广西林业科学2025，Vol.54Issue(2)：131-140,10.DOI:10.19692/j.issn.1006-1126.20250202
赤桉miR482f基因启动子克隆及其响应非生物胁迫和激素表达活性分析
Cloning of miR482f Gene Promoter of Eucalyptus camaldulensis and Analysis of its Expression Activity in Response to Abiotic Stresses and Hormones
摘要
Abstract
To understand regulatory mechanism of miR482f gene of Eucalyptus camaldulensis(Eca-miR482f)in response to abiotic stresses and hormones,Eca-miR482f gene promoter was cloned using PCR amplification.Cis-acting elements of Eca-miR482f gene promoter were predicted,and activity of Eca-miR482f gene promot-er was analyzed using transient expression assay of Nicotiana tabacum mediated by Agrobacterium,combined with luciferase reporter gene detection and GUS histochemical staining.Treatments included low temperature,salt and drought stresses as well as abscisic acid,auxin,salicylic acid and methyl jasmonate were set up to in-vestigate responsive characteristics of Eca-miR482f gene promoter to abiotic stresses and hormones.Results showed that Eca-miR482f gene promoter was 1 938 bp in length,and contained core elements such as TATA-box and CAAT-box as well as stress-responsive elements(low temperature,salt,drought and light)and hor-mone-responsive elements(abscisic acid,auxin,salicylic acid and methyl jasmonate).Luciferase reporter gene activity driven by Eca-miR482f gene promoter was significantly higher than that of negative control.Eca-miR482f gene promoter drove normal expression of GUS gene.Under abiotic stresses and hormonal treatments,activity of Eca-miR482f gene promoter significantly increased.Eca-miR482f gene promoter facilitated tran-scriptional expression of miR482f gene,which might be regulated by single factor or synergistic effects of multi-ple factors.关键词
荧光素酶报告基因检测/GUS染色/miR482f/启动子/赤桉Key words
luciferase reporter gene detection/GUS staining/miR482f/promoter/Eucalyptus camaldulensis分类
农业科技引用本文复制引用
周茜,谢怡玲,黄舒怡,林元震..赤桉miR482f基因启动子克隆及其响应非生物胁迫和激素表达活性分析[J].广西林业科学,2025,54(2):131-140,10.基金项目
国家自然科学基金面上项目(31470673) （31470673）
广东省重点领域研发计划项目林业专项(2020B020215002) （2020B020215002）
