植物与病原物互作中的生物膜研究

生物膜是一种附着于生命或非生命物体表面的有特殊结构和功能的微生物群落。它广泛地存在于自然界和人工环境中。本文对细菌生物膜的定义、形成过程、影响因素、抗性机制以及在植物与病原物互作中的研究作了简要综述。     

寄主——病原物互作

 植物病理学是研究两种生命形式之间关系的科学。所谓寄主——病原物互作,就是在一定条件下,植物发病过程中寄主和病原物相互作用,从而决定发病表型的生理、生化和遗传调控过程。它主要是从寄主——病原物相互关系的角度研究病理过程机制。     

植物受病原物诱导启动子概述

植物受病原物诱导启动子是一类能对病原物的侵染作出响应的启动子,其活性主要局限于被侵染之时及被侵染的位点。植物受病原物诱导启动子的这一特性赋予其在抗病基因工程中潜在的应用价值。相比植物庞大的启动子组,已发现的植物受病原物诱导启动子仍是少数,关于其作用机制的研究仍有待加强和深入,而其调控的基因与植物抗病性关系还需要引起足够的重视。作者在对植物受病原物诱导启动子进行归类的基础上,重点关注了病原物诱导启动子调控基因的编码产物与植物抗病性的关系,对植物受病原物诱导启动子的顺式调控元件作了简要分析,并对该领域的发展动向进行了讨论。     

植物之间互作效应及其生理机制

在分析不同植物互作效应的基础上,就互作效应在农林复合系统中对植物体内养分、土壤养分有效性及植物生物学产量等的影响进行了概述,(1)互作促进了植物根系生长,使植物根系表现出较强的竞争作用,也有利于根系间的养分竞争;(2)植物之间的他感作用普遍存在于自然生态系统及栽培生态系统中,在植物资源配置时应充分利用这种相生相克作用;(3)互作促使植物体内的养分转移,改善体内的养分状况,达到植物之间养分互补;(4)互作可以有效地提高土壤肥力,改善土壤的养分状况和根际养分吸收环境,维持植物生产力;(5)互作能更有效地利用光、热、水、土和养分等资源,并减轻病虫危害,表现出明显的产量优势。在豆科与非豆科植物互作中,豆科植物的生物固氮作用为其互作植物提供了氮素营养保证,这种形式互作的产量优势通常在缺氮情况下表现更为明显。     

木霉-植物互作机制的研究进展

木霉作为是一种重要的生防制剂已经成为化学农药替代化和减量化的主要产品,也是公认的最有前防治病害的目的。随着木霉-植物互作研究的深入,关于互作机制方面的研究已取得了明显的进展。木霉与植物的互作过程包括木霉对植物的感知、木霉在植物体表和体内的定殖并引起植物的一系列变化等复杂的过程,本文重点对木霉-植物互作过程中木霉在植物中的定殖、木霉和植物的生理生化变化以及激素信号传递模式进行综述,以明确木霉促进植物生长和防治植物病害的机理,明确木霉与植物的互作机制对进一步提高木霉的生防效果、扩大木霉的应用具有重要的意义。     

植物生长过程中水分、氮素、光照的互作效应

在综述前人研究成果的基础上，讨论了水分、氮素、光照在植物生长中的作用，认为其主要体现在对植物光合作用的影响上，并着重分析了水分、氮素、光照三因素中任两因素互作对植物生长的影响。     

昆虫内共生菌-昆虫-植物互作关系研究进展

在长期的协同进化过程中,昆虫与其体内的共生菌建立了密切的互利共生关系。昆虫内共生菌不仅能调控宿主昆虫的营养代谢和生殖代谢,还能协助昆虫抵御生物、非生物胁迫,提高昆虫对化学农药的抗性及对寄主植物的适应性等。因此,内共生菌是宿主昆虫生长发育及适应性的重要调控因子。目前,随着组学技术的不断发展,内共生菌在宿主昆虫和寄主植物中的原位功能不断被挖掘,通过对内共生菌-昆虫-植物互作模型的研究,将进一步揭示昆虫内共生菌与昆虫、植物的互作机理,加深对昆虫适应性机制的理解并推进新型害虫防控和靶标技术的研发。本文就昆虫内共生菌的起源、特点、分布和传递,昆虫内共生菌在昆虫-植物-环境互作中的作用,以及昆虫内共生菌研究的方法和新技术等方面进行了综述,并对未来昆虫内共生菌介导的防御效应及昆虫适应性机理等热点问题进行了展望。     

水稻IR26悬浮细胞与白叶枯病菌互作的过敏反应现象

 当水稻IR26悬浮细胞与白叶枯病菌的不亲和小种(JXOV)互作时,在数小时内,悬浮细胞死亡率可达60%、膜透性增加40%、呼吸升高一倍多,而水稻IR26悬浮细胞与白叶枯病菌的亲和小种(JXOⅢ)互作时,在数小时内,悬浮细胞死亡率只有20%、膜透性只增加20%、呼吸升高不足40%,这些数据表明在水稻悬浮细胞与白叶枯病菌互作系统中可能存在着和水稻植株与白叶枯病菌互作系统中相似的互作机制。     

一种新大豆豆荚炭疽病症状类型及其病原鉴定

通过病原菌分离、形态学观察、核糖体DNA ITS序列分析和致病性试验,研究引起鲜食大豆"台75"豆荚普遍出现条状不规则形病斑的原因.结果显示,条状不规则形病斑是由平头炭疽菌Colletotrichum truncatum引起的一种新的大豆豆荚炭疽病症状,该病原菌引起豆荚炭疽病症状主要有2种,一种是圆形斑,另一种是条状不规则斑即锈斑.在供试的7种大豆品种中,条状不规则斑主要出现在"台75"上,其余6个品种主要是圆形斑;"台75"条状不规则斑的病荚率为65.37%,其余品种上的病荚率分别为1.02%、1.87%、2.95%、3.56%、9.32%和12.25%.研究表明由平头炭疽菌引起的大豆豆荚炭疽病的症状类型与寄主品种相关.     

杧果与Fusarium mangiferae在转录水平上的互作机制

基于Illumina HiSeq~(TM)2000平台,对健康与感病杧果顶芽的转录组进行测序,采用BLAST软件将获得的Unigene与NCBI-nr、Swiss-Prot、KEGG和COG数据库进行比对,根据基因功能注释后分析杧果病健组织的差异表达基因(DEGs),并对DEGs进行GO和Pathway富集分析。结果表明:2个样品共获得119 815条Unigene,N50为1 546 bp,平均片段长度为880 bp;鉴定了29 878个DEGs,对随机挑选的11个差异表达基因进行了qRT-PCR验证,结果与转录组一致。以corrected P-value≤0.05为阈值的代谢途径有22条,其中大多数代谢途径与植物的抗逆响应密切相关;153个DEGs参与了淀粉和蔗糖代谢途径,DEGs主要是编码糖类异构酶、水解酶和转移酶等,参与葡萄糖水解、细胞碳水化合物、丙酮酸盐和核苷酸代谢等生物进程;在抗氧化生物过程中,编码活性氧代谢相关酶且log_2Ratio10或-10以上的DEGs有20个,14个属上调表达,表明活性氧代谢在杧果与病原互作过程中起到重要的调解作用;F.mangiferae侵染杧果后,以log_2Ratio10或-10为筛选条件,共获得酚类代谢相关差异表达基因53个,其中40个DEGs上调表达,推测杧果可能是通过合成加固细胞壁的木质素或生成抑菌作用的酚类化合物来提高对病原菌的抵抗能力;杧果与F.mangiferae互作过程中,钙信号传导、SA信号途径和丝裂原活化蛋白信号传导途径相关基因表达下调,造成了下游植物抗病基因RPM1的表达受到抑制,这可能是杧果畸形病的主要成因之一。     

Spatial eco-evolutionary feedback in plant-pathogen interactions

In recent years the potential for evolutionary change to drive ecological dynamics, and vice versa, has been widely recognized. However, the convincing examples of eco-evolutionary dynamics mainly stem from highly artificial experimental systems, with conspicuously few examples contributed by field systems. While rarely considered in the eco-evolutionary literature, the gene-for-gene hypothesis inherently recognizes the tight link between evolutionary and ecological dynamics. The boom-and-bust dynamics of some agricultural pathogens are an extreme demonstration of this. In this perspective, we place plant-pathogen systems in a spatial eco-evolutionary framework, which recognizes that ecology and evolution are tightly linked, take place at the same time scale and are strongly influenced by spatial structure. Specifically, we: i) exemplify how the ecological process of dispersal modifies rapid local coevolutionary dynamics and thereby shapes spatial variation in resistance, infectivity, and local adaptation; and ii) illustrate how the outcome of coevolution (spatial distribution in resistance, infectivity and local adaptation) drives ecological metapopulation processes. Overall, we conclude that both agricultural and wild pathosystems provide a unique illustration of the high relevance of spatial eco-evolutionary feedback in understanding species interactions.     

Protoplasts in the analysis of early plant-pathogen interactions: current applications and perspectives

Transient gene expression in protoplasts has been applied to the analysis of gene expression, promoter activity, subcellular localization of proteins, and genome editing. Its advantages have been realized in recent years in various physiological, biochemical, genetic, and molecular biological studies. The omics and systems biology studies of plant development and their interactions with the environment have revitalized this approach. Early plant-pathogen interaction represents one of the most unique signalling research areas. Here, we try to discuss some developments and current applications of the protoplast system. It is for sure that the system will significantly bridge the gap between high throughput approaches and functional analysis in the study of plant-pathogen interactions in the omics era.     

Programmed cell death pathways induced by early plant‐virus infection are determined by isolate virulence and stage of infection

Programmed cell death (PCD) pathways caused by Turnip mosaic virus (TuMV) infection before symptom appearance were studied by light microscopy and electrolyte leakage following sap inoculation of Brassica carinata (Ethiopian mustard) TZ‐SMN‐44‐6 plants. Leaf responses to inoculation with avirulent (TuMV‐avir) and virulent (TuMV‐vir) isolates, and mock‐inoculation, were compared at 2, 20 and 52 h after inoculation (hai). The phenotypes induced were localized resistance (TuMV‐avir) and systemic susceptibility (TuMV‐vir). No visible TuMV symptoms were recorded in any inoculated plants during the 2–52 hai sampling period, but appeared as chlorotic spots in inoculated leaves at 5 days after inoculation. With TuMV‐vir alone, they were followed by systemic infection (mosaic). Dead cell number, deformation, percentage area and percentage integrated intensity, and conductivity of electrolyte leakage data, were analysed to examine their possible roles in stimulating cell death pathways. At 2 hai, dead cell number and percentage area were significantly greater for TuMV‐avir than TuMV‐vir infection or mock‐inoculation. Overall, isolate TuMV‐vir caused significantly greater cell deformation than TuMV‐avir, whereas wounding by mock‐inoculation had negligible effects. By 52 hai, isolate TuMV‐avir caused significantly greater electrolyte leakage than isolate TuMV‐vir or mock‐inoculation. This suggests both isolates triggered morphological changes consistent with apoptotic‐like PCD and necrosis‐like PCD that depended upon isolate virulence and stage of infection, respectively. These findings highlight how quantification of dead cell deformation and electrolyte leakage offer a new understanding of compatible and incompatible plant responses to early virus infection in plants.     

双香豆素诱导斜纹夜蛾卵巢细胞程序性死亡的机理研究

为探索双香豆素(dicoumarin, DIC)抑制斜纹夜蛾Spodoptera litura卵巢细胞SL-221增殖及作用机制,首先,采用CCK-8法测定双香豆素等21个酚类化合物对SL-221细胞的毒性;其次,通过倒置显微镜观察4μg/mL双香豆素处理细胞48 h后其形态变化,并结合流式细胞术研究双香豆素对SL-221的细胞周期、线粒体膜电位及凋亡的影响;最后,通过实时荧光定量PCR技术(RT-qPCR)探索磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase, PI3K)/雷帕霉素靶蛋白(target of rapamycin, TOR)营养信号通路的下游关键基因的表达变化。结果表明,21个酚类化合物中双香豆素对SL-221细胞的增殖抑制活性最显著,且呈现明显的浓度依赖性,其48 h的IC₅₀为0.85μg/mL,对细胞的增殖抑制活性是阳性对照印楝素(IC₅₀为7.20μg/mL)的8.47倍。4μg/mL双香豆素处理SL-221细胞48 h后,显微镜下可见细胞肿胀、膜破裂及大量凋亡小体;流式细胞术分析发现,双...     

Hydroperoxides of fatty acids induce programmed cell death in tomato protoplasts

Arachidonic acid (AA) is a fatty acid elicitor abundant in the glycerolipids of the late blight pathogen Phytophthora infestans and related Oomycete species. Lipoxygenases (LOX), which catalyze the addition of molecular oxygen to the 1 or 5 position of a cis, cis -1,4- Z, Z -pentadiene system in polyunsaturated fatty acids, is induced in host plants such as tomato and potato during infection by P. infestans. Here it is reported that AA, the LOX metabolites of AA, 5- and 15-hydroperoxyeicosatetraenioc acid (5- and 15-HpETE), and the LOX metabolite of linoleic acid, 9-hydroperoxyoctadecadienoic acid (9-HpODE), are potent inducers of programmed cell death (PCD) in tomato protoplasts. 5- and 15-HpETE increased DNA fragmentation as detected by t erminal deoxynucleotidyl transferase-mediated d U TP-X n ick e nd l abeling (TUNEL) and increased DNA laddering as visualized by ligation-mediated PCR. Background levels of DNA laddering were decreased in intensity by Zn²⁺ and increased by Ca²⁺, effects that are consistent with the reported action of these cations on PCD-associated endonucleases in other systems. H₂O₂, methyl jasmonate, and linoleic and linolenic acids were not toxic to tomato protoplasts at concentrations up to 350 μ , and lipid peroxides (LD₁₀₀ = 80 μ ) were far more potent inducers of death than free AA within this same concentration range. These results indicate the potential of fatty acid peroxides and LOX-related metabolism to engage an apoptotic type of PCD in higher plant cells.     

Tissue specific colonization of Phytophthora capsici in Capsicum spp.: molecular insights over plant-pathogen interaction

Phytoparasitica - Root and crown rot (RCR) caused by Phytophthora capsici is present in all crop production areas of pepper and chili worldwide. This pathogen was recently reported at the Pacific...     

Involvement of nitric oxide generation in hypersensitive cell death induced by elicitin in tobacco cell suspension culture

Recent studies suggest that nitric oxide (NO), an important signaling and defense molecule in mammals, plays a key role in activating disease resistance in plants. We characterized NO production by tobacco Bright Yellow-2 cells pharmacologically after treatment with INF1, the major elicitin secreted by the late blight pathogen Phytophthora infestans, prepared from Escherichia coli. NO production rapidly occurred within 1h and reached a maximum level 3–6h after the addition of INF1. Carboxy-PTIO, a NO-specific scavenger, abolished INF1-induced NO production in a dose-dependent manner. Pretreatment of protein synthesis inhibitor cycloheximide and protein kinase inhibitor K252a blocked NO production 3–12h after INF1 treatment, indicating that NO production requires de novo protein synthesis and protein phosphorylation. In an investigation of the relations between NO generation and several defense responses induced by INF1, carboxy-PTIO completely suppressed activation of a 41-kDa protein kinase and cell death by INF1. Carboxy-PTIO also suppressed the induction of hypersensitive-related (hsr) genes HSR515 and HSR203J, the expression of which is strongly correlated with the hypersensitive response in plants. The results suggest that NO plays a crucial role in the induction of hypersensitive cell death.     

Apoptosis, programmed cell death and the hypersensitive response

Apoptosis is typically a morphologically identifiable form of programmed cell death in mammals that is regulated by genes with homologues in other animal Phyla. Although both plants and fungal plant pathogens exhibit forms of developmental programmed cell death, demonstrated morphological or genetic homologies with mammalian apoptosis are still generally lacking. Because of its ubiquity and the involvement of signal transduction pathways in its induction, a strong case is developing that the hypersensitive response is a specific form of plant programmed cell death evolved as a defense against microbial parasites. Data suggest that separate signalling pathways may lead to the cell death and the defense gene activation that characterize this response and that parasite-specific resistance genes represent only one of many types of genes involved in response regulation. However, despite some biochemical similarities between the hypersensitive response, forms of developmental programmed cell death in plants, and animal apoptosis, no unique and consistent markers for the hypersensitive response (or plant programmed cell death in general) have yet been found. Whether any of these forms of plant cell death should be called apoptosis depends on how the term is defined. Assuming the hypersensitive response is a form of programmed cell death and is the default state upon pathogen entry into a cell, it seems likely that intracellular biotrophic plant pathogens resemble some animal viruses in being able to suppress this response in susceptible hosts.