Expression And Function Of DGAT1in Camellia Oleifera

Camellia oleifera is one of the four most famous edible oil woody plants, and the grease extracted from the seed of camellia oleifera is called camellia seed oil. Camellia oleifera. is not only a excellent edible oil rich in unsaturated fatty acids, but also widely used to make cosmetics, medicine, candle, soap, and the substitutes of industrial oil. But the low oil content of camellia seed directly affect the development of camellia industry and economic benefits. TAG is the main ingredient in Camellia oleifera grease., DGAT is the only rate-limiting enzyme that acts in the final and committed step of TAG synthesis of the Kennedy way, its main function is to catalytic DAG plus acyl fatty acids form TAG, DGAT enzymes play an important role in TAG accumulation. So the research of Camellia oleifera DGAT1gene’s pression and function is important, to reveal the camellia relation gene expression patterns and functions in synthesis of lipids is of great significance. This thesis mainly studies as follows:1. DGAT1gene Southern hybridization analysis. Extract the examination and approval of national fine varieties’HUASHUO’leaves genomic DNA, through PCR preparation (DIG) digoxin labeled probe hybridization, with genomic DNA probe Southern hybridization analysis, the experimental results show that DGAT1gene in Camellia oleifera genomic DNA is a single copy genes.2. DGAT1gene subcellular localization research. Build fusion expression carrier pCXDG-CoDGATl, using gene gun to bombardment onion epidermal cells for transient expression, and watch it with fluorescent microscope. Results show that the Camellia oleifera DGAT1gene expression position is the cell membrane and cytoplasm, not expressed in the nucleus, and with DGAT1gene functional protein is not coincide with the transcription factor.3. Influence of DGAT1gene expression on oil content of Camellia oleifera seeds. Extracting camellia leaf, root, stamens and petals, calyx, ovary, stigma and flowers in different periods after the fruit and RNA, reverse transcription real-time fluorescent quantitative PCR analysis. Seed oil extract period at the same time, and analysis the oil content and fatty acid composition. Camellia seed oil content of up to50.47%, and the main composition of oil is oleic acid (80%). DGAT1gene expression in42weeks after flowering is in highest level, oil content in the biggest increase is42weeks later after flowering. From the increase trend of oil content and the different period of DGAT1gene expression analysis, DGAT1promote the synthesis of oils and fats. Relevant except fruit DGAT1gene expression in root amount is the highest. Through grey relevance analysis DGmjATl genes associated with oil content of seed number is0.9494, the analysis results suggest DGAT1gene in camellia oil has a vital role in the process of synthesis, and the number of associations with fatty acid composition changes basically is0.5, and illustrates that it will basically not affect the composition of fatty acids.4. DGAT1gene transgenic Arabidopsis thaliana and tobacco.Construction of plant expression vector pCAMBIA1304-CoDGAT1and pCXSN-CoDGATl, using strong carrier CaMV35s promoter gene over-expression purpose. Three positive strains were respectively to obtain transgenic arabidopsis thaliana plants, determination of transgenic arabidopsis seeds oil content is higher than the wild type9%, unsaturated fatty acid content in the composition of fatty acid increases slightly, transgenic plants growth is slower than the wild type, shape has not changed. Get5and6strains of transgenic tobacco strains. Tobacco genetically modified seeds, determination of oil content and fatty acid composition, seed oil content increase4%, fatty acid composition results showed no significant change. The experimental results show that DGAT1gene increase the use of oil content and change the fatty acid composition.