木薯MebZIP11基因克隆与原核表达分析  

Cloning and prokaryotic expression analysis of MebZIP11 gene in cassava(Manihot esculenta)

作　　者：雷鑫芳 潘子墨 林素妍 罗佳科 吴春来 曾坚 胡伟 LEI Xinfang;PAN Zimo;LIN Suyan;LUO Jiake;WU Chunlai;ZENG Jian;HU Wei(Guangdong Provincial Key Laboratory of Utilization and Conservation of Food and Medicinal Resources in Northern Region/Guangdong Engineering Research Center for Efficient Utilization of Water and Soil Resources in Northern Region/College of Biology and Agriculture,Shaoguan University,Shaoguan,Guangdong 512005,China;National Key Laboratory for Tropical Crop Breeding/Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Science,Haikou,Hainan 571101,China)

机构地区：[1]韶关学院广东省粤北食药资源利用与保护重点实验室/广东省粤北水土资源高效利用工程技术研究中心/生物与农业学院,广东韶关512005 [2]中国热带农业科学院热带作物生物育种全国重点实验室/热带生物技术研究所,海南海口571101

出　　处：《福建农林大学学报(自然科学版)》2025年第3期299-308,共10页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：广东省普通高校重点领域专项(2024ZDZX4018,2022ZDZX4047);广东省基础与应用基础研究基金项目(2025A1515010531,2023A1515010336);韶关学院重点项目(SZ2022KJ05);大学生创新创业训练计划项目(Sycxcy2024176)。

摘　　要：【目的】研究木薯A类碱性亮氨酸拉链11(Manihot esculenta basic leucine zipper11,MebZIP11)基因的功能,为探讨其转录因子在逆境胁迫中的作用提供依据。【方法】利用RT-PCR技术克隆木薯中的MebZIP11基因,并进行生物信息学分析。通过荧光定量PCR技术,分析MebZIP11基因在木薯不同组织和块根发育不同阶段中的表达模式,并评估其在多种逆境条件下的响应机制。【结果】MebZIP11基因位于木薯的第5号染色体,编码区全长1221 bp,具有典型的bZIP结构特征。序列比对显示,MebZIP11与麻风树和橡胶树的A类bZIP转录因子的序列同源性分别为80.00%和85.41%,确认其属于A类bZIP亚族。亚细胞定位试验显示,MebZIP11位于细胞核内。MebZIP11基因在储藏根的表达量最高。MebZIP11基因的表达量在甘露醇、NaCl和脱落酸处理下均显著升高,在H_(2)O_(2)和低温处理下其表达量均下降。通过原核表达系统获得了MebZIP11蛋白。【结论】木薯中的MebZIP11基因属于A类bZIP家族。MebZIP11基因在不同逆境和激素处理下表现出不同的响应程度。【Objective】This study aimed to investigate the function of Manihot esculenta basic leucine zipper11(MebZIP11)gene,and provide insights into the role of A-type bZIP transcription factor in M.esculenta(cassava)in response to abiotic stress.【Method】The MebZIP11 gene from cassava was cloned using RT-PCR technology,followed by bioinformatics analysis.The expression pattern of MebZIP11 was analyzed in various cassava tissues and at different stages of root tuber development via qRT-PCR,in combination with the evaluation on the gene's response to various stress conditions.【Result】The MebZIP11 gene is located on chromosome 5 of cassava,with a coding region of 1221 bp in length,exhibiting typical bZIP structural features.Sequence alignment showed that MebZIP11 shared 80.00%and 85.41%homology with the A-type bZIP sequences of Jatropha curcas and Hevea brasiliensis,respectively,confirming its belonging to the A-type bZIP subfamily.Subcellular localization assays demonstrated that the MebZIP11 protein was localized in the nucleus,and the MebZIP11 was most expressed in the storage roots by expression analysis.Under different treatment conditions,the expression of MebZIP11 was significantly induced by mannitol,NaCl and abscisic acid,while was suppressed by H_(2)O_(2)and cold treatment.Lastly,the MebZIP11 protein was successfully expressed and purified in prokaryotic system.【Conclusion】The MebZIP11 gene in cassava is identified as a member of A-type bZIP family.It exhibits distinct inducible responses under various stress and hormone treatments.

关 键 词：木薯 非生物胁迫 碱性亮氨酸拉链 逆境响应 原核表达 

分 类 号：S533[农业科学—作物学]