Isolation and functional characterization of cold-induced gene (AmCIP) promoter from Ammopiptanthus mongolicus

• Published in: Gene 2024-05, Vol.909, p.148311-148311, Article 148311
• Main Authors: Ge, Bohao, Dong, Kuo, Li, Rongchen, Bi, Xiaorui, Liu, Qianru, Zhang, Weiwei, Chen, Yuzhen, Lu, Cunfu
• Format: Article
• Language: English
• Subjects: Abiotic stress; acclimation; Ammopiptanthus mongolicus; Arabidopsis - metabolism; Arabidopsis thaliana; cold; cotyledons; Deletion analysis; DNA; drought; Fabaceae - genetics; gene expression; Gene Expression Regulation, Plant; genetically modified organisms; GUS-activity; mannitol; Plant Growth Regulators - metabolism; Plant Proteins - metabolism; Plants, Genetically Modified - genetics; Plants, Genetically Modified - metabolism; PrAmCIP promoter; promoter regions; Promoter Regions, Genetic; stress tolerance; Stress, Physiological - genetics; tobacco; transgenes; woody plants
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/j.gene.2024.148311

•The promoter sequence (1115-bp sequence) of a dehydrin gene, AmCIP, was amplified by the genome walking technique to obtain.•GUS fluorescence quantification and GUS histochemical staining analyses indicated that PrAmCIP is an induction type promoter that can be significantly induced by osmotic and cold stress.•5′ deletion analysis technique indicated that the MYC element is a key cis-element of the promoter in response to cold stress, and it was also found that the MYCATERD1 element present in the region of −189 bp ∼ -1bp may be a key cis-element of the promoter in response to osmotic stress. AmCIP is a dehydrin-like protein which involved in abiotic stress tolerance in xerophytes evergreen woody plant A. mongolicus. AmCIP could be induced in the cotyledon and radicle during cold acclimation. To further elucidate the regulation of the upstream region of the gene, we isolated and characterized the promoter of AmCIP. Herein, a 1115 bp 5′-flanking region of AmCIP genomic DNA was isolated and cloned by genome walking from A. mongolicus and the segment sequence was identified as “PrAmCIP” promoter. Analysis of the promoter sequence revealed the presences of some basic cis-acting elements, which were related to various environmental stresses and plant hormones. GUS histochemical staining of transgene tobacco showed that PrAmCIP was induced by 4℃, 55℃, NaCl, mannitol and ABA, whereas it could hardly drive GUS gene expression under normal conditions. Furthermore, we constructed three deletion fragments and genetically transformed them into Arabidopsis thaliana. GUS histochemical staining showed that the MYCATERD1 element of the CP7 fragment (-189 ∼ -1) may be a key element in response to drought. In conclusion, we provide an inducible promoter, PrAmCIP, which can be applied to the development of transgenic plants for abiotic stresse tolerance.