大豆谷胱甘肽还原酶基因的扩增及连作胁迫应答  

Amplification of glutathione reductase gene and its response to continuous cropping stress in soybean

作　　者：韩蓓 李晨 庞园园 方淑梅[1] 梁喜龙[2] HAN Bei;LI Chen;PANG Yuanyuan;FANG Shumei;LIANG Xilong(College of Life Science and Biotechnology,Heilongjiang Bayi Agricultural University,Daqing,Heilongjiang 163319,China;College of Agriculture,Heilongjiang Bayi Agricultural University,Daqing,Heilongjiang 163319,China)

机构地区：[1]黑龙江八一农垦大学生命科学技术学院,黑龙江大庆163319 [2]黑龙江八一农垦大学农学院,黑龙江大庆163319

出　　处：《干旱地区农业研究》2024年第3期60-67,97,共9页Agricultural Research in the Arid Areas

基　　金：国家级大学生创新创业训练计划项目(201910223012);国家自然科学基金项目(31201163)。

摘　　要：利用数据库Phytozome获得大豆胱甘肽还原酶(Glutathione reductase,GR)基因4个成员的编码序列(Coding sequence,CDS)序列,以大豆叶片和根中RNA为模板,经RT-PCR扩增获得Glyma10g03740和Glyma02g16010基因,二者大小均为1638 bp,基因结构相似;获得Glyma16g27210和Glyma02g08180基因,二者结构相似,基因大小均为1506 bp。系统进化分析显示,Glyma10g03740/Glyma02g16010和Glyma16g27210/Glyma02g08180蛋白聚类于不同分支,但分别都与豇豆、尖叶菜豆和芸豆亲缘关系最近。4个成员的结构域相同,均含有FAD/NAD-binding_dom结构域和Pyr_nucl-dis_OxRdtase_dimer结构域。三级结构显示Glyma16g27210和Glyma02g08180构象相似,Glyma10g03740和Glyma02g16010构象相似,4个蛋白均以二聚体结构存在,互作蛋白完全相同,包括2个硫氧还蛋白还原酶和8个硫氧还蛋白。RT-qPCR方法分析GRs基因对连作逆境的响应,结果显示,在连作胁迫下,敏感品种HF55的GRs基因表达在根中启动较早(出苗后15 d内),而叶片中启动较晚,出苗后45 d时表达仍呈上升趋势;对于抗性品种KX8,根和叶片中GRs基因各成员均在出苗后15~45 d表达量达到最高,30 d时根中GRs基因总表达量增加达19.03倍,叶片中GRs基因总表达量增加2.50倍。The CDS of four GR genes were obtained in the soybean database Phytozome.The GR genes were cloned by RT-PCR using RNA from leaf and root as templates,respectively.The sizes of Glyma10g03740 and Glyma02g16010 were both 1638 bp,indicating similar gene structures.The gene structures of Glyma16g27210 and Glyma02g08180 were similar with 1506 bp.Phylogenetic analysis showed that Glyma10g03740/Glyma02g16010 and Glyma16g27210/Glyma02g08180 were clustered in different branches,while they were all closely related to Vigna unguiculata,Phaseolus acutifolius and Phaseolus vulgaris,respectively.The four GRs had the same domains with FAD/NAD binding_dom and Pyr_Nucl dis_OxRdcase_Dimer.The tertiary structure showed that the conformations of Glyma16g27210 and Glyma02g08180 were similar,while those of Glyma10g03740 and Glyma02g16010 exhibited similarity as well.All four GRs existed in homodimers,with identical interacting proteins,including two thioredoxin reductases and eight thioredoxins.The RT-qPCR method was used to analyze the response of GRs to continuous cropping stress.The results showed that under continuous cropping stress,the GR s expression of sensitive variety HF55 was initiated earlier in roots,within 15 days after emergence,but it was activated later in leaves.The expression still showed an upward trend at 45 days after emergence.In the resistant variety KX8,the expression of GRs in roots and leaves reached their highest levels between 15 days and 45 days after emergence.At 30 days,the total expression of GRs in roots and leaves increased by 19.03 times and 2.50 times,respectively.

关 键 词：大豆 谷胱甘肽还原酶 连作逆境 基因扩增 基因表达 

分 类 号：S561.1[农业科学—作物学] S344.4[生物学—分子生物学] Q786