摘要
为探讨超氧化物歧化酶(superoxide dismutase,SOD)基因在玉米抗逆反应中的作用,研究选用2个抗旱性有明显差别的玉米品种为试验材料,采用RT-PCR、实时荧光定量PCR(qRT-PCR)及二维凝胶电泳(2-DE)耦联的基质辅助激光解吸电离/飞行时间质谱(MALDI-TOF)法,对ZmSOD基因的序列结构及其在干旱胁迫下的表达特性进行分析。结果表明:(1)从干旱敏感品种‘登海605’(DH605)和抗旱品种‘蠡玉35’(LY35)玉米叶片中分别成功克隆出ZmSOD1和ZmSOD2基因。DH605中ZmSOD1开放阅读框(ORF)全长456 bp,编码151个氨基酸,编码的蛋白等电点为5.76,分子量为15.05 kD。LY35中ZmSOD2 ORF全长459 bp,编码152个氨基酸,编码的蛋白等电点为5.65,分子量为15.11 kD;ZmSOD1和ZmSOD2是亲水性稳定蛋白,都含有Cu/Zn-SOD结构域,蛋白序列N端无信号肽及无跨膜结构域。同源性和系统进化分析表明,玉米ZmSOD和谷子Cu/Zn-SOD2的亲缘关系最近,相似性高达96.05%。(2)在干旱条件下,ZmSOD1在DH605中转录水平明显降低,LY35中ZmSOD2转录水平显著升高;2-DE耦联的质谱分析显示:ZmSOD1在DH605中表达量明显降低,LY35中ZmSOD2表达量无显著性变化,却检测到Mn-SOD蛋白表达量显著增加。(3)相关分析显示,干旱条件下,DH605叶片中ZmSOD1转录水平和其蛋白丰度及SOD酶活性呈极显著性正相关,LY35叶片中ZmSOD2转录水平与SOD酶活性呈显著性正相关。研究结果为进一步探索SOD基因在调节玉米抗性以及逆境胁迫应答过程中的作用机制提供了基础。
In order to explore the role of superoxide dismutase(SOD)gene in maize stress resistance,we selected two maize seedlings with distinct differences in drought resistance as experimental materials.The sequence structure and expression patterns of ZmSOD gene under drought stress were analyzed by RT-PCR,quantitative real-time PCR(qRT-PCR)and two-dimensional gel electrophoresis(2-DE)coupled with matrix assisted laser desorption ionization/time of flight mass spectrometry(MALDI-TOF).The results showed that:(1)The full-length cDNA sequences of SOD gene were cloned successfully from the leaves of drought-sensitive Denghai 605(DH605)and drought-resistant Liyu 35(LY35),respectively.The opening reading frame(ORF)of ZmSOD1 gene in DH605 was 456 bp in length,encoding 151 amino acids.The isoelectric point of ZmSOD1 protein was 5.76,and its molecular weight was 15.05 kD.The ORF of ZmSOD2 gene in LY35 was 459 bp in length,encoding 152 amino acids.The isoelectric point of ZmSOD2 protein was 5.65,and its molecular weight was 15.11 kD.ZmSOD1 and ZmSOD2 are hydrophilic stable proteins,both containing Cu/Zn-SOD domain without signal peptide and transmembrane domain at the N-terminal.Homology and phylogenetic analysis showed that ZmSOD was highly homologous to FmCu/Zn-SOD2,and the similarity was as high as 96.05%.(2)Under drought conditions,the transcription level of ZmSOD1 in DH605 was significantly decreased,and that of ZmSOD2 in LY35 was significantly increased.The 2-DE coupled mass spectrometry showed that the expression level of ZmSOD1 in DH605 was significantly decreased,the expression level of ZmSOD2 in LY35 was not significantly changed,but the expression level of Mn-SOD was significantly increased.(3)Correlation analysis showed that the transcription level of ZmSOD1 in DH605 leaves was marked positive correlated with its protein abundance and SOD activity,and the transcription level of ZmSOD2 in LY35 leaves was positively correlated with SOD activity.The results of this study provided a basis for further exploring the mechanism of SOD gene in regulating maize resistance and stress response.
作者
李玉华
赵振华
吴征
白佳田
王烁莹
王同朝
LI Yuhua;ZHAO Zhenhua;WU Zheng;BAI Jiatian;WANG Shuoying;WANG Tongchao(College of Life Sciences,Zhengzhou Normal University,Zhengzhou 450044,China;Collaborative Innovation Center of Henan Grain Crops,Agronomy College of Henan Agricultural University,Zhengzhou 450002,China)
出处
《西北植物学报》
CAS
CSCD
北大核心
2023年第7期1097-1106,共10页
Acta Botanica Boreali-Occidentalia Sinica
基金
河南省自然科学基金青年项目(212300410304)
河南省高等学校重点科研项目(21A210028)
大学生科研创新基金项目(2021007)。
