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| 橡胶树HbJAZ1.0基因启动子的克隆与活性分析 |
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杨 瑾1,2, 张世鑫2,3, 曹文卿1,2, 晁金泉2,3, 吴绍华2,3*, 胡景涛1*
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1. 重庆三峡学院 生物与食品工程学院, 重庆 万州 404100;2. 中国热带农业科学院橡胶研究所, 热带作物生物育种全国重点实验室,
农业农村部橡胶树生物学与遗传资源利用重点实验室, 海口571101;3. 中国热带农业科学院三亚研究院, 海南 三亚572025
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| 摘要: |
| JAZ(jasmonate ZIM-domain)是茉莉酸信号途径的阻遏蛋白,通过与转录因子互作抑制茉莉酸早期应答基因的表达。为探明HbJAZ1.0基因启动子的特征及活性,该研究以橡胶树品种‘热研7-33-97'为材料,通过PCR扩增、PlantCARE软件及拟南芥转基因技术对HbJAZ1.0基因启动子的顺式作用元件及表达活性进行了分析。结果表明:(1)从橡胶树基因组DNA中克隆了HbJAZ1.0基因上游的1 501 bp的启动子序列。(2)HbJAZ1.0基因上游的1 501 bp启动子序列中含有脱落酸响应顺式作用元件(ABRE)、水杨酸响应顺式作用元件(TCA-element)、光响应顺式作用元件(ACAT-motif、Box 4、G-box、GT1-motif、TCCC-motif、TCT-motif和chs-CMA1a)等。(3)构建了HbJAZ1.0启动子与GUS基因融合的植物表达载体,利用农杆菌介导转化拟南芥,转基因苗不同组织的GUS组织化学染色发现,HbJAZ1.0基因启动子能够驱动GUS基因在拟南芥根、茎、叶、花和果中表达。综上认为,HbJAZ1.0基因启动子具有组成型表达活性,可能受光信号及多种植物激素的调节。该研究结果为进一步研究HbJAZ1.0基因的功能奠定了基础。 |
| 关键词: 橡胶树, HbJAZ1.0启动子, 顺式作用元件, 克隆, 表达分析 |
| DOI：10.11931/guihaia.gxzw202406002 |
| 分类号:Q943 |
| 文章编号:1000-3142(2025)09-1702-09 |
| Fund project:国家自然科学基金(32360405); 海南省自然科学基金(323MS075, ZDYF2024XDNY224); 重庆三峡学院研究生科研创新项目(YJSKY23027)。 |
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| Cloning and activity analysis of HbJAZ1.0 genepromoter from rubber tree |
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YANG Jin1,2, ZHANG Shixin2,3, CAO Wenqing1,2, CHAO Jinquan2,3,
WU Shaohua2,3*, HU Jingtao1*
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1. College of Biology and Food Engineering, Chongqing Three Gorges University, Wanzhou 404100, Chongqing, China;2. State Key Laboratory
for Tropical Crop Breeding, Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture and Rural Affairs, Rubber
Research Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China;3. Sanya Research Institute,
Chinese Academy of Tropical Agricultural Sciences, Sanya 572025, Hainan, China
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| Abstract: |
| The JAZ(jasmonate ZIM-domain)is a repressor protein in the jasmonic acid signaling pathway, inhibiting the expression of early jasmonic acid response genes by interacting with related transcription factors. To elucidate the characteristics and activity of the HbJAZ1.0 gene promoter, this study used the rubber tree variety ‘CATAS 7-33-97' as material and analyzed the cis-acting elements and expression activity of the HbJAZ1.0 gene promoter through PCR amplification, PlantCARE software, and Arabidopsis thaliana transgenetic techniques. The results were as follows:(1)A total of 1 501 bp promoter sequence upstream of the HbJAZ1.0 gene was isolated from the genomic DNA of the rubber tree.(2)Several key cis-acting elements had been identified within the HbJAZ1.0 gene promoter, including ABRE for abscisic acid, TCA-element for salicylic acid, as well as ACAT-motif, Box 4, G-box, GT1-motif, TCCC-motif, TCT-motif and chs-CMA1a for light responsiveness.(3)A plant expression vector containing the GUS reporter gene under the control of the HbJAZ1.0 gene promoter had been constructed and transformed it into A. thaliana using Agrobacterium tumefaciens-mediated transformation. Histochemical GUS staining demonstrated extensive expression of the HbJAZ1.0 gene promoter throughout various tissues of Arabidopsis thaliana, including roots, stems, leaves, flowers, and fruits. In conclusion, the results indicate the HbJAZ1.0 gene promoter has a constitutive expression activity and might be regulated by light and various plant hormones. The results of the study may lay the foundation for further study on the function of HbJAZ1.0 gene. |
| Key words: Hevea brasiliensis(rubber tree), HbJAZ1.0 promoter, cis-acting element, cloning, expression analysis |
