| 甜菜M14品系BvM14-RIN4基因的克隆及应答盐胁迫分析 |
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| 引用本文： | 贾惠旭,李海英,端木慧子.甜菜M14品系BvM14-RIN4基因的克隆及应答盐胁迫分析[J].中国农学通报,2021,37(14):27-33. |
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| 作者姓名： | 贾惠旭 李海英 端木慧子 |
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| 作者单位： | 1.黑龙江大学农业微生物技术教育部工程研究中心,哈尔滨 1500500;2.黑龙江大学生命科学学院/黑龙江省普通高校分子生物学重点实验室,哈尔滨 150080 |
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| 基金项目： | 国家自然科学基金“转录因子BvGT-1调控乙二醛酶I基因参与甜菜单体附加系耐盐的分子机制”(32072122);国家自然基金青年项目“甜菜M14品系BvM14-RIN4蛋白盐胁迫响应蛋白互作网络的研究”(31801426);黑龙江省省属高等学校基本科研业务费基础研究项目(科技创新类专项重点项目)“甜菜M14品系与二倍体栽培甜菜的比较转录组学研究”(KJCXZD201717);中国博士后科学基金面上项目“甜菜M14品系BvM14-RIN4基因响应盐胁迫的分子机理研究”(2019M651314);黑龙江省普通高等学校青年创新人才培养计划“甜菜M14品系BvM14-RIN4基因耐盐功能及其蛋白调控机理研究”(UNPYSCT-2018008) |
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| 摘 要： | 为了研究甜菜单体附加系M14品系RIN4基因耐盐功能,本研究以甜菜M14品系为实验材料,通过PCR技术获得BvM14-RIN4基因cDNA全长序列,对BvM14-RIN4基因进行了生物信息学分析、组织特异性和应答盐胁迫分析。BvM14-RIN4基因ORF长度为264 bp,编码87个氨基酸,蛋白分子量为 9.67 kDa,理论等电点为9.64,初步鉴定该蛋白为亲水性蛋白;BvM14-RIN4蛋白质二级结构主要由延伸链和无规卷曲组成;BvM14-RIN4蛋白质三级结构预测出该蛋白的保守结构域和疏水区;系统进化树分析结果表明BvM14-RIN4蛋白与菠菜SoRIN4和藜麦CqRIN4蛋白亲缘关系最近;荧光定量PCR结果显示,BvM14 -RIN4基因在根中表达量是叶的2.26倍,说明其具有组织特异性;该基因在200 mmol/L NaCl处理后的根中上调表达,说明它参与甜菜M14品系应答盐胁迫过程。本研究在挖掘甜菜M14品系优质基因和开展甜菜遗传改良工作等方面具有重要意义,为后续该基因的功能研究提供了重要参考。
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| 关 键 词： | 甜菜M14品系 BvM14-RIN4 基因克隆 生物信息学分析 荧光定量PCR |
| 收稿时间： | 2020-12-29 |
BvM14-RIN4 Gene of Sugar Beet M14 Line: Cloning and Response to Salt Stress |
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| Jia Huixu,Li Haiying,Duanmu Huizi.BvM14-RIN4 Gene of Sugar Beet M14 Line: Cloning and Response to Salt Stress[J].Chinese Agricultural Science Bulletin,2021,37(14):27-33. |
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| Authors: | Jia Huixu Li Haiying Duanmu Huizi |
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| Institution: | 1.Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education, Heilongjiang University, Harbin 150500;2.Key Laboratory of Microbiology, College of Heilongjiang Province/School of Life Science, Heilongjiang University, Harbin 150080 |
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| Abstract: | To study the salt-resistant function of RIN4 gene in sugar beet monosomic additional line M14 line, sugar beet M14 line was used as experimental material, the full-length cDNA sequence of the BvM14-RIN4 gene was obtained by PCR, and the BvM14-RIN4 gene was analyzed by bioinformatics and its tissue specificity and response to salt stress were explored. The results showed that the ORF length of BvM14-RIN4 gene was 264 bp which encoded 87 amino acids, the molecular weight of the protein was 9.67 kDa, the theoretical pI was 9.64, and the protein was preliminarily identified as a hydrophilic protein. The secondary structure of BvM14-RIN4 protein was mainly composed of extension chain and random curl; the tertiary structure of BvM14-RIN4 protein predicted the conserved domain and hydrophobic region of the protein; the phylogenetic tree showed that BvM14-RIN4 protein was closely related to RIN4 protein of Spinacia oleracea L. and Chenopodium quinoa L.. The results of quantitative RT-PCR showed that the expression level of BvM14-RIN4 gene in roots leaves was 2.26 times that of leaves, which indicated that it had tissue specificity; the expression of BvM14-RIN4 gene was up-regulated in roots under 200 mmol/L NaCl, which indicated that it was involved the response of sugar beet M14 line to salt stress. This study is of certain significance in mining high quality genes of sugar beet M14 line and carrying out genetic improvement of sugar beet, the results could provide important references for studying the function of BvM14-RIN4 gene. |
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| Keywords: | sugar beet M14 line BvM14-RIN4 gene clone bioinformatics analysis quantitative RT-PCR |
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