本文選題：苧麻 + 木葡聚糖內(nèi)轉(zhuǎn)糖苷酶/水解酶�。� 參考：《華中農(nóng)業(yè)大學》2017年碩士論文

【摘要】：苧麻是我國特有的纖維作物,苧麻織物具有透氣、防霉、輕薄等諸多優(yōu)點,在紡織業(yè)中扮演著重要的角色。苧麻纖維是一種韌皮纖維,植物韌皮纖維發(fā)育涉及到韌皮部纖維細胞壁的擴展、加厚等生物學過程。對參與該過程的相關基因表達特性的研究能夠推動研究者對苧麻纖維發(fā)育進程的認識,有助于從分子水平上對苧麻纖維品質(zhì)進行改良。木葡聚糖內(nèi)轉(zhuǎn)糖苷酶/水解酶基因(XTH)在促進植物細胞壁延展以及響應逆境脅迫方面發(fā)揮著重要的作用。目前,在擬南芥、水稻、楊樹等模式植物中關于XTH家族的研究已較為成熟,而類似的研究在苧麻中還未見報道。本研究以苧麻優(yōu)質(zhì)品種“1504”為材料,基于本實驗室已有的3個轉(zhuǎn)錄組數(shù)據(jù)庫信息,克隆苧麻Bn XTH基因,分析基因的時空表達模式,并通過煙草亞細胞定位實驗研究基因在細胞中的表達和分布情況。主要實驗結(jié)果如下:(1)從轉(zhuǎn)錄組中篩選出20條苧麻Bn XTH基因,命名為Bn XTH1-Bn XTH20,并獲得其中12個基因的基因組DNA序列,分析基因的內(nèi)含子-外顯子結(jié)構(gòu)和功能域及構(gòu)建進化樹。(2)通過qRT-PCR測定基因在苧麻三季麻不同發(fā)育時期和部位的表達情況,研究20條基因時空表達模式,實驗結(jié)果表明:20條基因在不同發(fā)育時期莖的不同部位均有表達,多數(shù)基因在頭麻和二麻莖部的表達水平高于三麻,其中,Bn XTH11在頭麻莖部的表達水平高于二麻和三麻;分析Bn XTH基因在脅迫處理下的表達情況,發(fā)現(xiàn)20個基因均能響應干旱、高溫、低溫脅迫,在干旱和高溫脅迫下,Bn XTH11在莖部表達量極顯著上升,在低溫脅迫下,其表達量出現(xiàn)極顯著下降。(3)構(gòu)建Bn XTH11 GFP載體,轉(zhuǎn)入煙草,進行亞細胞定位分析,發(fā)現(xiàn)綠色熒光在細胞膜或細胞壁表達,表明該蛋白可能被轉(zhuǎn)運至細胞膜,最后分泌至細胞壁發(fā)揮作用。
[Abstract]:Ramie is a special fiber crop in China. Ramie fabric plays an important role in textile industry because of its many advantages, such as air permeability, mildew resistance, lightness and so on. Ramie fiber is a kind of phloem fiber. The development of plant phloem fiber is related to the extension and thickening of cell wall of phloem fiber. The study of the related gene expression characteristics involved in the process can promote the understanding of Ramie fiber development process and help to improve the Ramie fiber quality at the molecular level. Dextran glucosidase / hydrolase gene XTHplays an important role in promoting cell wall extension and responding to stress. At present, the studies of XTH family in Arabidopsis thaliana, rice, poplar and other model plants have been more mature, but similar studies have not been reported in Ramie. In this study, Ramie Bn XTH gene was cloned from three transcriptome databases of Ramie quality variety "1504", and the temporal and spatial expression pattern of the gene was analyzed. The expression and distribution of genes in tobacco cells were studied by subcellular localization experiment. The main results were as follows: (1) Twenty Bn XTH genes of Ramie were screened from the transcriptome, named Bn XTH1-Bn XTH20, and the genomic DNA sequences of 12 of them were obtained. The structure and functional domain of intron and exon of the gene were analyzed and the phylogenetic tree was constructed. QRT-PCR was used to determine the expression of the gene in different development stages and parts of Ramie, and the spatiotemporal expression patterns of 20 genes were studied. The results showed that 20 genes were expressed in different parts of stem at different developmental stages, and the expression level of most of them was higher in the stem part of head hemp than that in Sanma, and the expression level of Bn XTH11 in the stem part of head hemp was higher than that in the stem of head hemp and Sanma. By analyzing the expression of Bn XTH gene under stress, it was found that 20 genes were able to respond to drought, high temperature and low temperature stress, and the expression of Bn XTH11 in stem was significantly increased under drought and high temperature stress, and increased significantly under low temperature stress. Bn XTH11 GFP vector was constructed and transformed into tobacco for subcellular localization analysis. It was found that green fluorescence was expressed in the cell membrane or cell wall, indicating that the protein might be transported to the cell membrane. Finally secreted to the cell wall to play a role.
【學位授予單位】：華中農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S563.1

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