大豆GmFtsH2基因的克隆及表达分析  

Cloning and Expression Analysis of GmFtsH2 Genes in Soybean

作　　者：金勋[1] 杨柳[1] 潘红丽[1] 芮海英[1] 王丽娜[1] 李娜[1] 李泽宇[1] JIN Xun;YANG Liu;PAN Hong-li;RUI Hai-ying;WANG Li-na;LI Na;LI Ze-yu(Daqing Branch, Heilongjiang Academy of Agricultural Sciences,Daqing 163316,China)

机构地区：[1]黑龙江省农业科学院大庆分院,黑龙江大庆163316

出　　处：《黑龙江农业科学》2022年第1期6-13,19,共9页Heilongjiang Agricultural Sciences

摘　　要：为了进一步了解FtsH基因在大豆中的表达特性和功能,通过GenBank中拟南芥等其他植物FtsH基因序列设计1对简并引物,从盐处理的大豆叶片中克隆GmFtsH2的全长cDNA序列,并利用在线生物信息学软件对其进行分析。采用实时荧光定量PCR方法分析GmFtsH2在低温、盐、干旱和高温处理下的表达情况。从大豆中克隆到一条全长为2200 bp的mRNA序列,命名为GmFtsH2基因,开放阅读框2091 bp,编码696个氨基酸,相对分子量约为75.089 kDa,理论等电点为5.65,蛋白质不稳定系数为34.90,是稳定蛋白。大豆GmFtsH2与来源于三叶草、葡萄、水稻、玉米和拟南芥类的FtsH类同源蛋白的氨基酸序列比对发现,这些蛋白具有FtsH蛋白共同的保守结构域,包括N端两个跨膜域、AAA结构域、锌离子结合模块等。大豆GmFtsH2与其他19种植物FtsH蛋白的系统进化分析表明,与野生豆和菜豆两个蛋白的亲缘关系较近。RT-PCR分析表明,GmFtsH2在大豆的叶片、茎、根、茎尖的组织内均有表达,且在茎尖中表达量最高,茎中最低。大豆叶片在4℃低温和NaCl处理下,GmFtsH2的表达随时间延长逐渐被诱导,分别在处理后的9 h和12 h达到最高峰;而经干旱诱导的大豆叶片,GmFtsH2的表达在第3天达到最高峰,随后GmFtsH2的表达下调,第11天又达到一个小高峰。推测GmFtsH2在大豆抵御逆境胁迫中起作用。In order to further understand the expression characteristics and functions of FtsH gene in soybean,primers for cloning GmFtsH2 gene were designed in references with the FtsH gene sequences of Arabidopsis and other plants opened in GenBank.Full-length cDNA of GmFtsH2 was cloned from soybean leaves with salt treatment,and analyzed with bioinformatics software online.The expression of GmFtsH2 under the low temperature,salinity,drought and high temperature conditions were analyzed with real-time PCR.A full-length mRNA of 2200 bp was cloned from soybean,and named GmFtsH2 gene.The open reading frame was 2091 bp,encoding 696 amino acids.The relative molecular weight was about 75.089 kDa and the theoretical isoelectric point was 5.65.Its protein instability index was 34.90,indicting a stable protein.The alignment of GmFtsH2 in soybean and homologous protein amino acid sequence of FtsH from clover,grape,rice,corn and Arabidopsis sequence revealed that those proteins showed a common conservative domain structure,including membrane-spanning domains to N terminus,AAA-structure domain,Zinc ions in combination with module,etc.Phylogenetic analysis between GmFtsH2 in soybean and FtsH protein of the other 19 kinds of plants showed that it was resembled to wild beans and kidney beans.Real-time RT-PCR analysis showed that GmFtsH2 was expressed in soybean leaves,stem,root,stem tip,and the stem tip had the highest expression,the lowest expression was in the stem.The expression of GmFtsH2 in soybean leaves was induced gradually with time and to a maximum at 9 and 12 h respectively under 4℃low temperature and NaCl treatment.Under drought treatment GmFtsH2 expression reached to maximum after 3 days.Thereafter GmFtsH2 were down expressed subsequently and reached to a small peak again after 11 days treatment.It is suggested that the GmFtsH2 plays a role in the resistance of adversity stress in soybean.

关 键 词：大豆 FTSH 逆境胁迫 表达分析 

分 类 号：R73[医药卫生—肿瘤]