摘要
Clavaminate Synthase-Like(CSL)是生物体内的一种氧化还原酶,可催化氨基酸羟基化。为初步了解小麦CSL基因并探究其相关生物学功能,利用同源克隆的方法,以拟南芥At3g21360(GenBank Accession:NC_003074.8)基因CDS序列为探针搜索小麦基因组核酸序列数据库,得到小麦A、B、D三个亚基因组上的部分同源基因序列。对上述三条序列的全长与编码区进行克隆和测序,经生物信息学分析,发现这三个基因编码的蛋白属于CSL家族。命名该基因为TaCSL1,来自A、B、D三个亚基因组的部分同源基因分别命名为TaCSL1-3A、TaCSL1-3B、TaCSL1-3D。实时荧光定量PCR分析表明,这三个部分同源基因在小麦不同生长发育阶段和不同组织部位均有表达,除叶鞘组织外,其相对表达水平在各时期叶片中均高于其他组织。分析该基因在小麦不同生长发育时期叶片的相对表达水平发现,该基因的相对表达量在幼苗期至孕穗期持续提高,抽穗期显著下降,开花后再次提高,生育末期降低。就三个部分同源基因的表达水平而言,TaCSL1-3A相对表达量均较低,而TaCSL1-3B的相对表达量在孕穗期之前最高,TaCSL1-3D在孕穗期之后最高。本研究明确了TaCSL1的时空表达特征,可为深入研究小麦氨基酸羟基化过程和植物抗病机理研究提供新思路。
Clavaminate Synthase-Like(CSL)is an oxidoreductase in organisms,which can catalyze the hydroxylation of amino acids.In order to preliminarily understand wheat CSL gene and explore its related biological functions,using the method of homologous cloning,the CDS sequence of Arabidopsis At3g21360(GenBank Accession:NC_003074.8)was used as a probe to search the wheat genomic nucleic acid sequence database,and three homologous sequences located on wheat A,B and D subgenomes were obtained.The full length and coding region of the three homologs were obtained,and the proteins encoded by the three sequences were classified as CSL family by bioinformatics analysis.The gene was designated as TaCSL1,and the homologous genes from the three subgenomes A,B,and D were designated as TaCSL1-3A,TaCSL1-3B and TaCSL1-3D,respectively.Real-time fluorescence quantitative PCR analysis showed that TaCSL1-3A,TaCSL1-3B and TaCSL1-3D were expressed at different growth stages and in different tissues of wheat.Except for leaf sheath,the relative expression level of TaCSL1 in leaves was higher than that in other tissues.Results showed that the relative expression level of TaCSL1 increased from seedling stage to booting stage,but decreased significantly at heading stage,and then increased again after flowering,and finally decreased at the end of growth stage.The expression levels of the three homologs were showed that the relative expression level of TaCSL1-3A was always low;the relative expression level of TaCSL1-3B was highest before booting stage,and the relative expression level of TaCSL1-3D was highest after booting stage.To conclude,this study clarified the spatial-temporal expression characteristics of TaCSL1 and provided a new idea for the further study of plant amino acid hydroxylation process and plant disease resistance mechanism.
作者
陈恒
刘迎春
刘育秀
田仁美
周新颖
于利伟
谢彦周
高欣
CHEN Heng;LIU Yingchun;LIU Yuxiu;TIAN Renmei;ZHOU Xinying;YU Liwei;XIE Yanzhou;GAO Xin(College of Agronomy,Northwest A&F University,Yangling,Shaanxi 712100,China)
出处
《麦类作物学报》
CAS
CSCD
北大核心
2021年第8期921-929,共9页
Journal of Triticeae Crops
基金
中国博士后科学基金项目(2016M600817)
陕西省博士后基金项目(K3380218065)
大学生创新训练计划项目(S202010712388)。
