本文選題：不結球白菜 + 蘇州青��； 參考：《南京農(nóng)業(yè)大學》2016年碩士論文

【摘要】：抗壞血酸(L-Ascorbic acid,AsA),又稱維生素C,是動植物體中廣泛存在的一種水溶性抗氧化劑(Gallie 2013)。植物中合成的AsA不僅能使無法合成AsA的人類及一些動物獲得豐富的維生素C源,而且還能維持植物體的正常生長發(fā)育和代謝(安華明2004)。近年來廣泛通過基因工程技術提高農(nóng)作物的AsA含量,不僅有助于人體獲得較高水平的維生素C,同時也對增強植株自身的抗性,提高農(nóng)產(chǎn)品的營養(yǎng)品質具有重要影響。本文探究了脅迫和激素處理下不結球白菜抗壞血酸含量的變化與相關基因表達量之間的關聯(lián),為不結球白菜的優(yōu)質育種奠定理論基礎。本文主要研究結果如下:1.本研究旨在了解脅迫下不結球白菜抗壞血酸含量的變化,進一步找出調(diào)節(jié)抗壞血酸含量積累的關鍵基因。以'蘇州青'為材料,分別進行了過氧化氫脅迫和水淹脅迫處理,測定了其AsA含量以及合成代謝相關基因的表達量。結果表明,兩種處理下AsA含量均呈先上升后下降的趨勢,過氧化氫脅迫下GME和GPP基因表達量的變化趨勢與AsA含量變化趨勢相一致,而水淹脅迫下PMM、GME和VTC2基因表達量的變化趨勢與AsA含量變化趨勢一致,說明GME基因可能是兩種脅迫下均發(fā)揮作用的關健基因。2.以'蘇州青'為材料,分別以0、10、50、100和200 μmol·L-1的脫落酸和乙烯處理,旨在了解激素處理下不結球白菜抗壞血酸的含量變化,進一步找出調(diào)節(jié)AsA含量變化的關鍵基因。結果顯示:不同濃度ABA處理后AsA含量變化趨勢不同,基因表達量分析發(fā)現(xiàn)較低濃度ABA(10 μmol·L-1,50μmol·L-1和100 μmol·L-1)處理組PMM基因表達量變化趨勢與AsA含量變化趨勢一致。乙烯處理后AsA含量呈先升高后下降趨勢;10 μmol·L-1和50 μmol·L-1乙烯處理組GPP基因表達量變化趨勢與AsA含量變化趨勢一致,而100 μmol·L-1和200 μmol·L-1乙烯處理組PMM和GPP基因表達量變化趨勢與AsA含量變化趨勢一致。因此推測在脫落酸和乙烯處理下,PMM和GPP基因可能分別是其AsA生物合成途徑的關鍵基因。3.以不結球白菜'蘇州青'為材料,同源克隆了BcG E基因的全長,應用生物信息學分析了其氨基酸序列,并且構建了 BcGME-PCVA沉默載體。結果顯示,BcGME基因包含一個長度為1137 bp的開放式閱讀框(ORF),編碼379個氨基酸。推測其蛋白理論相對分子質量為42.97×103,理論等電點pI值為5.84,分子式為C1907H2949N519O570S22,屬于不穩(wěn)定蛋白�？偲骄杷笖�(shù)為-0.440,屬于親水蛋白,亞細胞定位預測該蛋白分布在細胞質中。BcGME氨基酸序列與十字花科大部分植物的相似性為90%以上,不結球白菜BcGME與十字花科的大白菜、甘藍型油菜、擬南芥相應蛋白的進化距離較近。并且不結球白菜和擬南芥GME蛋白三級結構相似,推測BcGME基因在抗壞血酸合成途徑中起到關鍵作用。
[Abstract]:L-Ascorbic ascorbic acid, also known as vitamin C, is a kind of water-soluble antioxidant, Gallie 2013, which is widely found in animals and plants. The AsA synthesized in plants can not only make the human and some animals that can't synthesize AsA obtain rich vitamin C sources, but also maintain the normal growth and metabolism of plants (Anhua 2004. In recent years, increasing the AsA content of crops by genetic engineering technology is not only helpful to obtain higher levels of vitamin C in human body, but also has an important effect on enhancing the resistance of plants and improving the nutritional quality of agricultural products. In this paper, the relationship between ascorbic acid content and gene expression in non-heading cabbage under stress and hormone treatment was investigated, which laid a theoretical foundation for quality breeding of non-heading Chinese cabbage. The main results of this paper are as follows: 1: 1. The purpose of this study was to find out the changes of ascorbic acid content in non-heading Chinese cabbage under stress and to find out the key genes to regulate the accumulation of ascorbic acid content. The content of AsA and the expression of genes related to metabolism were determined by hydrogen peroxide stress and water flooding stress. The results showed that the AsA content increased first and then decreased under both treatments, and the change trend of GME and GPP gene expression under hydrogen peroxide stress was consistent with that of AsA content. However, the change trend of GME and VTC2 gene expression was the same as that of AsA content under waterlogging stress, indicating that GME gene may be the key gene. 2. In order to understand the changes of ascorbic acid content in Chinese cabbage without hormone treatment and to find out the key genes to regulate the change of AsA content, 'Suzhou Qing' was treated with abscisic acid (Abscisic acid) and ethylene (200 渭 mol L ~ (-1), respectively, in order to understand the changes of ascorbic acid content in Chinese cabbage without hormone treatment. The results showed that the change trend of AsA content was different after different concentration of ABA-treated. The results of gene expression analysis showed that the change trend of AsA gene expression was the same as that of AsA content in the low concentration ABA 10 渭 mol L ~ (-1) 50 渭 mol ~ (-1) and 100 渭 mol L ~ (-1) groups. After ethylene treatment, the AsA content increased first and then decreased. The change trend of GPP gene expression in ethylene treated with 10 渭 mol L-1 and 50 渭 mol L-1 was consistent with that of AsA content. However, the change trend of PMM and GPP gene expression in ethylene treated with 100 渭 mol L-1 and 200 渭 mol L-1 was the same as that of AsA content. Therefore, it is suggested that PMM and GPP genes may be the key genes of AsA biosynthesis pathway under abscisic acid and ethylene treatment, respectively. The full length of BcGE gene was cloned homologous from 'Suzhou Qing', and its amino acid sequence was analyzed by bioinformatics, and BcGME-PCVA silencing vector was constructed. The results showed that the BcGME gene contained an open reading frame of 1137 BP encoding 379 amino acids. The theoretical molecular weight of the protein is 42.97 脳 10 ~ 3, the theoretical isoelectric point Pi is 5.84, and the molecular formula is C1907H2949N519O570S22, which is an unstable protein. The total average hydrophobic index was -0.440, which belonged to hydrophilic protein. The similarity between the amino acid sequence of BcGME and most plants of Cruciferae was more than 90%, and the similarity between BcGME and Brassica campestris was more than 90%, the subcellular localization predicted that the amino acid sequence of BcGME was more than 90% similar to that of Brassica chinensis. In Brassica napus, the corresponding proteins of Arabidopsis thaliana were close to each other. The tertiary structure of GME protein in non-heading Chinese cabbage and Arabidopsis thaliana is similar. It is suggested that BcGME gene plays a key role in ascorbic acid synthesis pathway.
【學位授予單位】：南京農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：S634.3

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本文編號：2003318