首页|铁皮石斛DobHLH33转录因子启动子的克隆及表达分析

铁皮石斛DobHLH33转录因子启动子的克隆及表达分析

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为初步探究碱性螺旋-环-螺旋(basic helix-loop-helix,bHLH)家族成员DobHLH33基因在铁皮石斛(Dendrobium officinale Kimura et Migo)中的生物学功能,本研究从铁皮石斛中克隆了 DobHLH33基因的启动子序列.结果显示,克隆出DobHLH33基因起始密码子上游启动子序列1821 bp,经PlantCARE预测分析该序列包含逆境胁迫顺式作用元件、光响应元件、激素响应元件、TATA-box、CAAT-box和CAAT-box核心启动子元件等.构建了 DobHLH33启动子的β-葡萄糖苷酸酶(β-glucuronidase,GUS)融合表达载体并通过花序浸染获得拟南芥转基因植株.GUS染色结果显示,该启动子能够驱动GUS基因在幼苗及各器官部位叶、花、果、柱头、花粉和叶脉中表达,其中在幼苗中表达最强;转基因幼苗在甘露醇和氯化钠胁迫下,DobHLH33启动子活性受诱导,转基因幼苗GUS染色明显加深.综上所述,DobHLH33可能参与铁皮石斛生长发育以及高盐和渗透胁迫响应,为铁皮石斛bHLH家族的功能研究提供基础和参考依据.
Analysis on cloning and expression of DobHLH33 transcription factor promoter from Dendrobium officinale
To investigate the biological function of DobHLH33 gene in Dendrobium officinale,a member of the ba-sic helix-loop-helix(bHLH)family,the promoter sequence of DobHLH33 gene was cloned from Dendrobium officinale.The results showed that the 1 821 bp upstream promoter sequence of the start codon of DobHLH33 gene was cloned,which was predicted by PlantCARE to contain stress cis-acting elements,light responsive elements,hormone responsive elements,TATA-box,CAAT-box and CAAT-box core promoter elements.The GUS fusion expression vector of Dob-HLH33 promoter was constructed and the transgenic Arabidopsis thaliana plants were obtained by inflorescence impreg-nation.GUS staining results showed that the promoter could activate GUS gene expressed in seedlings and various organ parts such as leaves,flowers,fruits,stigma,pollen and leaf veins,with the strongest expression in seedlings.Under the stress of mannitol and NaCl,the promoter activity of DobHLH33 was induced and GUS staining of transgenic seedlings was significantly enhanced.In conclusion,DobHLH33 may be involved in the growth and development of Dendrobium officinale,as well as its response to high salt and osmotic stress.This study provides the basis and reference for the func-tional research of the bHLH family of Dendrobium officinale.

Dendrobium officinalepromoterDobHLH33histochemical stainingadversity stress

杨莎、黄园、陈春伶、王健、陈晓凯、陈宏宇、孙庆文

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贵州中医药大学药学院,贵州贵阳 550025

贵州省中药民族药材种质资源保存及评价工程研究中心,贵州贵阳 550025

贵州中医药大学药用植物抗逆种质研究中心,贵州贵阳 550025

铁皮石斛 启动子 DobHLH33基因 组织化学染色 逆境胁迫

国家自然科学基金项目贵州省科技计划项目2019年博士启动基金项目贵州省梵净山地区生物多样性保护与利用重点实验室开放基金2020年博士启动基金项目2023年省级财政种业发展项目

82260740黔科合基础-ZK[2021]522贵中医博士启动[2019]12号[2020]2003贵中医博士启动[2020]08号

2024

生物资源
武汉大学,武汉市科学技术情报研究所

生物资源

CSTPCD
影响因子:0.485
ISSN:1006-8376
年,卷(期):2024.46(1)
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