本文選題：山核桃 + 油脂合成��； 參考：《浙江農(nóng)林大學》2017年碩士論文

【摘要】：山核桃(Carya cathayensis Sarg.)是我國特有的特色優(yōu)質(zhì)干果,具有極高的營養(yǎng)價值,富含優(yōu)質(zhì)脂肪酸和多種微量元素,是高油酸木本油料作物。在成油方面,通過對山核桃果實發(fā)育階段粗脂肪含量和脂肪酸組分的測定,結(jié)合山核桃果實發(fā)育階段的轉(zhuǎn)錄組數(shù)據(jù),分析油脂的來源、合成及降解,構(gòu)建了山核桃成油網(wǎng)絡,揭示了山核桃油脂積累過程中的基因表達規(guī)律。本研究運用實驗和生物信息學的研究手段對山核桃油脂合成過程中的脂肪酸去飽和酶基因家族做了全面分析,以期揭示山核桃高不飽和脂肪酸油脂組分機理,此外,通過轉(zhuǎn)基因手段驗證油脂合成候選基因功能,以期從分子水平解釋山核桃高油機制,篩選出提高含油率和特定脂肪酸含量的關(guān)鍵基因,為選育獲得高油和高品質(zhì)的山核桃品種提供理論依據(jù)。主要結(jié)果如下:1、山核桃油主要由棕櫚酸(C16:0)、硬脂酸(C18:0)、油酸(C18:1)、亞油酸(C18:2)以及亞麻酸(C18:3)五種組分組成,成熟階段的山核桃不飽和脂肪酸含量為92.45%,山核桃是高油酸植物,成熟階段的油酸占總不飽和脂肪酸的比例可達85.57%,此外亞油酸和亞麻酸含量也較為豐富,ω-6型脂肪酸/ω-3型脂肪酸比值為1.69:1,是多不飽和脂肪酸含量的最佳比值,有很高的營養(yǎng)價值。2、擴增得到山核桃SAD家族成員3個,分別是:CcSAD-1;CcSAD-2,CcSAD-3,都含有典型的Ferritin-like superfamily功能結(jié)構(gòu)域,含有1個保守基序(DLLNKYLY),同時,定量結(jié)果表明SAD基因表達在山核桃果實成熟過程中存在差異,隨著CcSADs基因表達上升,山核桃油中油酸含量增加。3、擴增得到山核桃FAD家族成員9個,分別是:CcFAD2;CcFAD3,3個拷貝,記為CcFAD3-1、CcFAD3-2、CcFAD3-3;CcFAD6;CcFAD7;CcFAD8,3個拷貝,記為CcFAD8-1、CcFAD8-2、CcFAD8-3,都含有典型的Membrance-FADS-like super family功能結(jié)構(gòu)域,含有2個保守的組氨酸基序(Hisbox),同時,定量結(jié)果表明FAD基因表達在山核桃果實成熟過程中存在差異,隨著CcFADs基因表達上升,山核桃油中不飽和脂肪酸比例增加。4、構(gòu)建3個油脂合成候選基因轉(zhuǎn)基因體系:35Spro-Uni11、35Spro-Uni72、35SproUni58,35Spro-Uni11轉(zhuǎn)基因擬南芥表型較野生型葉型小、葉片數(shù)多、抽薹率高、側(cè)枝多,但種子油含量未明顯增加;35Spro-Uni72和35Spro-Uni58轉(zhuǎn)基因擬南芥T1代表型較野生型擬南芥?zhèn)戎Χ?株型更飽滿,但是從T2代轉(zhuǎn)基因擬南芥中未再檢測到相應基因,猜測原因是擬南芥的自我保護機制,切除了異源基因。
[Abstract]:Carya cathayensis Sarg.) It is a unique high quality dried fruit with high nutritional value, rich in high quality fatty acids and trace elements, and is a high oleic acid woody oil crop. In the aspect of oil formation, the crude fat content and fatty acid composition of pecan fruit were determined, and the source, synthesis and degradation of oil were analyzed according to the transcriptional data of pecan fruit development stage, and the oil forming network of pecan was constructed. The rule of gene expression during the accumulation of pecan oil was revealed. In this study, the fatty acid desaturase gene family in the synthesis of pecan oil was analyzed by means of experiments and bioinformatics, in order to reveal the mechanism of the fatty acid desaturase gene in pecan oil synthesis. In order to explain the mechanism of high oil content in pecans at molecular level and to screen out the key genes to increase oil content and specific fatty acid content, the function of candidate genes for oil synthesis was verified by transgenic methods. To provide theoretical basis for breeding high oil and high quality pecan varieties. The main results are as follows: the pecan oil is composed mainly of five components: palmitic acid C16: 0, stearic acid C18: 0, oleic acid C18: 1, linoleic acid C18: 2) and linolenic acid (C18: 3). The unsaturated fatty acid content of pecan is 92.455.The pecan is a high oleic acid plant. The ratio of oleic acid to total unsaturated fatty acid in mature stage is 85.57, and the content of linoleic acid and linolenic acid is also relatively rich. The ratio of 蠅 -6 fatty acid to 蠅 -3 fatty acid is 1.69: 1, which is the best ratio of polyunsaturated fatty acid content. There are three members of the pecan SAD family, namely: CcSAD-1, CcSAD-2CcSAD-3, which contain a typical functional domain of Ferritin-like superfamily, a conserved motif DLLNKYYYY, and a conserved motif DLNKYLYYY. The quantitative results showed that there were differences in the expression of SAD gene during fruit ripening. With the increase of CcSADs gene expression, oleic acid content in pecan oil increased by .3.Nine members of the FAD family of pecan were obtained by amplification, which were respectively: 1 / CcFAD2CcFAD3, 3 copies. CcFAD3-1, CcFAD3-2, CcFAD3-3, CcFAD6, CcFAD7, CcFAD8, and CcFAD8-2, CcFAD8-3, all contained typical Membrance-FADS-like super family functional domains and two conserved histidine sequence Hisboxbox. The quantitative results showed that the expression of FAD gene was different in the ripening process of pecan fruit. With the increase of CcFADs gene expression, the proportion of unsaturated fatty acids in pecan oil increased .4.The transgenic Arabidopsis thaliana transgenic with three candidate genes for oil synthesis was constructed, the phenotype of Arabidopsis thaliana was smaller than that of wild type leaves, the number of leaves was more, the bolting rate was higher, and the lateral branches were more than those of wild type Arabidopsis thaliana. However, the seed oil content was not significantly increased in transgenic Arabidopsis thaliana and 35Spro-Uni58 transgenic Arabidopsis thaliana, the T1 representative type was more than that of wild type Arabidopsis thaliana, but the corresponding gene was not detected from T2 generation transgenic Arabidopsis thaliana. The hypothetical reason is the self-protection mechanism of Arabidopsis thaliana and the excision of allogenic genes.
【學位授予單位】：浙江農(nóng)林大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S664.1

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