State Key Laboratory of Efficient Production of Forest Resources, National Engineering Research Center of Tree Breeding and Ecological Restoration, College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China.
State Key Laboratory of Efficient Production of Forest Resources, National Engineering Research Center of Tree Breeding and Ecological Restoration, College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China.
Gene. 2024 May 30;909:148311. doi: 10.1016/j.gene.2024.148311. Epub 2024 Feb 22.
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.
AmCIP 是一种脱水素样蛋白,参与了旱生常绿木本植物白刺适应非生物胁迫的过程。AmCIP 可以在低温驯化过程中诱导子叶和胚根表达。为了进一步阐明该基因上游区域的调控机制,我们分离并鉴定了 AmCIP 的启动子。在此,通过基因组步移法从白刺中分离并克隆了 AmCIP 基因组 DNA 的 1115bp5'侧翼区,并将该片段序列鉴定为"PrAmCIP"启动子。启动子序列分析揭示了一些与各种环境胁迫和植物激素相关的基本顺式作用元件的存在。转基因烟草的 GUS 组织化学染色表明,PrAmCIP 可被 4℃、55℃、NaCl、甘露醇和 ABA 诱导,但在正常条件下几乎不能驱动 GUS 基因表达。此外,我们构建了三个缺失片段并将其遗传转化到拟南芥中。GUS 组织化学染色表明,CP7 片段(-189∼-1)的 MYCATERD1 元件可能是响应干旱的关键元件。综上所述,我们提供了一个可诱导的启动子 PrAmCIP,可应用于开发具有非生物胁迫耐受性的转基因植物。