本文關鍵詞：李果實蘋果酸轉運體的克隆表達及其有機酸的關聯(lián)性分析　出處：《福建農林大學》2017年碩士論文　論文類型：學位論文

  更多相關文章： 李 蘋果酸轉運體 有機酸 表達分析 關聯(lián)分析

【摘要】：有機酸是決定果實品質的重要因子,參與了果實風味、色澤、香氣的形成,在果實中的組成、轉運與積累機理是果實品質研究的重要課題。果實細胞中,蘋果酸轉運蛋白tDT(tonoplast dicarboxylate transporter)和蘋果酸跨膜離子通道蛋白ALMT(almuminum-activated malate transporter)基因家族介導胞質中蘋果酸跨膜轉運,對果實有機酸積累與降解機理研究具有重要意義。本研究通過'皇冠李'、'西瓜李'、'黑琥珀'和'葡萄李'等四個代表性品種,應用超高效液相色譜(UPLC)進行果實有機酸組分與含量的分析;通過同源克隆了一個液泡膜蘋果酸轉運蛋白tDT-like基因和兩個蘋果酸轉運離子通道ALMT4-like、ALMT9-like基因,并應用實時熒光定量PCR技術檢測tDT-like、AMM74-like和ALMT9-like基因在4個代表性品種中的5個發(fā)育階段的基因表達水平,并通過pearson相關分析和回歸建模系統(tǒng)地分析了李果實細胞蘋果酸跨膜轉運基因與各有機酸組分的關聯(lián)性,主要研究結果如下:1.李果實生長發(fā)育過程中pH變化呈V型變化,TA呈倒V型變化。黑琥珀、皇冠李、西瓜李的pH變化V型趨勢明顯,葡萄李變化較小。pH值大小順序為:成熟期,葡萄李黑琥珀≈西瓜李皇冠李,完熟期,葡萄李黑琥珀皇冠李≈西瓜李;TA變化與pH相似,黑琥珀、皇冠李、西瓜李的T倒V趨勢明顯,葡萄李變化較小,成熟期和完熟期果實的總酸大小順序為:西瓜李皇冠李黑琥珀葡萄李。高酸和低酸品種的差異主要表現(xiàn)為低酸品種酸降較快或時間較長,反之則酸降較慢或時間較短。2.蘋果酸是李果實中主要的有機酸,占總酸含量的57%～84%,李屬于典型的蘋果酸型果實;其次是酒石酸,個別品種(如西瓜李)高達18%～36%;而果實中的檸檬酸、琥珀酸、草酸、乙酸、富馬酸只以少量或微量存在。4個測試品成熟期果實蘋果酸含量順序為:皇冠李西瓜李≈黑琥珀葡萄李。3.蘋果酸轉運體基因對不同品種果實蘋果酸的調控作用有所差異。tDT-like基因對高蘋果酸含量品種(如皇冠李、西瓜李)果實酸的積累起到一定程度的正向調控作用,對低蘋果酸含量品種(如葡萄李)的調控作用不明顯。ALMT9-like基因對不同品種李果實蘋果酸的積累均表現(xiàn)為負向調控作用。ALMT4-like基因對皇冠李、西瓜李、葡萄李的蘋果酸積累起到起促進作用,但對黑琥珀的蘋果酸積累起到負向調控作用。4.蘋果酸轉運體與有機酸組分的關聯(lián)性分析結果顯示,ALMT4-like基因與蘋果酸、琥珀酸顯著相關,ALMT9-like基因與蘋果酸、檸檬酸顯著相關,與草酸極顯著相關。表明果實細胞中ALMT4-like和ALMT9-like基因除轉運蘋果酸外,可能還參與琥珀酸、檸檬酸等組分的轉運。
[Abstract]:Organic acid is an important factor determining fruit quality. It is involved in the formation of fruit flavor, color and aroma. The mechanism of composition, transportation and accumulation in fruit is an important topic of fruit quality research. Fruit cell, malate transporter tDT (tonoplast dicarboxylate transporter) and malic acid transmembrane ion channel protein ALMT (almuminum-activated malate transporter) gene mediated by cytoplasmic malate transport across the membrane, has important significance to research on fruit organic acid accumulation and degradation of machine. The study by Lee 'crown' and 'Lee' and 'watermelon Black Amber' and 'Lee' four grape cultivars, using ultra performance liquid chromatography (UPLC) for analysis of organic acids in fruit and content; by homologous cloning of a vacuolar malate transporter gene and tDT-like two malate transporter ALMT4-like ion channel, ALMT9-like gene, 5 stages of development and application of real time fluorescent quantitative PCR detection of tDT-like, AMM74-like and ALMT9-like genes in 4 representative varieties of gene expression level, and through Pearson correlation analysis and regression modeling and systematic analysis of the plum fruit cell malic acid Association cross membrane transporter genes and different organic acids, the main results are as follows: a V type change in the process of growth and development of pH 1. changes of plum fruit, TA presents an inverted V curve. The trend of pH change of Black Amber, crown plum and watermelon plum is obvious, and the change of grape plum is smaller. The size of the pH value of the order: the mature period, Li Li is watermelon Grape Black Amber crown plum, ripe stage, Li Li is Grape Black Amber crown watermelon Lee; the change of TA and pH is similar to that of Black Amber plum, plum, crown watermelon T inverted V trend is obvious, grape Li small changes, the total acid sequence of the mature stage and ripe stage of fruit is watermelon: Li Li Li Grape Black Amber crown. The difference of high acid and low acid variety mainly showed that the acid of low acid decreased faster or longer, and on the contrary, the acid decreased slowly or the time was shorter. 2. malic acid was the main organic acid of plum fruit, the total acid content of 57% ~ 84%, Li belongs to the typical type of fruit malic acid; followed by tartaric acid, individual species (such as watermelon Lee) up to 18% ~ 36%; and in the fruit of citric acid, succinic acid, oxalic acid, acetic acid, fumaric acid in small or trace amounts. 4 test product maturity fruit malic acid content in the order: crown Li Li Li is Black Amber grape watermelon. 3. the regulation of malic acid transporter gene on malic acid in different varieties was different. TDT-like gene played a positive regulatory role in the accumulation of acid in fruits of high malic acid content (such as crown plum, watermelon and plum), and the regulation effect on low malic acid varieties (such as grape and plum) was not obvious. The ALMT9-like gene showed negative regulation on the accumulation of malic acid in different varieties of plum fruit. ALMT4-like gene played a role in promoting malic acid accumulation in crown plum, watermelon plum and grape plum, but played a negative role in malic acid accumulation in Black Amber. 4. the correlation analysis between malic acid transporter and organic acid components showed that ALMT4-like gene was significantly correlated with malic acid and succinic acid. ALMT9-like gene was significantly correlated with malic acid and citric acid, and highly correlated with oxalate. It is suggested that the ALMT4-like and ALMT9-like genes in the fruit cells may also be involved in the transport of succinic and citric acid components, except for the transport of malic acid.
【學位授予單位】：福建農林大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S662.3

【參考文獻】

相關期刊論文 前10條

1 劉玉蓮;車飛;王海;陳佰鴻;陳年來;;蘋果果實中糖、酸和花青苷的組分及含量特征分析[J];西北林學院學報;2016年06期

2 趙宇;蔡麗;陳忠敏;;茶葉中草酸的含量測定[J];食品與發(fā)酵科技;2016年05期

3 鄭惠文;張秋云;李文慧;章世奎;席萬鵬;;新疆杏果實發(fā)育過程中可溶性糖和有機酸的變化[J];中國農業(yè)科學;2016年20期

4 周亮;楊文俠;曾芳;魏珊珊;姚鋒先;鄧利珍;鐘晰;鐘八蓮;;紐荷爾臍橙果實發(fā)育過程中糖、酸和維生素C含量的變化[J];中國南方果樹;2015年02期

5 黃群;陳丹;于麗麗;蔡韋煒;連峗芳;;芙蓉李抗氧化有效部位的篩選及抗氧化活性的比較[J];福建中醫(yī)藥大學學報;2014年05期

6 楊道富;林旗華;謝鴻根;蔡秋英;陳源;卓玉輝;;火龍果果實生長過程有機酸與可溶性糖變化研究[J];福建農業(yè)學報;2012年10期

7 張義;劉敏;;不同顏色李果實成熟期果皮色素的變化及與糖酸含量的相關性[J];北方園藝;2012年10期

8 鐘云;蔣儂輝;姜波;劉巖;曾繼吾;易干軍;;楊桃草酸的分布及其果實草酸含量變化的研究[J];熱帶作物學報;2011年01期

9 胡玲;李麗萍;王友升;;桃李果實風味物質的研究進展[J];安徽農業(yè)科學;2010年07期

10 冒愛榮;;間接原子吸收光譜法測定食品中的草酸[J];食品科學;2009年22期

相關博士學位論文 前3條

1 黃明;刺梨高含量抗壞血酸積累的分子機理研究[D];華中農業(yè)大學;2013年

2 張燕子;不同蘋果糖酸組成及蘋果酸轉運體功能研究[D];西北農林科技大學;2010年

3 劉威生;李種質資源遺傳多樣性及主要種間親緣關系的研究[D];中國農業(yè)大學;2005年

相關碩士學位論文 前4條

1 李鵬;李果實物理化學特性及其抗氧化活性研究[D];福建農林大學;2016年

2 郭忠仁;我國南方中國李種質資源收集和利用研究[D];南京農業(yè)大學;2006年

3 崔艷濤;李果實發(fā)育中內源激素與果實色澤形成的關系[D];河北農業(yè)大學;2006年

4 程和禾;黃金梨果實生長發(fā)育及貯藏生理特性研究[D];河北農業(yè)大學;2004年

，

本文編號：1343859