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Article(id=1316714775158047618, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1316714454184739470, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20260203, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1773158400000, receivedDateStr=2026-03-11, revisedDate=null, revisedDateStr=null, acceptedDate=1776700800000, acceptedDateStr=2026-04-21, onlineDate=1791858865774, onlineDateStr=2026-10-13, pubDate=1791043200000, pubDateStr=2026-10-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1791858865774, onlineIssueDateStr=2026-10-13, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1791858865774, creator=13701087609, updateTime=1791858865774, updator=13701087609, issue=Issue{id=1316714454184739470, tenantId=1146029695717560320, journalId=1192105938417971205, year='2026', volume='66', issue='10', pageStart='4661', pageEnd='5310', issueExtLink='null', onlineDate='null', pubDate='1791043200000', pubDateStr='2026-10-04', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=null, createTime=1791858789249, creator='13701087609', updateTime=1791885400527, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1316826070066774161, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1316714454184739470, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1316826070066774162, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1316714454184739470, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=4900, endPage=4919, ext={EN=ArticleExt(id=1316714775359374211, articleId=1316714775158047618, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Mechanisms of the response of EsrISR system of Corynebacterium glutamicum to hyperosmotic stress and maintenance of cell wall integrity, columnId=1192149543992045670, journalTitle=Acta Microbiologica Sinica, columnName=Research Article, runingTitle=null, highlight=null, articleAbstract=

[Objective] To elucidate the signal sensing logic of the three-component EsrISR system, which is composed of the accessory protein EsrI, the histidine kinase EsrS, and the response regulator EsrR, in Corynebacterium glutamicum under hyperosmotic stress and its molecular mechanism for maintaining cell wall homeostasis. [Methods] The core stress-responsive gene cluster was identified through RNA-seq. A series of deletion mutants and overexpression strains were constructed through homologous recombination technology. The transcriptional fusion reporter system, field emission scanning electron microscopy/transmission electron microscopy (FESEM/TEM), electrophoretic mobility shift assay (EMSA), and real-time quantitative reverse transcription PCR, (RT-qPCR) were employed to systematically investigate the sensing threshold, stress response patterns, and global target gene networks of this system. [Results] RNA-seq showed that the esrISR gene cluster was significantly upregulated under 1.7 mol/L NaCl stress. Under the same stress conditions, the deletion of the regulator EsrR led to a significant decrease in biomass, with the maximum OD600 dropping to 47.8% of that of the wild type. Moreover, severe damage and collapse of the cell wall were observed via electron microscopy. The construction and analysis of reporter plasmids showed that this system was driven by an internal promoter, peaking at 1.7 mol/L NaCl, with a maximum fluorescence intensity of 880 RFU/OD600. The proposed regulatory model indicated that in the absence of external stress, EsrS mainly exerted phosphatase activity to repress the transcription of esrI, thereby maintaining the repressed state of the system (deletion of esrS led to an approximately 15.0-fold upregulation of esrI). Upon sensing hyperosmotic signals, EsrS changed its activity, promoting the phosphorylation of the regulator EsrR. Subsequently, the phosphorylated EsrR specifically bound to the core regulatory region, thereby activating the positive feedback regulatory loop. Transcriptomic analysis revealed significant alterations in the expression of key genes associated with cell wall homeostasis in the esrR deletion mutant (ΔesrR). RT-qPCR analysis confirmed that EsrR functioned as a crucial regulator of cell wall biosynthesis primarily by positively activating key pathway genes. Specifically, the peptidoglycan synthesis genes murB and CGL_RS01830 were downregulated by 95.0% in ΔesrR and upregulated by 2.0-6.5 fold in esrR-OE; the trehalose biosynthesis genes treS, treY, and treZ were downregulated by 75.0%-83.0% in ΔesrR and upregulated by 2.5-4.2 fold in esrR-OE; cmrA and gatC were significantly repressed in ΔesrR and upregulated by 3.5-folds in esrR-OE, thereby effectively maintaining the physical homeostasis of the cell wall. [Conclusion] This study provides new insights into the regulation of cell wall homeostasis by the three-component EsrISR system in Corynebacterium, providing critical genetic targets for the robustness engineering of industrial chassis strains.

, authors=Aoyi JI, Zhihao FAN, Xintao LUO, Hui GAO, Meijuan XU*, authorsList=Aoyi JI, Zhihao FAN, Xintao LUO, Hui GAO, Meijuan XU, authorCompany=null, correspAuthors=Meijuan XU, authorNote=null, correspAuthorsNote=
*E-mail:
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【目的】 解析由辅助蛋白EsrI、组氨酸激酶EsrS与响应调节子EsrR组成的三组分EsrISR系统在谷氨酸棒杆菌(Corynebacterium glutamicum)应对高渗胁迫中的信号感应逻辑及其维持细胞壁稳态的分子机制。 【方法】 通过转录组测序(RNA sequencing, RNA-seq)筛选锁定应激核心基因簇，利用同源重组技术构建了一系列缺失突变体及过表达株，并结合转录融合报告系统、电子显微镜成像、凝胶迁移实验(electrophoretic mobility shift assay, EMSA)以及实时荧光定量逆转录PCR (real-time quantitative reverse transcription PCR, RT-qPCR)定量分析深入探讨了该系统的感应阈值、应激模式及全局靶基因网络。 【结果】 转录组学测序显示，在1.7 mol/L NaCl胁迫下esrISR基因簇呈显著上调。在同等胁迫下，响应调节子EsrR的缺失会导致生物量显著降低，最大OD600下降至野生型的47.8%；且电镜观察到细胞壁发生严重的破损塌陷现象。通过构建、分析报告质粒，发现此系统受到内部启动子驱动；在1.7 mol/L NaCl胁迫时esrISR基因表达呈峰值，最大荧光强度达880 RFU/OD600。本研究提出的调控模型表明，在无外界胁迫时EsrS主要发挥磷酸酶活性以维持系统的抑制状态(缺失esrS导致esrI表达上调约15.0倍)；感知高渗信号后EsrS活性发生改变，促使响应调节子EsrR发生磷酸化。随后，磷酸化后的EsrR特异性结合至核心调控区，进而开启正反馈调控回路。转录组学分析表明，ΔesrR中涉及细胞壁稳态的关键基因表达发生改变，尤其是肽聚糖合成基因(如murB、CGL_RS01830等)显著下调。RT-qPCR实验证实，EsrR具有调节细胞壁合成的功能，其主要通过正向激活细胞壁合成途径的关键基因响应高渗胁迫。其中，肽聚糖合成基因murB与CGL_RS01830在ΔesrR中下调约95.0%，在esrR过表达(esrR-OE)菌株中上调2.0-6.5倍；海藻糖合成基因treS、treY与treZ在缺失株中表达量下降了75.0%-83.0%，在过表达株中上调2.5-4.2倍；cmrA与gatC在缺失株中表达量下调并在过表达株中上调3.5倍，从而有效维持了细胞壁的物理稳态。 【结论】 本研究为三组分EsrISR系统参与棒状菌属细胞壁稳态的调控提供了新的见解，并为工业底盘菌株的鲁棒性改造提供了关键遗传靶点。

, authors=季傲艺, 范志豪, 罗薪涛, 高惠, 徐美娟*, authorsList=季傲艺, 范志豪, 罗薪涛, 高惠, 徐美娟, authorCompany=null, correspAuthors=徐美娟, authorNote=

作者贡献声明

季傲艺：数据收集和处理、论文撰写和修改；范志豪：协助实验操作、参与论文讨论；罗薪涛：数据收集和处理；高惠：研究构思和设计、论文审阅和修改；徐美娟：研究构思和设计、项目经费支持与实验平台资源、论文审阅与修改。

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articleId=1316714775158047618, language=CN, orderNo=3, keyword=细胞壁结构), Keyword(id=1316829803051860439, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=CN, orderNo=4, keyword=高渗胁迫)], refs=[Reference(id=1316829808085025274, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2026, volume=16, issue=2, pageStart=195, pageEnd=null, url=null, language=null, rfNumber=[1], rfOrder=0, authorNames=Qi L, He H, Xiang SH, Cao H, journalName=Catalysts, refType=null, unstructuredReference=Qi L, He H, Xiang SH, Cao H. A two-enzyme entry module triggers an endogenous biocatalytic cascade for green biosynthesis of pyridoxal 5′-phosphate in Corynebacterium glutamicum [J]. Catalysts, 2026, 16(2): 195., articleTitle=A two-enzyme entry module triggers an endogenous biocatalytic cascade for green biosynthesis of pyridoxal 5′-phosphate in Corynebacterium glutamicum, refAbstract=null), Reference(id=1316829808194077179, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2020, volume=8, issue=6, pageStart=866, pageEnd=null, url=null, language=null, rfNumber=[2], rfOrder=1, authorNames=Walter T, Al Medani N, Burgardt A, Cankar K, Ferrer L, Kerbs A, Lee JH, Mindt M, Risse JM, Wendisch VF, journalName=Microorganisms, refType=null, unstructuredReference=Walter T, Al Medani N, Burgardt A, Cankar K, Ferrer L, Kerbs A, Lee JH, Mindt M, Risse JM, Wendisch VF. Fermentative N-methylanthranilate production by engineered Corynebacterium glutamicum [J]. Microorganisms, 2020, 8(6): 866., articleTitle=Fermentative N-methylanthranilate production by engineered Corynebacterium glutamicum, refAbstract=null), Reference(id=1316829808298934780, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2012, volume=3, issue=4, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[3], rfOrder=2, authorNames=Zahoor A, Lindner SN, Wendisch VF, journalName=Computational and Structural Biotechnology Journal, refType=null, unstructuredReference=Zahoor A, Lindner SN, Wendisch VF. Metabolic engineering of Corynebacterium glutamicum aimed at alternative carbon sources and new products[J]. Computational and Structural Biotechnology Journal, 2012, 3(4): e201210004., articleTitle=Metabolic engineering of Corynebacterium glutamicum aimed at alternative carbon sources and new products, refAbstract=null), Reference(id=1316829808361849341, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2024, volume=77, issue=null, pageStart=108475, pageEnd=null, url=null, language=null, rfNumber=[4], rfOrder=3, authorNames=Li MH, Li H, Zhang X, Liang YC, Li C, Sun ML, Li K, Liu CG, Sinskey AJ, journalName=Biotechnology Advances, refType=null, unstructuredReference=Li MH, Li H, Zhang X, Liang YC, Li C, Sun ML, Li K, Liu CG, Sinskey AJ. Metabolic engineering of Corynebacterium glutamicum: unlocking its potential as a key cell factory platform for organic acid production[J]. Biotechnology Advances, 2024, 77: 108475., articleTitle=Metabolic engineering of Corynebacterium glutamicum: unlocking its potential as a key cell factory platform for organic acid production, refAbstract=null), Reference(id=1316829808433152510, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2025, volume=21, issue=12, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[5], rfOrder=4, authorNames=Schalck T, Katyal M, De Graeve S, Roba L, Baldoma JV, Swings T, Van den Bergh B, Michiels J, journalName=PLoS Genetics, refType=null, unstructuredReference=Schalck T, Katyal M, De Graeve S, Roba L, Baldoma JV, Swings T, Van den Bergh B, Michiels J. Reprogramming the EnvZ-OmpR two-component system confers ethanol tolerance in Escherichia coli by stabilizing the outer membrane and altering ferric homeostasis[J]. PLoS Genetics, 2025, 21(12): e1011707., articleTitle=Reprogramming the EnvZ-OmpR two-component system confers ethanol tolerance in Escherichia coli by stabilizing the outer membrane and altering ferric homeostasis, refAbstract=null), Reference(id=1316829808512844287, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2014, volume=151, issue=null, pageStart=120, pageEnd=127, url=null, language=null, rfNumber=[6], rfOrder=5, authorNames=Yang LB, Zhan XB, Zheng ZY, Wu JR, Gao MJ, Lin CC, journalName=Bioresource Technology, refType=null, unstructuredReference=Yang LB, Zhan XB, Zheng ZY, Wu JR, Gao MJ, Lin CC. A novel osmotic pressure control fed-batch fermentation strategy for improvement of erythritol production by Yarrowia lipolytica from glycerol[J]. Bioresource Technology, 2014, 151: 120-127., articleTitle=A novel osmotic pressure control fed-batch fermentation strategy for improvement of erythritol production by Yarrowia lipolytica from glycerol, refAbstract=null), Reference(id=1316829808605118976, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2026, volume=16, issue=null, pageStart=1728984, pageEnd=null, url=null, language=null, rfNumber=[7], rfOrder=6, authorNames=Khider MLK, Irla M, López MG, Gispert A, Konjetzko T, Meliawati M, Schmid J, Brautaset T, Brito LF, journalName=Frontiers in Microbiology, refType=null, unstructuredReference=Khider MLK, Irla M, López MG, Gispert A, Konjetzko T, Meliawati M, Schmid J, Brautaset T, Brito LF. CRISPR-Cas9-driven genome editing in Bacillus methanolicus MGA3[J]. Frontiers in Microbiology, 2026, 16: 1728984., articleTitle=CRISPR-Cas9-driven genome editing in Bacillus methanolicus MGA3, refAbstract=null), Reference(id=1316829808689005057, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2022, volume=7, issue=1, pageStart=533, pageEnd=540, url=null, language=null, rfNumber=[8], rfOrder=7, authorNames=Mohedano MT, Konzock O, Chen Y, journalName=Synthetic and Systems Biotechnology, refType=null, unstructuredReference=Mohedano MT, Konzock O, Chen Y. Strategies to increase tolerance and robustness of industrial microorganisms[J]. Synthetic and Systems Biotechnology, 2022, 7(1): 533-540., articleTitle=Strategies to increase tolerance and robustness of industrial microorganisms, refAbstract=null), Reference(id=1316829808768696834, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2017, volume=12, issue=10, pageStart=1700014, pageEnd=null, url=null, language=null, rfNumber=[9], rfOrder=8, authorNames=Gong ZW, Nielsen J, Zhou YJ, journalName=Biotechnology Journal, refType=null, unstructuredReference=Gong ZW, Nielsen J, Zhou YJ. Engineering robustness of microbial cell factories[J]. Biotechnology Journal, 2017, 12(10): 1700014., articleTitle=Engineering robustness of microbial cell factories, refAbstract=null), Reference(id=1316829808865165827, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2024, volume=73, issue=null, pageStart=108366, pageEnd=null, url=null, language=null, rfNumber=[10], rfOrder=9, authorNames=Guan AL, He ZX, Wang X, Jia ZJ, Qin JF, journalName=Biotechnology Advances, refType=null, unstructuredReference=Guan AL, He ZX, Wang X, Jia ZJ, Qin JF. Engineering the next-generation synthetic cell factory driven by protein engineering[J]. Biotechnology Advances, 2024, 73: 108366., articleTitle=Engineering the next-generation synthetic cell factory driven by protein engineering, refAbstract=null), Reference(id=1316829809167155716, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2025, volume=109, issue=null, pageStart=255, pageEnd=null, url=null, language=null, rfNumber=[11], rfOrder=10, authorNames=Takahashi Y, Hirasawa T, journalName=Applied Microbiology and Biotechnology, refType=null, unstructuredReference=Takahashi Y, Hirasawa T. Enhancement of ergothioneine production in Corynebacterium glutamicum by increasing osmotic pressure[J]. Applied Microbiology and Biotechnology, 2025, 109: 255., articleTitle=Enhancement of ergothioneine production in Corynebacterium glutamicum by increasing osmotic pressure, refAbstract=null), Reference(id=1316829809569808901, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2025, volume=63, issue=3, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[12], rfOrder=11, authorNames=Jeon S, Sohn YJ, Lee H, Park JY, Kim D, Lee ES, Park SJ, journalName=Journal of Microbiology, refType=null, unstructuredReference=Jeon S, Sohn YJ, Lee H, Park JY, Kim D, Lee ES, Park SJ. Recent advances in the design-build-test-learn (DBTL) cycle for systems metabolic engineering of Corynebacterium glutamicum [J]. Journal of Microbiology, 2025, 63(3): e2501021., articleTitle=Recent advances in the design-build-test-learn (DBTL) cycle for systems metabolic engineering of Corynebacterium glutamicum, refAbstract=null), Reference(id=1316829809997627910, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2023, volume=21, issue=null, pageStart=1955, pageEnd=1965, url=null, language=null, rfNumber=[13], rfOrder=12, authorNames=Kim GY, Kim J, Park G, Kim HJ, Yang JN, Seo SW, journalName=Computational and Structural Biotechnology Journal, refType=null, unstructuredReference=Kim GY, Kim J, Park G, Kim HJ, Yang JN, Seo SW. Synthetic biology tools for engineering Corynebacterium glutamicum [J]. Computational and Structural Biotechnology Journal, 2023, 21: 1955-1965., articleTitle=Synthetic biology tools for engineering Corynebacterium glutamicum, refAbstract=null), Reference(id=1316829810429641223, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2022, volume=168, issue=4, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[14], rfOrder=13, authorNames=De Sousa-d’Auria C, Constantinesco F, Bayan N, Constant P, Tropis M, Daffé M, Graille M, Houssin C, journalName=Microbiology, refType=null, unstructuredReference=De Sousa-d’Auria C, Constantinesco F, Bayan N, Constant P, Tropis M, Daffé M, Graille M, Houssin C. Cg1246, a new player in mycolic acid biosynthesis in Corynebacterium glutamicum [J]. Microbiology, 2022, 168(4): 001171., articleTitle=Cg1246, a new player in mycolic acid biosynthesis in Corynebacterium glutamicum, refAbstract=null), Reference(id=1316829811801178634, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2021, volume=9, issue=2, pageStart=409, pageEnd=null, url=null, language=null, rfNumber=[15], rfOrder=14, authorNames=Kumagai Y, Hirasawa T, Wachi M, journalName=Microorganisms, refType=null, unstructuredReference=Kumagai Y, Hirasawa T, Wachi M. Requirement of the LtsA protein for formation of the mycolic acid-containing layer on the cell surface of Corynebacterium glutamicum [J]. Microorganisms, 2021, 9(2): 409., articleTitle=Requirement of the LtsA protein for formation of the mycolic acid-containing layer on the cell surface of Corynebacterium glutamicum, refAbstract=null), Reference(id=1316829812174471692, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2025, volume=23, issue=4, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[16], rfOrder=15, authorNames=Isbilir B, Yeates A, Alva V, Bharat TAM, journalName=PLoS Biology, refType=null, unstructuredReference=Isbilir B, Yeates A, Alva V, Bharat TAM. Mapping the ultrastructural topology of the corynebacterial cell surface[J]. PLoS Biology, 2025, 23(4): e3003130., articleTitle=Mapping the ultrastructural topology of the corynebacterial cell surface, refAbstract=null), Reference(id=1316829812262552077, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2023, volume=63, issue=1, pageStart=106, pageEnd=123, url=null, language=null, rfNumber=[17], rfOrder=16, authorNames=楼洁, 胡晓, 梁延群, 朱怡铃, 音建华, journalName=微生物学报, refType=null, unstructuredReference=楼洁, 胡晓, 梁延群, 朱怡铃, 音建华. 肽聚糖的生物合成及其调控机制研究进展[J]. 微生物学报, 2023, 63(1): 106-123., articleTitle=肽聚糖的生物合成及其调控机制研究进展, refAbstract=null), Reference(id=1316829812338049550, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2023, volume=63, issue=1, pageStart=106, pageEnd=123, url=null, language=null, rfNumber=[17], rfOrder=17, authorNames=Lou J, Hu X, Liang YQ, Zhu YL, Yin JH, journalName=Acta Microbiologica Sinica, refType=null, unstructuredReference=Lou J, Hu X, Liang YQ, Zhu YL, Yin JH. Peptidoglycan biosynthesis and the regulatory mechanism[J]. Acta Microbiologica Sinica, 2023, 63(1): 106-123 (in Chinese)., articleTitle=null, refAbstract=null), Reference(id=1316829812556153359, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2006, volume=41, issue=2, pageStart=473, pageEnd=476, url=null, language=null, rfNumber=[18], rfOrder=18, authorNames=Liu HJ, Liu DH, Zhong JJ, journalName=Process Biochemistry, refType=null, unstructuredReference=Liu HJ, Liu DH, Zhong JJ. Quantitative response of trehalose and glycerol syntheses by Candida krusei to osmotic stress of the medium[J]. Process Biochemistry, 2006, 41(2): 473-476., articleTitle=Quantitative response of trehalose and glycerol syntheses by Candida krusei to osmotic stress of the medium, refAbstract=null), Reference(id=1316829812648428048, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2020, volume=33, issue=3, pageStart=462, pageEnd=473, url=null, language=null, rfNumber=[19], rfOrder=19, authorNames=MacIntyre AM, Barth JX, Pellitteri Hahn MC, Scarlett CO, Genin S, Allen C, journalName=Molecular Plant-Microbe Interactions®, refType=null, unstructuredReference=MacIntyre AM, Barth JX, Pellitteri Hahn MC, Scarlett CO, Genin S, Allen C. Trehalose synthesis contributes to osmotic stress tolerance and virulence of the bacterial wilt Pathogen Ralstonia solanacearum [J]. Molecular Plant-Microbe Interactions®, 2020, 33(3): 462-473., articleTitle=Trehalose synthesis contributes to osmotic stress tolerance and virulence of the bacterial wilt Pathogen Ralstonia solanacearum, refAbstract=null), Reference(id=1316829812996555281, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2025, volume=13, issue=5, pageStart=1120, pageEnd=null, url=null, language=null, rfNumber=[20], rfOrder=20, authorNames=Zou Y, Huang DN, Liu XX, Yang YK, Liu CL, Li Y, Bai ZH, journalName=Microorganisms, refType=null, unstructuredReference=Zou Y, Huang DN, Liu XX, Yang YK, Liu CL, Li Y, Bai ZH. Two-component MprAB system regulates the expression of genes involved in cell envelope biosynthesis in Corynebacterium glutamicum [J]. Microorganisms, 2025, 13(5): 1120., articleTitle=Two-component MprAB system regulates the expression of genes involved in cell envelope biosynthesis in Corynebacterium glutamicum, refAbstract=null), Reference(id=1316829813109801490, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2024, volume=10, issue=3, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[21], rfOrder=21, authorNames=Barretto LAF, Van PT, Fowler CC, journalName=Microbial Genomics, refType=null, unstructuredReference=Barretto LAF, Van PT, Fowler CC. Conserved patterns of sequence diversification provide insight into the evolution of two-component systems in Enterobacteriaceae [J]. Microbial Genomics, 2024, 10(3): 001215., articleTitle=Conserved patterns of sequence diversification provide insight into the evolution of two-component systems in Enterobacteriaceae, refAbstract=null), Reference(id=1316829813344682515, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2017, volume=8, issue=null, pageStart=2104, pageEnd=null, url=null, language=null, rfNumber=[22], rfOrder=22, authorNames=Liu YX, Rose J, Huang SJ, Hu YB, Wu Q, Wang D, Li CG, Liu ML, Zhou P, Jiang L, journalName=Nature Communications, refType=null, unstructuredReference=Liu YX, Rose J, Huang SJ, Hu YB, Wu Q, Wang D, Li CG, Liu ML, Zhou P, Jiang L. A pH-gated conformational switch regulates the phosphatase activity of bifunctional HisKA-family histidine kinases[J]. Nature Communications, 2017, 8: 2104., articleTitle=A pH-gated conformational switch regulates the phosphatase activity of bifunctional HisKA-family histidine kinases, refAbstract=null), Reference(id=1316829813424374292, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2015, volume=15, issue=null, pageStart=59, pageEnd=null, url=null, language=null, rfNumber=[23], rfOrder=23, authorNames=Mitchell SL, Ismail AM, Kenrick SA, Camilli A, journalName=BMC Microbiology, refType=null, unstructuredReference=Mitchell SL, Ismail AM, Kenrick SA, Camilli A. The VieB auxiliary protein negatively regulates the VieSA signal transduction system in Vibrio cholerae [J]. BMC Microbiology, 2015, 15: 59., articleTitle=The VieB auxiliary protein negatively regulates the VieSA signal transduction system in Vibrio cholerae, refAbstract=null), Reference(id=1316829813529231893, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2011, volume=157, issue=2, pageStart=373, pageEnd=386, url=null, language=null, rfNumber=[24], rfOrder=24, authorNames=Fritsch F, Mauder N, Williams T, Weiser J, Oberle M, Beier D, journalName=Microbiology, refType=null, unstructuredReference=Fritsch F, Mauder N, Williams T, Weiser J, Oberle M, Beier D. The cell envelope stress response mediated by the LiaFSR Lm three-component system of Listeria monocytogenes is controlled via the phosphatase activity of the bifunctional histidine kinase LiaS Lm[J]. Microbiology, 2011, 157(2): 373-386., articleTitle=The cell envelope stress response mediated by the LiaFSR Lm three-component system of Listeria monocytogenes is controlled via the phosphatase activity of the bifunctional histidine kinase LiaS Lm, refAbstract=null), Reference(id=1316829813801861654, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2017, volume=106, issue=5, pageStart=719, pageEnd=741, url=null, language=null, rfNumber=[25], rfOrder=25, authorNames=Kleine B, Chattopadhyay A, Polen T, Pinto D, Mascher T, Bott M, Brocker M, Freudl R, journalName=Molecular Microbiology, refType=null, unstructuredReference=Kleine B, Chattopadhyay A, Polen T, Pinto D, Mascher T, Bott M, Brocker M, Freudl R. The three-component system EsrISR regulates a cell envelope stress response in Corynebacterium glutamicum [J]. Molecular Microbiology, 2017, 106(5): 719-741., articleTitle=The three-component system EsrISR regulates a cell envelope stress response in Corynebacterium glutamicum, refAbstract=null), Reference(id=1316829813931885079, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2023, volume=50, issue=8, pageStart=3562, pageEnd=3574, url=null, language=null, rfNumber=[26], rfOrder=26, authorNames=陈晓婷, 乾莲, 陈卫国, 孙士正, 白宗凯, 赵纯静, 武有聪, journalName=微生物学通报, refType=null, unstructuredReference=陈晓婷, 乾莲, 陈卫国, 孙士正, 白宗凯, 赵纯静, 武有聪. 表皮葡萄球菌vraSR-srrAB突变株的构建及生物学表型[J]. 微生物学通报, 2023, 50(8): 3562-3574., articleTitle=表皮葡萄球菌vraSR-srrAB突变株的构建及生物学表型, refAbstract=null), Reference(id=1316829814284206616, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2023, volume=50, issue=8, pageStart=3562, pageEnd=3574, url=null, language=null, rfNumber=[26], rfOrder=27, authorNames=Chen XT, Qian L, Chen WG, Sun SZ, Bai ZK, Zhao CJ, Wu YC, journalName=Microbiology China, refType=null, unstructuredReference=Chen XT, Qian L, Chen WG, Sun SZ, Bai ZK, Zhao CJ, Wu YC. Construction and biological phenotype characterization of vraSR-srrAB deletion mutant of Staphylococcus epidermidis [J]. Microbiology China, 2023, 50(8): 3562-3574 (in Chinese)., articleTitle=null, refAbstract=null), Reference(id=1316829814741385753, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2021, volume=4, issue=null, pageStart=306, pageEnd=null, url=null, language=null, rfNumber=[27], rfOrder=28, authorNames=Wenzel M, Dekker MP, Wang BW, Burggraaf MJ, Bitter W, van Weering JRT, Hamoen LW, journalName=Communications Biology, refType=null, unstructuredReference=Wenzel M, Dekker MP, Wang BW, Burggraaf MJ, Bitter W, van Weering JRT, Hamoen LW. A flat embedding method for transmission electron microscopy reveals an unknown mechanism of tetracycline[J]. Communications Biology, 2021, 4: 306., articleTitle=A flat embedding method for transmission electron microscopy reveals an unknown mechanism of tetracycline, refAbstract=null), Reference(id=1316829816335221274, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2025, volume=20, issue=12, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[28], rfOrder=29, authorNames=Kim JH, Kang YJ, Kang YW, Park SE, Jin HB, Hong E, Park HH, journalName=PLoS One, refType=null, unstructuredReference=Kim JH, Kang YJ, Kang YW, Park SE, Jin HB, Hong E, Park HH. A putative membrane-associated YdjB-like protein from Salmonella enterica exhibits a non-canonical ACT-like fold[J]. PLoS One, 2025, 20(12): e0336436., articleTitle=A putative membrane-associated YdjB-like protein from Salmonella enterica exhibits a non-canonical ACT-like fold, refAbstract=null), Reference(id=1316829816729485851, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2011, volume=38, issue=9, pageStart=1385, pageEnd=1392, url=null, language=null, rfNumber=[29], rfOrder=30, authorNames=张清仪, 王阶平, 程钢, 刘钟慧, 范文瑾, 何进, journalName=微生物学通报, refType=null, unstructuredReference=张清仪, 王阶平, 程钢, 刘钟慧, 范文瑾, 何进. 苏云金芽孢杆菌双组分信号转导系统的生物信息学分析[J]. 微生物学通报, 2011, 38(9): 1385-1392., articleTitle=苏云金芽孢杆菌双组分信号转导系统的生物信息学分析, refAbstract=null), Reference(id=1316829816842732060, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2011, volume=38, issue=9, pageStart=1385, pageEnd=1392, url=null, language=null, rfNumber=[29], rfOrder=31, authorNames=Zhang QY, Wang JP, Cheng G, Liu ZH, Fan WJ, He J, journalName=Microbiology China, refType=null, unstructuredReference=Zhang QY, Wang JP, Cheng G, Liu ZH, Fan WJ, He J. Bioinformatics analysis of two-component signal transduction systems of Bacillus thuringiensis [J]. Microbiology China, 2011, 38(9): 1385-1392 (in Chinese)., articleTitle=null, refAbstract=null), Reference(id=1316829817203442205, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2025, volume=11, issue=12, pageStart=850, pageEnd=null, url=null, language=null, rfNumber=[30], rfOrder=32, authorNames=Wang MJ, Gu SY, Guo J, Wu JY, Wang XH, Noman M, Wang JY, Li L, journalName=Journal of Fungi, refType=null, unstructuredReference=Wang MJ, Gu SY, Guo J, Wu JY, Wang XH, Noman M, Wang JY, Li L. BcHK71 and BcHK67, two-component histidine kinases, regulate conidial morphogenesis, glycerol synthesis, and virulence in Botrytis cinerea [J]. Journal of Fungi, 2025, 11(12): 850., articleTitle=BcHK71 and BcHK67, two-component histidine kinases, regulate conidial morphogenesis, glycerol synthesis, and virulence in Botrytis cinerea, refAbstract=null), Reference(id=1316829817597706783, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2023, volume=290, issue=4, pageStart=1060, pageEnd=1077, url=null, language=null, rfNumber=[31], rfOrder=33, authorNames=Abhishek S, Deeksha W, Rajakumara E, journalName=The FEBS Journal, refType=null, unstructuredReference=Abhishek S, Deeksha W, Rajakumara E. Mechanistic insights into allosteric regulation of methylated DNA and histone H3 recognition by SRA and SET domains of SUVH5 and the basis for di-methylation of lysine residue[J]. The FEBS Journal, 2023, 290(4): 1060-1077., articleTitle=Mechanistic insights into allosteric regulation of methylated DNA and histone H3 recognition by SRA and SET domains of SUVH5 and the basis for di-methylation of lysine residue, refAbstract=null), Reference(id=1316829817987777056, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2024, volume=32, issue=9, pageStart=1381, pageEnd=1393.e4, url=null, language=null, rfNumber=[32], rfOrder=34, authorNames=Watanabe N, Savchenko A, journalName=Structure, refType=null, unstructuredReference=Watanabe N, Savchenko A. Molecular insights into the initiation step of the Rcs signaling pathway[J]. Structure, 2024, 32(9): 1381-1393.e4., articleTitle=Molecular insights into the initiation step of the Rcs signaling pathway, refAbstract=null), Reference(id=1316829818373653025, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2025, volume=16, issue=null, pageStart=6491, pageEnd=null, url=null, language=null, rfNumber=[33], rfOrder=35, authorNames=Sewgoolam B, Jim KK, de Bakker V, Bock FP, Gibson PS, Veening JW, journalName=Nature Communications, refType=null, unstructuredReference=Sewgoolam B, Jim KK, de Bakker V, Bock FP, Gibson PS, Veening JW. Genome-wide antibiotic-CRISPRi profiling identifies LiaR activation as a strategy to resensitize fluoroquinolone-resistant Streptococcus pneumoniae [J]. Nature Communications, 2025, 16: 6491., articleTitle=Genome-wide antibiotic-CRISPRi profiling identifies LiaR activation as a strategy to resensitize fluoroquinolone-resistant Streptococcus pneumoniae, refAbstract=null), Reference(id=1316829818491093538, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2021, volume=9, issue=10, pageStart=2026, pageEnd=null, url=null, language=null, rfNumber=[34], rfOrder=36, authorNames=Guffey AA, Loll PJ, journalName=Microorganisms, refType=null, unstructuredReference=Guffey AA, Loll PJ. Regulation of resistance in vancomycin-resistant enterococci: the VanRS two-component system[J]. Microorganisms, 2021, 9(10): 2026., articleTitle=Regulation of resistance in vancomycin-resistant enterococci: the VanRS two-component system, refAbstract=null), Reference(id=1316829818843415075, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2020, volume=5, issue=4, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[35], rfOrder=37, authorNames=Cassiano MHA, Silva-Rocha R, journalName=mSystems, refType=null, unstructuredReference=Cassiano MHA, Silva-Rocha R. Benchmarking bacterial promoter prediction tools: potentialities and limitations[J]. mSystems, 2020, 5(4): e00439-20., articleTitle=Benchmarking bacterial promoter prediction tools: potentialities and limitations, refAbstract=null), Reference(id=1316829818923106852, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2018, volume=9, issue=null, pageStart=1433, pageEnd=null, url=null, language=null, rfNumber=[36], rfOrder=38, authorNames=Landry BP, Palanki R, Dyulgyarov N, Hartsough LA, Tabor JJ, journalName=Nature Communications, refType=null, unstructuredReference=Landry BP, Palanki R, Dyulgyarov N, Hartsough LA, Tabor JJ. Phosphatase activity tunes two-component system sensor detection threshold[J]. Nature Communications, 2018, 9: 1433., articleTitle=Phosphatase activity tunes two-component system sensor detection threshold, refAbstract=null), Reference(id=1316829820756017701, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, doi=null, pmid=null, pmcid=null, year=2018, volume=206, issue=null, pageStart=177, pageEnd=185, url=null, language=null, rfNumber=[37], rfOrder=39, authorNames=Cao Q, Feng FF, Wang H, Xu XJ, Chen HC, Cai XW, Wang XR, journalName=Microbiological Research, refType=null, unstructuredReference=Cao Q, Feng FF, Wang H, Xu XJ, Chen HC, Cai XW, Wang XR. Haemophilus parasuis CpxRA two-component system confers bacterial tolerance to environmental stresses and macrolide resistance[J]. Microbiological Research, 2018, 206: 177-185., articleTitle=Haemophilus parasuis CpxRA two-component system confers bacterial tolerance to environmental stresses and macrolide resistance, refAbstract=null)], funds=[Fund(id=1316829805442613747, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, awardId=32570026, language=EN, fundingSource=National Natural Science Foundation of China(32570026), fundOrder=null, country=null), Fund(id=1316829805509722612, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, awardId=32570026, language=CN, fundingSource=国家自然科学基金(32570026), fundOrder=null, country=null), Fund(id=1316829805601997301, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, awardId=2023YFD1300700, language=EN, fundingSource=National Key Research and Development Program of China(2023YFD1300700), fundOrder=null, country=null), Fund(id=1316829807527182839, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, awardId=2023YFD1300700, language=CN, fundingSource=国家重点研发计划(2023YFD1300700), fundOrder=null, country=null), Fund(id=1316829807699149304, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, awardId=KLIB-KF202504, language=EN, fundingSource=Program of Key Laboratory of Industrial Biotechnology of Ministry of Education in Jiangnan University(KLIB-KF202504), fundOrder=null, country=null), Fund(id=1316829807820784121, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, awardId=KLIB-KF202504, language=CN, fundingSource=江南大学工业生物技术教育部重点实验室项目(KLIB-KF202504), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1316829799243432372, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, xref=null, ext=[AuthorCompanyExt(id=1316829799302152629, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, companyId=1316829799243432372, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China), AuthorCompanyExt(id=1316829799327318454, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, companyId=1316829799243432372, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=江南大学 生物工程学院，工业生物技术教育部重点实验室，江苏 无锡)])], figs=[ArticleFig(id=1316829803257381336, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=EN, label=Figure 1, caption=Effects of hyperosmotic stress under varying NaCl concentrations on the growth of Corynebacterium glutamicum. Growth curves of Corynebacterium glutamicum under various osmotic pressures (0.0-2.1 mol/L NaCl) were measured over time, with a control group (without added stress) for comparison. The 0.0 mol/L NaCl group is the control group. Data represent the mean of three independent replicates., figureFileSmall=LNdH52QBX/i1LSv94wetvg==, figureFileBig=PAd4Jjwy09QabJwAZJPLww==, tableContent=null), ArticleFig(id=1316829803332878809, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=CN, label=图1, caption=不同程度的渗透压胁迫对谷氨酸棒杆菌生长的影响, figureFileSmall=LNdH52QBX/i1LSv94wetvg==, figureFileBig=PAd4Jjwy09QabJwAZJPLww==, tableContent=null), ArticleFig(id=1316829803420959194, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=EN, label=Figure 2, caption=Structural domain architecture and amino acid sequence alignment of the EsrISR system proteins. A: Schematic diagram of the domain structures of EsrI (novel sensor protein), EsrS (histidine kinase), and EsrR (response regulator) (Numbers indicate the amino acid positions of conserved domains); B: Multiple amino acid sequence alignments of EsrS with homologous kinase proteins (top), and EsrR with homologous response regulators (bottom). The blue box indicates the conserved histidine (His) phosphorylation site, and the red box indicates the conserved aspartate (Asp) phosphorylation site. The letters highlighted with a red background represent highly conserved amino acid residues across different species., figureFileSmall=Y7yhzU99cuh01WrsjJgxfg==, figureFileBig=Ifq++SEo7ghgawYF0Qh2QQ==, tableContent=null), ArticleFig(id=1316829803488068059, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=CN, label=图2, caption=EsrISR结构域预测及多序列比对分析磷酸化位点, figureFileSmall=Y7yhzU99cuh01WrsjJgxfg==, figureFileBig=Ifq++SEo7ghgawYF0Qh2QQ==, tableContent=null), ArticleFig(id=1316829803580342748, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=EN, label=Figure 3, caption=Transcriptional orientation and specific fluorescence intensity of the EsrISR system promoters under hyperosmotic stress. A: Schematic diagrams of the P esrIS and P esrR promoters; B: specific fluorescence intensity (fluorescence/OD600 ratio) of the P esrIS and P esrR reporter systems under hyperosmotic stress conditions (1.3-2.1 mol/L NaCl) in the wild-type pXMJ19 and reporter strains (P19-P esrIS and P19-P esrR)., figureFileSmall=PKudThcbK64zguV95QzY8A==, figureFileBig=+day1rFDfDHq+2MKuo5tuA==, tableContent=null), ArticleFig(id=1316829803660034525, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=CN, label=图3, caption=EsrISR三组分系统启动子转录方向及在高渗胁迫下的荧光强度, figureFileSmall=PKudThcbK64zguV95QzY8A==, figureFileBig=+day1rFDfDHq+2MKuo5tuA==, tableContent=null), ArticleFig(id=1316829803735531998, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=EN, label=Figure 4, caption=Regulatory mechanism of the EsrISR system. A: Relative messenger RNA (mRNA) levels of the esrISR genes in mutant strains; B: EMSA analysis of EsrR binding to the promoter regions of P esrIS, P esrR, and the negative control P treZ . The + and - denote the presence or absence of the EsrR protein, respectively. The 25× and 150× indicate the molar ratios of the added EsrR protein to the DNA probe in the reaction mixtures., figureFileSmall=j9ST66e38udsR5Gm/OcF5g==, figureFileBig=cHoG/SaUd3Hp7Sk076DDTg==, tableContent=null), ArticleFig(id=1316829803811029471, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=CN, label=图4, caption=EsrISR三组分系统的调控机制, figureFileSmall=j9ST66e38udsR5Gm/OcF5g==, figureFileBig=cHoG/SaUd3Hp7Sk076DDTg==, tableContent=null), ArticleFig(id=1316829803873944032, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=EN, label=Figure 5, caption=Effects of the EsrISR system deletions on the hyperosmotic stress tolerance and growth of Corynebacterium glutamicum. A: Spot assays of the wild-type (WT) and mutant ΔesrS under different hyperosmotic stress conditions (1.1-1.7 mol/L NaCl); B: Growth curves of the WT and mutants under normal conditions (left panel) and 1.7 mol/L NaCl hyperosmotic stress (right panel)., figureFileSmall=U5xQw1RK1SJ38h67QKs0zw==, figureFileBig=f02rRRZZfgdVGsX8CnYZEA==, tableContent=null), ArticleFig(id=1316829803941052897, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=CN, label=图5, caption=EsrISR组分系统缺失对谷氨酸棒杆菌高渗耐受性与生长的影响, figureFileSmall=U5xQw1RK1SJ38h67QKs0zw==, figureFileBig=f02rRRZZfgdVGsX8CnYZEA==, tableContent=null), ArticleFig(id=1316829804020744674, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=EN, label=Figure 6, caption=Effects of the EsrISR system deletions on the cell morphology of Corynebacterium glutamicum under normal conditions. A: Surface morphology of the wild-type (WT); B: Surface morphology of the mutant ΔesrS; C: Surface morphology of the mutant ΔesrR. The cells were observed using field emission scanning electron microscopy (FESEM)., figureFileSmall=4wuK95C8sdmHqhxoaN5ZTQ==, figureFileBig=Bu1Bx4eSe9mtU0vrcYGn5w==, tableContent=null), ArticleFig(id=1316829804083659235, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=CN, label=图6, caption=正常培养条件下EsrISR系统缺失对谷氨酸棒杆菌细胞形态的影响, figureFileSmall=4wuK95C8sdmHqhxoaN5ZTQ==, figureFileBig=Bu1Bx4eSe9mtU0vrcYGn5w==, tableContent=null), ArticleFig(id=1316829804167545316, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=EN, label=Figure 7, caption=Transcriptomic analysis of the ΔesrR mutant under normal conditions. A: A volcano plot showing differentially expressed genes (DEGs) between the wild-type (WT) and the ΔesrR mutant (A total of 703 significant DEGs are highlighted, including 193 up-regulated and 510 down-regulated genes); B: KEGG pathway enrichment analysis of the DEGs (The color of the bubbles represents the q-value, with greener colors indicating smaller q-values and greater statistical significance, while the size of the bubbles represents the number of genes)., figureFileSmall=L4DjSj9auyoM4sYayYiSqQ==, figureFileBig=BcvLnRhM4t85I0xokXyJRg==, tableContent=null), ArticleFig(id=1316829804255625701, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=CN, label=图7, caption=正常培养条件下ΔesrR 突变体的转录组学分析, figureFileSmall=L4DjSj9auyoM4sYayYiSqQ==, figureFileBig=BcvLnRhM4t85I0xokXyJRg==, tableContent=null), ArticleFig(id=1316829804335317478, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=EN, label=Figure 8, caption=RT-qPCR analysis of target gene expression in ΔesrR and esrR-OE strains across different growth phases. Relative mRNA levels of genes involved in peptidoglycan synthesis, stress response, and cell envelope modification (murA, murB, CGL_RS01830, treS, treY, treZ, otsA, otsB, cmrA, gatC, pks13) in the esrR knockout mutant (ΔesrR) and esrR overexpression (esrR-OE) strains were determined relative to the wild-type strain across multiple growth phases. A: OD600=1.0; B: OD600=2.0; C: OD600=3.0. The 16S rRNA gene was used as an internal reference gene for normalization. The horizontal dashed line at y=1 represents the baseline relative expression level of the wild-type strain. Error bars represent the standard deviations of three independent biological replicates., figureFileSmall=GIMk33KrJN2xjHzSVWGn2g==, figureFileBig=QWZIBPs2xyXwMRjVDvs/lQ==, tableContent=null), ArticleFig(id=1316829804419203559, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=CN, label=图8, caption=ΔesrR 及 esrR-OE菌株中靶基因随 OD600 变化的RT-qPCR分析, figureFileSmall=GIMk33KrJN2xjHzSVWGn2g==, figureFileBig=QWZIBPs2xyXwMRjVDvs/lQ==, tableContent=null), ArticleFig(id=1316829804498895336, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=EN, label=Figure 9, caption=Proposed working model of the regulatory mechanism mediated by the EsrISR three-component system in response to cell envelope stress. Under non-stress conditions (upper panel), the auxiliary membrane protein EsrI interacts with the sensor kinase EsrS, inhibiting its autophosphorylation and keeping the response regulator EsrR in an inactive state. Upon exposure to cell envelope stress signals such as high osmolarity (lower panel), the inhibitory effect of EsrI is relieved. EsrS autophosphorylates and subsequently transfers the phosphoryl group to EsrR. The phosphorylated EsrR (EsrR-P) then binds to the promoter regions of target genes, activating the transcription of its own esrISR operon (positive feedback loop) as well as genes involved in peptidoglycan (murA/murB), trehalose (treS/treY/treZ/otsA/otsB), and mycolic acid (cmrA/gatC/pks13) biosynthesis, thereby reinforcing the cell envelope to combat the environmental stress., figureFileSmall=f9dPloIsG2dhbAfQ9O/SMQ==, figureFileBig=DR6zDpOuvlAy6eTzkCXZYw==, tableContent=null), ArticleFig(id=1316829804561809897, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=CN, label=图9, caption=EsrISR三组分系统响应胁迫的调控机制模式图, figureFileSmall=f9dPloIsG2dhbAfQ9O/SMQ==, figureFileBig=DR6zDpOuvlAy6eTzkCXZYw==, tableContent=null), ArticleFig(id=1316829804641501674, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=EN, label=Table 1, caption=

The strains and plasmids used in this study

, figureFileSmall=null, figureFileBig=null, tableContent=
Plasmid/StrainFeature
Plasmid
pET28aShuttle vector between E. coli and C. glutamicum, KanR, P lac
pXMJ19Shuttle vector between E. coli and C. glutamicum, ChlR, P tac
pK18mobsacBSuicide plasmid carrying sacB for gene deletion in C. glutamicum, KanR
pET28a-esrRpET28a harboring esrR, KanR
pXMJ19-esrIpXMJ19 harboring esrI, ChlR
pXMJ19-esrSpXMJ19 harboring esrS, ChlR
pXMJ19-esrRpXMJ19 harboring esrR, ChlR
pXMJ19-P esrIS-mKate2pXMJ19 harboring P esrIS and mKate2, ChlR
pXMJ19-P esrR-mKate2pXMJ19 harboring P esrR and mKate2, ChlR
pK18-ΔesrIpK18mobsacB for esrI deleting
pK18-ΔesrSpK18mobsacB for esrS deleting
pK18-ΔesrRpK18mobsacB for esrR deleting
pK18-ΔCg0424pK18mobsacB for Cg0424 deleting
pK18-ΔmurBpK18mobsacB for murB deleting
pK18-ΔtreYpK18mobsacB for treY deleting
pK18-ΔtreSpK18mobsacB for treS deleting
pK18-ΔtreZpK18mobsacB for treZ deleting
pK18-ΔgatCpK18mobsacB for gatC deleting
Strain
E. coli BL21(DE3)The expressing host for recombinant protein
E. coli DH5αThe expressing host for recombinant protein
CA00C. glutamicum ATCC 13032
ES01BL21 harboring pET28a
ES02BL21 harboring pXMJ19
ES03BL21 harboring pK18mobsacB
ES04BL21 harboring pET28a-esrR
ES05DH5α harboring pXMJ19-esrI
ES06DH5α harboring pXMJ19-esrS
ES07DH5α harboring pXMJ19-esrR
ES08DH5α harboring pXMJ19-P esrIS -mKate2
ES09DH5α harboring pXMJ19-P esrR -mKate2
ES10BL21 harboring pK18-ΔesrI
ES11BL21 harboring pK18-ΔesrS
ES12BL21 harboring pK18-ΔesrR
CA01CA00 harboring pXMJ19-esrI
CA02CA00 harboring pXMJ19-esrS
CA03CA00 harboring pXMJ19-esrR
CA04CA00 harboring pXMJ19-P esrIS -mKate2
CA05CA00 harboring pXMJ19-P esrR -mKate2
CA06CA00ΔesrI
CA07CA00ΔesrS
CA08CA00ΔesrR
), ArticleFig(id=1316829804712804843, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=CN, label=表1, caption=

本研究所用质粒和菌株

, figureFileSmall=null, figureFileBig=null, tableContent=
Plasmid/StrainFeature
Plasmid
pET28aShuttle vector between E. coli and C. glutamicum, KanR, P lac
pXMJ19Shuttle vector between E. coli and C. glutamicum, ChlR, P tac
pK18mobsacBSuicide plasmid carrying sacB for gene deletion in C. glutamicum, KanR
pET28a-esrRpET28a harboring esrR, KanR
pXMJ19-esrIpXMJ19 harboring esrI, ChlR
pXMJ19-esrSpXMJ19 harboring esrS, ChlR
pXMJ19-esrRpXMJ19 harboring esrR, ChlR
pXMJ19-P esrIS-mKate2pXMJ19 harboring P esrIS and mKate2, ChlR
pXMJ19-P esrR-mKate2pXMJ19 harboring P esrR and mKate2, ChlR
pK18-ΔesrIpK18mobsacB for esrI deleting
pK18-ΔesrSpK18mobsacB for esrS deleting
pK18-ΔesrRpK18mobsacB for esrR deleting
pK18-ΔCg0424pK18mobsacB for Cg0424 deleting
pK18-ΔmurBpK18mobsacB for murB deleting
pK18-ΔtreYpK18mobsacB for treY deleting
pK18-ΔtreSpK18mobsacB for treS deleting
pK18-ΔtreZpK18mobsacB for treZ deleting
pK18-ΔgatCpK18mobsacB for gatC deleting
Strain
E. coli BL21(DE3)The expressing host for recombinant protein
E. coli DH5αThe expressing host for recombinant protein
CA00C. glutamicum ATCC 13032
ES01BL21 harboring pET28a
ES02BL21 harboring pXMJ19
ES03BL21 harboring pK18mobsacB
ES04BL21 harboring pET28a-esrR
ES05DH5α harboring pXMJ19-esrI
ES06DH5α harboring pXMJ19-esrS
ES07DH5α harboring pXMJ19-esrR
ES08DH5α harboring pXMJ19-P esrIS -mKate2
ES09DH5α harboring pXMJ19-P esrR -mKate2
ES10BL21 harboring pK18-ΔesrI
ES11BL21 harboring pK18-ΔesrS
ES12BL21 harboring pK18-ΔesrR
CA01CA00 harboring pXMJ19-esrI
CA02CA00 harboring pXMJ19-esrS
CA03CA00 harboring pXMJ19-esrR
CA04CA00 harboring pXMJ19-P esrIS -mKate2
CA05CA00 harboring pXMJ19-P esrR -mKate2
CA06CA00ΔesrI
CA07CA00ΔesrS
CA08CA00ΔesrR
), ArticleFig(id=1316829804805079532, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=EN, label=Table 2, caption=

The primers used in this study

, figureFileSmall=null, figureFileBig=null, tableContent=
Primer namesPrimer sequences (5′→3′)
pET28A-FAGCAACCGCACCTGTGGC
pET28A-RGCTTCCTTTCGGGCTTTGTTAGC
pET28A-esrR-FCGCGGATCCATGGTTGATGTGTTTTTGGTCG
pET28A-esrR-RCGACGGAGCTCGAATTCTTAGTCAAG
F-pET28A-esrR-FCACTCGAGGGATCTTGACTAAGAATTCGAGCTCC
F-pET28A-esrR-RCAACCATGGATCCGCGACCCATTTG
P19-esrI-FCGAAAGGAGATGAGTACTTTTCAGGAGC
P19-esrI-RGATCCTCTAGAGCTAGTCATTCTGATCAGC
F-P19-esrI-RCTCATCTCCTTTCGACCTGCAGGCATG
FF-P19-esrI-FCTAGCTCTAGAGGATCCCCGGGTACC
P19-esrS-FCAGGTCGAAAGGAGGTGCCTTTATATCC
P19-esrS-RGATCCTCTAGAGCTACACATCCATGGTG
F-P19-esrS-RCTCCTTTCGACCTGCAGGCATG
F-P19-esrS-FGTAGCTCTAGAGGATCCCCGGGTAC
P19-esrR-FCTTGCAATGCATCATCATCATCATCACGTTGATGTGTTTTTG
P19-esrR-RCTGCAGGCATTAGTCAAGATCCCTCGAGTGAG
P19-F-esrR-FGAGGGATCTTGACTAATGCCTGCAGGTCG
P19-F-esrR-RGATGATGCATTGCAAGCTTAATTAATTCTGTTTCC
p19-P esrIS -mKate2-FCAGGTCGACGTGAACGTGGCCAAG
p19-P esrIS -mKate2-RTCCTCCTTTATCCTGCACCGCGGTC
p19-P esrIS -mKate2-F-FGTCGACCTGCAGGCATGCAAGC
p19-P esrIS -mKate2-F-RTCTAGAGGATCCCCGGGTACCGAGC
p19-P esrR -mKate2-FGGAGGACAACCATGCTATCCGAATTG
p19-P esrR -mKate2-RGGAAGCTTTTATCGATGTCCGAGTTTAGAC
p19-P esrR -mKate2-F-FGATAAAAGCTTCCCCGCGAAAGCGGGGTTTTTTTTTTTG
p19-P esrR -mKate2-F-RCTTAATTAATTCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTC
esrI-S-FCATGATTACGAATTCAGACTCTATTGTGGAGTGTTGAGG
esrI-S-RGCAGGCGTCACAGGCACGATAC
esrI-X-FCCTGTGACGCCTGCCATACGAG
esrI-X-RCGAGCTCCCCGACCCTATGTGAG
pk18-F-esrI-FGTCGGGGAGCTCGGTACCCG
pk18-F-esrI-RGAGTCTGAATTCGTAATCATGTCATAGCTGTTTCC
esrS-S-FCAGCTATGACATGATTACGAATTCCTTCAAAGTAGATGGGATCG
esrS-S-RGTACGACCAACGCTTGCCCCGCAG
esrS-X-FCTGCGGGGCAAGCGTTGGTCGTAC
esrS-X-RGGTCGACTCTAGAGGATCCCATTTCGATCAACTTGAGGTTTGCCC
pk18-F-esrS-FGGATCCTCTAGAGTCGACCTGCAGGCATGCAAGCTTGGCACTGG
pk18-F-esrS-RGAATTCGTAATCATGTCATAGCTGTTTCCTGTGTGAAATTG
esrR-1-FCATGATTACGAATTCGAAAAAGGGGAGTGGGG
esrR-1-RCTTTTTAGCCGCCAGCCTTTCAGGTTAC
esrR-2-FGAAAGGCTGGCGGCTAAAAAGAGTGGCG
esrR-2-RGTACCGAGCTCCTGCTACACCGGGTGG
pk18-F-esrR-FGTGTAGCAGGAGCTCGGTAC
pk18-F-esrR-RCGAATTCGTAATCATGTCATAGCTGTTTCC
esrIS 5′ UTR-FGCTTGGCAGGGATGCGTGCGG
esrIS 5′ UTR-RGATATAAAGGCACCGCTTGCCCCGCAG
esrR 5′ UTR-FCCTGAAAAGTACTCATAGACTCTATTGTGGAGTGTTGAGG
esrR 5′ UTR-RCGCTTGCCCCGCAGGCTG
), ArticleFig(id=1316829804914131437, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=CN, label=表2, caption=

本研究所用引物

, figureFileSmall=null, figureFileBig=null, tableContent=
Primer namesPrimer sequences (5′→3′)
pET28A-FAGCAACCGCACCTGTGGC
pET28A-RGCTTCCTTTCGGGCTTTGTTAGC
pET28A-esrR-FCGCGGATCCATGGTTGATGTGTTTTTGGTCG
pET28A-esrR-RCGACGGAGCTCGAATTCTTAGTCAAG
F-pET28A-esrR-FCACTCGAGGGATCTTGACTAAGAATTCGAGCTCC
F-pET28A-esrR-RCAACCATGGATCCGCGACCCATTTG
P19-esrI-FCGAAAGGAGATGAGTACTTTTCAGGAGC
P19-esrI-RGATCCTCTAGAGCTAGTCATTCTGATCAGC
F-P19-esrI-RCTCATCTCCTTTCGACCTGCAGGCATG
FF-P19-esrI-FCTAGCTCTAGAGGATCCCCGGGTACC
P19-esrS-FCAGGTCGAAAGGAGGTGCCTTTATATCC
P19-esrS-RGATCCTCTAGAGCTACACATCCATGGTG
F-P19-esrS-RCTCCTTTCGACCTGCAGGCATG
F-P19-esrS-FGTAGCTCTAGAGGATCCCCGGGTAC
P19-esrR-FCTTGCAATGCATCATCATCATCATCACGTTGATGTGTTTTTG
P19-esrR-RCTGCAGGCATTAGTCAAGATCCCTCGAGTGAG
P19-F-esrR-FGAGGGATCTTGACTAATGCCTGCAGGTCG
P19-F-esrR-RGATGATGCATTGCAAGCTTAATTAATTCTGTTTCC
p19-P esrIS -mKate2-FCAGGTCGACGTGAACGTGGCCAAG
p19-P esrIS -mKate2-RTCCTCCTTTATCCTGCACCGCGGTC
p19-P esrIS -mKate2-F-FGTCGACCTGCAGGCATGCAAGC
p19-P esrIS -mKate2-F-RTCTAGAGGATCCCCGGGTACCGAGC
p19-P esrR -mKate2-FGGAGGACAACCATGCTATCCGAATTG
p19-P esrR -mKate2-RGGAAGCTTTTATCGATGTCCGAGTTTAGAC
p19-P esrR -mKate2-F-FGATAAAAGCTTCCCCGCGAAAGCGGGGTTTTTTTTTTTG
p19-P esrR -mKate2-F-RCTTAATTAATTCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTC
esrI-S-FCATGATTACGAATTCAGACTCTATTGTGGAGTGTTGAGG
esrI-S-RGCAGGCGTCACAGGCACGATAC
esrI-X-FCCTGTGACGCCTGCCATACGAG
esrI-X-RCGAGCTCCCCGACCCTATGTGAG
pk18-F-esrI-FGTCGGGGAGCTCGGTACCCG
pk18-F-esrI-RGAGTCTGAATTCGTAATCATGTCATAGCTGTTTCC
esrS-S-FCAGCTATGACATGATTACGAATTCCTTCAAAGTAGATGGGATCG
esrS-S-RGTACGACCAACGCTTGCCCCGCAG
esrS-X-FCTGCGGGGCAAGCGTTGGTCGTAC
esrS-X-RGGTCGACTCTAGAGGATCCCATTTCGATCAACTTGAGGTTTGCCC
pk18-F-esrS-FGGATCCTCTAGAGTCGACCTGCAGGCATGCAAGCTTGGCACTGG
pk18-F-esrS-RGAATTCGTAATCATGTCATAGCTGTTTCCTGTGTGAAATTG
esrR-1-FCATGATTACGAATTCGAAAAAGGGGAGTGGGG
esrR-1-RCTTTTTAGCCGCCAGCCTTTCAGGTTAC
esrR-2-FGAAAGGCTGGCGGCTAAAAAGAGTGGCG
esrR-2-RGTACCGAGCTCCTGCTACACCGGGTGG
pk18-F-esrR-FGTGTAGCAGGAGCTCGGTAC
pk18-F-esrR-RCGAATTCGTAATCATGTCATAGCTGTTTCC
esrIS 5′ UTR-FGCTTGGCAGGGATGCGTGCGG
esrIS 5′ UTR-RGATATAAAGGCACCGCTTGCCCCGCAG
esrR 5′ UTR-FCCTGAAAAGTACTCATAGACTCTATTGTGGAGTGTTGAGG
esrR 5′ UTR-RCGCTTGCCCCGCAGGCTG
), ArticleFig(id=1316829804981240302, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=EN, label=Table 3, caption=

Transcriptome data of the three-component system esrISR at a 1.7 mol/L NaCl concentration

, figureFileSmall=null, figureFileBig=null, tableContent=
GeneFPKM values (WT vs. 1.7 mol/L NaCl)Fold change
esrI30.9 vs. 3 324.3107.3↑
esrS28.0 vs. 349.712.5↑
esrR76.3 vs. 402.95.3↑
), ArticleFig(id=1316829805044154863, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=CN, label=表3, caption=

三组分系统 esrISR 在1.7 mol/L NaCl浓度下的转录组数据

, figureFileSmall=null, figureFileBig=null, tableContent=
GeneFPKM values (WT vs. 1.7 mol/L NaCl)Fold change
esrI30.9 vs. 3 324.3107.3↑
esrS28.0 vs. 349.712.5↑
esrR76.3 vs. 402.95.3↑
), ArticleFig(id=1316829805119652336, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=EN, label=Table 4, caption=

Transcriptional levels of key genes involved in cell wall homeostasis in wild-type and mutant strains

, figureFileSmall=null, figureFileBig=null, tableContent=
Gene locusGeneFunctionWT FPKMΔesrR FPKM
NCgl2470murAUDP-N-acetylglucosamine 1-carboxyvinyltransferase299.5476.6
NCgl0346murBUDP-N-acetylmuramate dehydrogenase804.6N.D.
NCgl0345CGL_RS018303-phosphoshikimate 1-carboxyvinyltransferase593.40.2
NCgl2221treSMaltose alpha-D-glucosyltransferase13.92.0
NCgl2037treYMalto-oligosyltrehalose synthase63.3103.7
NCgl2045treZMalto-oligosyltrehalose trehalohydrolase99.5133.4
NCgl2534otsATrehalose-6-phosphate synthase389.1370.7
NCgl2537otsBTrehalose-6-phosphate phosphatase508.7528.1
NCgl2385cmrAMycolate reductase CmrA90.6113.7
NCgl1198gatCAsp-tRNA (Asn)/Glu-tRNA (Gln) amidotransferase subunit GatC158.865.5
NCgl2773pks13Polyketide synthase Pks13141.3237.5
), ArticleFig(id=1316829805199344113, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714775158047618, language=CN, label=表4, caption=

野生型及突变株中细胞壁稳态相关关键基因的转录水平

, figureFileSmall=null, figureFileBig=null, tableContent=
Gene locusGeneFunctionWT FPKMΔesrR FPKM
NCgl2470murAUDP-N-acetylglucosamine 1-carboxyvinyltransferase299.5476.6
NCgl0346murBUDP-N-acetylmuramate dehydrogenase804.6N.D.
NCgl0345CGL_RS018303-phosphoshikimate 1-carboxyvinyltransferase593.40.2
NCgl2221treSMaltose alpha-D-glucosyltransferase13.92.0
NCgl2037treYMalto-oligosyltrehalose synthase63.3103.7
NCgl2045treZMalto-oligosyltrehalose trehalohydrolase99.5133.4
NCgl2534otsATrehalose-6-phosphate synthase389.1370.7
NCgl2537otsBTrehalose-6-phosphate phosphatase508.7528.1
NCgl2385cmrAMycolate reductase CmrA90.6113.7
NCgl1198gatCAsp-tRNA (Asn)/Glu-tRNA (Gln) amidotransferase subunit GatC158.865.5
NCgl2773pks13Polyketide synthase Pks13141.3237.5
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谷氨酸棒杆菌EsrISR系统响应高渗胁迫与维持细胞壁完整性的机制研究
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季傲艺 , 范志豪 , 罗薪涛 , 高惠 , 徐美娟 *
微生物学报 | 研究报告 2026,66(10): 4900-4919
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微生物学报 |研究报告 2026 , 66 (10) : 4900 -4919
谷氨酸棒杆菌EsrISR系统响应高渗胁迫与维持细胞壁完整性的机制研究
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季傲艺, 范志豪, 罗薪涛, 高惠, 徐美娟*
作者信息
  • 江南大学 生物工程学院，工业生物技术教育部重点实验室，江苏 无锡
通讯作者:
徐美娟
作者简介:

作者贡献声明

季傲艺：数据收集和处理、论文撰写和修改；范志豪：协助实验操作、参与论文讨论；罗薪涛：数据收集和处理；高惠：研究构思和设计、论文审阅和修改；徐美娟：研究构思和设计、项目经费支持与实验平台资源、论文审阅与修改。

Mechanisms of the response of EsrISR system of Corynebacterium glutamicum to hyperosmotic stress and maintenance of cell wall integrity
Aoyi JI, Zhihao FAN, Xintao LUO, Hui GAO, Meijuan XU*
Affiliations
  • Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China
    • Corresponding Author:
    *E-mail:
出版时间: 2026-10-04 doi: 10.13343/j.cnki.wsxb.20260203
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【目的】 解析由辅助蛋白EsrI、组氨酸激酶EsrS与响应调节子EsrR组成的三组分EsrISR系统在谷氨酸棒杆菌(Corynebacterium glutamicum)应对高渗胁迫中的信号感应逻辑及其维持细胞壁稳态的分子机制。 【方法】 通过转录组测序(RNA sequencing, RNA-seq)筛选锁定应激核心基因簇，利用同源重组技术构建了一系列缺失突变体及过表达株，并结合转录融合报告系统、电子显微镜成像、凝胶迁移实验(electrophoretic mobility shift assay, EMSA)以及实时荧光定量逆转录PCR (real-time quantitative reverse transcription PCR, RT-qPCR)定量分析深入探讨了该系统的感应阈值、应激模式及全局靶基因网络。 【结果】 转录组学测序显示，在1.7 mol/L NaCl胁迫下esrISR基因簇呈显著上调。在同等胁迫下，响应调节子EsrR的缺失会导致生物量显著降低，最大OD600下降至野生型的47.8%；且电镜观察到细胞壁发生严重的破损塌陷现象。通过构建、分析报告质粒，发现此系统受到内部启动子驱动；在1.7 mol/L NaCl胁迫时esrISR基因表达呈峰值，最大荧光强度达880 RFU/OD600。本研究提出的调控模型表明，在无外界胁迫时EsrS主要发挥磷酸酶活性以维持系统的抑制状态(缺失esrS导致esrI表达上调约15.0倍)；感知高渗信号后EsrS活性发生改变，促使响应调节子EsrR发生磷酸化。随后，磷酸化后的EsrR特异性结合至核心调控区，进而开启正反馈调控回路。转录组学分析表明，ΔesrR中涉及细胞壁稳态的关键基因表达发生改变，尤其是肽聚糖合成基因(如murB、CGL_RS01830等)显著下调。RT-qPCR实验证实，EsrR具有调节细胞壁合成的功能，其主要通过正向激活细胞壁合成途径的关键基因响应高渗胁迫。其中，肽聚糖合成基因murB与CGL_RS01830在ΔesrR中下调约95.0%，在esrR过表达(esrR-OE)菌株中上调2.0-6.5倍；海藻糖合成基因treS、treY与treZ在缺失株中表达量下降了75.0%-83.0%，在过表达株中上调2.5-4.2倍；cmrA与gatC在缺失株中表达量下调并在过表达株中上调3.5倍，从而有效维持了细胞壁的物理稳态。 【结论】 本研究为三组分EsrISR系统参与棒状菌属细胞壁稳态的调控提供了新的见解，并为工业底盘菌株的鲁棒性改造提供了关键遗传靶点。

谷氨酸棒杆菌  /  三组分系统  /  细胞壁结构  /  高渗胁迫

[Objective] To elucidate the signal sensing logic of the three-component EsrISR system, which is composed of the accessory protein EsrI, the histidine kinase EsrS, and the response regulator EsrR, in Corynebacterium glutamicum under hyperosmotic stress and its molecular mechanism for maintaining cell wall homeostasis. [Methods] The core stress-responsive gene cluster was identified through RNA-seq. A series of deletion mutants and overexpression strains were constructed through homologous recombination technology. The transcriptional fusion reporter system, field emission scanning electron microscopy/transmission electron microscopy (FESEM/TEM), electrophoretic mobility shift assay (EMSA), and real-time quantitative reverse transcription PCR, (RT-qPCR) were employed to systematically investigate the sensing threshold, stress response patterns, and global target gene networks of this system. [Results] RNA-seq showed that the esrISR gene cluster was significantly upregulated under 1.7 mol/L NaCl stress. Under the same stress conditions, the deletion of the regulator EsrR led to a significant decrease in biomass, with the maximum OD600 dropping to 47.8% of that of the wild type. Moreover, severe damage and collapse of the cell wall were observed via electron microscopy. The construction and analysis of reporter plasmids showed that this system was driven by an internal promoter, peaking at 1.7 mol/L NaCl, with a maximum fluorescence intensity of 880 RFU/OD600. The proposed regulatory model indicated that in the absence of external stress, EsrS mainly exerted phosphatase activity to repress the transcription of esrI, thereby maintaining the repressed state of the system (deletion of esrS led to an approximately 15.0-fold upregulation of esrI). Upon sensing hyperosmotic signals, EsrS changed its activity, promoting the phosphorylation of the regulator EsrR. Subsequently, the phosphorylated EsrR specifically bound to the core regulatory region, thereby activating the positive feedback regulatory loop. Transcriptomic analysis revealed significant alterations in the expression of key genes associated with cell wall homeostasis in the esrR deletion mutant (ΔesrR). RT-qPCR analysis confirmed that EsrR functioned as a crucial regulator of cell wall biosynthesis primarily by positively activating key pathway genes. Specifically, the peptidoglycan synthesis genes murB and CGL_RS01830 were downregulated by 95.0% in ΔesrR and upregulated by 2.0-6.5 fold in esrR-OE; the trehalose biosynthesis genes treS, treY, and treZ were downregulated by 75.0%-83.0% in ΔesrR and upregulated by 2.5-4.2 fold in esrR-OE; cmrA and gatC were significantly repressed in ΔesrR and upregulated by 3.5-folds in esrR-OE, thereby effectively maintaining the physical homeostasis of the cell wall. [Conclusion] This study provides new insights into the regulation of cell wall homeostasis by the three-component EsrISR system in Corynebacterium, providing critical genetic targets for the robustness engineering of industrial chassis strains.

Corynebacterium glutamicum  /  three-component system  /  cell wall structure  /  hyperosmotic stress
季傲艺, 范志豪, 罗薪涛, 高惠, 徐美娟. 谷氨酸棒杆菌EsrISR系统响应高渗胁迫与维持细胞壁完整性的机制研究. 微生物学报, 2026 , 66 (10) : 4900 -4919 . DOI: 10.13343/j.cnki.wsxb.20260203
Aoyi JI, Zhihao FAN, Xintao LUO, Hui GAO, Meijuan XU. Mechanisms of the response of EsrISR system of Corynebacterium glutamicum to hyperosmotic stress and maintenance of cell wall integrity[J]. Acta Microbiologica Sinica, 2026 , 66 (10) : 4900 -4919 . DOI: 10.13343/j.cnki.wsxb.20260203
正文
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谷氨酸棒杆菌(Corynebacterium glutamicum)是一种非致病性、无芽孢的革兰氏阳性土壤细菌，属于棒状菌属中的生物安全菌[1]。同时也是一种重要的高产氨基酸工业底盘菌株，被广泛应用于大规模生物制造，如生产L-谷氨酸、L-亮氨酸等[2-4]。在工业发酵过程中，由于补料、机械搅拌及产物积累，胞外环境常处于剧烈波动中，其中渗透压胁迫(osmotic stress)是影响菌株生长与生产水平的核心限制因素[4-7]。底盘细胞在面对上述渗透压胁迫时能否通过高效的信号转导与代谢重排来维持生理机能的稳定，直接决定了菌株的鲁棒性(即在多变压力下维持代谢稳态与生理平衡的能力)[8-10]。因此，深入解析谷氨酸棒杆菌感应并应对高渗环境的分子机制，不仅是提升底盘细胞鲁棒性的核心途径，对于构建高强度工业底盘也具有重要的理论与实践意义[11]。特别是近年来，各类合成生物学工具(如新型生物传感器的设计以及基于CRISPRi的动态代谢调控等)在谷氨酸棒杆菌中的广泛应用[12-13]，使得挖掘天然的环境响应网络并将其转化为智能调控元件成为了工业底盘改造的前沿热点。
作为棒状菌属的代表菌种之一，谷氨酸棒杆菌拥有极其复杂的细胞壁(cell wall)结构。该屏障由内层的肽聚糖(peptidoglycan)、中层的阿拉伯半乳聚糖(arabinogalactan)以及外层特有的分枝菌酸层(mycolic acids)共同组成[14-17]，这3种物质构成了抵御外界物理化学损伤的第一道防线。研究表明，细胞壁的结构完整性与化学组成是菌株鲁棒性的物质基础，当细胞面临高渗胁迫时需要迅速激活特定的响应系统，通过加强细胞壁的物理强度、积累海藻糖等相容性溶质以抵御胞内渗透压失衡引发的生长抑制[18-19]。
为了应对多变的环境胁迫并维持细胞壁稳态，细菌形成了高度复杂的信号转导网络，以精准感知外界或自身的压力信号。虽然经典的双组分信号转导系统(two-component system, TCS)在感知渗透压、pH值等外部刺激并重编排生理功能中发挥了基础作用，但在应对更为复杂的工业发酵环境时，细菌通常依赖于更加精细的调控机制[20-22]。近年来，三组分系统逐渐引起关注，其中EsrISR三组分系统便是典型代表。研究表明，该系统中的第三组分(辅助蛋白)通常通过与组氨酸激酶(histidine kinase, HK)或响应调节因子(response regulator, RR)发生直接物理互作来调控磷酸化级联反应[23-24]。本文对EsrISR三组分系统的定义并不是指3条独立的信号传导链，而是根据esrISR基因簇的形式，以及辅助蛋白EsrI与组氨酸激酶(HK)、响应调节因子(RR)之间的转录调控关联所定义的复合调控系统。
为了实现这种系统级的控制，谷氨酸棒杆菌中存在一个进化保守且与外界胁迫应激反应高度相关的三组分基因簇esrISR。该模块编码一个假定的组氨酸激酶(EsrS)、一个响应调节因子(EsrR)以及一个小分子辅助蛋白(EsrI)。在无胁迫条件下，辅助蛋白EsrI与组氨酸激酶EsrS之间存在显著的转录调控关联，并对系统发挥抑制作用。然而，尽管已知该系统参与了菌株的抗逆响应，但在环境胁迫发生时EsrI确切的作用机制以及它是如何在转录层面与EsrS-EsrR协同调控以解除抑制并驱动下游靶基因的表达，至今尚未完全阐明[25-26]。
本研究以EsrISR系统为切入点，全面解析三组分系统内部的转录调控及响应高渗胁迫的分子调控网络。本研究旨在探究在无胁迫条件下EsrI对EsrS在转录水平上的关联及其对系统的潜在抑制作用，并进一步考察面临高渗胁迫时EsrS向响应调节因子EsrR传递信号的动态过程及激活机制。此外，本研究还将评估被激活的EsrISR信号枢纽对肽聚糖、分枝菌酸及海藻糖协同合成的驱动作用，阐明该过程在代谢流量重分配和细胞壁屏障加固中的生理功能。在此基础上进一步解析谷氨酸棒杆菌复杂信号转导网络，以填补相关理论空白，并为高鲁棒性工业底盘细胞的理性设计提供具有潜力的基因元件与干预靶点。
1 材料与方法
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1.1 菌株、质粒及引物
本研究所使用的菌株和质粒如表1所示，所用引物信息如表2所示。
1.2 培养基和培养条件
LB培养基(g/L)：NaCl 10.0，蛋白胨10.0，酵母粉5.0。固体LB培养基(g/L)：NaCl 10.0，蛋白胨10.0，酵母粉5.0，琼脂粉17.0。大肠杆菌感受态制备试剂：A液-CaCl2 0.1 mol/L；B液-CaCl2 0.1 mol/L，15%甘油。BHI培养基(g/L)：脑心浸出肉汤38.5。固体BHI培养基(g/L)：脑心浸出肉汤38.5，琼脂粉17.0。LBGS培养基(g/L)：酵母提取物5.0，NaCl 10.0，蛋白胨10.0，葡萄糖7.0，20%蔗糖。谷氨酸棒杆菌感受态培养基(g/L)：脑心浸出肉汤38.5，甘氨酸3.0，0.01%吐温-80。TB培养基(g/L)：酵母粉24.00，蛋白胨12.00，甘油4.00，K2HPO4 12.54，KH2PO4 2.31。
LB于121 ℃灭菌15 min，TB于115 ℃灭菌20 min。
谷氨酸棒杆菌培养条件为30 ℃、220 r/min。培养时抗生素终浓度为：氨苄青霉素100 μg/mL、卡那霉素50 μg/mL、氯霉素20 μg/mL。
1.3 缺失菌株的构建
利用pK18mobsacB穿梭质粒携带的蔗糖致死基因sacB，使菌株进行二次同源重组，达到敲除基因的目的。
1.4 生长曲线的测定
将待测菌株保菌管分别划线于相应BHI抗性固体培养基上，在30 ℃恒温培养箱中倒置培养24 h，进行菌种活化。培养结束后，从平板上挑取单菌落，在超净台中进行无菌操作，接种至装有10 mL液体BHI相应抗性的培养基中，30 ℃、200 r/min培养12 h，至对数生长中后期OD600为4.0，获得活化种子液。将上述制备的种子液按体积分数为1%的接种量，转接至装有50 mL新鲜BHI相应抗性液体培养基的250 mL不带挡板的三角瓶中，30 ℃、200 r/min培养。接种后的初始OD600值为0.1。自接种起，每隔2 h取样1次，每次吸取200 μL培养液，以未接种的空白培养基作为对照，使用紫外-可见分光光度计测定波长600 nm处的吸光度值(OD600)。
在菌液浓度超出仪器准确测量范围(OD600=0.1-0.8)的情况下，使用无菌生理盐水对样液进行适当倍数的稀释后再行测定。每个重组菌株及野生型对照均设置3个生物学重复。最终以培养时间为横坐标、对应的OD600值为纵坐标，利用GraphPad Prism软件绘制并比较各菌株的生长曲线。
1.5 突变株制样及形态观察
使用无菌刮刀从BHI培养基平板上刮取单菌落，转移至已灭菌的1.5 mL离心管中，以4 ℃、2 500 r/min离心1 min，收集到约黄豆大小的菌体沉淀。沿管壁缓慢加入2.5%戊二醛固定液将沉淀完全浸没，于4 ℃静置固定过夜。弃去固定液后，加入0.1 mol/L PBS轻柔漂洗3次，每次10 min。随后使用梯度浓度的无水乙醇进行脱水。处理完成后的样品使用电子显微镜(Hitachi公司)进行观察检测[27]。
1.6 突变株高渗耐受性点板实验
将相应的谷氨酸棒杆菌培养至对数生长期(OD600为4.0)，然后以10倍稀释梯度连续稀释至10-7，每个稀释梯度各取1.5 µL稀释液，在含有相应氯化钠浓度胁迫的BHI平板上进行点板，于30 ℃下培养24 h至菌落长出，观察结果。
1.7 RNA的提取、反转录
RNA提取方法见RNA simple提取试剂盒[天根生化科技(北京)有限公司]说明书。实时荧光定量逆转录PCR (real-time quantitative reverse transcription PCR, RT-qPCR)使用ChamQ SYBR-qPCR Master Mix试剂盒(武汉爱博泰克生物科技有限公司)，体系详见说明书。
1.8 转录组测序分析
将谷氨酸棒杆菌接种于BHI培养基中，30 ℃培养至OD600为0.6时，4 ℃、12 000 r/min离心2 min，收集菌体沉淀，用液氮保存。选择在对数前期取样，主要是为了排除后期由于营养受限带来的次级干扰，从而反映突变株早期的基因表达状态。经NanoDrop ND-1000核酸检测仪(LabTech公司)检测RNA的浓度与纯度。基于Illumina二代高通量测序平台(HiSeq 4000)测序，由金唯智科技有限公司完成。
1.9 实时荧光定量逆转录PCR
选择了14个相关基因，以高水平、恒定表达的16S rRNA基因为内参基因，以逆转录后的cDNA为模板，使用ChanQ Universal SYBR qPCR Master Mix进行RT-qPCR分析，检测mRNA相对表达量。qPCR结果与转录组测序数据结果均以|log2 fold change|表示。
1.10 凝胶迁移实验(electrophoretic mobility shift assay, EMSA)
以谷氨酸棒杆菌ATCC 13032基因组为模板，使用表2中的引物进行PCR扩增。PCR反应体系(25.0 μL)：2×KeyPo SE Master Mix 12.5 µL，上、下游引物(10 µmol/L)各1.0 µL，DNA模板0.5 µL，ddH2O 10.0 µL。PCR反应条件：94 ℃预变性2 min；98 ℃变性10 s，58 ℃退火30 s，68 ℃延伸30 s，共35个循环；72 ℃终延伸10 min。切胶纯化后获得目的基因的5′-UTR启动子探针。将EsrR蛋白、EMSA结合缓冲液与探针于25 ℃条件下孵育30 min后，加入上样缓冲液，并在6%非变性聚丙烯酰胺凝胶上4 ℃、100 V电泳2 h，使用光谱成像系统可视化结合带。
2 结果与分析
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2.1 谷氨酸棒杆菌在高渗胁迫下的生长抑制程度及 esrISR 核心靶点的确立
本研究首先通过生长曲线测定，量化评估了不同程度的渗透压胁迫对野生型(WT)菌株生长的抑制程度。生长曲线(图1)显示，随着NaCl浓度升高，WT菌株的倍增时间显著延长，最大OD600值在1.7 mol/L NaCl条件下较对照组下降约32%。1.9 mol/L NaCl对生长的抑制更为显著，OD600仅为5.5左右，而当胁迫浓度进一步提升至2.1 mol/L时，菌株的生长受到了极大的抑制，最大OD600仅在1.5左右，当2.3 mol/L时菌株生长则被完全抑制。
表3结果显示，编码三组分系统的基因簇esrISR转录水平显著上调。数据显示，在1.7 mol/L NaCl胁迫下，基因esrI的FPKM值从无胁迫时的30.9显著升至3 324.3，差异倍数高达107.3；基因esrS与基因esrR的FPKM值也分别由28.0和76.3显著升至349.7和402.9，上调倍数分别为12.5和5.3。由于该组分系统显著的上调表达，该系统被确认为本研究的核心靶点。
2.2 EsrISR三组分系统的结构域鉴定与序列保守性分析
本研究利用UniProt数据库(https://www.uniprot.org/)对其编码蛋白的结构域进行分析，并采用TMHMM Server v.2.0 (http://www.cbs.dtu.dk/services/TMHMM-2.0/)对蛋白跨膜区进行预测[28-29]。结构域预测分析结果如图2A所示，esrISR基因簇编码3种核心蛋白。其中，传感器激酶EsrS在N端(6-59 aa)存在PspC结构域，其C端(293-379 aa)含有典型组氨酸激酶催化与ATP结合结构域(HATPase_dom)，负责消耗ATP进行自身磷酸化，是信号向下游传递的基础[25,30]。信号执行由响应调节因子EsrR完成，其N端(3-117 aa)包含REC结构域，负责接收来自EsrS的磷酸基团；其C端(163-228 aa)拥有的HTH_luxR型结构域，预示其具备特异性结合靶基因启动子并驱动防御网络的能力[31-32]。该系统区别于典型双组分系统的显著特征是额外编码了辅助跨膜蛋白EsrI。EsrI包含N端PspC结构域(51-98 aa)与C端LiaF样结构域(300-385 aa)。现有的同源蛋白功能研究表明，N端的PspC结构域在多种细菌的噬菌体休克系统中广泛存在，并被证实能够直接感知细胞被膜完整性的破坏或跨膜电位的异常波动，是触发细胞壁应激响应的核心传感模块；而C端的LiaF样结构域在经典的LiaFSR三组分包膜应激系统中已被证实为关键的抑制性元件。在无外界胁迫的状态下，LiaF样结构域能够通过与组氨酸激酶发生直接的物理互作，将其锁定在无活性的状态，从而对整个系统发挥负向调节作用。这表明EsrI可通过这两大结构域实现对应激信号的敏感捕捉与感应阈值的精确控制[24,33-34]。
多重序列比对结果如图2B所示，EsrS的H207磷酸化位点与EsrR的D51磷酸化受体位点在多种细菌的同源系统中高度保守，暗示该系统具备精密的信号传导潜力。基于上述结构特征与保守性分析，本研究随后对其启动子区域架构及应激环境下的转录活性展开了深入分析。
2.3 esrISR 基因座的双向启动子架构鉴定及其对梯度渗透胁迫的差异性响应
本研究通过使用BPROM进行了生物信息学分析，预测出该基因簇存在着复杂的双向转录模式[35]。如图3A所示，调节基因esrI与传感器基因esrS相邻且转录方向相反，两者共享一段上游基因间区。该区域包含一个共享启动子区P esrIS，其能够沿正向(5′→3′)驱动esrS的转录，并沿反向驱动esrI的转录。此外，预测结果表明在esrS的编码序列(coding sequence, CDS)内部存在一个内部启动子P esrR，用于沿正向独立驱动下游响应调节因子esrR的表达。
为验证这种独特的转录架构，本研究构建了2个转录融合报告质粒：将esrIS启动子P esrIS 与预测的esrR启动子P esrR 分别融合至红色荧光蛋白mKate2的编码基因。这些报告菌株在1.3-2.1 mol/L的NaCl浓度梯度中培养，通过比荧光强度(RFU/OD600)评估启动子活性。如图3B所示，携带空载体pXMJ19的对照菌株在所有测试条件下均呈现微弱荧光，排除了背景干扰的可能性。P esrIS 和P esrR 启动子均呈现显著的应激诱导激活的特性，但两者展现出截然不同的响应特征。P esrIS 启动子的活性呈现渐进式增长，在1.7-1.9 mol/L NaCl浓度区间达到稳定期。P esrR 启动子在1.7 mol/L NaCl的胁迫条件下出现明显而强烈的诱导峰值，其最大比荧光强度约达880 RFU/OD600。与1.3 mol/L NaCl胁迫条件下相比，表达水平明显上调，这表明响应调节基因esrR对该特定渗透阈值具有高度敏感性。在1.9 mol/L的渗透胁迫条件下，P esrIS 活性始终保持较高水平，而P esrR 活性则明显下降。这表明P esrR 启动子在1.7 mol/L浓度下促进菌株的快速适应，而P esrIS 启动子则在极端近致死条件下(1.9 mol/L)仍能维持信号感知模块(EsrS和EsrI)的关键活性水平。然而，当盐浓度升至2.1 mol/L时，2个启动子的活性均急剧下降，这可能是由于生长受到严重抑制，超出了生理耐受极限。
报告基因检测证实了EsrISR系统的转录激活与应激强度紧密相关。esrR不仅作为操纵子的一部分被驱动，还受一个高度敏感的独立内部启动子调控，这表明其调控机制具有复杂的层次结构。这种精密且复杂的转录输出受控于传感器激酶EsrS与调节子EsrR之间的动态反馈逻辑。
2.4 基于缺失突变与EMSA的EsrISR系统反馈调控机制解析
为了深入探究P esrIS 与P esrR 启动子响应高渗胁迫的分子机制，本研究构建了相关基因的缺失突变体并进行了RT-qPCR检测。在负调控方面，之前的假设认为EsrI可能是主要的抑制因子。然而，机制分析表明，在非应激条件下，传感器激酶EsrS主要作为磷酸酶发挥作用，通过持续不断地对EsrR去磷酸化防止信号通路过早激活。esrS的缺失会导致磷酸酶活性的丧失，可能使EsrR极易受内源性磷酸供体或交叉作用的磷酸化影响，导致系统被持续激活。RT-qPCR结果有力地证实了这一机制特征：与EsrI作为主要抑制因子的假设不同，ΔesrS突变体展现出了显著的转录激活，其esrI转录本上调了约15.0倍(图4A)。关于正向调控，RT-qPCR结果证实该基因簇的双向转录高度依赖于响应调节子EsrR。在ΔesrR突变体中，esrI基因的转录水平基本丧失，表明EsrR对激活驱动esrI基因的核心启动子至关重要。
为验证该正向调控是否是直接作用，本研究进一步开展了凝胶迁移实验。结果显示，当纯化的磷酸化EsrR (EsrR-P)蛋白与包含esrI-esrS基因共有启动子区以及esrR自身启动子区的DNA探针共同孵育时，随着EsrR-P蛋白浓度的梯度增加，游离的DNA探针信号逐渐减弱，并出现了一条明显的迁移率滞后(shift)条带。本研究引入了转录组分析中受EsrR影响较小的基因treZ的启动子区P treZ 作为阴性对照探针，结果显示即使在最高蛋白浓度下也未观察到结合条带(图4B)。靶基因启动子呈现的实验结果与阴性对照的未结合结果相互验证，在一定程度上协助排除了蛋白非特异性结合的可能，进一步支持了EsrR-P能够特异性结合在该基因簇核心调控区的结论。上述结果揭示了强效正反馈回路：激活的EsrR通过结合该核心枢纽驱动双向转录，既增强抑制元件esrI的表达，又强化调控模块(esrS-esrR)的功能。
基于这些发现，我们提出EsrISR系统的磷酸酶激活模型。(1)基础抑制：正常状态下，EsrS通过其磷酸酶活性维持EsrR处于未磷酸化的状态，从而抑制系统的激活；(2)胁迫触发：当感知到外界渗透胁迫时，该磷酸酶活性被抑制或消除，使EsrR通过接受内源性磷酸基团而以磷酸化形式进行积累。(3)反馈调控的放大效应：活化的EsrR-P结合靶基因的核心区，同时启动esrI基因与esrS-esrR模块的转录。在理清该三组分系统内部的自调控逻辑后，需进一步解析EsrR调控的全局防御网络[36-37]，从而评估该系统对维持细菌高渗耐受性的全局性贡献。
2.5 EsrISR系统高渗胁迫下的生长表型分析
本研究在NaCl诱导的高渗胁迫条件下对野生型和esrS缺失突变体进行了点板实验，实验结果如图5A所示，WT菌株仍能维持较强的耐受性，生长良好；在1.3 mol/L NaCl胁迫下WT与敲除株相比并无明显差异。在1.5 mol/L NaCl胁迫下，ΔesrS菌株生长落后于WT。在1.7 mol/L NaCl胁迫下表型差异最为明显，ΔesrS菌株形成的菌落明显小于WT。这些结果表明EsrISR系统对谷氨酸棒杆菌在高渗透压条件下生存至关重要，尤其在胁迫强度较高时。
在标准BHI培养基中的生长分析显示，野生型菌株表现出强劲的生长能力，其延滞期约为4 h，对数期持续4-24 h，最终在32 h达到约13.3的最大OD600值(图5B)。相比之下，突变体表现出不同程度的生长缺陷。ΔesrI突变体生物量降低，OD600峰值仅为9.1，约为WT的68.4%。值得注意的是，分别缺失传感器激酶和响应调节因子导致生物量更严重下降：ΔesrS和ΔesrR突变体的OD600峰值分别仅达8.0和4.6 (分别仅为WT的60.2%和34.6%)。这些结果表明EsrISR系统在细菌的正常生长繁殖中发挥着关键作用，且响应调节因子的缺失比传感器激酶的缺失造成更严重的损害。
在高渗胁迫条件下(1.7 mol/L NaCl)，各菌株的生长相较于BHI培养基呈现全面抑制。生长曲线分析显示，渗透胁迫显著延长了菌株的延滞期(从4 h延长至约12 h)，对数生长期相应推迟至12-32 h。其中，野生型菌株依旧表现出最强耐受性(最大OD600为9.0)，而缺失突变体则呈现出与基因型相关的生长缺陷。与正常培养条件下的趋势一致，ΔesrR突变体的受损最为严重，OD600峰值仅为4.3 (约为WT的47.8%)，ΔesrS突变体可达7.0 (约为WT的77.8%)。综合非胁迫与胁迫条件下的数据可知，尽管高盐环境对总体生长具有普遍的抑制作用，但关键调控元件缺失带来的影响依然显著：野生型表现出最高耐受性，而ΔesrS与ΔesrR突变株始终表现出严重的生长缺陷，且响应调节因子的缺失(ΔesrR)造成的损害在任何条件下均为最大。这证实了随着环境压力加剧，EsrISR系统在维持细菌增殖中的重要性日益凸显。由于生长能力的丧失通常源于细胞壁膜等物理屏障的破坏，本研究进一步利用扫描与透射电子显微镜对高渗胁迫下细胞超微结构的变化进行了深入观察。
2.6 ΔesrS 与ΔesrR 突变体的电镜分析揭示细胞表面的结构缺陷
基于先前研究的生长表型，本研究推测EsrISR系统的缺失直接导致了菌体物理防御屏障能力的受损。为直观验证这一假设，本研究选取ΔesrS和ΔesrR突变体，利用冷场发射扫描电镜(field emission scanning electron microscopy, FESEM)技术，对正常培养条件下的菌体表面形态及内部超微结构进行了深入观察。
如图6所示，扫描电镜(FESEM)分析揭示了各菌株之间显著的表面形态差异。野生型细胞形态饱满、表面光滑，是典型的短杆状结构(图6A)。相比之下，突变株细胞则呈现不同程度的结构塌陷，且其受损程度与其生长表型的缺陷相一致。其中ΔesrS突变体表面出现了不规则的皱褶和凹陷(图6B)；而ΔesrR突变株的形态受损最为严重，细胞明显萎缩变形，表面可见破裂，短杆状结构完整性丧失(图6C)。
这些直观的形态学观察结果表明，即使在正常培养条件下突变体的细胞外围物理屏障也已发生了显著的结构性破损。这种基础防御结构的丧失从物理层面解释了为何这些突变体在高渗胁迫下无法正常增殖这一现象，进一步证实了响应调节因子EsrR在维持基础细胞壁完整性中发挥着不可或缺的决定性作用[35,37]。
2.7 ΔesrR 突变体的转录组分析及细胞壁合成靶基因网络鉴定
为识别EsrR调节子对维持细胞壁稳态的靶基因网络，本研究在正常条件下进行了WT与ΔesrR的转录组学分析。比较分析揭示，即使在无外部胁迫的情况下，EsrR的缺失仍然导致了显著的转录影响。如图7所示，野生型与ΔesrR突变株之间共存在703个差异表达基因(differentially expressed genes, DEGs)。其中，相较于野生型菌株，突变体中有510个基因下调表达，而193个基因上调表达。下调基因数量远超上调基因这一现象，揭示了EsrR在全局调控中主要作为转录激活因子发挥作用，维持着对细胞稳态至关重要的庞大基因网络的基础表达水平。进一步的KEGG通路富集分析显示，这些差异基因主要富集于“微生物在不同环境中的代谢(microbial metabolism in diverse environments)” “双组分系统” “硫代谢” “碳代谢”及“ABC转运蛋白”等核心途径。在细菌的代谢网络中，细胞壁合成依托这些基础代谢的协同作用完成。“碳代谢”等途径负责为肽聚糖和海藻糖的合成提供充足的糖基前体与能量。基础代谢的广泛受阻在本质上直接切断了细胞壁大分子组装的物质来源，导致细胞壁合成受阻。这表明EsrISR系统在细胞信号网络中处于枢纽位置，广泛负责协调基础代谢流。
结合电镜观察与前体代谢数据，本研究从转录组中梳理了细胞壁合成与修饰相关的差异基因网络(表4)。数据显示，在ΔesrR突变株中肽聚糖合成途径基因显著下调，UDP-N-乙酰胞壁酸脱氢酶基因murB在转录本中FPKM为N.D. (not detected)，另一前体合成基因CGL_RS01830也显著被抑制(FPKM从593.4骤降至0.2)。与此同时，海藻糖与分枝菌酸合成基因(如treS、treY、pks13等)呈现出微弱的上调趋势，这可能是突变体在面临致命的细胞壁结构崩塌时为勉强维持细胞基础完整性而激发的短暂代偿反应。
2.8 RT-qPCR验证EsrR对肽聚糖合成、分枝菌酸修饰及海藻糖代谢通路的多靶点正向调控
为了探究在细菌的持续生长过程中这些关键基因的转录水平是否会发生变化，并进一步确证EsrR的核心调控功能，本研究随后在OD600为1.0、2.0、3.0三个阶段采用RT-qPCR对上述调控网络进行定量验证(图8)。结果显示，在ΔesrR突变株中不同代谢通路随OD600值的变化呈现出明显的转录差异。负责维持内层肽聚糖骨架强度的关键合成基因murB及CGL_RS01830在OD600为1.0-3.0范围内其表达量显著被抑制，并在整个生长周期内持续处于极低水平(图8A)。2.7节的转录组数据显示，在OD600为0.6时海藻糖合成和分枝菌酸合成途径中的多数关键靶基因尚能维持正常的转录水平；随着生长推进(OD600为1.0时)，海藻糖合成簇(treS、treY、treZ)及分枝菌酸合成与转运靶基因(cmrA、gatC)的表达量开始微有下降；但OD600为2.0及3.0时，上述基因急剧下降了75%-83%，导致这2条代谢途径最终被全面抑制(图8B)。相反，在esrR-OE中这些基因在整个实验周期内(OD600为1.0-3.0)均保持着一定强度的转录。海藻糖合成与分枝菌酸合成及转运的多个基因在3个生长时期均稳定保持着1.2-4.8倍的显著上调；肽聚糖合成基因murB及CGL_RS01830的表达量较野生型也有2.0-6.5倍的显著提升；而pks13的转录水平在2株重组菌的各个生长阶段均未表现出明显波动(图8C)。
综合转录组学与RT-qPCR数据，突变株中相关代谢途径基因的转录水平随时间推移逐渐下降，而过表达株中相关基因则呈现持续激活，充分证明了EsrR在驱动并维持细胞壁骨架中发挥着不可或缺的作用。这种受控于EsrR的细胞壁合成基因转录机制，不仅从分子层面解释了突变体细胞壁受损的原因，更阐明了其在持续生长的胁迫环境下，维持工业底盘细胞形态完整性与鲁棒性的重要生理功能。
3 讨论与结论
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本研究整合表型分析、转录组测序及RT-qPCR等验证实验，系统解析了谷氨酸棒杆菌EsrISR系统应对高渗胁迫的全局调控机制。
结合上述结果，本研究提出了EsrISR三组分系统的调控机制模式图(图9)。在非胁迫条件下，辅助跨膜蛋白EsrI与传感器激酶EsrS协同作用，EsrS主要发挥磷酸酶功能，维持系统的基础稳态。这种调控在避免能量无效消耗、维持细胞正常生长的同时，保证了细胞壁的完整性。当遭遇高渗胁迫时，该静息稳态被打破，EsrS激酶活性被激活并发生自身磷酸化，随后将信号传递给响应调节因子EsrR；叠加EsrR自身的正反馈放大效应，促使细胞大量表达抗逆相关基因以应对极端环境造成的损伤。进一步的靶基因分析表明，EsrR是驱动抗逆代谢重编程的核心枢纽。在ΔesrR突变株中，murB等靶基因转录受到高达95%的显著抑制，为突变株即使在正常条件下也呈现出细胞壁破损表型提供了直接的证据。
综上所述，本研究鉴定的靶基因调控网络为工业底盘菌株的抗逆改造提供了明确的遗传操作位点。该系统所展现出的磷酸酶特性为合成生物学中动态调控线路的构建提供了极具潜力的天然元件。相较于传统的静态表达系统，这种依赖环境感知的机制能够有效平衡细胞生长与产物合成之间的代谢负担。例如，未来可借鉴此机制，通过对EsrS的感应结构域进行工程化改造，创建能够特异性响应酸胁迫、醇胁迫等其他复杂工业发酵应激信号的合成启动子。基于此，后续可通过对EsrR及其靶启动子进行工程化改造，构建更加智能的人工响应网络，进而综合提升谷氨酸棒杆菌在复杂发酵环境中的鲁棒性与转化率。此外，尽管本研究在转录层面证实了EsrI与EsrS存在紧密的遗传学联系，但受限于实验方法，尚未在蛋白水平直接验证二者的物理结合。在未来的研究中，我们将进一步利用细菌双杂交与Pull-down等生化实验明确EsrI与EsrS的蛋白互作关系，并检测ΔesrI菌株中EsrR的磷酸化状态，从而更全面地揭示该三组分系统的精密物理调控网络。
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  • 国家自然科学基金(32570026)
  • 国家重点研发计划(2023YFD1300700)
  • 江南大学工业生物技术教育部重点实验室项目(KLIB-KF202504)
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参考文献 引证文献
排序方式：
[1]
Qi L, He H, Xiang SH, Cao H. A two-enzyme entry module triggers an endogenous biocatalytic cascade for green biosynthesis of pyridoxal 5′-phosphate in Corynebacterium glutamicum [J]. Catalysts, 2026, 16(2): 195.
[2]
Walter T, Al Medani N, Burgardt A, Cankar K, Ferrer L, Kerbs A, Lee JH, Mindt M, Risse JM, Wendisch VF. Fermentative N-methylanthranilate production by engineered Corynebacterium glutamicum [J]. Microorganisms, 2020, 8(6): 866.
[3]
Zahoor A, Lindner SN, Wendisch VF. Metabolic engineering of Corynebacterium glutamicum aimed at alternative carbon sources and new products[J]. Computational and Structural Biotechnology Journal, 2012, 3(4): e201210004.
[4]
Li MH, Li H, Zhang X, Liang YC, Li C, Sun ML, Li K, Liu CG, Sinskey AJ. Metabolic engineering of Corynebacterium glutamicum: unlocking its potential as a key cell factory platform for organic acid production[J]. Biotechnology Advances, 2024, 77: 108475.
[5]
Schalck T, Katyal M, De Graeve S, Roba L, Baldoma JV, Swings T, Van den Bergh B, Michiels J. Reprogramming the EnvZ-OmpR two-component system confers ethanol tolerance in Escherichia coli by stabilizing the outer membrane and altering ferric homeostasis[J]. PLoS Genetics, 2025, 21(12): e1011707.
[6]
Yang LB, Zhan XB, Zheng ZY, Wu JR, Gao MJ, Lin CC. A novel osmotic pressure control fed-batch fermentation strategy for improvement of erythritol production by Yarrowia lipolytica from glycerol[J]. Bioresource Technology, 2014, 151: 120-127.
[7]
Khider MLK, Irla M, López MG, Gispert A, Konjetzko T, Meliawati M, Schmid J, Brautaset T, Brito LF. CRISPR-Cas9-driven genome editing in Bacillus methanolicus MGA3[J]. Frontiers in Microbiology, 2026, 16: 1728984.
[8]
Mohedano MT, Konzock O, Chen Y. Strategies to increase tolerance and robustness of industrial microorganisms[J]. Synthetic and Systems Biotechnology, 2022, 7(1): 533-540.
[9]
Gong ZW, Nielsen J, Zhou YJ. Engineering robustness of microbial cell factories[J]. Biotechnology Journal, 2017, 12(10): 1700014.
[10]
Guan AL, He ZX, Wang X, Jia ZJ, Qin JF. Engineering the next-generation synthetic cell factory driven by protein engineering[J]. Biotechnology Advances, 2024, 73: 108366.
[11]
Takahashi Y, Hirasawa T. Enhancement of ergothioneine production in Corynebacterium glutamicum by increasing osmotic pressure[J]. Applied Microbiology and Biotechnology, 2025, 109: 255.
[12]
Jeon S, Sohn YJ, Lee H, Park JY, Kim D, Lee ES, Park SJ. Recent advances in the design-build-test-learn (DBTL) cycle for systems metabolic engineering of Corynebacterium glutamicum [J]. Journal of Microbiology, 2025, 63(3): e2501021.
[13]
Kim GY, Kim J, Park G, Kim HJ, Yang JN, Seo SW. Synthetic biology tools for engineering Corynebacterium glutamicum [J]. Computational and Structural Biotechnology Journal, 2023, 21: 1955-1965.
[14]
De Sousa-d’Auria C, Constantinesco F, Bayan N, Constant P, Tropis M, Daffé M, Graille M, Houssin C. Cg1246, a new player in mycolic acid biosynthesis in Corynebacterium glutamicum [J]. Microbiology, 2022, 168(4): 001171.
[15]
Kumagai Y, Hirasawa T, Wachi M. Requirement of the LtsA protein for formation of the mycolic acid-containing layer on the cell surface of Corynebacterium glutamicum [J]. Microorganisms, 2021, 9(2): 409.
[16]
Isbilir B, Yeates A, Alva V, Bharat TAM. Mapping the ultrastructural topology of the corynebacterial cell surface[J]. PLoS Biology, 2025, 23(4): e3003130.
[17]
楼洁, 胡晓, 梁延群, 朱怡铃, 音建华. 肽聚糖的生物合成及其调控机制研究进展[J]. 微生物学报, 2023, 63(1): 106-123.
Lou J, Hu X, Liang YQ, Zhu YL, Yin JH. Peptidoglycan biosynthesis and the regulatory mechanism[J]. Acta Microbiologica Sinica, 2023, 63(1): 106-123 (in Chinese).
[18]
Liu HJ, Liu DH, Zhong JJ. Quantitative response of trehalose and glycerol syntheses by Candida krusei to osmotic stress of the medium[J]. Process Biochemistry, 2006, 41(2): 473-476.
[19]
MacIntyre AM, Barth JX, Pellitteri Hahn MC, Scarlett CO, Genin S, Allen C. Trehalose synthesis contributes to osmotic stress tolerance and virulence of the bacterial wilt Pathogen Ralstonia solanacearum [J]. Molecular Plant-Microbe Interactions®, 2020, 33(3): 462-473.
[20]
Zou Y, Huang DN, Liu XX, Yang YK, Liu CL, Li Y, Bai ZH. Two-component MprAB system regulates the expression of genes involved in cell envelope biosynthesis in Corynebacterium glutamicum [J]. Microorganisms, 2025, 13(5): 1120.
[21]
Barretto LAF, Van PT, Fowler CC. Conserved patterns of sequence diversification provide insight into the evolution of two-component systems in Enterobacteriaceae [J]. Microbial Genomics, 2024, 10(3): 001215.
[22]
Liu YX, Rose J, Huang SJ, Hu YB, Wu Q, Wang D, Li CG, Liu ML, Zhou P, Jiang L. A pH-gated conformational switch regulates the phosphatase activity of bifunctional HisKA-family histidine kinases[J]. Nature Communications, 2017, 8: 2104.
[23]
Mitchell SL, Ismail AM, Kenrick SA, Camilli A. The VieB auxiliary protein negatively regulates the VieSA signal transduction system in Vibrio cholerae [J]. BMC Microbiology, 2015, 15: 59.
[24]
Fritsch F, Mauder N, Williams T, Weiser J, Oberle M, Beier D. The cell envelope stress response mediated by the LiaFSR Lm three-component system of Listeria monocytogenes is controlled via the phosphatase activity of the bifunctional histidine kinase LiaS Lm[J]. Microbiology, 2011, 157(2): 373-386.
[25]
Kleine B, Chattopadhyay A, Polen T, Pinto D, Mascher T, Bott M, Brocker M, Freudl R. The three-component system EsrISR regulates a cell envelope stress response in Corynebacterium glutamicum [J]. Molecular Microbiology, 2017, 106(5): 719-741.
[26]
陈晓婷, 乾莲, 陈卫国, 孙士正, 白宗凯, 赵纯静, 武有聪. 表皮葡萄球菌vraSR-srrAB突变株的构建及生物学表型[J]. 微生物学通报, 2023, 50(8): 3562-3574.
Chen XT, Qian L, Chen WG, Sun SZ, Bai ZK, Zhao CJ, Wu YC. Construction and biological phenotype characterization of vraSR-srrAB deletion mutant of Staphylococcus epidermidis [J]. Microbiology China, 2023, 50(8): 3562-3574 (in Chinese).
[27]
Wenzel M, Dekker MP, Wang BW, Burggraaf MJ, Bitter W, van Weering JRT, Hamoen LW. A flat embedding method for transmission electron microscopy reveals an unknown mechanism of tetracycline[J]. Communications Biology, 2021, 4: 306.
[28]
Kim JH, Kang YJ, Kang YW, Park SE, Jin HB, Hong E, Park HH. A putative membrane-associated YdjB-like protein from Salmonella enterica exhibits a non-canonical ACT-like fold[J]. PLoS One, 2025, 20(12): e0336436.
[29]
张清仪, 王阶平, 程钢, 刘钟慧, 范文瑾, 何进. 苏云金芽孢杆菌双组分信号转导系统的生物信息学分析[J]. 微生物学通报, 2011, 38(9): 1385-1392.
Zhang QY, Wang JP, Cheng G, Liu ZH, Fan WJ, He J. Bioinformatics analysis of two-component signal transduction systems of Bacillus thuringiensis [J]. Microbiology China, 2011, 38(9): 1385-1392 (in Chinese).
[30]
Wang MJ, Gu SY, Guo J, Wu JY, Wang XH, Noman M, Wang JY, Li L. BcHK71 and BcHK67, two-component histidine kinases, regulate conidial morphogenesis, glycerol synthesis, and virulence in Botrytis cinerea [J]. Journal of Fungi, 2025, 11(12): 850.
[31]
Abhishek S, Deeksha W, Rajakumara E. Mechanistic insights into allosteric regulation of methylated DNA and histone H3 recognition by SRA and SET domains of SUVH5 and the basis for di-methylation of lysine residue[J]. The FEBS Journal, 2023, 290(4): 1060-1077.
[32]
Watanabe N, Savchenko A. Molecular insights into the initiation step of the Rcs signaling pathway[J]. Structure, 2024, 32(9): 1381-1393.e4.
[33]
Sewgoolam B, Jim KK, de Bakker V, Bock FP, Gibson PS, Veening JW. Genome-wide antibiotic-CRISPRi profiling identifies LiaR activation as a strategy to resensitize fluoroquinolone-resistant Streptococcus pneumoniae [J]. Nature Communications, 2025, 16: 6491.
[34]
Guffey AA, Loll PJ. Regulation of resistance in vancomycin-resistant enterococci: the VanRS two-component system[J]. Microorganisms, 2021, 9(10): 2026.
[35]
Cassiano MHA, Silva-Rocha R. Benchmarking bacterial promoter prediction tools: potentialities and limitations[J]. mSystems, 2020, 5(4): e00439-20.
[36]
Landry BP, Palanki R, Dyulgyarov N, Hartsough LA, Tabor JJ. Phosphatase activity tunes two-component system sensor detection threshold[J]. Nature Communications, 2018, 9: 1433.
[37]
Cao Q, Feng FF, Wang H, Xu XJ, Chen HC, Cai XW, Wang XR. Haemophilus parasuis CpxRA two-component system confers bacterial tolerance to environmental stresses and macrolide resistance[J]. Microbiological Research, 2018, 206: 177-185.
2026年第66卷第10期
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doi: 10.13343/j.cnki.wsxb.20260203
  • 接收时间：2026-03-11
  • 首发时间：2026-10-13
  • 出版时间：2026-10-04
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  • 收稿日期：2026-03-11
  • 录用日期：2026-04-21
基金
National Natural Science Foundation of China(32570026)
国家自然科学基金(32570026)
National Key Research and Development Program of China(2023YFD1300700)
国家重点研发计划(2023YFD1300700)
Program of Key Laboratory of Industrial Biotechnology of Ministry of Education in Jiangnan University(KLIB-KF202504)
江南大学工业生物技术教育部重点实验室项目(KLIB-KF202504)
作者信息
    江南大学 生物工程学院，工业生物技术教育部重点实验室，江苏 无锡

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2种不同金属材料的力学参数
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Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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