Cloning And Promoter Analysis Of BnaFIL Gene In Brassica Napus

Brassica napus(AACC,2 n = 38),as one of the most important oil crops in the world,widely cultivated in China In China and around the world,rape is the fifth largest after rice soybean corn and wheat crops,and cooking oil and important source of protein Rapeseed besides has the edible value at the same time,in the chemical medicine and biological energy industry also has a huge demand for rape Timely flowering and morphological differentiation of flower not only affects the yield and quality of rapeseed,leaf determines its adaptability so ecological area.Thus,flowering plant and flower of morphological development is one of the important target traits of rapeseed breeding Early use of whole genome sequencing and early high density gene chips mining related genes,find aFIL genes related to flowering sooner or later,in the early and late flower varieties FIL genes exist in a SNP loci located on the fifth introns of the gene,the late by genetic engineering means from the separation of Brassica napus L.were used to obtain BnaFIL gene and its promoter In this paper,we make the following four aspects of research:1.Genome-wide association analysis of flowering related genes in rapeseeds from 203semi-wintery inbred lines in China was conducted using Brassica 60 K SNP microarrays.A SNP locus on FIL gene was detected,which was located on chromosome A03 and was significantly correlated with flowering time.Fluorescent quantitative PCR results showed that the expression of BnaFIL gene was different between the early and late flowering materials.The expression of BnaFIL gene was low in the late flowering materials,while the expression of BnaFIL was high in the early flowering materials.2.A total length of 2312 bp BnaFIL gene was cloned from 15 different flowering stages of Brassica napus L.By comparison,we found that there was a SNP site between the early flower and the late flower,which was located in the fifth intron.Bioinformatics analysis of the cloned BnaFIL gene showed that it was most likely to be located in the nucleus.BnaFIL is most closely related to Arabidopsis thaliana,a member of the cruciferous family.3.The promoter of this gene was cloned,and the comparison analysis showed that there were cis-reaction elements such as CAAT-box element and TATA-box element and two SNP sites in the early and late flowering materials.4.Build p BnaFIL-GUS promoter expression vectors of BnaFIL gene,the recombinant plasmid into soil agrobacterium GV3101,inflorescence impregnation method agrobacterium containing a recombinant plasmid into arabidopsis thaliana,the recombinant plasmid expressed in arabidopsis thaliana.The results showed that the expression intensity of GUS gene was higher under the promoter of early flowering material than that under the promoter of late flowering material.