Analysis Of FT Gene Expression And Study On A7-FT Transcriptional Regulation In Brassica Napus.L

Shortening the growth period of Brassica napus has important strategic significance and economic value,making the research of early maturing rape breeding become a key point.Because the flowering period of rape is closely related to the maturation period,the molecular mechanism of rape blossom has also become a research hotspot.The flowering key gene FT(FLOWERING LOCUS T)is a signal integration factor of the photoperiod pathway,the vernalization pathway,and the autonomous pathway.However,the research on the regulation of FT gene expression in Brassica napus is imperfect.Due to the multiple copies of the FT gene from Brassica napus,the A7-FT gene was studied in this paper.1.Real-time fluorescence quantitative analysis(RT-qPCR)was used to study the expression pattern of FT gene in different varieties(strains)at different periods.The results showed that the FT gene expression patterns of different varieties(strains)of Brassica napus were basically the same,showing a trend of increasing first and then decreasing.,And in the flowering period,the FT gene expression was the largest;The expression levels of FT gene in weak winter materials 103,104,and 1358 at the seedling stage,flower bud differentiation stage,bud stage,and flowering stage were all higher than those of Xiangyou 15 and Zhongshen 11 with stronger winter traits.Expression levels and the ecotype of Brassica napus have no obvious rules.2.The A7-FT gene promoter sequence was obtained from the DNA of Brassica napus L.Xiangyou 15 by PCR.Using PLACE and PlantCARE online tools to predict the sequence,the results showed that in addition to the promoter core elements TATA-box and CAAT-box,the A7-FT gene promoter also has photoresponses,hormones(abscisic acid,salicylic acid).The cis-acting elements associated with endosperm expression,stress resistance,and physiological control.3.Based on the predicted distribution of cis-acting elements,specific primers were designed and clones with different fragment lengths were cloned.The vectors of 5′deleted fragment were constructed with pCAMBIA1303 vector and designated as M1,M2,M3,M4,and M5.Through Agrobacterium-mediated Arabidopsis thaliana,GUS staining and decolorization results showed that there may be some negative regulatory element binding sites in-549 ~-238,and-238 ~ +1 region is the core of the promoter.