本文選題：甘藍(lán)型油菜 + 擬南芥�。� 參考：《華中農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：維生素E,包括生育酚和生育三烯酚,是一種重要的脂溶性有機(jī)物,它具有很強(qiáng)的抗氧化性,在動(dòng)植生命中有著不可替代的重要地位。人類對(duì)維生素E的需求量巨大,但其中90%左右靠人工合成。天然維生素E主要來(lái)源于植物葉片和油料作物種子,人工合成的維生素E在生物活性上與天然維生素E相差甚遠(yuǎn)。因此提高綠色植物特別是油料作物種子中維生素E的含量就顯得尤為重要。油菜是世界上最重要的油料作物之一,油菜種子富含維生素E且種植面積大,因此油菜維生素E育種日漸成為研究的熱點(diǎn)之一,越來(lái)越受到育種工作者的關(guān)注。GWAS分析表明,油菜BnA9CHLSYN位點(diǎn)對(duì)種子中生育酚含量有著很強(qiáng)的效應(yīng)。因此猜測(cè)油菜種子中生育酚合成的“瓶頸”在于葉綠素合成酶對(duì)生育酚合成的前體物質(zhì)PDP進(jìn)行“回收”,重新合成葉綠素,導(dǎo)致PDP濃度過(guò)低。想要打破這一“瓶頸”,就有必要研究葉綠素合成酶(Chlorophyll synthetase,CHLSYN)與生育酚合成之間的關(guān)系。本研究借助擬南芥這一載體研究種子中CHLSYN的表達(dá)量與種子中生育酚合成之間的關(guān)系,主要研究結(jié)果如下:1.不同甘藍(lán)型油菜的BnA9CHLSYN位點(diǎn)對(duì)種子中生育酚含量的效應(yīng)不同,據(jù)此將這些材料分為兩組,H組和L組(分別對(duì)應(yīng)BnA9CHLSYN位點(diǎn)對(duì)種子中生育酚含量呈正向效應(yīng)和負(fù)向效應(yīng)),H組材料種子中BnA9CHLSYN的表達(dá)水平低于L組材料種子中該基因的表達(dá)水平,生育酚含量相反。2.野生型背景下,采用特異性啟動(dòng)子干擾擬南芥種子中CHLSYN的表達(dá)之后,種子中生育酚含量提高,最多可提高33.69%,種子特異性超表達(dá)CHLSYN,最多可使擬南芥種子中生育酚含量降低44.73%,即擬南芥種子中CHLSYN的表達(dá)水平與生育酚含量呈負(fù)相關(guān)關(guān)系。擬南芥vte5純合突變體背景下也得到相同的結(jié)論。3.采用相同種子特異性啟動(dòng)子干擾擬南芥種子中CHLSYN的同時(shí)超表達(dá)AtHPT,種子中生育酚含量最多能提高125.38%,而在種子中特異性超表達(dá)AtHPT最多可使生育酚含量提高123.56%,兩者未達(dá)到顯著差異。4.更換超表達(dá)AtHPT的啟動(dòng)子,采用不同種子特異性啟動(dòng)子干擾擬南芥種子中CHLSYN的同時(shí)超表達(dá)AtHPT,種子中生育酚含量最多能提高161.71%,而使用新的啟動(dòng)子超表達(dá)AtHPT之后,最多只能使生育酚含量提高95.99%。
[Abstract]:Vitamin E, including tocopherol and reproductive trienol, is an important lipophilic organic substance, which has strong antioxidant activity and plays an irreplaceable role in plant life. Human demand for vitamin E is enormous, but about 90% of it is artificially synthesized. The natural vitamin E mainly comes from plant leaves and oil crop seeds. The synthetic vitamin E is far from natural vitamin E in biological activity. Therefore, it is very important to increase the content of vitamin E in green plants, especially oil crop seeds. Rape is one of the most important oil crops in the world. Rape seeds are rich in vitamin E and have large planting area. The BnA9CHLSYN site of rape has a strong effect on the content of tocopherol in seeds. Therefore, it is speculated that the bottleneck of tocopherol synthesis in rapeseed seeds is that chlorophyll synthetase "recovers" the precursor of tocopherol synthesis, PDP, and resynthesizes chlorophyll, which leads to the low concentration of PDP. In order to break this bottleneck, it is necessary to study the relationship between chlorophyll synthetase Chlorophyll synthetic enzyme CHLSYN and tocopherol synthesis. In this study, Arabidopsis thaliana was used to study the relationship between the expression of CHLSYN and the synthesis of tocopherol in seeds. The main results were as follows: 1. The effect of BnA9CHLSYN locus on the content of tocopherol in different Brassica napus was different. These materials were divided into two groups: group H and group L (the positive and negative effects of BnA9CHLSYN locus on the content of tocopherol in seeds). The expression level of BnA9CHLSYN in H group was lower than that in L group. The content of tocopherol is opposite. Under the wild type background, the content of tocopherol in Arabidopsis thaliana seeds increased after the specific promoter was used to interfere with the expression of CHLSYN in Arabidopsis thaliana seeds. At most, the content of tocopherol in Arabidopsis thaliana seeds was decreased by 44.73%. The expression level of CHLSYN in Arabidopsis thaliana seeds was negatively correlated with the content of tocopherol. The same conclusion was obtained under the background of Arabidopsis vte5 homozygous mutants. The content of tocopherol in Arabidopsis thaliana seeds could be increased by using the same seed specific promoter to interfere with the simultaneous overexpression of AtHPT.The content of tocopherol in seeds could be increased by 125.38%, but the content of tocopherol in seeds could be increased by 123.56 at most, but there was no significant difference between the two. 4. When the promoter of AtHPT was replaced with different seed specific promoters which interfered with CHLSYN in Arabidopsis thaliana seeds, the content of tocopherol in seeds increased at most 161.71%, but after the new promoter was used to express AtHPT in Arabidopsis thaliana seeds, the content of tocopherol in seeds was increased by 161.71%. At most, the content of tocopherol increased 95.99%.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S565.4

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