大豆耐胁迫相关蛋白编码基因GmSAMDC1的克隆及表达分析  

Cloning and Expression Analysis of Soybean GmSAMDC1-encoding Gene

作　　者：任锐[1,2] 杨凤玺 王涛 高乐[1] 智海剑[1] 李凯[1] REN Rui;YANG Feng-xi;WANG Tao;GAO Le;ZHI Hai-jian;LI Kai(Soybean Research Institute of Nanjing Agricultural University/Key Laboratory of Biology and Genetic Improvement of Soybean, Ministry of Agriculture /National Center for Soybean Improvement/National Key Laboratory for Crop Genetics and Germplasm Enhancement, Nanjing 210095, China;Environmental Horticulture Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China;Institute of Cereal and Oil Crops, Handan Academy of Agricultural Sciences, Handan 056001, China)

机构地区：[1]南京农业大学大豆研究所/农业部大豆生物学与遗传育种重点实验室/国家大豆改良中心/作物遗传与种质创新国家重点实验室,江苏南京210095 [2]广东省农业科学院环境园艺研究所,广东广州510640 [3]邯郸市农业科学院粮油作物研究所,河北邯郸056001

出　　处：《大豆科学》2018年第5期672-680,共9页Soybean Science

基　　金：国家重点研发计划项目(2017YFD0101504);中央高校基本科研业务费(Y0201600115);国家自然科学基金(31671718;31371646;31571690);现代农业产业技术体系(CARS-04);转基因生物新品种培育科技重大专项(2016ZX08004-004);江苏省现代作物生产协同创新项目(JCIC-MCP)

摘　　要：S-腺苷甲硫氨酸脱羧酶(S-adenosine methionine decarboxylase,SAMDC)是亚精胺和精胺合成的关键酶,更是多胺生物合成的限速酶之一,在植物耐胁迫反应中发挥重要调控作用。为研究大豆SAMDC编码基因的结构和表达特性,本研究从抗病大豆品种科丰1号中克隆了位于大豆基因组2号染色体上的GmSAMDC1(Glyma. 02G128000)基因,分析结果表明:其完整ORF长度为1 068 bp,编码一个由355个氨基酸组成的包含1个SAM_decarbox结构域的蛋白; GmSAMDC1基因所编码蛋白的理论等电点为4. 86,相对分子质量为38 987. 13 Da,为亲水性蛋白,不含跨膜区;大豆中共有8个GmSAMDC1的同源基因,GmSAMDC1与红车轴草(PNY09439. 1,82. 64%)和拟南芥(At SAMDC3,67. 22%)中的SAMDC蛋白编码基因亲缘关系最近; GmSAMDC1启动子序列包含防卫和胁迫响应元件、植物激素应答元件、光应答元件等许多顺式作用元件; GmSAMDC1在花中的表达量最高,与其大豆同源基因Glyma. 01G071300(序列相似性为94. 6%)、Glyma. 18G278800(66. 8%)和Glyma. 08G25580(66. 5%)的组织特异性表达模式比较相似; Glyma. 02G128000-GFP融合蛋白在细胞膜和细胞质上表达。本研究结果为进一步阐明大豆GmSAMDC1基因在大豆耐胁迫过程中的作用提供了理论依据。S-adenosine methionine decarboxylase（ SAMDC）, the key enzyme of spermine biosynthesis, is one of the speed-limit enzymes in the biosynthesis of polyamine playing an important regulatory role in plant tolerance to stress. To explore the structure and expression characteristies of soybean SAMDC-encoding genes, the gene GmSAMDC1 （Glyma. 02G128000） loca-ted on chromosome 2 was cloned from the resistance soybean cultivar Kefeng 1 in this study. Results showed that the gene Gm-SAMDC1, of which complete ORF length was 1 068 bp, and encoded a protein of 355 amino acids containing a ＇ SAM_decar-box＇ domain. For this protein, its isoelectric point （pI） was 4. 86, and the relative molecular mass was 38 987. 13 Da, con-taining no transmembrane region as the hydrophilic protein. There were 8 GmSAMDC1 homologous in soybean, and the genetic relationship of the SAMDC protein-coding genes in red clover （ PNY09439. 1, 82. 64% ） and Arabidopsis thaliana （ AtSAM-DC3, 67. 22% ） was closest. The GmSAMDC1 promoter sequence contains many cis-acting elements such as defense and stress response elements, plant hormone response elements, and light response components. The expression of GmSAMDC1 in flowers was the highest, which was similar to the tissue specific expression pattern of Glyma. 01 GO71300 （sequence similarity is 94. 6% ）, Glyma. 18G278800 （66. 8% ） and Glyma. 08G25580 （66. 5% ）. Fusion protein Glyma. 02G128000-GFP was expressed in cell membrane and cytoplasm. The results of this study lay a theoretical basis for further elucidation of the role of soybean GmSAMDC1 in the orocess of soybean tolerance to stresses.

关 键 词：大豆 GmSAMDC1 耐胁迫 顺式作用元件 亚细胞定位 

分 类 号：S565.1[农业科学—作物学]