植物抗病分子机制研究进展

近十年来,植物抗病分子机制研究取得显著进展。综述了植物抗病基因的克隆及其结构分析、病原菌无毒基因及其相关致病因子的克隆与研究、信号传导相关因子的克隆及其结构分析以及植物-病原菌的相互作用研究,重点介绍了以植物特异抗病基因为介导的诱导防卫作用机制(包括抗病基因编码毒素蛋白,进而抑制病原菌的繁殖;显性基因编码病原菌致病性的靶标物;抗病基因表达产物直接引发抗病反应和基因对基因的抗病作用机制等)的研究进展,以期为植物抗病育种提供有益的信息。     

植物抗病基因克隆与功能研究进展

植物抗病基因（R基因）是分子植物病理学和植物基因工程研究的热点之一,R基因的克隆及其在抗病反应中的功能研究为揭示植物抗病机制和有效－控制植物病害奠定了基础,本文介绍了R基因的成功克隆方法和克隆新策略,对R基因编码产物的功能进行了分类分析,并对通过遗传工程途径发展R基因介导的抗病植物新品种进行了展望。     

寄主植物与病原菌免疫反应的分子遗传基础

近年来,大量的植物抗病基因和病原菌无毒基因被克隆,抗病基因和无毒基因的结构、功能及其互作关系的研究也取得重大进展。在植物中,由病原菌模式分子(pathogen-associated molecular patterns, PAMPs)引发的免疫反应(PAMP-triggered immunity, PTI)和由效应因子引发的免疫反应(effector-triggered immunity, ETI)是植物在长期进化过程中形成的两类抵抗病原物的机制。PTI反应主要通过细胞表面受体(patternrecognition receptors, PRRs)识别并结合PAMPs从而激活下游免疫反应,而在ETI反应中,则通过植物R基因(resistance gene,R)与病原菌无毒基因(avirulence gene, Avr)产物间的直接或间接相互作用来完成免疫反应。本文对植物PTI反应和ETI反应分别进行了概述,重点探讨了植物R基因与病原菌Avr基因之间的互作遗传机理,并对目前植物抗性分子遗传机制研究和抗病育种中的问题进行了探讨和展望。     

植物抗病基因结构、功能及其进化机制研究进展

植物与病原菌在长期的共进化和相互选择的过程中,逐渐形成了组织障碍、非寄主抗性和小种专化抗性等有效的防御机制。小种专化抗性(基因对基因抗性)主要是由植物抗病基因识别相应的病原菌无毒基因并激活植物体内抗病信号进而抵御病原菌的侵染。从目前已克隆的 70 多个抗病基因来看,它们在结构上具有高度保守性,主要包括核苷酸结合位点(NBS),亮氨酸重复结构(LRR), 蛋白激酶结构域(PK), 果蝇蛋白 Toll 和哺乳动物蛋白质白细胞介素 1 受体[interleukin(IL)-1 receptor]类似结构域(TIR), 双螺旋结构(CC)或亮氨酸拉链(LZ)和跨膜结构域(TM)等,其在抗病基因与病原菌无毒(效应)蛋白互作以及植物内部免疫信号传导中起着重要的作用。同时,抗病基因又通过基因复制、遗传重组等进化机制形成多基因家族,为植物抗病的专化性和多样性提供了重要的遗传基础。本文主要讨论了近来已克隆抗病基因的结构特征、功能以及抗病基因进化机制研究的进展。     

植物抗病基因与病原菌无毒基因互作的分子基础

近10年来,大量的植物抗病基因和病原菌无毒基因被克隆,抗病基因和无毒基因的结构、功能及其互作关系的研究也取得重大进展。通过介绍抗病基因与无毒基因互作的两种模式,从抗病基因与病原菌无毒基因互作角度探讨了抗病基因在植物抗病育种和农作物生产中直用的问题,提出抗细菌和真菌单基因转化很难赋予农作物切实抗性。     

昆虫病原菌(Isaria fumosorosea)查尔酮异构酶基因CHI克隆及生物信息学分析

本研究旨在对昆虫病原菌玫烟色棒束孢(Isaria fumosorosea)查尔酮异构酶基因CHI进行克隆、测序及相关生物信息学分析.序列分析结果显示:IfCHI基因编码区全长为1152bp,编码383个氨基酸,理论等电点(pI)D为9.72,属不稳定水溶性蛋白,其二级结构为混合型,蛋白质溶剂可及性主要分为三类,三级结构由α-螺旋、β-折叠、β-转角和无规则卷曲组成,同源序列分析及多重序列比对分析存在明显的差异.该昆虫病原菌玫烟色棒束孢IfCHI编码基因的成功克隆及生物信息学分析,为进一步研究病原菌CHI基因的遗传特性和表达机制以及为寻找更多的查尔酮类化合物提供了理论基础.     

水稻抗稻瘟病近等基因系的cDNA微阵列分析

应用cDNA微阵列对来源于中156／谷梅2号重组自交系的水稻抗稻瘟病近等基因系G205和G71的稻瘟病菌胁迫基因表达谱进行了分析,发现有3个cDNA克隆的表达仅在抗病基因系G205接种病原菌12h后受到诱导,其中两个为功能已知基因,另一个为功能未知的新基因。另有35个差异表达克隆在两个近等基因系中均检测到,其中17个克隆的表达在G205和G71均受到病原菌的诱导,另外18个克隆的表达则在G205和G71均受到病原菌原抑制。序列分析表明,这些稻瘟病菌应答基因分别与防卫反应,信号传递,逆境胁迫和光合作用及糖代谢等功能相关,为植物抗病机制提供了相关信息。另外,Northern还证实了编码富含甘氨酸蛋白基因（Glycinerich protein Grp)的表达受稻瘟病病原菌的诱导,是一个稻瘟病诱导相关基因。     

植物抗病基因同源序列及其在抗病基因克隆与定位中的应用

近10年来已有20多个植物抗病基因被克隆,测序,这些抗病基因所编码的蛋白中大多含有核苷酸结合位点,富含亮氨酸重复序列,蛋白激酶,亮氨酸拉链结构,跨膜结构域,Toll白介素－1区域等保守结构域。利用这些保守结构域合成PCR引物,已扩增出大量的植物抗病基因同源序列（RGA）。对RGA与抗病基因的关系进行了分析,讨论了RGA在研究抗病基因进化中的作用,指出RGA在抗病基因定位和转基因中具有重要意义。     

水稻抗稻瘟病基因资源与分子育种策略

稻瘟病是水稻主要病害之一。利用抗病品种是防治稻瘟病最经济、有效和安全的措施。近年来,随着植物先天免疫机制、抗病分子生物学以及水稻和稻瘟菌基因组学研究的不断深入,一系列参与病原菌识别和防卫信号传导与应答,以及外源抗菌蛋白、病原菌激发子等抗病相关基因陆续被鉴定和克隆,为提高水稻抗稻瘟病能力提供了一些新的基因资源、育种策略和技术。本文概述了国内外近年来克隆的主要抗稻瘟病基因资源及其在分子育种研究中应用的进展,提出了通过转基因手段整合不同防卫反应关键调控基因的抗稻瘟病聚合育种策略。     

同源序列法克隆植物抗病基因研究进展及其在野生稻中的应用

随着各种测序计划的完成及生物信息学的发展,植物抗病基因的克隆取得了很大进展,至今有几十个基因已被克隆。研究发现,大多抗病基因都存在特异的保守序列,如富亮氨酸重复序列、核苷酸结合位点、丝氨酸-苏氨酸激酶等。抗病基因的结构特征不仅预示了植物抗病反应中可能的作用机制,而且为分离克隆植物抗病基因提供了一条可行的新途径,如基于同源序列的候选基因法,又称为同源序列法。我们简要综述了已克隆抗病基因的结构、同源序列法克隆抗病基因的研究进展,并对其在野生稻中的应用做了展望。     

水稻抗稻瘟病基因的标记辅助选择及定位克隆

水稻抗稻瘟病基因-稻瘟病菌无毒基因相互作用体系是当今植物分子病理学和抗病育种学研究领域的模式体系之一,其中抗病基因的分子定位与克隆及其标记辅助选择已成为该体系的重要内容。本文就这方面的研究进展作一简要综述,以期为水稻抗病育种提供有益的信息。     

The Genetic and Molecular Basis of Plant Resistance to Pathogens

Plant pathogens have evolved numerous strategies to obtain nutritive materials from their host,and plants in turn have evolved the preformed physical and chemical barriers as well as sophisticated two-tiered immune system to combat pathogen attacks.Genetically, plant resistance to pathogens can be divided into qualitative and quantitative disease resistance,conditioned by major gene(s) and multiple genes with minor effects,respectively.Qualitative disease resistance has been mostly detected in plant defense against biotrophic pathogens,whereas quantitative disease resistance is involved in defense response to all plant pathogens,from biotrophs,hemibiotrophs to necrotrophs.Plant resistance is achieved through interception of pathogen-derived effectors and elicitation of defense response.In recent years,great progress has been made related to the molecular basis underlying host-pathogen interactions.In this review,we would like to provide an update on genetic and molecular aspects of plant resistance to pathogens.     

植物病原物无毒基因及其功能

植物抗病基因与病原物无毒基因产物间直接或间接相互作用导致产生的基因对基因抗性是植物抗病性的重要形式。无毒基因已在多种植物病原物 ,包括真菌、细菌、病毒和卵菌等中得到克隆。绝大多数已克隆无毒基因之间 ,及其与已知蛋白之间 ,均无显著序列同源性。然而 ,多数已克隆植物抗病基因有较高序列一致性 ,产物往往具有相似的结构域。由序列一致性很高的抗病基因产物与没有明显序列同源性的无毒基因产物相互作用 ,介导产生的过敏性细胞坏死和抗病性 ,在产生速度、强度和组织特异性等方面均可能有显著差异。无毒基因具有双重功能 :在含互补抗病基因植物中表现无毒效应 ,而在不含互补抗病基因植物中显示小种、菌株、致病型、或种特异性毒性效应     

Lectin-mediated resistance impairs plant virus infection at the cellular level

Plants possess a multilayered defense response, known as plant innate immunity, to infection by a wide variety of pathogens. Lectins, sugar binding proteins, play essential roles in the innate immunity of animal cells, but the role of lectins in plant defense is not clear. This study analyzed the resistance of certain Arabidopsis thaliana ecotypes to a potexvirus, plantago asiatica mosaic virus (PlAMV). Map-based positional cloning revealed that the lectin gene JACALIN-TYPE LECTIN REQUIRED FOR POTEXVIRUS RESISTANCE1 (JAX1) is responsible for the resistance. JAX1-mediated resistance did not show the properties of conventional resistance (R) protein-mediated resistance and was independent of plant defense hormone signaling. Heterologous expression of JAX1 in Nicotiana benthamiana showed that JAX1 interferes with infection by other tested potexviruses but not with plant viruses from different genera, indicating the broad but specific resistance to potexviruses conferred by JAX1. In contrast with the lectin gene RESTRICTED TEV MOVEMENT1, which inhibits the systemic movement of potyviruses, which are distantly related to potexviruses, JAX1 impairs the accumulation of PlAMV RNA at the cellular level. The existence of lectin genes that show a variety of levels of virus resistance, their targets, and their properties, which are distinct from those of known R genes, suggests the generality of lectin-mediated resistance in plant innate immunity.     

Plant disease resistance genes: recent insights and potential applications

Plant disease resistance genes (R genes) encode proteins that detect pathogens. R genes have been used in resistance breeding programs for decades, with varying degrees of success. Recent molecular research on R proteins and downstream signal transduction networks has provided exciting insights, which will enhance the use of R genes for disease control. Definition of conserved structural motifs in R proteins has facilitated the cloning of useful R genes, including several that are functional in multiple crop species and/or provide resistance to a relatively wide range of pathogens. Numerous signal transduction components in the defense network have been defined, and several are being exploited as switches by which resistance can be activated against diverse pathogens.     

植物抗病基因的进化

植物抗病基因在进化中形成了几种共有的进化形式。植物祖先抗病基因的复制创造了新基因座。基因间和基因内重组导致了变异,也导致了新特异性抗病基因的产生。另外,与特异性识别相关的富含亮氨酸重复区顺应于适应性选择。同样,类转座元件在抗病基因座中的插入加速了抗病基因的进化。随着抗病基因的进化,抗病反应也呈现出多样化,代表着植物与病原物动态进化的不同阶段。     

Two TIR:NB:LRR genes are required to specify resistance to Peronospora parasitica isolate Cala2 in Arabidopsis

Resistance responses that plants deploy in defence against pathogens are often triggered following a recognition event mediated by resistance (R) genes. The encoded R proteins usually contain a nucleotide-binding site (NB) and a leucine-rich repeat (LRR) domain. They are further classified into those that contain an N-terminal coiled coil (CC) motif or a Toll interleukin receptor (TIR) domain. Such R genes, when transferred into a susceptible plant of the same or closely related species, usually impart full resistance capability. We have used map-based cloning and mutation analysis to study the recognition of Peronospora parasitica (RPP)2 (At) locus in Arabidopsis accession Columbia (Col-0), which is a determinant of specific recognition of P. parasitica (At) isolate Cala2. Genetic mapping located RPP2 to a 200-kb interval on chromosome 4, which contained four adjacent TIR:NB:LRR genes. Mutational analysis revealed three classes of genes involved in specifying resistance to Cala2. One class, which resulted in pleiotropic effects on resistance to other P. parasitica (At) isolates, was unlinked to the RPP2 locus; this class included AtSGT1b. The other two classes were mapped within the interval and were specific to Cala2 resistance. Representatives of each of these classes were sequenced, and mutations were found in one or the other of two (RPP2A and RPP2B) of the four TIR:NB:LRR genes. RPP2A and RPP2B complemented their specific mutations, but failed to impart resistance when present alone, and it is concluded that both genes are essential determinants for isolate-specific recognition of Cala2. RPP2A has an unusual structure with a short LRR domain at the C-terminus, preceded by two potential but incomplete TIR:NB domains. In addition, the RPP2A LRR domain lacks conserved motifs found in all but three other TIR:NB:LRR class proteins. In contrast, RPP2B has a complete TIR:NB:LRR structure. It is concluded that RPP2A and RPP2B cooperate to specify Cala2 resistance by providing recognition or signalling functions lacked by either partner protein.     

Molecular communication between host plant and the fungal tomato pathogenCladosporium fulvum

Host genotype specificity in interactions between biotrophic fungal pathogens and plants in most cases complies with the gene-for-gene model. Success or failure of infection is determined by absence or presence of complementary genes, avirulence and resistance genes, in the pathogen and the host plant, respectively. Resistance, expressed by the induction of a hypersensitive response followed by other defence responses in the host, is envisaged to be based on recognition of the pathogen, mediated through direct interaction between products of avirulence genes of the pathogen (the so-called race-specific elicitors) and receptors in the host plant, the putative products of resistance genes. The interaction between the biothrophic fungusCladosporium fulvum and its only host tomato is a model system to study fungus-plant gene-for-gene relationships. Here we report on isolation, characterization and biological function of putative pathogenicity factors ECP1 and ECP2 and the race-specific elicitors AVR4 and AVR9 ofC. fulvum and cloning and regulation of their encoding genes. Disruption ofecp1 andecp2 genes has no clear effect on pathogenicity ofC. fulvum. Disruption of theavr9 gene, which codes for the race-specific 28 amino acid AVR9 elicitor, in wild type avirulent races, leads to virulence on tomato genotypes carrying the complementary resistance geneCf9. The avirulence geneavr4 encodes a 105 amino acid race-specific elicitor. A single basepair change in the avirulence geneavr4 leads to virulence on tomato genotypes carrying theCf4 resistance gene.