Screening,Gene Clone And Expression Of Rich In PUFAs Of Ribes L.in Northeast Area

Ribes L.belongs to the Saxifragaceae,also known as gooseberry.Fruit products of Ribes L.taste good,rich in carbohydrates,vitamin C,and other kinds of nutrition.Fruit can be processed into jam,juice,wine,with high economic and medicinal value.Besides unsaturated fatty acids has medical value to improve human’s immunity,reduce blood fat and improve arteriosclerosis,etc.This study mainly focuses on the molecular regulatory network of the PUFAs of Ribes L.,screening eight varieties of Ribes L.in northeast area which rich in polyunsaturated fatty acids,and analyze of function relationship and the expression pattern of temporal and spacial of PUFAs genes in Ribes L.,such as:RmD6E,RmD8B,RmFAD3,RrABI3,RrNFYB and RrNFYC2.so that we can lay a solid foundation for explaining the molecular mechanism of PUFAs synthesis.The main results obtained are as follows:1.Determination of unsaturated fatty acids of 8 varieties Ribes L.in northeast areaUsing Soxhlet extraction and gas chromatography method to measure the crude fat content and the fatty acid composition of eight varieties Ribes L..In eight cultivars,the crude fat rate of Ribes nigrum L is 21.19%with highest rank,but that of ’Heifeng’ is only 13.94%with lowest rank.The contents of fatty acid and the unsaturated fatty acid of Ribes rubrum L.are higher than those of other kinds,but both contents of Ribes nigrum L.are lowest.2.The full-length cDNA sequences cloning of PUFAs genes in Ribes L.Use RACE to clone the full-length cDNA of PUFAs genes in Ribes L.We achieved six genes which are RmD6E with the length of 1766bp(accession number:KX056120).RmD8B with the length of 1666bp(accession number:KX061751)、RmFAD3 with the length of l539bp(accession number:KX061752).RrAB13 with the length of 2228bp(accession number:KX061754)、RrNF－YB with the length of 96lbp(accession number:KX061753)、RrNF-YC2 with the length of 1382bp(accession number:KX061755).3.The phylogenetic analysis of PUFAs genes of the Ribes L.The study analyzes the genetic RmD6Et RmD8Bt RmFAD3f RrABI3,RrNFYB and RrNFYC2 under the help of various online softwares of biology,and then predicts primary and secondary structures of protein in six genes followed by systemic phylogenetic analysis.RmD6E,RmD8B gene and blackcurrant of D6D,D8B gene has closest relationship,and participate to the composition and regulation of fatty acid,it in shown in the phylogenetic analysis that they have comparatively close genetic relationship with the species eontaining oil.RmFAD3 protein is not similar to other species,own a single genus.RrABI3 protein has closet relationship with cattle and humans in mammals,and has closet relationship with tomato and potato in plant.RrNFYB,RrNFYC2 proteins belong to a hydrophilic protein.RrNFYB of Ribes L.,Arabidopsis and Medicago truncatula genetic relationship is closer;RrNFYC2 genes of Ribes L.close genetic relationship with Populus euphratica.4.The expression patterns analysis of temporal and spacial of Ribes L.PUFAs genes by using Realtime PCR methodThe expression patterns analysis of temporal and spacial of RmD6E,RmD8B9 RmFAD3,RrABI3,RrNFYB and RrNFYC2 gene by using Realtime PCR method in four stages 8 species:It is found that there is the direct proportion between fruit size and expression quantity of RmD6E,RmD8B and RmABI3,but the relation between fruit size and expression quantity of RmFAD3 is inverse proportion.The expression levels of RrNFYC2 and RrNFYB genes has not obvious contact with the fruit size.