大豆GmNIC1基因克隆及非生物胁迫应答分析  

Cloning of soybean GmNIC1 gene and analysis of abiotic stress response

作　　者：张琦 刘兴宇 任馨原 刘雅心 隋京芮 严红静 赵晓菊 徐杰飞 ZHANG Qi;LIU Xing-yu;REN Xin-yuan;LIU Ya-xin;SUI Jing-rui;YAN Hong-jing;ZHAO Xiao-ju;XU Jie-fei(Biological Engineering,Daqing Normal University,Daqing,Heilongjiang 163712,China;Jiamusi Branch of Heilongjiang Academy of Agricultural Sciences/Jiamusi Comprehensive Experimental Station of National Soybean Industry Technology System,Jiamusi,Heilongjiang 154002,China)

机构地区：[1]大庆师范学院生物工程学院,黑龙江大庆163712 [2]黑龙江省农业科学院佳木斯分院/国家大豆产业技术体系佳木斯综合试验站,黑龙江佳木斯154002

出　　处：《西南农业学报》2024年第12期2573-2581,共9页Southwest China Journal of Agricultural Sciences

基　　金：黑龙江省省属高等学校基本科研业务费项目(2023-KYYWF-0025);大庆市指导性科技计划项目(zd-2024-62);黑龙江省自然科学基金项目(LH2021C001)。

摘　　要：【目的】NAD+是植物生长发育过程中的重要调节物质,NIC(烟酰胺酶)是NAD+合成过程中的关键酶,研究大豆中NIC的功能,以期创制大豆抗逆新品种。【方法】利用生物信息学,探究大豆GmNIC1基因的结构与功能,并通过RT-PCR进行基因表达模式分析。【结果】大豆GmNIC1基因CDS区长度为243 bp,共编码80个氨基酸。其编码蛋白的分子式为C_(396)H_(666)N_(114)O_(110)S_(3),相对分子质量为8880.52 Da,理论等电位点(pI)为10.18,亲水性平均值和脂肪系数分别为-0.198和97.62,该蛋白的不稳定指数是47.10,它是不存在跨膜结构域的非分泌性亲水蛋白。该蛋白质二级结构主要由61.25%的α-螺旋、31.25%的无规则卷曲以及6.25%的β-转角和1.25%的片层构成,且不存在跨膜结构,定位于细胞膜上,其中信号肽位于1~22位氨基酸区域。GmNIC1基因启动子区域含有6种激素响应元件和4种逆境响应元件,且与GmNIC1的互作蛋白中含有AP2/ERF结构域和Ser/Thr蛋白激酶家族,其具有参与非生物胁迫应答调控的功能。GmNIC1蛋白的氨基酸序列与野生大豆(Glycine max)同源蛋白的关系最近。RTPCR检测发现,GmNIC1基因在大豆叶片中表达量最高,同时叶面外施烟酰胺能促进GmNIC1基因的表达,但在非生物胁迫下GmNIC1基因表达量明显降低。【结论】本研究成功克隆大豆GmNIC1基因,并对其编码蛋白的理化性质及表达模式进行分析。发现烟酰胺含量的增加能提高烟酰胺酶活性从而促进GmNIC1基因表达,且GmNIC1基因在调控大豆非生物胁迫方面具有重要作用。[Objective]NIC(Nicotinamide Enzyme)is a key enzyme in the synthesis of NAD+,which is an important regulatory substance in plant growth and development.The present paper aimed to study the function of NIC in soybean(Glycine max)so as to create new soybean varieties with stress resistance.[Method]The bioinformatics methods were used to investigate the structure and function of soybean GmNICI gene,and analyze gene expression patterns through RT-PCR.[Result]It was found that the length of the CDS region of the soybean GmNICI gene was 243 bp,encoding a total of 80 amino acids.The molecular formula of the encoded protein was C_(396)H_(666)N_(114)O_(110)S_(3),the theoretical molecular weight was 8880.52 Da,the theoretical isoelectric point(PI)was 10.18,the average hydrophilicity and fat coefficient were-0.198 and 97.62,the instability index of the protein was 47.10,and it was a non-secretory hydrophilic protein without a transmembrane do-main.The secondary structure of GmNIC1 was mainly composed of alpha helices(61.25%),irregular coils(31.25%),beta turns(6.25%),and lamellar structure(1.25%),and there was no transmembrane structure,localized on the cell membrane,with the signal peptide located in the 1-22 amino acid region.The interacting protein with GmNIC1 contained the AP2/ERF domain and Ser/Thr protein kinase family,which had the function of participating in the regulation of abiotic stress response.The expression level of GmNICI gene was the high-est in soybean leaves,and foliar application of nicotinamide could promote the expression of GmNICl gene.However,under abiotic stress,the expression level of GmNICl gene was significantly reduced.[Conclusion]GmNICl gene was successfully cloned from soybean,and its expres-sion as well as the physicochemical properties of its encoded protein were analyzed.The increase in nicotinamide content can enhance nicotin-amide enzyme activity and promote the expression of GmNICl gene,which plays an important role in regulating soybean abiotic stress.

关 键 词：大豆 GmNIC1 生物信息学 非生物胁迫 

分 类 号：S565.1[农业科学—作物学]