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epsps regulates cell elongation by disrupting the balance of lignin and flavonoid biosynthesis in cotton

Journal of Integrative Agriculture 2024年第10期23卷 3437-3456页

作者： Qingdi Yan Wei Hu Chenxu Gao Lan Yang Jiaxian Yang Renju Liu Masum Billah Yongjun Lin Ji Liu Pengfei Miao Zhaoen Yang Fuguang Li Wenqiang Qin State Key Laboratory of Cotton Bio‑breeding and Integrated Utilization/Institute of Cotton Research Chinese Academy ofAgricultural SciencesAnyang 455000China National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research Huazhong AgriculturalUniversityWuhan 430070China State Key Laboratory of Cotton Bio-breeding and Integrated Utilization/Zhengzhou University Zhengzhou 450066China National Nanfan Research Institute(Sanya) Chinese Academy of Agricultural SciencesSanya 572025China

epsps is a key gene in the shikimic acid synthesis pathway that has been widely used in breeding crops with herbicide ***,its role in regulating cell elongation is poorly *** the overexpression of epsps genes,we generated lines resistant to glyphosate that exhibit an unexpected dwarf phenotype.A representative line,DHR1,exhibits a stable dwarf phenotype throughout its entire growth *** for plant height,the other agronomic traits of DHR1 are similar to its transgenic explants *** section observations showed that DHR1 internodes are shortened due to reduced elongation and division of the internode *** hormones confirmed that DHR1 is not a classical brassinolide(BR)-or gibberellin(GA)-related dwarfing *** analysis and fine mapping confirmed that the epsps gene is the causal gene for dwarfism,and the phenotype can be inherited in different *** and metabolome analyses showed that genes associated with the phenylpropanoid synthesis pathway are enriched in DHR1 compared with *** metabolites are enriched in DHR1,whereas lignin metabolites are *** enhancement of flavonoids likely results in differential expression of auxin signal pathway genes and alters the auxin response,subsequently affecting cell *** study provides a new strategy for generating dwarfs and will accelerate advancements in light simplification in the cultivation and mechanized harvesting of cotton.

关键词： epsps cotton T-DNA dwarfism herbicide resistance

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月季epsps基因的克隆及生物信息学分析

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分子植物育种 2024年第8期22卷 2488-2494页

作者：张金炜刘峰张椿芳王天康宁波城市职业技术学院景观生态学院宁波315100

月季是园林绿化中常用的花卉,因此十分有必要对其抗性基因进行研究。5-烯醇丙酮酸莽草酸3-磷酸合酶(EPSP;3-磷酸莽草酸1-羧基乙烯基转移酶;EC 2.***.1.19)为莽草酸途径中的关键酶,也是除草剂草甘膦的靶向酶。为了探究月季epsps基因的结构... 详细信息

月季是园林绿化中常用的花卉,因此十分有必要对其抗性基因进行研究。5-烯醇丙酮酸莽草酸3-磷酸合酶(EPSP;3-磷酸莽草酸1-羧基乙烯基转移酶;EC 2.***.1.19)为莽草酸途径中的关键酶,也是除草剂草甘膦的靶向酶。为了探究月季epsps基因的结构和功能,本研究利用月季基因组信息设计特异引物克隆得到月季的epsps基因,命名为Rcepsps。经RT-PCR克隆后得到Rcepsps基因的ORF全长为1 566 bp,共编码521个氨基酸,分子质量为55.19 kD,等电点为7.5,亲水系数为0.009。初步研究得知Rcepsps属于epsps I型蛋白,与同属蔷薇科的野草莓的序列同源性极高。研究结果可为后续抗性基因的利用提供理论基础。

关键词：月季 epsps 生物信息学分析

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Generation of insect-resistant and glyphosate-tolerant rice by introduction of a T-DNA containing two Bt insecticidal genes and an epsps gene

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2015年第10期16卷 824-831页

作者： Qi-chao ZHAO Ming-hong LIU Xian-wen ZHANG Chao-yang LIN Qing ZHANG Zhi-cheng SHEN State Key Laboratory of Rice Biology Institute of Insect Sciences Zhejiang University

Insect resistance and glyphosate tolerance have been two of the most important traits in the genetic improvement of various crops. In this study, two Bacillus thuringiensis （Bt） insecticidal genes, CrylAc and Cryllg, and a modified glyphosate-tolerant 5-enolpyruvylshikimate-3-phosphate synthase （epsps） gene （GIO） were combined into a single transferred DNA （T-DNA） fragment and introduced into rice by Agrobacterium-mediated transformation. A transgenic line with single-copy T-DNA insertion named GAI-14 was found to be highly resistant to striped stem borer and rice leaf roller, and tolerant to glyphosate. Analysis of T-DNA border sequence suggested that the transgenes were inserted at the chromosome 3 and appeared to have not interrupted any known or putative genes. A field trial observed no significant difference in the basic agronomic traits between GAI-14 and the recipient rice.

关键词： Bt gene stacking Bt resistance management epsps Transgenic rice

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Transcriptomic analysis reveals the transcription factors involved in regulating the expression of epsps gene,which confers glyphosate resistance of goosegrass (Eleusine indica)

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Journal of Integrative Agriculture 2021年第8期20卷 2180-2194页

作者： ZHANG Chun YU Chao-jie ZHANG Tai-jie GUO Wen-lei TIAN Xing-shan Guangdong Provincial Key Laboratory of High Technology for Plant Protection Institute of Plant ProtectionGuangdong Academy of Agricultural SciencesGuangzhou 510640P.R.China

Glyphosate inhibits the enzyme 5-enolpyruvylshikimate-3-phosphate synthase(epsps)and overexpression of the epsps gene is one of the molecular mechanisms conferring glyphosate resistance in weeds.A regulatory sequence of epsps gene was isolated previously,and an alteration in its 5´-untranslated region(UTR)pyrimidine(Py)-rich stretch element is involved in the regulation of epsps expression in glyphosate-resistant(GR)Eleusine ***,the transcription factors involved in this regulatory sequence remain to be *** this study,we investigated the regulatory network of epsps overexpression associated genes in a GR *** population by *** differentially expressed transcript analyses revealed that glyphosate treatment caused an increase in the expression of 2752 unigenes and a decrease in the expression of 4025 unigenes in the GR ***,compared to the glyphosatesusceptible(GS)*** them,1373 unigenes were identified to be co-expressed with the epsps gene in GR *** and KEGG pathway analyses showed that the up-regulated unigenes were mainly enriched in chloroplasts and associated with the shikimate biosynthesis pathway,chlorophy II and peroxisome metabolism ***,the expression of a Shikimate kinase which catalyzed the conversion of Shikimate to Shikimate 3-phosphate(S3P,a substrate of epsps),was also *** transcription factors were identified as likely to be involved in the regulation of the epsps expression,and three of them(ARF2,ARF8 and BPC6)showed more binding sites because of a(CT)n insertion of the 5´-UTR Py-rich stretch element in ***,the yeast one-hybrid assay illustrated that ARF8 and BPC6 could bind to the 5´-UTR Py-rich stretch element of wild type epsps,but could not bind to the mutated *** data suggests that the transcriptional regulation of epsps expression is complex and was significantly altered in GR *** discoveries provide new references for further study of the

关键词： transcriptomic epsps 5´-UTR Py-rich stretch element transcription factor glyphosate resistance

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The epsps Pro106Ser substitution solely accounts for glyphosate resistance in a goosegrass(Eleusine indica) population from Tennessee, United States

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Journal of Integrative Agriculture 2016年第6期15卷 1304-1312页

作者： Janel L Huffman Chance W Riggins Lawrence E Steckel Patrick J Tranel Department of Crop Sciences University of Illinois Urbana IL 61801 USA Department of Plant Sciences University of Tennessee Jackson TN 38301 USA

Previous studies have documented the occurrence of glyphosate-resistant （GR） goosegrass （Eleusine indica （L.） Gaertn.） and, in at least some cases, resistance is due to an altered target site. Research was performed to determine if an altered target site was responsible for GR in a Tennessee, United States goosegrass population （TennGR）. DNA sequencing revealed a mutation in TennGR plants conferring the Prol06Ser 5-enolpyruvylshikimate-3-phosphate synthase （epsps） substitution previously identified in other GR populations. F2 populations were derived from TennGR plants crossed with plants from a glyphosate-susceptible population （TennGS） and analyzed for their response to glyphosate and genotyped at the epsps locus. Plants from the F2 populations segregated 1：2：1 sensitive：intermediate：resistant in response to a selec- tive dose of glyphosate, and these responses co-segregated with the epsps genotypes （PP106, PS106, and SS106）. To separately investigate the effect of the Prol06Ser substitution on GR, glyphosate dose-response curves and 50% effective dose （EDso） values were compared among the three genotypes and the two parental populations. The SS106 genotype was 3.4-fold resistant relative to the PP106 genotype, identical to the resistance level obtained when comparing the resistant and susceptible parental populations. We conclude that the mutation conferring a Prol06Ser epsps mutation is solely responsible for GR in the TennGR goosegrass population.

关键词： herbicide resistance monogenic trait glyphosate PCR amplification of specific alleles (PASA) epsps target-site mutation

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Reconstruction of enzymatic activity from split genes encoding glyphosate-tolerant epsps protein of Psedomonas fluorescens G2 strain by intein mediated protein complementation

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Chinese Science Bulletin 2006年第13期51卷 1652-1654页

作者： DUN Baoqing LU Wei ZHANG Wei PING Shuzhen WANG Xujing CHEN Ming XU Yuquan JIN Dan WANG Jin ZHAO Zhonglin LIANG Aimin HOU Songna XU Ming-Qun LIN Min Biotechnology Research Institute Chinese Academy of Agricultural Sciences Beijing 100081 China New England Biolabs Inc. Beverly Massachusetts 01915 USA

A mutagenesis library was constructed using GPS-LS system to insert a random 5 aa into the 5-enolpyruvylshikimate-3-phosphate synthase (epsps) encoded by aroA gene. Active epsps pro- teins were identified by the ability to rescue growth of aroA-deleted mutant ER2799 on M9 minimal media. 12 unique sites, which can tolerate a 5-aa insertion, were identified. In all of the 12 sites, only F295/T296 site was found to split the G2-epsps properly by co-transformation of plasmids into E. coli ER2799. The G2-epsps gene was then divided into N-termi- nal and C-terminal from F295/T296 site which were fused to the N-terminal and C-terminal of *** intein, respectively, creating two plasmids pMEPS- N295IN and pKEPSc296Ic. Co-transformation of plasmids, pMEPSN295IN and pKEPSc296Ic, rescu- ed growth of ER2799 in M9 minimal media, indicating that the intein splicing domains were bringing the epsps fragments together to generate activity. Re- consituted activity of splitted G2-epsps enzyme was 4.48 U/mg.

关键词：断裂基因蛋白质再生转基因 epsps

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转cry1Ab和epsps基因玉米C0030.3.5对土壤固氮细菌丰度和群落结构的影响

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环境科学 2018年第8期39卷 3885-3893页

作者：王蕊朱珂李刚刘惠芬王晶修伟明赵建宁杨殿林农业部环境保护科研监测所天津300191 天津农学院农学与资源环境学院天津300384

为评估转基因玉米种植对土壤氮周转功能微生物的潜在风险,以转cry1Ab和epsps基因玉米C0030.3.5(TM)及其亲本玉米DBN318(PM)为研究对象,于2015年拔节期、抽雄期、乳熟期、完熟期采集根际土和非根际土进行试验,并采用荧光定量PCR(quantita... 详细信息

为评估转基因玉米种植对土壤氮周转功能微生物的潜在风险,以转cry1Ab和epsps基因玉米C0030.3.5(TM)及其亲本玉米DBN318(PM)为研究对象,于2015年拔节期、抽雄期、乳熟期、完熟期采集根际土和非根际土进行试验,并采用荧光定量PCR(quantitative real-time PCR,q PCR)和末端限制性片段长度多态性(terminal restriction fragment length polymorphism,TRFLP)技术分析土壤固氮微生物nif H基因丰度和多样性.结果表明,TM和PM根际土和非根际土固氮细菌nif H基因丰度随生长时期整体呈现先升高后降低的变化趋势,无论是根际土还是非根际土,同一生长时期2种玉米nif H基因丰度间差异均不显著.相关分析显示,土壤固氮细菌nif H基因丰度与有机质含量呈极显著正相关.T-RFLP结果表明,所获得的14种TRFs中,43 bp和155 bp片段所代表的固氮细菌为共有优势种群,无论是根际土还是非根际土,同一生长时期各T-RFs的相对丰度在TM和PM间差异同样不显著.土壤固氮细菌的Shannon指数和Evenness指数随生长期整体呈现出先升高后降低的变化趋势,无论是根际土还是非根际土,同一生长时期TM和PM的Shannon指数间及Evenness指数间均无显著差异.主成分分析(principal component analysis,PCA)表明TM和PM土壤固氮细菌群落结构组成无显著差异.冗余分析(redundancy analysis,RDA)显示土壤铵态氮和p H对固氮细菌群落结构组成影响显著.

关键词： cry1Ab epsps 转基因玉米 nifH基因丰度群落结构

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过量表达epsps转基因对栽培稻与杂草稻杂种后代稻瘟病抗病性的影响

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植物生理学报 2020年第9期56卷 1872-1880页

作者：林凯青韩光煜罗琼何平南蓬复旦大学生命科学学院生物多样性与生态工程教育部重点实验室上海200438 云南农业大学植物保护学院农业生物多样性与病害控制教育部重点实验室昆明650201

5-烯醇式丙酮酰莽草酸-3-磷酸合酶(5-enolpyruvylshikimate-3-phosphate synthase,epsps)是植物体莽草酸途径中除草剂草甘膦的靶标酶。本研究利用过量表达epsps转基因栽培稻×杂草稻杂种后代(WH1)进行转录组测序研究,发现水杨酸信号转... 详细信息

5-烯醇式丙酮酰莽草酸-3-磷酸合酶(5-enolpyruvylshikimate-3-phosphate synthase,epsps)是植物体莽草酸途径中除草剂草甘膦的靶标酶。本研究利用过量表达epsps转基因栽培稻×杂草稻杂种后代(WH1)进行转录组测序研究,发现水杨酸信号转导途径发生显著富集;稻瘟病抗病性鉴定实验结果表明,过量表达epsps转基因栽培稻×杂草稻杂种阳性后代(WH1+)与阴性后代(WH1–)的稻瘟病抗病性没有显著差异;代谢物含量检测表明,转基因阳性植株与阴性植株的水杨酸含量基本一致。由此可见,过量表达epsps转基因事件不会通过改变水杨酸水平来影响栽培稻×杂草稻杂种后代植株的稻瘟病抗病性。

关键词： epsps 稻瘟病抗病性转录组水杨酸 LC-MS

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epsps外源基因对陆地棉农艺性状和纤维品质的影响

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棉花学报 2015年第5期27卷 489-493页

作者：燕树锋刘海芳铁双贵祝水金浙江大学农业与生物技术学院农学系浙江杭州310085 河南省农业科学院粮食作物研究所河南郑州450002 济源市农业局河南济源454650

以G6-1至G6-19等19个转基因抗草甘膦除草剂棉花种质为材料,以其非转基因背景亲本中棉所49为对照,研究epsps外源基因导入对陆地棉农艺性状和纤维品质的影响,结果表明外源基因的导入对棉花纤维上半部平均长度、马克隆值、断裂伸长率和断... 详细信息

以G6-1至G6-19等19个转基因抗草甘膦除草剂棉花种质为材料,以其非转基因背景亲本中棉所49为对照,研究epsps外源基因导入对陆地棉农艺性状和纤维品质的影响,结果表明外源基因的导入对棉花纤维上半部平均长度、马克隆值、断裂伸长率和断裂比强度等品质性状有显著影响;对于棉花衣分和铃重等农艺性状亦有显著的影响,衣分变异范围为30.5%~42.7%,T1和T2相关性分析表明衣分的变异为可遗传性变异,铃重最高为6.5 g,最低为4.6 g,两者相差40%。因此,转基因棉花育种研究不仅要注重目标性状的转育,还要注重非目标性状的筛选。

关键词：棉花 epsps 外源基因农艺性状纤维品质

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外源抗草甘膦epsps基因在大豆基因组中的整合与定位

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作物学报 2010年第3期36卷 365-375页

作者：王晓波蒋凌雪魏利刘林陆伟李文欣王俊陶波常汝镇邱丽娟中国农业科学院作物科学研究所/农作物基因资源与基因改良国家重大科学工程/农业部作物种质资源引用与生物技术重点开放实验室北京100081 东北农业大学黑龙江哈尔滨150030 中国农业科学院生物技术研究所北京100081

通过基因特异引物扩增和免疫层析试纸法分别对转基因大豆中外源epsps基因和其编码的蛋白进行检测,结果表明,epsps基因不仅已整合到大豆基因组中,而且epsps蛋白可以正常表达。利用染色体步移法获得了转基因大豆插入位点的侧翼序列,序列... 详细信息

通过基因特异引物扩增和免疫层析试纸法分别对转基因大豆中外源epsps基因和其编码的蛋白进行检测,结果表明,epsps基因不仅已整合到大豆基因组中,而且epsps蛋白可以正常表达。利用染色体步移法获得了转基因大豆插入位点的侧翼序列,序列比对表明35S上游的大豆DNA序列起始于Gm02:7912740,NOS下游的大豆DNA序列起始于Gm02:7777705。在本研究检测序列范围内发现插入位点两侧不同位置有3个不同的未知片段、2个大豆基因组序列倒位,同时发现2个大豆基因组片段丢失。外源基因不是以点插入方式整合,而是导致大豆基因组约135kb片段的移位;NOS下游大豆基因组序列发生重排,导致一个编码HEC1和HEATrepeat两个功能域的基因(Glyma02g09790)结构受到影响,首次发现该基因在ABA和PEG处理时下调表达,推测该基因通过ABA信号通路参与胁迫应答。本研究通过对抗除草剂epsps基因在大豆基因组中的插入位点分析,明确了外源epsps基因在大豆基因组中的整合、定位及其侧翼序列,为转基因大豆安全评价提供了依据。

关键词：转基因大豆插入位点基因组 epsps 抗草甘膦定位侧翼序列

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