本文選題：草莓 + 堿性/中性轉化酶��； 參考：《甘肅農(nóng)業(yè)大學》2017年碩士論文

【摘要】：糖分不僅作為能源物質參與植物生長發(fā)育、形態(tài)建成,還被認為是一種信號分子響應環(huán)境脅迫、調(diào)控植物生長發(fā)育,進而影響果實品質形成。草莓是世界著名的漿果,果實中糖分積累對品質具有極其重要的影響。本研究通過研究不同草莓品種中糖代謝及轉運相關基因與果實糖含量和組分、糖代謝酶活性及與糖含量相關性,及其進化、表達和對多種激素、外源糖及病原真菌的響應,明確其對草莓糖分積累的分子機制。主要結果如下:(1)草莓果實可溶性糖由葡萄糖、果糖和蔗糖構成,其中糖分積累以蔗糖為主;有機酸主要由檸檬酸和蘋果酸構成,其中檸檬酸的含量是蘋果酸的3-4倍;果實發(fā)育過程中蔗糖的積累與堿性/中性轉化酶活性呈顯著負相關;(2)草莓堿性/中性轉化酶基因(Fa.A/N-Invs)家族由8個成員組成,在本研究分析的樣品中檢測到其中4個成員(Fa.A/N-Inv A,Fa.A/N-Inv B,Fa.A/N-Inv C,Fa.A/N-Inv E)轉錄表達;(3)Fa.A/N-Invs在各個組織中均有表達,成熟葉片中表達最顯著,表明堿性/中性轉化酶能夠參與植物多種發(fā)育過程;果實發(fā)育過程中,Fa.A/N-Invs表達模式與酶活性變化吻合,說明該家族基因轉錄水平的調(diào)控是果實品質調(diào)節(jié)的重要環(huán)節(jié)之一;(4)Fa.A/N-Invs能響應外源IAA、GA和6-BA;在蔗糖堿性/中性轉化酶酶促反應中,蔗糖和葡萄糖分別作為底物和產(chǎn)物,對Fa.A/N-Invs表達起促進和反饋抑制作用,蔗糖類似物松二糖也能誘導Fa.A/N-Invs差異表達,說明草莓堿性/中性轉化酶基因參與多種激素和糖分相關信號通路;(5)草莓葉片受炭疽病病原菌C.fructicola侵染后Fa.A/N-Invs的表達量上調(diào)、A/N-Inv活性升高,其中Fa.A/N-Inv B/E對病原菌的早期響應最敏感,說明草莓堿性/中性轉化酶能通過影響碳水化合物分配參與草莓對病原真菌的防衛(wèi)響應;(6)"紅顏"果實自白果期開始蔗糖含量迅速增加,"申陽"果實蔗糖積累緩慢,果實成熟時"紅顏"果實蔗糖含量(50.74mg·g-1FW)約為"申陽"果實的2倍(23.18mg·g-1FW),進行蔗糖轉運蛋白家族基因(FaSUTs)的表達分析表明,"紅顏"草莓隨著果實發(fā)育FaSUT1、FaSUT6的表達量呈現(xiàn)上升趨勢,FaSUT2穩(wěn)定表達,FaSUT7僅在成熟果實中表達,"申陽"草莓中FaSUTs在成熟果實中幾乎不表達,說明FaSUT1-2、FaSUT6-7在成熟期果實的轉錄表達差異可能是導致兩個品種果實蔗糖含量差異的原因之一。本研究表明:草莓堿性/中性轉化酶在幼果期通過分解光合產(chǎn)物促進碳水化合物再分配進而影響果實發(fā)育;果實進入膨大期堿性/中性轉化酶活性下降,果實糖分開始迅速積累;果實成熟時,蔗糖代謝酶凈活性趨于零,蔗糖轉運蛋白家族基因轉錄產(chǎn)物積累上調(diào),調(diào)控果實蔗糖輸入,促進果實蔗糖積累。
[Abstract]:Sugar not only participates in plant growth and morphogenesis as energy material, but also is considered as a signal molecule in response to environmental stress, regulating plant growth and development, and then affecting fruit quality formation. Strawberry is a famous berry in the world. The accumulation of sugar in the fruit has an extremely important effect on the quality. In this study, we studied the relationship between sugar metabolism and transport related genes, sugar content and components, activities of sugar metabolism enzymes and their correlation with sugar content, their evolution, expression and response to many hormones, exogenous sugar and pathogenic fungi in different strawberry varieties. The molecular mechanism of sugar accumulation in strawberry was clarified. The main results are as follows: (1) the soluble sugar in strawberry fruit is composed of glucose, fructose and sucrose, the sugar accumulation is mainly sucrose, the organic acid is mainly composed of citric acid and malic acid, the content of citric acid is 3-4 times that of malic acid. There was a significant negative correlation between the accumulation of sucrose and alkaline / neutral invertase activity during fruit development. (2) Strawberry alkaline / neutral invertase gene (Fa.A / N-Invs) family was composed of eight members. Four members of Fa.Ar N-Inv Fa.ArN-Inv / Fa.Ar / N-Inv / Fa.ArN-Inv / Fa.ArN-Inv Evs were found to be expressed in all tissues, and the most significant expression was found in mature leaves, indicating that alkaline / neutral invertase can participate in many plant development processes. During fruit development, the expression pattern of Fa.A / N-Invs coincided with the change of enzyme activity, which indicated that the regulation of transcription level of genes in this family was one of the important links in fruit quality regulation, and Fa.ArN-Invs could respond to exogenous IAAGA and 6-BA.In the enzymatic reaction of sucrose alkaline / neutral invertase, Sucrose and glucose, as substrates and products, promoted and inhibited the expression of Fa.A/N-Invs, and sucrose analogue, loose disaccharide, also induced differential expression of Fa.A/N-Invs. The results showed that alkaline / neutral invertase gene was involved in many hormone and sugar related signaling pathways. (5) the expression of Fa.A/N-Invs in strawberry leaves was up-regulated by anthracnose pathogen C.fructicola, and Fa.A/N-Inv B / E was the most sensitive to the early response of the pathogen. The results showed that alkaline / neutral invertase could participate in the defense response of strawberry to pathogenic fungi by affecting carbohydrate distribution. The sucrose content of "Hongyan" fruit increased rapidly at the beginning of the fruiting period, but the accumulation of sucrose in "Shenyang" fruit was slow. The sucrose content of "Hongyan" fruit at maturity was about 2 times that of "Shenyang" fruit, and the expression analysis of sucrose transporter family gene FaSUTsshowed that the expression of FaSUT6 in "Hongyan" strawberry increased with the development of fruit. The stable expression of FaSUT2 was only expressed in mature fruit, but FaSUTs in "Shenyang" strawberry was hardly expressed in mature fruit. It was suggested that the difference of transcription expression of FaSUT1-2 and FaSUT6-7 in mature fruit might be one of the reasons leading to the difference of sucrose content between the two varieties. The results showed that alkaline / neutral invertase promoted carbohydrate redistribution by decomposition of photosynthates in young fruits and decreased the activity of alkaline / neutral invertase at the stage of fruit expansion. The sugar content began to accumulate rapidly and the net activity of sucrose metabolism enzyme tended to be zero when fruit matured. The accumulation of transcription products of sucrose transporter family was up-regulated which regulated the sucrose input and promoted the accumulation of sucrose in fruit.
【學位授予單位】：甘肅農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S668.4

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