【摘要】：生物柴油是一種可再生的生物能源,可以作為化石燃油的替代品,在緩解能源危機(jī)和減少環(huán)境污染方面有重要作用,通過(guò)基因工程方法調(diào)控植物體中脂肪酸的合成可以提高植物中的含油量,從而為制造生物柴油提供更多的原材料。油體是植物細(xì)胞中主要的儲(chǔ)油細(xì)胞器,而油體鈣蛋白(Caleosin)作為油體結(jié)構(gòu)的重要組成成分,與油體的形成、油體的數(shù)量、油體結(jié)構(gòu)的穩(wěn)定以及植物細(xì)胞中油脂的積累有密切的關(guān)系。煙草具有繁殖快速,生物量大,遺傳轉(zhuǎn)化體系成熟等優(yōu)點(diǎn),通過(guò)基因工程手段提高煙草葉片中的油脂含量可以用來(lái)提取油脂制備生物柴油,為此本研究在做了以下試驗(yàn)：1.通過(guò)逆轉(zhuǎn)錄PCR方法克隆了油料作物蓖麻的油體鈣蛋白(Caleosin)基因,并對(duì)該基因在蓖麻不同組織器官以及不同發(fā)育時(shí)期種子中的表達(dá)情況進(jìn)行了半定量表達(dá)分析,結(jié)果表明,蓖麻Caleosin基因在嫩葉、莖、幼根、開(kāi)花后20d、30d、40d的種子中檢測(cè)到了表達(dá),在種子中的表達(dá)量要高于在其他組織中的表達(dá)量,并且主要集中在種子發(fā)育中后期(油脂積累期)表達(dá),說(shuō)明Caleosin基因的表達(dá)與油脂積累過(guò)程是同步的。2.將克隆的蓖麻Caleosin基因與植物表達(dá)載體pCAMBIA 1303連接,構(gòu)建了含有蓖麻Caleosin基因的重組植物表達(dá)載體pCAMBIA-RcClo。利用農(nóng)桿菌介導(dǎo)的葉盤法將重組表達(dá)載體轉(zhuǎn)入煙草,獲得了轉(zhuǎn)基因煙草植株。3.對(duì)獲得的轉(zhuǎn)基因煙草植株的葉片進(jìn)行脂肪酸含量測(cè)定,結(jié)果表明轉(zhuǎn)基因煙草葉片的脂肪酸含量比野生型煙草葉片提高了8.1%。
[Abstract]:Biodiesel is a renewable bio-energy source that can be used as an alternative to fossil fuels and plays an important role in alleviating the energy crisis and reducing environmental pollution, Controlling the synthesis of fatty acids in plants by genetic engineering can increase the oil content in plants and provide more raw materials for the manufacture of biodiesel. Oil body is the main organelle of oil storage in plant cells. As an important component of oil body structure, (Caleosin) is closely related to the formation of oil body, the quantity of oil body, the stability of oil body structure and the accumulation of oil in plant cells. Tobacco has the advantages of rapid reproduction, large biomass and mature genetic transformation system. Increasing the oil content in tobacco leaves by genetic engineering can be used to extract oil to produce biodiesel. In this study, the following experiments were carried out: 1. The oil troponin (Caleosin) gene of oil crop castor was cloned by reverse transcription PCR. The expression of the gene in different tissues and organs of castor and seeds at different developmental stages was analyzed by semi-quantitative analysis. The expression of ricin Caleosin gene was detected in young leaves, stems, roots and seeds 20 days after anthesis. The expression level in seeds was higher than that in other tissues, and it was mainly expressed in the middle and late stage of seed development (oil accumulation stage). It is suggested that the expression of Caleosin gene is synchronized with the accumulation of oil. 2. The recombinant plant expression vector pCAMBIA-RcCloin containing castor plant Caleosin gene was constructed by ligating the cloned ricin Caleosin gene with plant expression vector pCAMBIA 1303. The recombinant expression vector was transformed into tobacco by Agrobacterium tumefaciens mediated leaf disc method, and transgenic tobacco plant. 3. 3 was obtained. The fatty acid content of transgenic tobacco leaves was measured. The results showed that the fatty acid content of transgenic tobacco leaves was 8.1% higher than that of wild tobacco leaves.
【學(xué)位授予單位】：山西農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：Q943.2;S565.6

【相似文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前2條

1 李傳寶;蓖麻Caleosin基因的克隆及功能驗(yàn)證[D];山西農(nóng)業(yè)大學(xué);2016年

2 丁勇;甘藍(lán)型油菜caleosin基因克隆與表達(dá)[D];華中農(nóng)業(yè)大學(xué);2006年

，

本文編號(hào)：2165007