-03-14 11:50

【摘要】：作为植物一大类次生代谢产物,黄酮类化合物在植物的生长发育中起重要作用,参与植物花、果实、种子的着色并参与植物与环境之间的互作。同时,黄酮类化合物对人体的身体健康有促进作用。黄酮代谢通路上有许多酶将底物催化成诸多衍生物,例如查尔酮异构酶(chalcone isomerase CHI)。CHI的位置处于苯丙烷代谢通路的一个分支点,催化查尔酮到黄烷酮的反应。该反应产生的底物在之后的代谢通路上生成许多衍生物,这些衍生物在植物的生长发育和与外界互作的过程中起重要的作用。本文从中间锦鸡儿中克隆到了一个CHI基因并对其功能进行了研究,具体结果如下:1.中间锦鸡儿CiCHIL基因gDNA全长1434bp、cDNA全长为941bp、ORF为630bp,5'-UTR长164bp,3'-UTR长147bp,共编码209个氨基酸,其中包含一段12个腺苷酸组成的Poly A尾,共含4个外显子和3个内含子。2.构建过表达载体p35S::CiCHIL,并转化野生型拟南芥,得到纯合体株系。3.检测过表达植株当中总黄酮含量,过表达CiCHI基因的拟南芥中总黄酮含量高于野生型拟南芥。4.利用实时荧光定量PCR检测了 UV-B处理下CiCHIL的表达情况,发现CiCHIL受紫外诱导。对野生型和过表达植株进行紫外处理,发现紫外处理后过表达株系表现出较野生型更强的质膜稳定性。
[Abstract]:As a class of secondary metabolites of plants, flavonoids play an important role in the growth and development of plants, participate in the coloring of plant flowers, fruits and seeds, and participate in the interaction between plants and the environment. At the same time, flavonoids can promote the health of human body. There are many enzymes in flavonoids metabolism pathway which catalyze substrate to many derivatives, such as chalcone isomerase (chalcone isomerase CHI). Chi is located at a branch point of phenylpropanone metabolism pathway, and catalyzes the reaction of chalcone to flavanone. The substrate produced by this reaction produces many derivatives on the subsequent metabolic pathway, which play an important role in the growth and development of plants and in the process of interaction with the outside world. In this paper, a CHI gene was cloned from Caragana intermedium and its function was studied. The results are as follows: 1. The full-length gDNA of CiCHIL gene of Caragana intermedium is 941bp, 630bp, 164bp and 147bp respectively. It encodes 209 amino acids, including a Poly A tail consisting of 12 adenylates, which is composed of 12 adenylates, and the total length is 1434bp.ORF is 630bp.UTR is 164bp and 147bp.UTR is 1434bp.ORF is 630bp. There are 4 exons and 3 introns. 2. The expression vector p35S / CiCHIL was constructed and transformed into wild type Arabidopsis thaliana to obtain a homozygous strain. The content of total flavonoids in over-expressed plants was higher than that in wild-type Arabidopsis thaliana. 4. The expression of CiCHI gene in Arabidopsis thaliana was higher than that in wild-type plants. Real-time fluorescence quantitative PCR was used to detect the expression of CiCHIL under UV-B treatment. It was found that CiCHIL was induced by ultraviolet. After UV treatment of wild and over-expressed plants, it was found that the over-expressed lines showed stronger plasmalemma stability than that of wild plants.
【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

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