[1]陈杰,杨天宇,黄新元.水稻丝氨酸羟甲基转移酶基因OsSHMT5克隆与功能分析[J].江苏农业学报,2025,(02):209-220.[doi:doi:10.3969/j.issn.1000-4440.2025.02.001]
 CHEN Jie,YANG Tianyu,HUANG Xinyuan.Cloning and functional analysis of serine hydroxymethyltransferase gene OsSHMT5 in rice[J].,2025,(02):209-220.[doi:doi:10.3969/j.issn.1000-4440.2025.02.001]
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水稻丝氨酸羟甲基转移酶基因OsSHMT5克隆与功能分析()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2025年02期
页码:
209-220
栏目:
遗传育种·生理生化
出版日期:
2025-02-28

文章信息/Info

Title:
Cloning and functional analysis of serine hydroxymethyltransferase gene OsSHMT5 in rice
作者:
陈杰12杨天宇2黄新元2
(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)
Author(s):
CHEN Jie12YANG Tianyu2HUANG Xinyuan2
(1.Institute of Vegetable Crops, Jiangsu Academy of Agricultural Sciences/Jiangsu Key Laboratory for Horticultural Crop Genetic Improvement, Nanjing 210014, China;2.State Key Laboratory of Crop Genetics and Germplasm Enhancement & Utilization/College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing 210095, China)
关键词:
水稻丝氨酸羟甲基转移酶OsSHMT5基因硫元素硒元素
Keywords:
riceserine hydroxymethyltransferaseOsSHMT5 genesulphurselenium
分类号:
S511
DOI:
doi:10.3969/j.issn.1000-4440.2025.02.001
文献标志码:
A
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。
Abstract:
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.

参考文献/References:

[1]MCNEIL J B, BOGNAR A L, PEARLMAN R E. In vivo analysis of folate coenzymes and their compartmentation in Saccharomyces cerevisiae[J]. Genetics,1996,142(2):371-381.
[2]LI R, MOORE M, KING J. Investigating the regulation of one-carbon metabolism in Arabidopsis thaliana[J]. Plant & Cell Physiology,2003,44(3):233-241.
[3]赵玉琪,周志林,唐君,等. 甘薯、番茄和拟南芥中丝氨酸羟甲基转移酶基因家族功能研究[J]. 合肥工业大学学报(自然科学版),2022,45(12):1705-1714.
[4]马莉,陈丽梅. 植物丝氨酸羟甲基转移酶基因研究进展[J]. 生物技术通报,2008(2):15-19.
[5]VOLL L M, JAMAI A, RENN P, et al. The photorespiratory Arabidopsis shm1 mutant is deficient in SHM1[J]. Plant Physiology,2006,140(1):59-66.
[6]ZHANG Y, SUN K H, SANDOVAL F J, et al. One-carbon metabolism in plants:characterization of a plastid serine hydroxymethyltransferase[J]. The Biochemical Journal,2010,430(1):97-105.
[7]WANG D K, LIU H Q, LI S J, et al. Characterization and molecular cloning of a serine hydroxymethyltransferase 1 (OsSHM1) in rice[J]. Journal of Integrative Plant Biology,2015,57(9):745-756.
[8]WU J X, ZHANG Z G, ZHANG Q, et al. The molecular cloning and clarification of a photorespiratory mutant,oscdm1,using enhancer trapping[J]. Frontiers in Genetics,2015,6:226.
[9]FU S, WANG K, MA T T, et al. An evolutionarily conserved C4HC3-type E3 ligase regulates plant broad-spectrum resistance against pathogens[J]. Plant Cell,2022,34(5):1822-1843.
[10]PAN T, JIN H M, ZHOU C H, et al. Rice serine hydroxymethyltransferases:evolution,subcellular localization,function and perspectives[J]. Plants,2024,13(8):1116.
[11]MISHRA P, JAIN A, TAKABE T, et al. Heterologous expression of serine hydroxymethyltransferase-3 from rice confers tolerance to salinity stress in E. coli and Arabidopsis[J]. Frontiers in Plant Science,2019,10:217.
[12]HUANG X Y, CHAO D Y, KOPRIVOVA A, et al. Nuclear localised more sulphur accmulation 1 epigenetically regulates sulphur homeostasis in Arabidopsis thaliana[J]. PLoS Genetics,2016,12(9):e1006298.
[13]CHEN J, HUANG X Y, SALT D E, et al. Mutation in OsCADT1 enhances cadmium tolerance and enriches selenium in rice grain[J]. New Phytologist,2020,226(3):838-850.
[14]YAN M, PAN T, ZHU Y, et al. Floury endosperm20 encoding SHMT4 is required for rice endosperm development[J]. Plant Biotechnology Journal,2022,20(8):1438-1440.
[15]MATSUSAKA H, FUKUDA M, ELAKHDAR A, et al. Serine hydroxymethyltransferase participates in the synthesis of cysteine-rich storage proteins in rice seed[J]. Plant Science:An International Journal of Experimental Plant Biology,2021,312:111049.
[16]ELLIS D R, SALT D E. Plants,selenium and human health[J]. Current Opinion in Plant Biology,2003,6(3):273-279.
[17]BOYD R. Selenium stories[J]. Nature Chemistry,2011,3(7):570.
[18]HARTHILL M. Review:micronutrient selenium deficiency influences evolution of some viral infectious diseases[J]. Biological Trace Element Research,2011,143(3):1325-1336.
[19]RAYMAN M P. Selenium intake,status,and health:a complex relationship[J]. Hormones (Athens,Greece),2020,19(1):9-14.
[20]Food and Agriculture Organization of the United Nations. Human vitamin and mineral requirements:report of a joint FAO/WHO expert consultation[R]. Bangkok:Food and Agriculture Organization of the United Nations,2002.
[21]TAN L C, NANCHARAIAH Y V, VAN HULLEBUSCH E D, et al. Selenium:environmental significance,pollution,and biological treatment technologies[J]. Biotechnology Advances,2016,34(5):886-907.
[22]SCHIAVON M, NARDI S, DALLA VECCHIA F, et al. Selenium biofortification in the 21 century:status and challenges for healthy human nutrition[J]. Plant and Soil,2020,453(1/2):245-270.
[23]SUN G X, LIU X, WILLIAMS P N, et al. Distribution and translocation of selenium from soil to grain and its speciation in paddy rice (Oryza sativa L.)[J]. Environmental Science & Technology,2010,44(17):6706-6711.
[24]WILLIAMS P N, LOMBI E, SUN G X, et al. Selenium characterization in the global rice supply chain[J]. Environmental Science & Technology,2009,43(15):6024-6030.
[25]ZHAO F J, MCGRATH S P. Biofortification and phytoremediation[J]. Current Opinion in Plant Biology,2009,12(3):373-380.
[26]SPARKES I A, RUNIONS J, KEARNS A, et al. Rapid,transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants[J]. Nature Protocols,2006,1(4):2019-2025.
[27]MA X L, ZHANG Q Y, ZHU Q L, et al. A robust CRISPR/Cas9 system for convenient,high-efficiency multiplex genome editing in monocot and dicot plants[J]. Molecular Plant,2015,8(8):1274-1284.
[28]陈杰. 水稻和拟南芥镉耐性相关基因OsSHM4和PP2A-4C的克隆和功能解析[D]. 南京:南京农业大学,2019.
[29]ENGEL N, EWALD R, GUPTA K J, et al. The presequence of Arabidopsis serine hydroxymethyltransferase SHM2 selectively prevents import into mesophyll mitochondria[J]. Plant Physiology,2011,157(4):1711-1720.

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备注/Memo

备注/Memo:
收稿日期:2024-09-09基金项目: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)作者简介:陈杰(1992-),男,江苏丹阳人,博士,助理研究员,主要从事植物矿质元素吸收与积累机制研究。(E-mail)jiechen@jaas.ac.cn通讯作者:黄新元,(E-mail)xinyuan.huang@njau.edu.cn
更新日期/Last Update: 2025-03-27