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

Abstract

Abstract 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 1048 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 showed that PrAmCIP was induced by 4℃, 55℃, NaCl, drought 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(-122~-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 under abiotic stresses.