橡胶树白粉菌Oidium heveae侵染寄主拟南芥的筛选

为筛选出橡胶树白粉菌Oidium heveae Steimnann新的寄主载体,通过接种野生型拟南芥Col-0、突变体Sr1-4D及eds1,观察了橡胶树白粉菌侵染不同拟南芥突变体的过程,PCR扩增测序法验证相关致病基因,构建橡胶树白粉菌-拟南芥互作体系。结果表明,在Col-0、Sr1-4D叶片上面,橡胶树白粉菌产生部分菌丝后停止生长,不能形成典型的橡胶树白粉病症状;但能成功侵染eds1叶片,在叶片的正、背面有银白色辐射状菌丝,后期在病斑上出现一层粉层,表现橡胶树白粉病的典型症状。组织染色和显微观察结果显示,在突变体eds1叶片上橡胶树白粉菌完成了侵染过程。PCR扩增测序结果表明接种后突变体eds1叶片上及组织中病菌均为橡胶树白粉菌;使用橡胶树白粉菌3个致病相关基因作为靶标,验证了eds1和橡胶树上白粉菌基因组中的3个致病相关基因相似度均达到99%~100%。表明橡胶树白粉菌可侵染拟南芥突变体eds1。     

OsDCL1基因沉默突变体诱导稻瘟病抗性的转录组分析

稻瘟病是严重危害水稻生长的真菌病害,每年给水稻生产带来重大经济损失。与野生型日本晴NPB相比,OsDCL1基因沉默突变体增强了水稻对稻瘟菌的广谱抗病性。为了解水稻OsDCL1-RNAi突变体的抗病机制,我们分析和比较了它和野生型日本晴NPB在稻瘟菌侵染前后转录组水平的变化。响应稻瘟菌侵染的7 129个差异表达基因(DEGs)中,NPB和OsDCL1-RNAi突变体分别有5 382和5 180个基因表达发生了变化,参与生物胁迫反应、信号通路、蛋白代谢和RNA调控4个通路的基因明显被富集。以野生型未受稻瘟菌侵染的基因表达为对照,在OsDCL1-RNAi突变体中共发现1 318个DEGs,且超过70%是上调的,其中很多基因参与了生物胁迫反应(26.9%)和信号通路(11%),上调表达基因中与生物胁迫反应相关的主要为PR基因和细胞壁相关基因。另外,还发现10个茉莉酸相关基因和11个水杨酸相关基因分别在OsDCL1-RNAi突变体中显著下调和上调表达。在OsDCL1-RNAi突变体中还发现WRKY和MYB等一些转录因子家族以及部分受体类激酶被诱导表达。用qRT-PCR方法验证部分差异表达的基因,其结果与DEGs基本一致。OsDCL1-RNAi突变体中转录组变化的分析,为深入了解该突变体抗瘟性增强的机制奠定了基础。     

用DNA异质性分析稻瘟菌细胞质专化小种形成趋势

 随着中国以"三系"配套为基础的杂交稻的大面积推广,水稻雄性不育细胞质遗传基础单一化是否会导致稻瘟菌产生细胞质专化致病小种的问题日益为人们所关注。本研究以随机扩增多态型DNA技术为手段,通过分析不同来源的水稻雄性不育细胞质的遗传相似性以及中国南方杂交稻主栽区的稻瘟菌不同生理小种同对野败型专化致病菌株90-2的遗传相似性进行分析,探查寄主及病原菌两方面的遗传基础,并在此上研究分析产生细胞质专化小种的寄主、环境条件及病原菌变异的趋势。     

稻瘟病菌拮抗细菌bio-2的抑菌作用及其鉴定

稻瘟病是水稻的重要病害,可引起大幅度减产.目前,利用水稻抗瘟性品种和使用化学农药仍是防治稻瘟病行之有效的方法,但因抗瘟品种的单一化、稻瘟病菌生理小种组成的复杂性和变异性、化学农药的毒性以及病原菌的抗药性,使水稻稻瘟病的防治受到一定的制约,因此,生物防治越来越受到人们的关注.     

氮肥对水稻品种-稻瘟菌小种相对寄生适合度的影响

 选择具有不同抗病性水稻品种-稻瘟菌小种组合,在室内、田间分别设7个、3个不同氮肥水平,测定不同组合不同氮肥水平下品种-小种相对寄生适合度,方差分析结果表明,施氮量对品种小种相对寄生适合度的影响较小。相对寄生适合度可以作为一项较为稳定的参量用于水稻品种与稻瘟病小种互作研究。     

稻瘟菌MoHYR1蛋白的原核表达与纯化

正稻瘟菌(Magnaporthe oryzae)是高等丝状子囊真菌,由其侵染水稻引发的稻瘟病是水稻第一大病害,生产上造成了巨大的经济损失。在植物与病原菌的互作过程中,植物会产生防御应答反应,其中包括活性氧自由基(reactive oxygen species,ROS)。ROS在植物抵御病原菌的侵入以及病原     

浅谈品种布局在综合治理稻瘟病中的作用

1981年广东省稻瘟大流行后,韶关地区开始研究稻瘟菌生理小种的分布和优势种群,并据此更换了一批品种,1984年稻瘟再度流行时避免了损失。1985年起,广东省农作物病虫测报站连续3年对16个县作生理小种测定,发现ZA群小种出现频次在增加,     

水稻抗瘟性与过氧化物酶同工酶的初步研究

本研究采用垂直板聚丙烯酰胺凝胶电泳的方法,测定了11个稻瘟菌小种接种的18个水稻品种(品系)的过氧化物酶同工酶。结果表明,受侵染植株与对照间的同工酶谱带数目及其相对迁移率(Rf)无明显差异、无新谱带出现。说明受侵染植株与对照间过氧化物酶同工酶的代谢途径没有变化;但过氧化物酶同工酶的活性与水稻抗瘟性有密切关系。接种7天后植株体内该酶的活性非亲和组合显著大于亲和组合。     

氮肥对水稻品种-稻瘟菌小种相对寄生适合度的影响

选择具有不同抗病性水稻品种 -稻瘟菌小种组合 ,在室内、田间分别设 7个、3个不同氮肥水平 ,测定不同组合不同氮肥水平下品种 -小种相对寄生适合度 ,方差分析结果表明 ,施氮量对品种小种相对寄生适合度的影响较小。相对寄生适合度可以作为一项较为稳定的参量用于水稻品种与稻瘟病小种互作研究。     

稻瘟病菌生理小种变化与水稻品种抗瘟性丧失之间的关系

本文根据近年稻瘟病菌生理小种鉴定研究结果,对生理小种变化与水稻品种抗瘟性丧失之间的关系进行了初步分析。(1)我省稻瘟病菌生理小种组成发生了变化。优势小种由1979—1982年的G群变为1983年后的B群小种;(1)品种的抗性随潜在致病力强的生理小种出现频率上升而降低;(3)伴随抗病品种种植面积比率的扩大,潜在致病小种迅速增殖,丧失抗性的危险性加大;(4)品种抗性随抗优势种群频率下降而降低。     

Systemic resistance induced in Arabidopsis thaliana by Trichoderma asperellum SKT-1, a microbial pesticide of seedborne diseases of rice

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     

Assessment of blast disease resistance in transgenic PRms rice using a gfp-expressing Magnaporthe oryzae strain

Rice blast caused by the fungus Magnaporthe oryzae (anamorph Pyricularia grisea ) is one of the most devastating diseases of cultivated rice worldwide. In this study, a green fluorescent protein ( gfp )-expressing M. oryzae strain was generated and used to investigate the infection process in a commercial rice cultivar. Expression of the gfp gene did not affect the pathogenicity of the M. oryzae transformants. Confocal microscopy allowed in vivo imaging of this pathogen during infection of rice tissues. Magnaporthe oryzae pathogenicity was examined on both leaf and root tissues. In roots of wild-type plants, the fungus penetrated into epidermal and cortical cells, and colonized the central cylinder and xylem vessels. However, the dimorphic growth pattern typically observed during the biotrophic and necrotrophic stages of leaf colonization was not observed during colonization of root tissues. Furthermore, events occurring during infection of rice plants constitutively expressing the maize pathogenesis-related PRms gene were characterized and compared with those occurring during the interaction of this pathogen with untransformed rice plants. Fungal penetration was drastically reduced and delayed in tissues of PRms plants compared to untransformed plants. These results indicated that the gfp -expressing M. oryzae represents a strategic tool for the assessment of blast disease resistance in transgenic rice which can be also applied to the analysis of the M. oryzae interaction with other cultivars or mutants of important crop species.     

乙烯信号传导途径因子OsEIL 6调控水稻抗稻瘟病反应

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。     

Induced systemic resistance in arabidopsis thaliana against Pseudomonas syringae pv. tomato by 2,4-diacetylphloroglucinol-producing Pseudomonas fluorescens

Pseudomonas fluorescens strains that produce the polyketide antibiotic 2,4-diacetylphloroglucinol (2,4-DAPG) are among the most effective rhizobacteria that suppress root and crown rots, wilts, and damping-off diseases of a variety of crops, and they play a key role in the natural suppressiveness of some soils to certain soilborne pathogens. Root colonization by 2,4-DAPG-producing P. fluorescens strains Pf-5 (genotype A), Q2-87 (genotype B), Q8r1-96 (genotype D), and HT5-1 (genotype N) produced induced systemic resistance (ISR) in Arabidopsis thaliana accession Col-0 against bacterial speck caused by P. syringae pv. tomato. The ISR-eliciting activity of the four bacterial genotypes was similar, and all genotypes were equivalent in activity to the well-characterized strain P. fluorescens WCS417r. The 2,4-DAPG biosynthetic locus consists of the genes phlHGF and phlACBDE. phlD or phlBC mutants of Q2-87 (2,4-DAPG minus) were significantly reduced in ISR activity, and genetic complementation of the mutants restored ISR activity back to wild-type levels. A phlF regulatory mutant (overproducer of 2,4-DAPG) had ISR activity equivalent to the wild-type Q2-87. Introduction of DAPG into soil at concentrations of 10 to 250 μM 4 days before challenge inoculation induced resistance equivalent to or better than the bacteria. Strain Q2-87 induced resistance on transgenic NahG plants but not on npr1-1, jar1, and etr1 Arabidopsis mutants. These results indicate that the antibiotic 2,4-DAPG is a major determinant of ISR in 2,4-DAPG-producing P. fluorescens, that the genotype of the strain does not affect its ISR activity, and that the activity induced by these bacteria operates through the ethylene- and jasmonic acid-dependent signal transduction pathway.     

No role for bacterially produced salicylic Acid in rhizobacterial induction of systemic resistance in Arabidopsis

ABSTRACT The role of bacterially produced salicylic acid (SA) in the induction of systemic resistance in plants by rhizobacteria is far from clear. The strong SA producer Pseudomonas fluorescens WCS374r induces resistance in radish but not in Arabidopsis thaliana, whereas application of SA leads to induction of resistance in both plant species. In this study, we compared P. fluorescens WCS374r with three other SA-producing fluorescent Pseudomonas strains, P. fluorescens WCS417r and CHA0r, and P. aeruginosa 7NSK2 for their abilities to produce SA under different growth conditions and to induce systemic resistance in A. thaliana against bacterial speck, caused by P. syringae pv. tomato. All strains produced SA in vitro, varying from 5 fg cell(-1) for WCS417r to >25 fg cell(-1) for WCS374r. Addition of 200 muM FeCl(3) to standard succinate medium abolished SA production in all strains. Whereas the incubation temperature did not affect SA production by WCS417r and 7NSK2, strains WCS374r and CHA0r produced more SA when grown at 33 instead of 28 degrees C. WCS417r, CHA0r, and 7NSK2 induced systemic resistance apparently associated with their ability to produce SA, but WCS374r did not. Conversely, a mutant of 7NSK2 unable to produce SA still triggered induced systemic resistance (ISR). The possible involvement of SA in the induction of resistance was evaluated using SA-nonaccumulating transgenic NahG plants. Strains WCS417r, CHA0r, and 7NSK2 induced resistance in NahG Arabidopsis. Also, WCS374r, when grown at 33 or 36 degrees C, triggered ISR in these plants, but not in ethylene-insensitive ein2 or in non-plant pathogenesis- related protein-expressing npr1 mutant plants, irrespective of the growth temperature of the bacteria. These results demonstrate that, whereas WCS374r can be manipulated to trigger ISR in Arabidopsis, SA is not the primary determinant for the induction of systemic resistance against bacterial speck disease by this bacterium. Also, for the other SAproducing strains used in this study, bacterial determinants other than SA must be responsible for inducing resistance.     

稻瘟病菌效应蛋白MoIEEP1功能初探

由稻瘟病菌引起的稻瘟病是全球最严重的植物真菌病害之一, 严重威胁水稻生产安全?效应蛋白是病原菌与植物对抗过程中分泌产生的一种蛋白质, 可作为毒力因子促进侵染或作为无毒因子触发防御反应?本研究对实验室前期筛选到的在稻瘟病菌侵染早期诱导表达的效应蛋白(infection-induced expression effector protein 1, MoIEEP1)进行了功能验证与分析?结果表明: MoIeep1 在稻瘟病菌侵染初期8 h表达量最高; 信号肽和亚细胞定位分析验证该蛋白N端含有17个氨基酸的信号肽且在水稻原生质体中呈明亮点状的定位信号, 初步推测该定位信号为过氧化物酶体或线粒体等细胞器; 与野生型菌株Guy11相比, MoIeep1 基因敲除突变体菌丝生长无明显差异, 但其致病性受到影响?以上研究结果为进一步探究该蛋白质的作用机理打下良好基础, 也为揭示其他效应蛋白在稻瘟病菌侵染水稻过程中的功能提供新的理论依据?     

钙离子响应蛋白GCaMP6s在稻瘟菌中的异源表达

稻瘟病严重威胁全球水稻的产量,钙离子信号通路参与了稻瘟菌生长、发育和致病,了解这些过程中菌株细胞质中钙离子浓度的变化对稻瘟病的防治具有重要意义.GCaMP6s作为一种细胞质钙离子感受蛋白在动物和植物中已有大量应用.本研究利用不同稻瘟菌菌株构建了 GCaMP6s基因的异源表达菌株,在0.01 g/mL钙离子处理下,GCa...