Functional Analysis of A7-FT Promoter in Brassica napus L.

doi:10.7668/hbnxb.201751081

Abstract

Abstract: Based on the whole genome sequence of Brassica napus, the sequence of A7-FT gene promoter was obtained by PCR. The promoter online prediction software was used to predict the function and structure of the promotor, and five promoter sequences with different fragment lengths were obtained from the 5'-end deletion according to the distribution of predicted cis-elements. The GUS gene expression vector containing promoters with different fragment lengths was constructed and T₂ seedlings were obtained by Agrobacterium -mediated Arabidopsis thaliana. After GUS staining and decolorization, the function of A7-FT gene promoter was explored to study the flowering of B. napus. The study of regulatory mechanisms provided a theoretical basis.The A7-FT gene promoter sequence was obtained from the DNA of Brassica napus L. Xiangyou 15 by PCR. Using the PLACE and PlantCARE online tools to predict the sequence, it was found that in addition to the promoter core elements CAATbox and TATAbox,the A7-FT gene promoter also had photo responses, hormone response components, expression response element of endosperm,response element of stress resistance and physiological control. Based on the predicted distribution of cis-acting elements, specific primers were designed to clone the promoter with different fragment lengths. A 5' segment deletion vector was constructed with pCAMBIA1303 vector and designated as M1, M2, M3, M4, and M5, respectively. Through Agrobacterium -mediated Arabidopsis thaliana, GUS staining and decolorization results showed that there might be some negative regulatory element binding sites between-1 549 and-238, and the core region of the promoter was from-238 to +1.

Key words: Brassica napus L., FT gene, Expression, A7-FT promoter, Arabidopsis thaliana

CLC Number:

S635.03
Q78

ZHANG Xiyu, HE Hui, LI Zhen, XING Man, HONG Bo, GUAN Chunyun. Functional Analysis of A7-FT Promoter in Brassica napus L.[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(2): 59-65. doi: 10.7668/hbnxb.201751081.

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