Effects Of Rapeseed Genes BnGLIPL And BnGLIP7 On Seed Oil Content

Rapeseed oil is traditional edible oil in China.Because of its high nutritional value,rape-seed oil gets the favor of the people.However,the current annual imports of rapeseed oil accounted for more than 30% of demand.It is an effective means to improve the self-sufficiency rate of rape-seed oil in China by increasing the oil content of rapeseed.The general way to increase the oil content is to increase the accumulation of oil in seed by changing the expression of certain genes in the fatty acid synthesis pathway,and reducingdecomposition of TAG in the crop is also important.GDSL is a kind of hydrolytic enzyme that is widely found in plants and animals,and there aredifferent kinds of substrates for it,such as sulfide ester,aromatic ester,phospholipid and amino acid.This studycharacterized the function of AT1G71250(gdsl-lipase like,GLIPL),AT5G15720(gdsl-motif lipase 7,GLIP7)and itshomologous genes(Bn GLIPL,Bn GLIP7)in Arabidopsis thaliana,the resultsare as follows:1)The complete CDS and promoter sequence of Bn GLIPL and Bn GLIP7 were cloned in Brassica napus.2)It was found that GLIPL and GLIP7 were highly conserved in different plants of the genus Brassica,and the sequence similarity was 90%,and there were four conserved structures in each gene.The results showed that GLIPL and GLIP7 were highly conserved in plants,which indicates similar functions for them.3)Homozygous mutantgliplwere obtained in Arabidopsis thaliana;The Bn GLIPL and Bn GLIP7 overexpression vector containing the double 35 s promoter were constructed and the transgenic lines were obtained.4)q RT-PCR was performed on roots,stems,leaves,buds and siliques of Arabidopsis thaliana,and GLIPL was found to be expressed in all organs and its expressing level in siliques of 4 DAF or older is higher than other organs.By GUS staining,Bn GLIP7 expressing signal was strongly detected in germinated seeds,flower organs and developmental seeds.Bn GLIP1 was expressed in seedlings,inflorescence and young pods.5)We characterizedoil content of overexpression lines and homozygous T-DNA insert mutants,and found the oil content was decreased by 9.54% in overexpressing lines Bn GLIPL than the wild type(significantly higher),and oil content of the mutants increased by 7.63% compared with the wild type(significantly higher);The oil content of overexpression of GLIP7 was 4.66% lower than that of wild type(significantly higher).