转录因子AtWRKY28在拟南芥与甘蓝链格孢菌(Alternaria brassicicola)亲和性互作中的功能分析

WRKY是植物特有的一类转录因子,是一个超级大家族,与植物多种逆境胁迫应答密切相关,在植物防御病原菌的侵染中发挥十分重要的作用。拟南芥转录因子WRKY28属于WRKY家族的成员,包括一个WRKYGQK核心序列和一个Cys2His2锌指型结构。为了鉴定拟南芥WRKY28的功能,构建了反义抑制表达株系和转基因过表达株系并进行抗病性鉴定。研究表明,反义抑制表达株系降低了对腐生型真菌甘蓝链格孢菌(Alternaria brassicicola)的抗性,而转基因过表达株系提高了对甘蓝链格孢菌的抗性。由此可见,转录因子WRKY28参与了拟南芥对甘蓝链格孢菌的免疫应答,是其抗病反应中的一个正调控因子。     

Pre- and postinvasion defenses both contribute to nonhost resistance in Arabidopsis

Nonhost resistance describes the immunity of an entire plant species against nonadapted pathogen species. We report that Arabidopsis PEN2 restricts pathogen entry of two ascomycete powdery mildew fungi that in nature colonize grass and pea species. The PEN2 glycosyl hydrolase localizes to peroxisomes and acts as a component of an inducible preinvasion resistance mechanism. Postinvasion fungal growth is blocked by a separate resistance layer requiring the EDS1-PAD4-SAG101 signaling complex, which is known to function in basal and resistance (R) gene-triggered immunity. Concurrent impairment of pre- and postinvasion resistance renders Arabidopsis a host for both nonadapted fungi.     

Broad-spectrum mildew resistance in Arabidopsis thaliana mediated by RPW8

Plant disease resistance (R) genes control the recognition of specific pathogens and activate subsequent defense responses. We show that the Arabidopsis thaliana locus RESISTANCE TO POWDERY MILDEW8 (RPW8) contains two naturally polymorphic, dominant R genes, RPW8.1 and RPW8.2, which individually control resistance to a broad range of powdery mildew pathogens. Although the predicted RPW8.1 and RPW8.2 proteins are different from the previously characterized R proteins, they induce localized, salicylic acid-dependent defenses similar to those induced by R genes that control specific resistance. Apparently, broad-spectrum resistance mediated by RPW8 uses the same mechanisms as specific resistance.     

葡萄MrRPV1蛋白LRR结构域的功能研究

[目的]明确葡萄霜霉病抗性蛋白(MrRPV1)LRR结构域对葡萄霜霉菌特异性识别的影响,为进一步研究葡萄与霜霉菌间的互作机理打下基础.[方法]将MrRPV1和白粉病抗性蛋白(MrRUN1)两个蛋白的LRR结构域互换,构建重组表达载体后转化至感霜霉病和白粉病的欧亚种西拉葡萄胚性愈伤组织中以获得转基因植株,并通过DNA、RNA分子水平检测和室内接种试验进行抗病性鉴定.[结果]共获得7个RGA10-8LRR阳性转基因(重组了MrRPV1蛋白LRR结构域)株系和6个RGA8-10LRR阳性转基因(重组了MrRUN蛋白LRR结构域)株系.在7个RGA10-8LRR阳性转基因株系中,仅RGA10-8-13阳性植株表现出与MrRPV1转基因苗S8-1一致的坏死斑点,肉眼观察不到白色霜霉状物,但7个阳性转基因系均未表现出明显的白粉病抗性.在6个RGA8-10LRR阳性转基因株系中,RGA8-10-3和RGA8-10-28两个株系表现出对白粉病的抗性,有35%左右的细胞表现出细胞程序性坏死(PCD);RGA8-10LRR转基因苗均不抗霜霉病.[结论]MrRPV1的LRR结构域可能介导对葡萄霜霉菌的特异性识别,而MrRUN1的LRR结构域可能介导对白粉病菌的特异性识别.     

Cleavage of Arabidopsis PBS1 by a bacterial type III effector

Plant disease-resistance (R) proteins are thought to function as receptors for ligands produced directly or indirectly by pathogen avirulence (Avr) proteins. The biochemical functions of most Avr proteins are unknown, and the mechanisms by which they activate R proteins have not been determined. In Arabidopsis, resistance to Pseudomonas syringae strains expressing AvrPphB requires RPS5, a member of the class of R proteins that have a predicted nucleotide-binding site and leucine-rich repeats, and PBS1, a protein kinase. AvrPphB was found to proteolytically cleave PBS1, and this cleavage was required for RPS5-mediated resistance, which indicates that AvrPphB is detected indirectly via its enzymatic activity.     

黄瓜基因组中抗霜霉病候选基因的生物信息学及其表达特异性

【目的】寻找黄瓜抗霜霉病基因,研究黄瓜抗霜霉病机制。【方法】利用拟南芥和甜瓜抗霜霉病蛋白质序列,搜索黄瓜基因组数据库;通过生物信息学分析候选基因特征,以黄瓜抗霜霉病自交系M801-3-1为试材,分析候选基因在叶片组织中的表达状态。【结果】共获得187个黄瓜抗霜霉病候选基因;确定了候选基因染色体位置和排列特点、序列相似性特征以及系统进化关系;大部分黄瓜R基因在未接种霜霉病菌的抗病自交系和感病叶片组织中都有一定的表达量,在接种霜霉病菌后在抗病自交系M801-3-1中Csa001907和Csa002921表达量下调,感病自交系M302-3 中Csa001907和Csa002921变化不明显。【结论】在黄瓜基因组中存在185个与拟南芥抗霜霉病基因同源的R基因,2个与甜瓜的抗霜霉病基因At1和At2同源的eR基因,通过聚类分析这些基因可以分为6类,初步认定Csa001907和Csa002921为黄瓜抗霜霉病的R基因,这2个基因在叶片组织中有一定表达量,其表达量减少可诱发抗病黄瓜品种的抗病反应。     

3个小麦新种质的选育与鉴定

针对目前山西省小麦抗病性差的现状,以携带有抗病基因的小麦-簇毛麦6VS/6AL易位系92R137和92R149为亲本,与感病小麦品种忻96-74、矮多1号、黑粒小麦76杂交,进行抗病、丰产、优质和矮秆基因的重组,选育出3个抗病新种质G10,S06和S25,并综合应用农艺性状、细胞学、分子标记等方法对其进行鉴定。结果表明,G10为半矮秆、特大穗,对小麦白粉病和条锈病免疫;S06为矮秆,对白粉病免疫;S25具有5+10优质亚基、紫粒,对白粉病和条锈病免疫。G10,S06和S25是小麦抗病、优质育种的新种质材料。     

山东黄瓜主要种质资源多抗性鉴定

利用单一接种法,分别接种鉴定了24个黄瓜种质材料对黄瓜白粉病、霜霉病、褐斑病和枯萎病的抗性。结果表明,有3份材料高抗白粉病,9份材料抗白粉病;2份材料高抗霜霉病,8份材料抗霜霉病;1份材料高抗褐斑病,12份材料达到抗病级;在枯萎病的接种试验中,没有发现高抗和抗病材料,仅得到了8份中抗材料。综合4种病原菌的接种结果,发现材料H、U、B、E、L、P对4种病害都达到中抗以上水平;材料V除了不抗枯萎病对其他3种叶部病害都达到高抗水平;材料H高抗霜霉病;材料R与材料J对白粉病达高抗水平。     

Pathogen effectors target Arabidopsis EDS1 and alter its interactions with immune regulators

Plant resistance proteins detect the presence of specific pathogen effectors and initiate effector-triggered immunity. Few immune regulators downstream of resistance proteins have been identified, none of which are known virulence targets of effectors. We show that Arabidopsis ENHANCED DISEASE SUSCEPTIBILITY1 (EDS1), a positive regulator of basal resistance and of effector-triggered immunity specifically mediated by Toll-interleukin-1 receptor-nucleotide binding-leucine-rich repeat (TIR-NB-LRR) resistance proteins, forms protein complexes with the TIR-NB-LRR disease resistance proteins RPS4 and RPS6 and with the negative immune regulator SRFR1 at a cytoplasmic membrane. Further, the cognate bacterial effectors AvrRps4 and HopA1 disrupt these EDS1 complexes. Tight association of EDS1 with TIR-NB-LRR-mediated immunity may therefore derive mainly from being guarded by TIR-NB-LRR proteins, and activation of this branch of effector-triggered immunity may directly connect to the basal resistance signaling pathway via EDS1.     

NBS-LRR Proteins and Their Partners:Molecular Switches of Plant Defense

Specificity of the plant innate immune system is often conferred by resistance（R）proteins.Most plant disease resistance （R）proteins contain a series of leucine-rich repeats（LRRs）,a nucleotide-binding site（NBS）,and a putative amino-terminal signaling domain.They are termed NBS-LRR proteins.The LRRs are mainly involved in recognition,and the amino-terminal domain determines signaling specificity,whereas the NBS domain presumably functions as a molecular switch.During the past years,the most important discoveries are the role of partners in NBS-LRR gene mediated defenses,mounting support for the so-called＂guard hypothesis＂of R gene function,and providing evidence for intramolecular interactions and intermolecular interactions within NBS- LRR proteins as a mode of signaling regulation.The outcome of these interactions determines whether a plant activates its defense responses.     

木薯25个WRKY家族转录因子在生物胁迫下的表达分析

根据拟南芥和水稻的WRKY基因和木薯基因组序列,利用生物信息学方法预测木薯Me WRKY转录因子家族成员,并对其进行系统进化关系和保守结构域的分析。通过病原菌接种处理,分析不同Me WRKY转录因子的表达差异。结果显示,木薯共编码25个与抗病相关WRKY蛋白,在病原菌胁迫条件下,其中16个Me WRKY基因的表达受到显著影响,表明这些WRKY蛋白可能参与木薯的防御应答反应。     

参与植物防御反应的LRR型蛋白结构与功能

植物含有多种富含亮氨酸重复（LRRs）结构的蛋白质,它们在植物生长、发育和抗病反应等方面发挥着重要作用。综述了这类具有LRRs结构蛋白质家族的结构特征及其参与植物防御反应的功能。参与植物防御反应LRR型蛋白质家族包括：抗病基因编码蛋白质、类受体蛋白激酶、多聚半乳糖醛酸酶抑制蛋白和伸展蛋白家族。这四大蛋白质家族成员主要通过LRRs结构识别并结合病原物蛋白质,参与抗病信号传递,诱导植物防卫基因的表达,使植物获得系统抗性。其中LRRs序列中氨基酸的不同和单位重复数目的差异决定了蛋白识别的特异性和结合能力。     

Genomic insights into the immune system of the sea urchin

Comparative analysis of the sea urchin genome has broad implications for the primitive state of deuterostome host defense and the genetic underpinnings of immunity in vertebrates. The sea urchin has an unprecedented complexity of innate immune recognition receptors relative to other animal species yet characterized. These receptor genes include a vast repertoire of 222 Toll-like receptors, a superfamily of more than 200 NACHT domain-leucine-rich repeat proteins (similar to nucleotide-binding and oligomerization domain (NOD) and NALP proteins of vertebrates), and a large family of scavenger receptor cysteine-rich proteins. More typical numbers of genes encode other immune recognition factors. Homologs of important immune and hematopoietic regulators, many of which have previously been identified only from chordates, as well as genes that are critical in adaptive immunity of jawed vertebrates, also are present. The findings serve to underscore the dynamic utilization of receptors and the complexity of immune recognition that may be basal for deuterostomes and predicts features of the ancestral bilaterian form.     

SA、MeJA、ACC和苹果轮纹病病原菌诱导湖北海棠MhWRKY1基因的表达

 【目的】从湖北海棠叶片中克隆MhWRKY1转录因子的全长cDNA序列,分析该基因在各种组织中(叶、茎、根)的表达特性,并分析SA、MeJA、ACC在叶、茎、根中诱导MhWRKY1基因的表达模式以及苹果轮纹病病原菌诱导条件下湖北海棠叶片中该基因的表达特性。【方法】利用电子克隆技术和RT-PCR验证相结合的方法,从SA诱导的湖北海棠全长cDNA文库中,克隆MhWRKY1转录因子的全长序列;利用生物信息学的方法对其进行结构和功能的初步分析;利用实时荧光定量RT-PCR技术分析该基因在不同组织中的表达以及在SA、MeJA、ACC和苹果轮纹病病原菌诱导下的表达特性。【结果】克隆了MhWRKY1基因的全长cDNA序列为1 338 bp,GenBank数据库登录号为FJ598139。生物信息学分析表明,该基因最大开放阅读框为993 bp,编码330个氨基酸。推导的氨基酸序列与杨树WRKY26、杨树WRKY20、大豆WRKY,马铃薯WRKY2、烟草WRKY、拟南芥WRKY7、水稻WRKY53的同源性分别为68%、68%、66%、60%,59%,49%和43%。该转录因子含有1个WRKY结构域,其N端含有1个WRKYGQK结构域,C端含有1个C2H2锌脂结构,属于第Ⅱ类型的转录因子。表达分析结果表明,MhWRKY1在叶和茎中的表达量较大,分别是根的4.81和3.75倍。在湖北海棠叶、茎、根中,SA、MeJA、ACC都可以诱导该基因的表达。另外,在所研究的72 h内,苹果轮纹病病原菌可以诱导该基因表达,且表达量在12 h达到最大,是未处理之前的9倍左右。【结论】MhWRKY1转录因子可能参与SA、MeJA和ET介导的植物抗病防卫反应的基本信号通路,并且参与对苹果轮纹病病原菌引起的防卫反应,在湖北海棠的的抗病过程中可能起着非常重要的作用。     

茉莉酸和真菌病原诱导的水稻WRKY30转录因子基因的分离及表达特征

 【目的】分离水稻WRKY30,为阐明WRKY转录因子在水稻抗病反应中的调节作用提供依据,为抗病性的合理利用和品种遗传改良提供基因材料。【方法】采用RT-PCR获得包含最大开放阅读框（ORF）的WRKY30 cDNA序列,运用生物信息学手段进行序列分析,通过Southern杂交确定基因在基因组中的拷贝数,利用Northern分析或RT-PCR方法,研究基因的器官特异性表达和病原、激素以及非生物胁迫诱导表达的特征。【结果】 WRKY30 包含一个长2 022 bp的完整ORF,编码674个氨基酸,具有1个核定位信号和2个典型的WRKY保守结构域,锌指纹组成为C2H2,归类于WRKY第一组,与大麦、高粱和短柄草等单子叶植物WRKY的氨基酸序列同一性较高。WRKY30在基因组中以单拷贝存在,在茎、叶和颖果中表达丰度最高,根和穗中次之,花中最低。WRKY30受稻瘟菌和纹枯菌快速诱导。抗病相关信号分子水杨酸（SA）和茉莉酸甲酯（MeJA）也能诱导该基因表达迅速上调,而乙烯（ET）对其表达无明显影响,脱落酸抑制其表达。除伤引起WRKY30表达上调外,其它非生物胁迫如冷、高盐和干旱对其表达无明显影响。【结论】WRKY30可能参与水稻对稻瘟菌和纹枯菌防御反应的调控,这种调节作用依赖于SA和JA介导的信号途径,而与ET信号途径无关。     

黑麦Imperial与KingⅡ中白粉病抗性基因的染色体定位

[目的]将黑麦Imperial及KingⅡ所带的白粉病抗性基因定位在染色体上,为其在小麦育种中的应用奠定基础。[方法]以2套小麦-黑麦附加系(中国春×Imperial和Holdfast×KingⅡ)为研究材料,研究黑麦Imperial及KingⅡ中白粉病抗性基因的染色体位置。[结果]中国春×Imperial的附加系6R和Holdfast×KingⅡ的附加系3R和6R对白粉病具有免疫力。这说明黑麦Imperial的白粉病抗性基因位于6R染色体上,而黑麦KingⅡ中白粉病抗性基因位于3R和6R染色体上。黑麦KingⅡ的3R染色体上的白粉病抗性基因可能是新基因,为小麦育种提供了新的白粉病抗原。这说明3R和6R染色体上的白粉病抗性基因可在小麦中表达。[结论]该研究为将黑麦白粉病抗性基因导入小麦提供了依据。     

5种植物免疫诱导剂在黄瓜上应用效果

[目的]研究5种植物免疫诱导剂对黄瓜光合生理变化和抗病性的影响,为植物免疫诱导剂在黄瓜上应用提供科学依据.[方法]采用盆栽试验,测定5种植物免疫诱导剂对黄瓜光合生理变化、抗病性和防御酶活性的影响.[结果]那氏齐齐发诱导剂可提高叶绿素含量、促进黄瓜植株花期提前,产量相比对照提高3.5倍;植物免疫诱抗蛋白提高黄瓜植株光合性...     

青花菜转录因子基因BoWRKY3的克隆与表达分析

以青花菜为材料,从叶片中分离WRKY转录因子基因BoWRKY3(登录号:JN120758),并进行序列分析;利用逆转录聚合酶链式反应(RT‐PCR)对霜霉菌侵染前后叶片BoWRKY3的表达模式进行分析.测序结果表明:BoWRKY3的基因组DNA全长为1136bp,有3个内含子,长度分别为81、101和96bp;编码区全长为858bp,编码285个氨基酸,具WRKYGQK保守区和CX5CX23HX1H锌指结构.RT‐PCR结果表明,BoWRKY3的表达受霜霉菌诱导,表达量在接种6~36h时最高,暗示该基因与青花菜霜霉病抗性相关.序列比对结果表明,BoWRKY3与十字花科同源基因的差异最小,而与禾本科作物的序列差异最大,关系最远.     

A conserved p38 MAP kinase pathway in Caenorhabditis elegans innate immunity

A genetic screen for Caenorhabditis elegans mutants with enhanced susceptibility to killing by Pseudomonas aeruginosa led to the identification of two genes required for pathogen resistance: sek-1, which encodes a mitogen-activated protein (MAP) kinase kinase, and nsy-1, which encodes a MAP kinase kinase kinase. RNA interference assays and biochemical analysis established that a p38 ortholog, pmk-1, functions as the downstream MAP kinase required for pathogen defense. These data suggest that this MAP kinase signaling cassette represents an ancient feature of innate immune responses in evolutionarily diverse species.     

油菜菌核病相关的WRKY转录因子研究进展

介绍了WRKY转录因子的结构特点,综述了WRKY转录因子在植物防卫反应中的作用以及在油菜菌核病防卫调控机制中的研究,在此基础上,提出了研究WRKY转录因子可能为油菜菌核病防治及抗病育种提供新的思路。