Cloning of CiNAC038 Gene Promoter in Caragana intermedia and Profiling of Its Responses to Hormones

doi:10.13560/j.cnki.biotech.bull.1985.2019-1078

Abstract

Abstract: The NAC transcription factor family is one of the large specific families in plants and play essential roles in plant development. To understand the molecular mechanism of CiNAC038 expression regulation would lay the foundation for the study of CiNAC038 function in Caragana intermedia. Genome walking method was used to clone the promoter sequence of CiNAC038 gene from genomic DNA of C. intermedia,and Cis-acting elements of the promoter were analyzed. The GUS expression vector was constructed and transformed into Arabidopsis thaliana,and the tissue-specific expression analysis of A. thaliana was performed. Transgenic plants were induced with ABA and the relationship between ABA and CiNAC038 was investigated. The result showed that the cloned fragment was 1 800 bp and contained multiple cis-elements. The vectors ProCiNAC038∶GUS was constructed successfully,and transformed into wild-type A. thaliana by floral dip method. The results of GUS histochemical staining showed that the roots of transgenic Arabidopsis seedlings were darkly stained while the hypocotyls were unstained. The mature veins,fruit pod ends,petals,anthers and other tissues of the transgenic Arabidopsis were darkly stained while the stems were unstained. The GUS reporter gene driven by the CiNAC038 promoter was mainly expressed in tissues and organs of plant leaves,roots and flowers. ABA-induced expression revealed that GUS stains became lighter with increasing concentration. CiNAC038 promoter might be an ABA-inhibitive promoter.

Key words: Caragana intermedia, CiNAC038, promoter, GUS staining

HONG Ge-riqiqige, WANG Yan-fei, GAO Xian-ling, PANG Cai-xia, SHANG Xiao-rui, LI Guo-jing, WANG Rui-gang. Cloning of CiNAC038 Gene Promoter in Caragana intermedia and Profiling of Its Responses to Hormones[J]. Biotechnology Bulletin, 2020, 36(7): 55-61.

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