發(fā)布時間：2018-05-11 08:09

  本文選題：脫落酸 + 果實成熟�。� 參考：《中國農(nóng)業(yè)大學》2017年博士論文

【摘要】：果實是果樹類園藝作物的重要經(jīng)濟器官,是人們?nèi)粘ｏ嬍车闹匾�組成部分和營養(yǎng)物質(zhì)來源。但是由于果實的成熟與衰老無法有效的控制,導致果品在生產(chǎn)、貯藏和流通過程中造成巨大的經(jīng)濟損失和資源浪費。因此,我們應該積極開展對果實成熟衰老的生理生化及其分子機制的研究,探索調(diào)控果實成熟和延長貨架期的有效方法,以滿足人們對高質(zhì)量果品供應的需求。脫落酸(ABA)是重要的植物激素,在果實成熟過程中起著十分重要的調(diào)控作用。然而ABA對果實成熟調(diào)控機制還缺少分子水平的證據(jù)。本文以非呼吸躍變型果實櫻桃和躍變型果實模式植物番茄為試材,利用基因沉默技術(shù),研究了 ABA代謝與信號轉(zhuǎn)導途徑關(guān)鍵基因?qū)�果實成熟的調(diào)控作用,為闡明ABA調(diào)控果實發(fā)育的分子機制提供了大量證據(jù)。取得的主要結(jié)果如下:1.本文利用病毒誘導的基因沉默(VIGS)技術(shù)鑒定了櫻桃果實中ABA降解途徑關(guān)鍵酶ABA 8'-羥化酶基因PacCYP707A1的功能。PacCYP707A1沉默的櫻桃果實成熟過程沒有受到影響,ABA代謝基因與花青素合成基因的表達也沒有明顯的變化。但是在離體果實失水的過程中,PacCYP707A1-VIGS果實積累更高水平的ABA,導致果實的失水率降低。這些結(jié)果表明PacCYP707A1調(diào)控果實的內(nèi)源ABA含量進而影響櫻桃離體果實的失水率。2.本研究利用VIGS鑒定了番茄果實中ABA生物合成關(guān)鍵酶9-順式-環(huán)氧類胡蘿卜素雙加氧酶基因SlNCED1與ABA 8'-羥化酶基因SlCYP707A2的功能。研究發(fā)現(xiàn),SlNCED1與SlCYP707A2是調(diào)控番茄果實ABA合成與降解的關(guān)鍵基因,且對果實成熟分別具有正調(diào)控和負調(diào)控作用。這些結(jié)果表明通過ABA的生物合成與降解途徑調(diào)控果實中ABA的含量可以影響番茄果實的成熟。3.本研究利用RNA干擾(RNAi)技術(shù)研究了番茄中ABA葡萄糖基轉(zhuǎn)移酶基因SlUGT75C1的生理功能。SlUGT75C1沉默的番茄果實中ABA含量升高而ABA-GE的含量降低,表明SlUGT75c1能在植物體內(nèi)通過ABA糖基化途徑調(diào)控ABA的內(nèi)穩(wěn)態(tài)。SlUGT75C1-RNAi果實由于ABA含量的升高導致成熟提前,但果實中類胡蘿卜素的含量降低。這些結(jié)果表明SlUGT75C1介導的ABA糖結(jié)合途徑在調(diào)控的果實成熟過程中起著重要的作用。4.本文篩選出了參與番茄ABA信號轉(zhuǎn)導的關(guān)鍵2C型蛋白磷酸酶成員S1PP2C3。利用RNAi在植株中抑制SlPP2C3的表達導致植株在種子休眠與萌發(fā)、幼苗初生根生長,響應干旱脅迫等方面都表現(xiàn)出典型的ABA高度敏感表型,表明S1PP2C3是番茄ABA信號的負調(diào)控因子。SlPP2C3-RNAi番茄果實成熟顯著提前,且乙烯釋放量明顯升高。通過基因表達分析發(fā)現(xiàn)S1PP2C3可能通過調(diào)節(jié)乙烯合成基因ACOs與乙烯受體基因ETRs的表達調(diào)控果實成熟。此外,轉(zhuǎn)錄組數(shù)據(jù)表明S1PP2C3介導的ABA信號可能參與調(diào)控果實的角質(zhì)層形成。綜上所述,本研究證明了櫻桃與番茄中多個ABA代謝途徑與信號轉(zhuǎn)導關(guān)鍵基因?qū)�果實的成熟具有重要的調(diào)控功能,同時證明了通過生物技術(shù)操縱ABA相關(guān)基因調(diào)控果實成熟和品質(zhì)的可能性。本研究的結(jié)果為揭示果實發(fā)育與成熟調(diào)控網(wǎng)絡奠定了基礎(chǔ),為人工調(diào)控果實的成熟提供了理論基礎(chǔ),也為通過定向分子育種改良果品提供了新思路。
[Abstract]:Fruit is an important economic organ of fruit tree horticultural crops. It is an important part of people's daily diet and a source of nutrients. However, due to the failure of the fruit to be effectively controlled, the fruit can cause huge economic loss and waste of resources in the process of production, storage and circulation. Therefore, we should actively carry out the problem. The physiological, biochemical and molecular mechanisms of fruit ripening and senescence are studied to explore effective methods to regulate fruit maturity and prolong shelf life to meet the demand for high quality fruit supply. Abscisic acid (ABA) is an important plant hormone and plays a very important regulatory role in fruit ripening. However, ABA is a regulatory machine for fruit ripening. In this paper, there is no evidence of molecular level. In this paper, the control effect of key genes of ABA metabolism and signal transduction pathway on fruit ripening was studied by using gene silencing technique. A lot of evidence was provided to elucidate the molecular mechanism of controlling fruit development by ABA. The main results are as follows: 1. in this paper, the virus induced gene silencing (VIGS) technique was used to identify the key enzyme of ABA degradation pathway ABA 8'- hydroxylase gene PacCYP707A1 in cherry fruit. The maturation process of cherry fruit ripening was not affected by the function of.PacCYP707A1 silencing, and the expression of the ABA metabolic gene and anthocyanin synthesis gene was not obvious. But in the process of water loss in the isolated fruit, the PacCYP707A1-VIGS fruit accumulates a higher level of ABA, which leads to the decrease of the water loss rate of the fruit. These results show that PacCYP707A1 regulates the endogenous ABA content of fruit and then affects the water loss rate of cherry in vitro.2.. This study used VIGS to identify the key enzyme of the key enzyme of ABA biosynthesis in tomato fruit, 9- The function of CIS CIS carotenoid bioxygenase gene SlNCED1 and ABA 8'- hydroxylase gene SlCYP707A2. It is found that SlNCED1 and SlCYP707A2 are key genes regulating the synthesis and degradation of tomato fruit ABA, and have positive regulation and negative regulation on fruit ripening. These results show that the biosynthesis and degradation routes of ABA are carried out. The content of ABA in fruit can affect the mature.3. of tomato fruit. The study of RNA interference (RNAi) technique to study the physiological function of ABA glucosyltransferase gene SlUGT75C1 in tomato, the content of ABA in tomato fruit with.SlUGT75C1 silencing is increased and the content of ABA-GE decreased, indicating that SlUGT75c1 can be carried through ABA glycosylation in plants. The endogenous.SlUGT75C1-RNAi fruit of ABA regulated the maturity of the fruit due to the increase of ABA content, but the content of carotenoid in the fruit decreased. These results showed that the SlUGT75C1 mediated ABA sugar binding pathway played an important role in the regulation of fruit ripening,.4. this paper screened the key to participate in the signal transduction of tomato ABA. The 2C type protein phosphatase member S1PP2C3. uses RNAi to inhibit the expression of SlPP2C3 in the plant, resulting in the seed dormancy and germination, the growth of the primary root of the seedlings, and the typical ABA highly sensitive phenotype in response to drought stress, which indicates that S1PP2C3 is a negative regulating factor of the ABA signal of tomato, and the fruit maturity of the tomato is significantly advanced in advance. By gene expression analysis, S1PP2C3 may regulate fruit ripening by regulating the expression of ethylene synthesis gene ACOs and ethylene receptor gene ETRs. In addition, the transcriptional data indicate that S1PP2C3 mediated ABA signals may be involved in regulating the formation of the cuticle in fruit. Multiple ABA metabolic pathways and key signal transduction genes in tomatoes have important regulatory functions on fruit ripening. At the same time, the possibility of regulating fruit maturity and quality by manipulating ABA related genes through biotechnology has been proved. The results of this study have laid a foundation for revealing the fruit development and maturation control network and for artificial regulation of fruit. Maturity provides a theoretical basis, and provides a new way of improving fruit by directional molecular breeding.

【學位授予單位】：中國農(nóng)業(yè)大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：S641.2;S662.5

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本文編號：1873142