【摘要】：作為植物一大類次生代謝產(chǎn)物,黃酮類化合物在植物的生長(zhǎng)發(fā)育中起重要作用,參與植物花、果實(shí)、種子的著色并參與植物與環(huán)境之間的互作。同時(shí),黃酮類化合物對(duì)人體的身體健康有促進(jìn)作用。黃酮代謝通路上有許多酶將底物催化成諸多衍生物,例如查爾酮異構(gòu)酶(chalcone isomerase CHI)。CHI的位置處于苯丙烷代謝通路的一個(gè)分支點(diǎn),催化查爾酮到黃烷酮的反應(yīng)。該反應(yīng)產(chǎn)生的底物在之后的代謝通路上生成許多衍生物,這些衍生物在植物的生長(zhǎng)發(fā)育和與外界互作的過(guò)程中起重要的作用。本文從中間錦雞兒中克隆到了一個(gè)CHI基因并對(duì)其功能進(jìn)行了研究,具體結(jié)果如下:1.中間錦雞兒CiCHIL基因gDNA全長(zhǎng)1434bp、cDNA全長(zhǎng)為941bp、ORF為630bp,5'-UTR長(zhǎng)164bp,3'-UTR長(zhǎng)147bp,共編碼209個(gè)氨基酸,其中包含一段12個(gè)腺苷酸組成的Poly A尾,共含4個(gè)外顯子和3個(gè)內(nèi)含子。2.構(gòu)建過(guò)表達(dá)載體p35S::CiCHIL,并轉(zhuǎn)化野生型擬南芥,得到純合體株系。3.檢測(cè)過(guò)表達(dá)植株當(dāng)中總黃酮含量,過(guò)表達(dá)CiCHI基因的擬南芥中總黃酮含量高于野生型擬南芥。4.利用實(shí)時(shí)熒光定量PCR檢測(cè)了 UV-B處理下CiCHIL的表達(dá)情況,發(fā)現(xiàn)CiCHIL受紫外誘導(dǎo)。對(duì)野生型和過(guò)表達(dá)植株進(jìn)行紫外處理,發(fā)現(xiàn)紫外處理后過(guò)表達(dá)株系表現(xiàn)出較野生型更強(qiáng)的質(zhì)膜穩(wěn)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.
【學(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：Q943.2

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