小麦泛素结合酶TaUBC16基因的克隆与功能分析
E2 泛素结合酶在调控植物生长发育和胁迫信号转导过程中发挥着重要作用.本研究以小麦抗旱品种晋麦 47的cDNA为模板克隆出E2 泛素结合酶TaUBC16,该基因全长 447 bp,编码 148 个氨基酸.顺式作用元件分析发现,TaUBC16 启动子区含有与分生组织发育、胁迫响应、植物激素应答相关的多种顺式作用元件.利用小麦 RNA-Seq转录组数据结合qRT-PCR验证分析发现,TaUBC16 在小麦不同组织器官和发育阶段普遍表达,其中在 30 d籽粒中的表达量较高,且均能被 PEG-6000、甘露醇和 ABA 显著诱导表达.烟草叶片和小麦原生质体亚细胞定位分析表明,TaUBC16 蛋白分布于...
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| Published in | 作物学报 Vol. 50; no. 8; pp. 1971 - 1988 |
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| Main Authors | , , , , , , |
| Format | Journal Article |
| Language | Chinese |
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甘肃农业大学生命科学技术学院,甘肃兰州 730070%省部共建干旱生境作物学国家重点实验室,甘肃兰州 730070
12.08.2024
省部共建干旱生境作物学国家重点实验室,甘肃兰州 730070 |
| Subjects | |
| Online Access | Get full text |
| ISSN | 0496-3490 |
| DOI | 10.3724/SP.J.1006.2024.31080 |
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| Abstract | E2 泛素结合酶在调控植物生长发育和胁迫信号转导过程中发挥着重要作用.本研究以小麦抗旱品种晋麦 47的cDNA为模板克隆出E2 泛素结合酶TaUBC16,该基因全长 447 bp,编码 148 个氨基酸.顺式作用元件分析发现,TaUBC16 启动子区含有与分生组织发育、胁迫响应、植物激素应答相关的多种顺式作用元件.利用小麦 RNA-Seq转录组数据结合qRT-PCR验证分析发现,TaUBC16 在小麦不同组织器官和发育阶段普遍表达,其中在 30 d籽粒中的表达量较高,且均能被 PEG-6000、甘露醇和 ABA 显著诱导表达.烟草叶片和小麦原生质体亚细胞定位分析表明,TaUBC16 蛋白分布于细胞质和细胞核.通过异源表达TaUBC16 转基因拟南芥进行生长发育表型分析发现,转基因株系开花时间早于野生型,其籽粒相比于野生型更为饱满,千粒重显著高于野生型.基于启动子区-388 bp 位点(T-A)的多态性,开发了TaUBC16 基因的竞争性等位基因特异性PCR(kompetitive allele-specific PCR,KASP)标记,鉴定了TaUBC16 的单倍型,发现TaUBC16-Hap I的千粒重、粒长和粒宽显著高于TaUBC16-Hap II,并在我国小麦育种进程中得到正向选择.本研究结果将为进一步揭示TaUBC16 基因参与调控小麦生长发育和响应逆境胁迫分子机理提供理论依据. |
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| AbstractList | E2 泛素结合酶在调控植物生长发育和胁迫信号转导过程中发挥着重要作用.本研究以小麦抗旱品种晋麦 47的cDNA为模板克隆出E2 泛素结合酶TaUBC16,该基因全长 447 bp,编码 148 个氨基酸.顺式作用元件分析发现,TaUBC16 启动子区含有与分生组织发育、胁迫响应、植物激素应答相关的多种顺式作用元件.利用小麦 RNA-Seq转录组数据结合qRT-PCR验证分析发现,TaUBC16 在小麦不同组织器官和发育阶段普遍表达,其中在 30 d籽粒中的表达量较高,且均能被 PEG-6000、甘露醇和 ABA 显著诱导表达.烟草叶片和小麦原生质体亚细胞定位分析表明,TaUBC16 蛋白分布于细胞质和细胞核.通过异源表达TaUBC16 转基因拟南芥进行生长发育表型分析发现,转基因株系开花时间早于野生型,其籽粒相比于野生型更为饱满,千粒重显著高于野生型.基于启动子区-388 bp 位点(T-A)的多态性,开发了TaUBC16 基因的竞争性等位基因特异性PCR(kompetitive allele-specific PCR,KASP)标记,鉴定了TaUBC16 的单倍型,发现TaUBC16-Hap I的千粒重、粒长和粒宽显著高于TaUBC16-Hap II,并在我国小麦育种进程中得到正向选择.本研究结果将为进一步揭示TaUBC16 基因参与调控小麦生长发育和响应逆境胁迫分子机理提供理论依据. |
| Abstract_FL | E2 ubiquitin-conjugating enzyme plays an important role in regulating plant growth and development,and stress signal transduction.In this study,the TaUBC16 gene encoding E2 ubiquitin-conjugating enzyme was cloned from the cDNA of the drought-tolerant wheat cultivar Jinmai 47.The gene was 447 bp in length and encoded 148 amino acids.The cis-acting element analysis showed that the promoter region of TaUBC16 contained various cis-acting elements related to meristem development,stress responses,and plant hormone responses.By the wheat RNA-seq transcriptome data analysis combined with qRT-PCR vali-dation,it was found that TaUBC16 was generally expressed in different tissues/organs and at different growth stages of wheat,whereas the highest expression level was exhibited in developing grains at 30 days after anthesis.The relative expression level of TaUBC16 was highly induced by PEG,mannitol,and ABA stresses.The subcellular localization in tobacco leaves and wheat pro-toplasts showed that TaUBC16 proteins were located in both cytoplasm and nucleus.The phenotypic analysis of the heterologous expression of TaUBC16 in transgenic Arabidopsis revealed that the transgenic lines had earlier flowering time than the wild type,and its seeds were more plumpness with higher 1000-grain weight than the wild type.Based on the polymorphism of the promoter region-388 bp site(T-A),a kompetitive allele-specific PCR(KASP)marker of TaUBC16 gene was developed and its haplotypes were identified.The haplotype TaUBC16-Hap I had higher thousand-kernel weight,kernel length and width than TaUBC16-Hap II,and had been subjected to positive selection in wheat breeding processes in China.The results of this study provide a theoreti-cal basis for further revealing the involvement of the TaUBC16 gene in the regulation of wheat growth and development and the molecular mechanism responding to adverse stresses. |
| Author | 陈涛 张沛沛 高维东 张龙 张艳艳 杨德龙 胡城祯 |
| AuthorAffiliation | 省部共建干旱生境作物学国家重点实验室,甘肃兰州 730070;甘肃农业大学生命科学技术学院,甘肃兰州 730070%省部共建干旱生境作物学国家重点实验室,甘肃兰州 730070 |
| AuthorAffiliation_xml | – name: 省部共建干旱生境作物学国家重点实验室,甘肃兰州 730070;甘肃农业大学生命科学技术学院,甘肃兰州 730070%省部共建干旱生境作物学国家重点实验室,甘肃兰州 730070 |
| Author_FL | GAO Wei-Dong ZHANG Long HU Chen-Zhen ZHANG Pei-Pei ZHANG Yan-Yan CHEN Tao YANG De-Long |
| Author_FL_xml | – sequence: 1 fullname: GAO Wei-Dong – sequence: 2 fullname: HU Chen-Zhen – sequence: 3 fullname: ZHANG Long – sequence: 4 fullname: ZHANG Yan-Yan – sequence: 5 fullname: ZHANG Pei-Pei – sequence: 6 fullname: YANG De-Long – sequence: 7 fullname: CHEN Tao |
| Author_xml | – sequence: 1 fullname: 高维东 – sequence: 2 fullname: 胡城祯 – sequence: 3 fullname: 张龙 – sequence: 4 fullname: 张艳艳 – sequence: 5 fullname: 张沛沛 – sequence: 6 fullname: 杨德龙 – sequence: 7 fullname: 陈涛 |
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| Keywords | TaUBC16 transgenesis subcellular localization 亚细胞定位 转基因 基因表达 wheat 小麦 gene expression KASP标记 KASP marker |
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| Title | 小麦泛素结合酶TaUBC16基因的克隆与功能分析 |
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