本文選題：玫瑰 + DFR��； 參考：《華中農(nóng)業(yè)大學(xué)》2016年博士論文

【摘要】：玫瑰(Rosa rugosa Thunb.)是一種具有重要經(jīng)濟(jì)價(jià)值的觀賞植物,其花色艷麗、香氣溢人。玫瑰花瓣是提取香料的重要材料,玫瑰精油因價(jià)格高產(chǎn)量低,被人們譽(yù)為液體黃金。除此之外,玫瑰花還具有一定的消炎,活血,抗氧化等。然而目前對(duì)玫瑰的研究主要局限栽培生理和新品種選育上,隨著玫瑰轉(zhuǎn)基因體系的成熟,利用生物技術(shù)提高玫瑰產(chǎn)品附加值成為可能。本課題以玫瑰為材料研究類黃酮合成的分子機(jī)制。我們從玫瑰花瓣中分離得到二氫黃酮醇還原酶(Rr DFR1)、黃酮醇合成酶(Rr FLS1)和花青素還原酶(Rr ANR)并對(duì)它們的功能進(jìn)行了深入研究。主要研究結(jié)果如下:1通過(guò)對(duì)玫瑰、野薔薇、桃、香石竹、杜鵑、山茶以及矮牽牛7個(gè)物種的紅花白花進(jìn)行色素成分分析,發(fā)現(xiàn)紅花和白花中都能檢測(cè)到黃酮醇,而花青素在白花中幾乎檢測(cè)不到。通過(guò)對(duì)紅花白花中類黃酮合成相關(guān)基因的表達(dá)分析,表明FLS和DFR的競(jìng)爭(zhēng)性表達(dá)與花青素和黃酮醇的積累有關(guān)。為了進(jìn)一步闡明這種調(diào)控規(guī)律的作用,我們從玫瑰、矮牽牛和桃中分離了FLS基因,從玫瑰和矮牽牛中分離DFR基因。煙草轉(zhuǎn)基因結(jié)果表明,超表FLS能夠增強(qiáng)黃酮醇的合成,減少花青素的積累,從而使轉(zhuǎn)基因煙草花色由紅色變?yōu)榘咨?相反,超量表達(dá)DFR基因,抑制內(nèi)源Nt FLS的表達(dá),促進(jìn)花青素合成。基于上述結(jié)果,我們認(rèn)為FLS和DFR對(duì)共同底物的競(jìng)爭(zhēng),分別實(shí)現(xiàn)黃酮醇和花青素的合成,從而決定紅白花顏色的分化。2利用FPNI-PCR技術(shù)從玫瑰中分離得到Rr ANR,構(gòu)建超量表達(dá)載體轉(zhuǎn)化煙草。轉(zhuǎn)基因煙草花色變白,原花青素和ABA含量增加,抗氧化能力增強(qiáng)。通過(guò)對(duì)Rr ANR轉(zhuǎn)基因煙草和未轉(zhuǎn)基因煙草轉(zhuǎn)錄組測(cè)序結(jié)果的分析,結(jié)果表明轉(zhuǎn)基因煙草中有1586個(gè)基因上調(diào)表達(dá),其中包括相關(guān)抗逆基因和次生代謝物合成的相關(guān)基因,特別是ABA途徑和抗氧化酶系統(tǒng)。為了進(jìn)一闡明Rr ANR提高植物抗逆性的分子機(jī)理,從煙草中分離了Nt ABF。超表達(dá)Nt ABF顯著提高植株抗逆性以及抗氧化酶相關(guān)基因的表達(dá)。外源PA和ABA處理同樣可以上調(diào)Nt ABF和抗氧化酶相關(guān)基因的表達(dá)。上述結(jié)果表明超量表達(dá)Rr ANR提高植物抗性是通過(guò)活性氧的清除和ABA信號(hào)實(shí)現(xiàn)的。
[Abstract]:Rosa rugosa Thunb. is an important ornamental plant with an important economic value. The rose petals are an important material for the extraction of spices. Rose essential oil is regarded as a liquid gold because of high price and low yield. In addition, the rose also has some anti-inflammatory, blood activating, antioxidant and so on. However, rose is currently on the rose. The research mainly limited the cultivation physiology and the selection of new varieties. With the maturity of the rose transgenic system, it is possible to use biotechnology to increase the added value of the Rose products. This topic studies the molecular mechanism of the flavonoid synthesis by the rose as the material. We isolated the two flavonol reductase (Rr DFR1) from the rose petals, and the synthesis of flavonol Enzyme (Rr FLS1) and anthocyanin reductase (Rr ANR) and their functions are deeply studied. The main results are as follows: 1 through the analysis of the pigment composition of rose, Rosa, peach, carnation, Rhododendron, camellia and Petunia, the flavonols can be detected in red flower and white flower, and anthocyanin is in white flower. The expression of the flavonoid synthesis related genes in safflower white flowers showed that the competitive expression of FLS and DFR was related to the accumulation of anthocyanin and flavonol. In order to further clarify the role of this regulation, we separated the FLS gene from roses, Petunia and peaches, and separated DFR from roses and Petunia. Gene. The results of tobacco transgenic show that FLS can enhance the synthesis of flavonol, reduce the accumulation of anthocyanin and make the color of transgenic tobacco change from red to white. On the contrary, overexpressing DFR gene, inhibiting the expression of endogenous Nt FLS and promoting the synthesis of anthocyanin. Based on the above results, we think that the competition of FLS and DFR to common substrates is divided. Do not achieve the synthesis of flavonols and anthocyanins, thus determining the differentiation of red and white flower color.2 using FPNI-PCR technology to separate Rr ANR from roses and construct a super expression vector to convert tobacco. The color of the transgenic tobacco is white, the content of proanthocyanidins and ABA is increased, and the antioxidant capacity is enhanced. Rr ANR transgenic tobacco and untransgenic tobacco are used. The results of the transcriptional sequence analysis showed that 1586 genes were up-regulated in transgenic tobacco, including related genes and related genes of secondary metabolites, especially the ABA pathway and antioxidant enzyme system. In order to clarify the molecular mechanism of Rr ANR to improve plant stress resistance, the Nt ABF. overexpression N was isolated from tobacco. T ABF significantly increased plant stress resistance and expression of antioxidant enzymes related genes. Exogenous PA and ABA treatments could also increase the expression of Nt ABF and antioxidant enzymes related genes. The above results showed that the overexpression of Rr ANR increased plant resistance by scavenging of reactive oxygen species and ABA signal.

【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：S685.12

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