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1.
荧光假单胞菌Pseudomonas fluorescens 2P24是根围促生细菌(PGPR),具有Ⅲ型分泌系统(T3SS)。为了在2P24中表达植物过敏反应激发子harpin,赋予生防菌诱导抗病性能力,本文选择可在2P24中表达的来自Pseudomonas syringae pv.tomato DC3000的avrPto1基因启动子与水稻细菌性条斑病菌Xanthomonasoryzae pv.oryzicola harpin蛋白编码基因hpa1进行融合,实现了harpin蛋白在2P24的表达。重组菌株通过T3SS分泌harpin蛋白,可激发烟草产生过敏反应(HR),激活HR途径的HIN1基因和HRS203J基因以及病程相关蛋白PR1a基因的转录表达。harpin重组菌株与2P24一样,对小麦赤霉病菌Fusarium graminearum和棉花枯萎病菌F.oxysporum f.sp.vasinfectum具有抑制作用。这为利用植物-病原物互作中激发植物产生抗病性的激发子来遗传改良生防微生物奠定了理论和实践基础。  相似文献   

2.
本研究通过DNA体外重组技术,将稻瘟病菌MoSlp1基因和拟南芥几丁质寡糖受体基因AtCERK1的跨膜结构域和胞内结构域基因融合,构成嵌合基因MoSlp1-AtCERK1。在烟草瞬时表达试验中,MoSlp1-AtCERK1嵌合蛋白诱导烟草产生以过敏性坏死为表现形式的过敏反应。以带有MoSlp1-AtCERK1蛋白的农杆菌注射烟草,可增强烟草对烟草花叶病毒的抗病性。同时提高烟草的PR-1基因转录表达水平,表明MoSlp1-AtCERK1嵌合蛋白基因通过水杨酸信号传导的途径激活烟草系统获得性抗病。  相似文献   

3.
诱导抗病性是实现植物病害绿色防控的重要途径,大丽轮枝菌蛋白激发子PevD1能激活植物免疫系统,提高本生烟对烟草花叶病毒(TMV)和烟草野火病病原菌Pseudomonas syringae pv.tabaci、棉花对大丽轮枝菌Verticillium dahliae的抗病性,但分子机制不清晰。前期转录组测序(RNA-Seq)分析结果显示,本生烟响应PevD1诱导的差异表达基因显著富集在倍半萜烯和三萜烯的合成途径中。本文进一步分析了这些差异表达基因的功能,并通过测定倍半萜植保素辣椒醇合成关键基因EAS和EAH的转录表达水平和辣椒醇积累量,证明PevD1能诱导本生烟产生植保素辣椒醇,明确了PevD1诱导植保素辣椒醇的产生是提高植物抗病性的重要机制之一。  相似文献   

4.
 水稻和稻瘟病菌互作是研究植物与病原菌互作的模式体系。本文利用3个水稻抗稻瘟病近等基因品系(CO39、C101LAC和C101A51)和2个特异性菌株(M209和M210)构成不同亲和程度的互作关系,研究了稻瘟病菌对3个水稻信号传导途径关键酶合成基因、5个防御反应病程相关蛋白基因和1个防御反应转录因子调控基因诱导表达的作用。结果表明:稻瘟病菌诱导了各种互作关系中水稻OsLOXOsAOSOsPAL酶合成基因的表达,水稻启动了茉莉酸和水杨酸防御反应信号传导途径。在不亲和的互作反应中,稻瘟病菌能不同程度地诱导水稻OsPR1aOsPR2OsPR3-1OsPR3-2OsPR4基因的表达,从而有效激活了防御反应系统,使水稻植株表现为抗病;而在亲和的互作反应中,多数OsPR基因的表达水平低、时间短或没有表达,水稻植株表现为感病。OsMyb基因在各种互作关系中有不同的诱导表达。说明这些防御相关基因的诱导表达可能与水稻抗稻瘟病性相关。  相似文献   

5.
 青霉菌灭活菌丝体(Dry Mycelium of Penicillium chrysogenum,DMP)是工业生产青霉素的残余副产物,研究发现DMP能够提高多种作物的抗病性。本文研究了DMP对烟草BY-2悬浮细胞防卫反应的诱导作用,并初步探索其诱导机制,结果表明:DMP处理烟草BY-2悬浮细胞后,产生了活性氧迸发,在处理后30 min达到峰值;DMP诱导烟草BY-2悬浮细胞胞外基质碱性化,该变化能被蛋白激酶抑制剂K252a部分抑制;苯丙氨酸解氨酶(PAL)和过氧化物酶(POD)的活性被诱导而明显升高,分别在处理后4 h和8 h达到峰值;DMP诱导了病程相关蛋白基因PR-1aPR-1b以及抗病信号传导途径关键基因NPR1的表达。说明DMP能够诱导烟草BY-2悬浮细胞产生抗病防卫反应,其抗病信号可能是通过水杨酸信号途径进行传导的,蛋白质磷酸化参与了该抗病信号的传导过程。  相似文献   

6.
 将编码大豆凝集素的lec-s基因插入植物表达载体pBI121中,构建植物重组表达质粒pBI121:: lec-s。由根癌土壤杆菌EHA105介导的叶盘法转化烟草,获得了转基因烟草株系。PCR和RT-PCR检测证明lec-s基因已转入烟草植株中。接种烟草花叶病毒(Tobacco mosaic virus,TMV)进行抗病性试验结果表明,转基因烟草叶片上的病斑数显著减少,说明转基因烟草表现出对TMV的抗性。定量RT-PCR检测发现,接种TMV后,抗病防卫基因(PR-1a、GST1、Pal和hsr515)在转基因烟草叶片中显著上调表达。这些结果表明,大豆凝集素基因lec-s转化烟草可对TMV产生抗性,其作用机制可能在于lec-s基因参与了植物的防卫信号通路,诱导了抗病防卫基因在转基因植株体内的表达,增强了植株对TMV的系统抗性。  相似文献   

7.
为明确毁灭炭疽菌Colletotrichum destructivum诱抗蛋白诱导烟草的抗病性及其作用,采用喷雾、摩擦接种方法及RT-PCR技术研究了诱抗蛋白的预防保护作用,以及烟草悬浮细胞经诱导后过氧化物酶(POD)、多酚氧化酶(PPO)、苯丙氨酸解氨酶(PAL)活性及脯氨酸(Pro)含量和病程相关基因表达的变化。结果表明,接种3、5和7 d后,该诱抗蛋白对烟草炭疽病的诱抗效果分别为58.00%、48.99%和49.65%,对烟草白粉病的诱抗效果分别为83.26%、80.76%和78.60%,并可以抑制烟草普通花叶病毒的复制及在寄主体内的扩增;经诱抗蛋白处理后,烟草悬浮细胞POD、PPO、PAL活性及Pro含量明显提高;诱抗蛋白能够诱导烟草病程相关蛋白基因PR-1a、PR-1b以及抗病信号传导途径关键基因NPR1的表达。表明毁灭炭疽菌诱抗蛋白可诱导烟草产生抗病性,可能与烟草悬浮细胞中POD、PAL、PPO的活性及Pro的含量提高以及相关病程基因表达有关。  相似文献   

8.
采用PCR方法从水稻细菌性条斑病菌RS105菌株中扩增harpinXooc编码基因hrf2,将其克隆到酵母表达载体pPICZαA的分泌信号肽基因下游,获得重组表达质粒pPICZαA-hrf2。重组表达质粒线性化后电击转化至毕赤酵母宿主菌X-33,经抗生素Zeocin筛选和PCR鉴定后,得到重组酵母菌X-33/pPICZαA-hrf2。用甲醇诱导重组酵母菌表达目标蛋白,发酵上清液经浓缩后进行SDS-PAGE电泳分析,在约18kD处有特异目标条带出现。Western blot检测表明表达产物具有良好的抗原性。生物活性检测表明酵母重组表达蛋白harpinXooc能够诱导烟草产生过敏反应和促进烟草生长,活性高于在大肠杆菌中表达的harpinXooc。  相似文献   

9.
乙烯信号传导途径因子OsEIL 6调控水稻抗稻瘟病反应   总被引:2,自引:0,他引:2  
稻瘟病(rice blast)是水稻生产上最严重的病害之一。抗病相关基因的挖掘对稻瘟病的防治具有重要意义。研究表明植物EIN3/EIL家族基因在抗病过程中发挥着重要作用。本研究采用RNAi技术探究OsEIL6参与的水稻抗稻瘟病反应。稻瘟菌侵染时基因表达谱检测结果表明,OsEIL6在水稻和稻瘟菌非亲和组合中受到诱导表达。稻瘟菌接种结果显示,水稻OsEIL6沉默株系和野生型植株‘TG394’相比抗性下降;实时荧光定量RT-PCR结果分析表明,OsEIL6的表达量下降导致乙烯合成途径中OsACO1和乙烯信号传导途径的OsERF063和OsERF073的转录水平下降。亚细胞定位研究发现该基因定位于水稻细胞质。OsEIL6沉默株系中ROS合成途径标记基因OsrbohA和OsrbohB的表达量均明显下调,表明该基因可能通过影响ROS的合成调控水稻抗稻瘟病反应。本研究结果将有助于进一步揭示OsEIL6参与的乙烯信号传导途径介导的水稻抗稻瘟病反应机制。  相似文献   

10.
转hrf1基因水稻对稻曲病抗性分析   总被引:3,自引:0,他引:3  
用注射接种法测定9个转hrf1基因水稻品系对稻曲病的抗性,结果表明B12-2m、B12、HTRP2和NJH12转基因系对稻曲病的抗性有显著提高,其中NJH12对稻曲病的抗性表现最突出。在江苏南京和安徽潜山两地的田间测定结果显示NJH12对稻曲病的抗性表现明显,与对照相比防效提高65%以上。用RT-PCR测定抗病转基因系中防卫基因的表达,在抗病转基因系中,Ospr1a、Ospr1b和PAL等防卫反应基因的表达明显增强。转hrf1基因水稻可能通过诱导防卫反应基因的表达提高水稻对稻曲病的抗性。  相似文献   

11.
Liu F  Liu H  Jia Q  Wu X  Guo X  Zhang S  Song F  Dong H 《Phytopathology》2006,96(10):1052-1059
ABSTRACT HpaG(Xooc), produced by Xanthomonas oryzae pv. oryzicola, is a member of harpin group of proteins that stimulate plant growth, hypersensitive cell death (HCD), and pathogen defense. The protein contains two copies of the glycine-rich motif (GRM), a characteristic of harpins, and a cysteine, which is absent in other harpins. Genetic modification generated the pro-tein mutants HpaG(Xooc)MG (MG) by deleting GRMs and HpaG(Xooc)C47T (C47T) by replacing cysteine with threonine. When applied to tobacco plants, C47T and MG were 1.2- and 1.7-fold stronger, respectively, than HpaG(Xooc) in inducing HCD, which occurred consistently with expression of the marker genes hin1 and hsr203. The proteins markedly alleviated infection of tobacco by Tobacco mosaic virus and Arabidopsis and tomato by Pseudomonas syringae. Treating tobacco plants with HpaG(Xooc), C47T, and MG decreased the viral infection by 58, 81, and 92%, respectively. In Arabidopsis and tomato plants treated with HpaG(Xooc), C47T, or MG, P. syringae multiplication was inhibited; bacterial population multiplied in 5 days in these plants were ca. 160-, 1,260-, or 15,860-fold smaller than that in control plants. So pathogen defense was induced in both plants. Defense-related genes Chia5, NPR1, and PR-1a were expressed consistently with resistance. In response to HpaG(Xooc), C47T, and MG, aerial parts and roots of tomato plants increased growth by 15 and 53%, 25 and 77%, and 46 and 106%, relative to controls. The expansin gene, EXP2, involved in the cell expansion and plant growth was expressed coordinately with plant growth promotion. These results suggest that the presence of GRM and cysteine in HpaG(Xooc) represses the effects of the protein in plants.  相似文献   

12.
Peng JL  Bao ZL  Ren HY  Wang JS  Dong HS 《Phytopathology》2004,94(10):1048-1055
ABSTRACT Harpin(Xoo), encoded by the hpaG(Xoo) gene of Xanthomonas oryzae pv. oryzae, is a member of the harpin group of proteins that induce pathogen resistance and hypersensitive cell death (HCD) in plants. We elaborated whether both processes are correlated in hpaG(Xoo)-expressing tobacco (HARTOB) plants, which produced harpin(Xoo) intracellularly. Resistance to fungal, bacterial, and viral pathogens increased in HARTOB, in correlation with the expression of hpaG(Xoo), the gene NPR1 that regulates several resistance pathways, and defense genes GST1, Chia5, PR-1a, and PR-1b that are mediated by different signals. However, reactive oxygen intermediate burst, the expression of HCD marker genes hsr203 and hin1, and cell death did not occur spontaneously in HARTOB, though they did in untransformed and HARTOB plants treated exogenously with harpin(Xoo). Thus, the transgenic expression of harpin(Xoo) confers nonspecific pathogen defense in the absence of HCD.  相似文献   

13.
 研究了一种α型elicitin-parasiticein对烟草微敏反应(microscopic hypersensitive response, micro-HR)和防卫反应分子表征基因表达的诱导作用。用150 nmol/L的parasiticein喷洒烟草叶片12 h后, 经曲利本蓝(trypan blue)染色, 光镜下可观察到有细胞坏死, 说明parasiticein引起了micro-HR, 而水和低浓度的parasiticein不能引起micro-HR。用parasiticein注射叶片可引起肉眼可见的HR, 注射用parasiticein浓度远比喷洒引起micro-HR的浓度低。Parasiticein还可诱导对TMV的抗性, 浓度在30 nmol/L时比150 nmol/L的效果好。用parasiticein注射烟草30 min, HR表征基因hsr203Jhin1开始表达, 在9 h内逐渐降低, 12~16 h肉眼可观察到HR。Parasiticein喷洒烟草后, 病程相关蛋白(pathogenesis-related, PR)基因PR-1b在诱导的第1 d到第5 d都有一定量的表达。可见, parasiticein能同步诱导过敏反 应和抗病 性及其分 子表征基 因的表达 。  相似文献   

14.
Chen L  Qian J  Qu S  Long J  Yin Q  Zhang C  Wu X  Sun F  Wu T  Hayes M  Beer SV  Dong H 《Phytopathology》2008,98(7):781-791
Harpin proteins from gram-negative plant-pathogenic bacteria can stimulate hypersensitive cell death (HCD) and pathogen defense as well as enhance growth in plants. Two of these diverse activities clearly are beneficial and may depend on particular functional regions of the proteins. Identification of beneficial and deleterious regions might facilitate the beneficial use of harpin-related proteins on crops without causing negative effects like cell death. Here, we report the identification and testing of nine functional fragments of HpaG(Xooc), a 137-amino-acid harpin protein from Xanthomonas oryzae pv. oryzicola, the pathogen that causes bacterial leaf streak of rice. Polymerase chain reaction-based mutagenesis generated nine proteinaceous fragments of HpaG(Xooc); these caused different responses following their application to Nicotiana tabacum (tobacco) and Oryza sativa (rice). Fragment HpaG62-137, which spans the indicated amino acid residues of the HpaG, induced more intense HCD; in contrast, HpaG10-42 did not cause evident cell death in tobacco. However, both fragments stimulated stronger defense responses and enhanced more growth in rice than the full-length parent protein, HpaG(Xooc). Of the nine fragments, the parent protein and one deletion mutant of HpaG(Xooc) tested, HpaG10-42, stimulated higher levels of rice growth and resulted in greater levels of resistance to X. oryzae pv. oryzae and Magnaporthe grisea. These pathogens cause bacterial leaf blight and rice blast, respectively, the two most important diseases of rice world-wide. HpaG10-42 was more active than HpaG(Xooc) in inducing expression of several genes that regulate rice defense and growth processes and activating certain signaling pathways, which may explain the greater beneficial effects observed from treatment with that fragment. Overall, our results suggest that HpaG10-42 holds promise for practical agricultural use to induce disease resistance and enhance growth of rice.  相似文献   

15.
从毛头鬼伞Coprinus comatus中提取的碱性糖蛋白Y3可以降低烟草花叶病毒(TMV)的侵染。克隆获得Y3蛋白cDNA后与真核表达载体pPIC-9k连接,重组载体pPIC-9k-Y3成功电转化入毕赤酵母Pichia pastoris后,转化子在28℃、250 r/min培养条件下,使用1.0%甲醇诱导表达6 d,成功实现了Y3蛋白的真核表达。300 μg/mL浓度的Y3蛋白对普通烟进行诱导处理后摩擦接种TMV结果表明,诱导处理后烟草体内除过氧化物酶POD外,多酚氧化酶PPO、苯丙氨酸解氨酶PAL和β-1,3葡聚糖酶活性都有不同程度提高。蛋白处理24 h后植株中PPO活性达到最大,为对照组的2倍。利用实时荧光定量PCR (qRT-PCR)研究抗性相关基因表达情况,Y3诱导后烟草植株碱性病程相关基因1(PR1-b)、病程相关基因非表达子1(NPR1)、PAL在转录水平上均显著上调(P<0.05)。推测真核表达产物Y3蛋白可通过提高水杨酸信号途径相关防御酶活性以及抗病基因转录来诱导烟草对TMV产生系统抗性,为Y3蛋白作为生物农药开发提供理论基础。  相似文献   

16.
Systemic acquired resistance (SAR) is induced following inoculation of Peronospora tabacina sporangia into the stems of Nicotiana tabacum plants highly susceptible to the pathogen. Previous results have shown that accumulation of acidic β-1,3-glucanases (PR-2's) following induction of SAR by P. tabacina may contribute to resistance to P. tabacina. We showed that up-regulation of the PR-2 gene, PR-2d, following stem inoculation with P. tabacina, is associated with SAR. Studies using plants transformed with GUS constructs containing the full length promoter from PR-2d or promoter deletions, provided evidence that a previously characterized regulatory element that is involved in response to salicylic acid (SA), may be involved in regulation of PR-2d following induction of SAR with P. tabacina. This work provides evidence that regulation of PR-2 genes during P. tabacina-induced SAR may be similar to regulation of these genes during infection of N-gene tobacco by TMV or following exogenous application of SA, and provides further support for the role of SA in regulation of genes during P. tabacina-induced SAR.  相似文献   

17.
激活蛋白PeaT1诱导烟草对TMV的系统抗性   总被引:4,自引:0,他引:4  
枯斑三生烟草(Samsun-NN)经激活蛋白PeaT1诱导后接种烟草花叶病毒(Tobacco mosaic virus,TMV),对TMV产生了明显的系统获得抗性,枯斑抑制率达54.15%,枯斑大小也受到一定程度的限制。研究结果表明,PeaT1处理烟草植株下位三片叶不同时间后,其上部叶片中PPO、POD和PAL 3种抗病防御酶活性均比对照提高,第4 d酶活性达到最高值。实时荧光定量PCR检测结果显示,经PeaT1诱导4 d后烟草叶片中抗病相关基因PR1a、PR1b、NPR1在转录表达水平上较未诱导对照都有不同程度的上调。由以上结果我们推测PeaT1诱导烟草产生了系统获得抗性,本研究为进一步阐明PeaT1诱导植物抗病的信号传导途径奠定了基础。  相似文献   

18.
BACKGROUND: Trichoderma asperellum SKT-1 is a microbial pesticide of seedborne diseases of rice. To investigate the mechanisms of disease suppression in SKT-1, the ability to induce systemic resistance by SKT-1, or its cell-free culture filtrate (CF), was tested using Arabidopsis thaliana Col-0 plants. RESULTS: Both SKT-1 and its CF elicit an induced systemic resistance against the bacterial leaf speck pathogen Pseudomonas syringae pv. tomato DC3000 in Col-0 plants. Involvement of plant hormones in the induced resistance by SKT-1 and CF was assessed using Arabidopsis genotypes such as the jasmonic acid (JA)-resistant mutant jar1, the ethylene (ET)-resistant mutant etr1, the plant impaired in salicylic acid (SA) signalling transgenic NahG and the mutant npr1 impaired in NPR1 activity. In soil experiments using SKT-1, no significant disease suppression effect was observed in NahG transgenic plants or npr1 mutant plants. Expression levels of SA-inducible genes such as PR-1, PR-2 and PR-5 increased substantially in the leaves of Col-0 plants. Expression levels of JA/ET-induced genes such as PDF1.2a, PR-3, PR-4 and AtVsp1 were also induced, but the levels were not as high as for SA-inducible genes. In a hydroponic experiment using CF from SKT-1, all Arabidopsis genotypes showed an induced systemic resistance by CF and increased expression levels of JA/ET- and SA-inducible genes in leaves of CF-treated plants. CONCLUSION: The SA signalling pathway is important in inducing systemic resistance to colonisation by SKT-1, and both SA and JA/ET signalling pathways combine in the signalling of induced resistance by CF. These results indicate that the response of A. thaliana is different from that found in root treatments with barley grain inoculum and CF from SKT-1. Copyright © 2011 Society of Chemical Industry  相似文献   

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