Identification and Functional Characterization of the RcFAH12 Promoter from Castor Bean in Arabidopsis thaliana

• Published in: Separations 2023-01, Vol.10 (1), p.2
• Main Authors: Di, Jianjun, Li, Guorui, Wang, Xiaoyu, Huang, Fenglan, Chen, Yongsheng, Wang, Yue, Sun, Jiaxin, Zhang, Chunlin, Zhang, Qingbo, Wang, Gang, Zhang, Lijun
• Format: Article
• Language: English
• Subjects: Bioaccumulation; Biosynthesis; castor bean; Cloning; Fatty acids; Gene expression; multiple transcriptional start sites; oleoyl-12-hydroxylase; Plasmids; promoter; RACE; Raw materials; Ricinoleic acid; RNA polymerase; Seeds; Triglycerides
• ISSN: 2297-8739; 2297-8739
• DOI: 10.3390/separations10010002

Castor (Ricinus communis L.) seed oil is the commercial source of ricinoleate, a valuable raw material used in many industries. Oleoyl-12-hydroxylase (RcFAH12) is a key enzyme in the biosynthesis of ricinoleate, accumulating nearly 90% of the triacylglycerol in castor seeds. Little is known about the transcriptional regulation of RcFAH12. We used rapid amplification of cDNA 5′ ends (5′RACE) to locate the transcription start site (TSS) of RcFAH12, and the sequence of a 2605 bp region, −2506~+99, surrounding the TSS was cloned. We then investigated these regions to promote β-glucuronidase (GUS) expression in transgenic Arabidopsis by the progressive 5′ and 3′ deletions strategies. The GUS staining showed that the GUS accumulation varied in tissues under the control of different deleted fragments of RcFAH12. In addition, the GUS expression driven by the RcFAH12 promoter markedly accumulated in transgenic seeds, which indicated that RcFAH12 might play an important role in the biosynthesis of ricinoleic acid. This study will lay a potential foundation for developing a tissue-specific promoter in oil-seed crops.