水稻丝氨酸羟甲基转移酶基因OsSHMT5克隆与功能分析  

Cloning and functional analysis of serine hydroxymethyltransferase gene OsSHMT5 in rice

作　　者：陈杰[1,2] 杨天宇[2] 黄新元 CHEN Jie;YANG Tianyu;HUANG Xinyuan(Institute of Vegetable Crops,Jiangsu Academy of Agricultural Sciences/Jiangsu Key Laboratory for Horticultural Crop Genetic Improvement,Nanjing 210014,China;State Key Laboratory of Crop Genetics and Germplasm Enhancement&Utilization/College of Resources and Environmental Sciences,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]江苏省农业科学院蔬菜研究所/江苏省高效园艺作物遗传改良重点实验室,江苏南京210014 [2]南京农业大学资源与环境科学学院/作物遗传与创制创新利用全国重点实验室,江苏南京210095

出　　处：《江苏农业学报》2025年第2期209-220,共12页Jiangsu Journal of Agricultural Sciences

基　　金：江苏省自然科学基金青年基金项目(BK20210389);国家自然科学基金青年基金项目(32202592)。

摘　　要：水稻中已克隆到OsSHMT4(OsCADT1),该基因编码丝氨酸羟甲基转移酶(SHMT),可调控硫酸盐/硒酸盐吸收和同化,其高度同源基因OsSHMT5的功能尚不清楚。本研究基于反向遗传学研究方法,对水稻OsSHMT5基因进行分子克隆,发现OsSHMT5与OsSHMT4(OsCADT1)的同源性高达84%。亚细胞定位结果表明,OsSHMT5蛋白定位于细胞核。利用CRISPR/Cas9技术构建OsSHMT5基因敲除突变体,发现OsSHMT5基因敲除突变体的地上部或地下部中总硫/总硒及各形态硒含量与野生型相比无显著差异。基于转录组水平分析发现,OsSHMT5对水稻硫酸盐/硒酸盐吸收和同化相关基因的表达无显著调控作用。在水稻地下部,OsSHMT5可能具有四吡咯结合、血红素结合、氧化还原酶活性、电子转移活性和铁离子结合等分子功能;在地上部,OsSHMT5可能具有转录调节因子活性、分子功能调节因子、内肽酶调节活性、肽酶抑制剂活性、铜离子结合等分子功能。本研究结果为水稻中OsSHMT家族功能的研究奠定了基础。In rice,OsSHMT4(OsCADT1)has been cloned.This gene encodes serine hydroxymethyltransferase(SHMT),which can regulate the absorption and assimilation of sulfate/selenate.However,the function of its highly homologous gene,OsSHMT5,remains unclear.Based on the reverse genetics research method,the OsSHMT5 gene was cloned in this study.It was found that the homology between OsSHMT5 and OsSHMT4(OsCADT1)was as high as 84%.The results of subcellular localization indicated that OsSHMT5 was localized in the nucleus.The OsSHMT5 gene knockout mutants were constructed using the CRISPR/Cas9 technology.It was found that there were no significant differences in the total sulfur/total selenium and various forms of selenium content in the root or shoot of the OsSHMT5 gene knockout mutants compared with the wild-type.Based on transcriptome-level analysis,it was found that OsSHMT5 had no significant regulatory effect on the expression of genes related to sulfate/selenate absorption and assimilation in rice.In the root,OsSHMT5 may have molecular functions such as tetrapyrrole binding,heme binding,oxidoreductase activity,electron transfer activity,and iron ion binding.In the shoot,OsSHMT5 may have molecular functions such as transcription regulator activity,molecular function regulator,endopeptidase regulatory activity,peptidase inhibitor activity,and copper ion binding.The results of this study lay a foundation for the research on the functions of the OsSHMT family in rice.

关 键 词：水稻 丝氨酸羟甲基转移酶 OsSHMT5基因 硫元素 硒元素 

分 类 号：S511[农业科学—作物学]