GT和GATA转录因子对甘蓝型油菜BnA5.FAD2和 BnC5.FAD2启动子功能的调控

doi:10.3864/j.issn.0578-1752.2018.24.002

Abstract

Abstract:

【Objective】 Fatty acid desaturase 2 gene (FAD2) is an important gene for controlling oleic acid content in rapeseed. And in this paper, the interaction between GATA family and BnA5.FAD2 and BnC5.FAD2 gene promoters or GT family transcription factors and BnA5.FAD2 and BnC5.FAD2 gene promoters was studied to make sure the transcription regulation mechanism to provide the theoretical basis for high oleic acid breeding. 【Method】 The potential cis-elements were predicted by deletion of promoter fragments and bioinformatics analysis, and candidate transcription factors were screened from PlantTFDB transcription factor database. Real-time PCR (qRT-PCR) was used to detect the expression pattern of transcription factors to further screen candidate transcription factors by comparison with BnA5.FAD2 and BnC5.FAD2 gene expression patterns. Transcription factors were co-transformed with BnA5.FAD2 and BnC5.FAD2 gene promoters into yeast and verified interaction relationship between promoter and transcription factors by yeast one-hybrid. The transcription factors were transformed into Arabidopsis thaliana with BnA5.FAD2 and BnC5.FAD2 gene promoter sequence, and the reporter gene GFP was detected by western blot to analyze the effects of transcription factors on promoters. 【Result】 After knocking out the GT or GATA cis-elements from BnA5.FAD2 and BnC5.FAD2 gene promoters respectively, the GFP protein abundance decreased, which indicate that GT and GATA cis-elements play important roles in regulating gene transcription. Yeast one-hybrid results showed that the GATA family transcription factors (Bna010243-1, Bna026124-1, Bna026124-2) and the GT family transcription factors (Bna010915 and Bna013749) can interact with BnA5.FAD2 and BnC5.FAD2 gene promoters. Western blot analysis showed that when the GATA family transcription factor and the promoter fragment containing GATA elements were co-transformed into Arabidopsis thaliana, the GFP protein content was significantly higher than that of no transcription factor. When the GATA element was knocked out, the GFP protein content did not change significantly. When the GT family transcription factor and the promoter fragment containing the GT element were co-transformed into Arabidopsis thaliana, the GFP protein content changed significantly. When the GT element was knocked out, the GFP protein content did not change significantly.【Conclusion】 The GATA family transcription factors Bna010243-1, Bna026124-1, Bna026124-2 can interact with the GATA elements in the BnA5.FAD2 and BnC5.FAD2 gene promoters to enhance gene expression levels. The GT family transcription factor Bna010915 interacts with the GT elements in the BnA5.FAD2 and BnC5.FAD2 gene promoters to positively regulate gene expression; Bna013749 interacts with the GT elements in the BnA5.FAD2 and BnC5.FAD2 gene promoters to negatively regulate gene expression.

Key words: Brassica napus, GATA transcription factor, GT transcription factor, BnA5.FAD2 promoter, BnC5.FAD2 promoter

LIU Fang,XIAO Gang,GUAN ChunYun. Regulation of GT and GATA Transcription Factors on Promoter Function of BnA5.FAD2 and BnC5.FAD2 Genes in Brassica napus[J].Scientia Agricultura Sinica, 2018, 51(24): 4603-4614.

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Regulation of GT and GATA Transcription Factors on Promoter Function of BnA5.FAD2 and BnC5.FAD2 Genes in Brassica napus

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