The Research Of The Effect Of PcG Gene On Oil Accumulation In Arabidopsis Leaves And Seeds

Lipids are the most efficient form of energy storage for plant seeds.Oil bodies are the smallest storage lipid organelles in plants(mainly triacylglycerol),which provide energy for life activities and metabolic processes.Oil bodies are present in plant tissues that accumulate oil and are rapidly hydrolyzed during seedling growth after seed germination and converted into carbohydrates as the seedlings grow.Polycomb-group proteins of Pc G can be modified by chromatin to make genes genetically silenced.In plants,Pc G protein is involved in the regulation of various developmental processes such as cell fate determination,vegetative growth and development,flowering time control and embryogenesis.The laboratory found in the previous research that the double mutant ring1 a ring1b,clf swn of Arabidopsis PRC complex with loss of function showed an embryonic structure during vegetative growth,and a large amount of neutral oil(TAG)accumulated in the embryonic structure.In this experiment,molecular biology,cell biology,biochemistry,and transcriptomics were used to conduct preliminary studies on the distribution and accumulation of oil and fat on the leaves and seeds of Arabidopsis mutants swn and ring.1.Through thin layer chromatography analysis,it was found that in the leaves,the mutant swn and ring leaves had higher oil content than the control wild type,and the ring was significantly higher than the wild type control;in the seeds,the ring seed oil content was higher than the wild type Seed control.2.Through gas chromatography analysis,it was found that the proportion of fatty acid components in the mutant swn leaves and seeds did not change much,and the proportion of fatty acids in oleic acid(C18:1)in the mutant ring leaves was significantly reduced compared with the wild type,and linoleic acid(C18:2)The proportion is significantly lower than that of wild type.Compared with wild-type seeds,the proportion of trace acid(C16:0)in fatty acids in ring seeds was significantly reduced,and the proportion of oleic acid(C18:1)in fatty acids was significantly increased.3.Through confocal laser microscopic observation of Arabidopsis oil bodies,it was found that the fluorescence signal of oil bodies in ring and swn epidermal cells was stronger than that of wild type,and the oil in the mutant was very small compared with the distribution of a very small amount of oil bodies in wild type The body distribution is more dense,and some oil bodies are distributed in pieces;the average number of wild-type oil bodies in seeds is 6.76±1.86,the average number of swn oil bodies is 7.24±1.31,and the average number of ring oil bodies is 7.93±1.83.The average number of oil bodies in ring seeds increased significantly.4.Transcription analysis of the leaves revealed that a total of 704 genes were up-regulated and 590 genes were down-regulated in the swn mutant.A total of 1786 genes were up-regulated and 1485 genes were down-regulated in the ring mutant,which was significantly different from wild-type gene expression.Among the mutants,the genes involved in oil transport,oil synthesis,fatty acid synthesis,and related transcription factors were significantly up-regulated,The up-regulation of these genes suggests that the SWN and RING genes are closely related to the accumulation of oil and the formation of oil bodies.?5.Through fluorescence quantitative PCR analysis,it was found that the expression of oil body membrane protein-related genes CALEOSIN3,GRP19,OLE3,etc.in the leaves were significantly higher than the wild type,and the fatty acid synthesis related genes GPAT9,PDAT1,GPAT3,FATB,KCS2 The two mutants showed an overall upregulation trend;the oil body membrane protein-related genes CALEOSIN3,CALEOSIN4,OBAP1 and fatty acid synthesis-related genes DGAT1,DGAT2,ACC1,PDAT1,and GPAT9 all showed a significant upregulation trend in the ring.Only the expression of oil body membrane protein-related genes was up-regulated.The quantitative results further confirmed that the loss of SWN and RING gene function can promote the accumulation of oil.In summary: the results of thin layer analysis,chromatography analysis,oil body observation,transcriptome and quantitative analysis showed that the loss of RING gene function can promote the accumulation of Arabidopsis leaf and seed oil;the loss of SWN gene function can promote The accumulation of oil in leaves is not obvious in seeds.