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1.
植物病原菌产生蛋白水解酶被认为是病原菌侵染寄主的一个重要策略。作者对水稻纹枯病菌(Rhizoctonia solani Kühn)产生的胞外蛋白酶的特性进行了研究。结果表明,水稻纹枯病菌产生一个分子量约为49.5ku的胞外蛋白酶。该酶对胰蛋白酶专一性底物Benz-Phe-Val-Arg-NA有较高的活性,并能从羧基端降解Arg和Lys。胰蛋白酶抑制剂对该酶活性有较强的抑制作用,表明该蛋白酶是一种类胰蛋白酶。在离体试验下,该蛋白酶可以降解水稻细胞壁蛋白并释放羟脯氨酸。SDS-PAGE电泳分析显示,至少有4个多肽被降解,分子量范围分别在14.4～20.1、20.1～31ku和31～43ku范围内。说明该胞外蛋白酶具有降解水稻细胞壁的能力,因此,该蛋白酶可能在水稻纹枯病菌的致病过程中起重要作用。 相似文献
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小麦纹枯病菌(Rhizoctonia cerealis Vander Hoeven)在以小麦细胞壁为唯一碳氮源的液体培养基中可产生多种蛋白酶,培养液经硫酸铵沉淀,SP-Sepharose Fast Flow离子交换层析,Phenyl(high-sub)-Sepharose Fast Flow疏水层析。Sephadex G-75分子筛层析,分离纯化到分子量约40kD的一种蛋白酶。该酶能水解胰蛋白酶专一性底物Benz-Phe-Val-Arg-NA,对凝乳弹性蛋白酶底物Sue-Ala-Ala-Pro-Phe-NA和枯草杆菌蛋白酶底物CBZ-Gly-Gly-Leu-NA无活性,能被胰蛋白酶抑制剂aprotinin、leupeptin和soybean trypsin inhibitor强烈抑制,丝氨酸蛋白酶抑制剂PMSF能部分抑制酶的活性。表明该蛋白酶是丝氨酸蛋白酶中的一种类胰蛋白酶。 相似文献
3.
玉米茎腐病菌产生的细胞壁降解酶的致病作用 总被引：23，自引：1，他引：22
本文通过酶活分析和超微结构观察证明了玉米茎腐病菌能产生一系列细胞壁降解酶(CWDE),并能明显浸解胚根组织。浸解的组织发生质壁分离及原生质外流。随细胞壁降解酶浓度增加,酶浸解能力增强,两者呈显著的正相关。相比较而言,玉米赤霉病Fusarium graminearum在病株内产生的细胞壁降解酶的浸解能力比玉米腐霉茎腐病Pythium aphanidermatum强。2种病菌活体外产生的果胶酶与纤维素酶(Cx)存在明显的协同作用。毒素不能提高细胞壁降解酶的浸解能力,反而有一定的抑制作用。 相似文献
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5.
温湿度调控对番茄灰霉病菌产生的细胞壁降解酶的影响 总被引：6，自引：0，他引：6
番茄灰霉病菌在致病过程中能够产生4种细胞壁降解酶,以PMG酶活性最高,其次是β-葡萄糖苷酶和PG酶,Cx最少。灰霉病菌在不同温度下侵染番茄叶片时产生的致病酶活性不同,4种酶在20℃时表现了最高的活性,15℃次之,当温度达到25℃时,各种酶的活性都急剧下降,随着温度的再升高,酶活更低。随着湿度的增高,病菌产生的细胞壁降解酶的活性也增加,当相对湿度达到90%以上时,4种酶的活性也达到最高。温湿度对番茄灰霉病菌产生细胞壁降解酶的影响趋势,与其对发病的影响趋势是一致的。 相似文献
6.
玉米茎腐病菌产生的细胞壁降解酶种类及其活性分析 总被引：20，自引：0，他引：20
玉米茎腐病菌Pythium aphanidermatum、Fusarium graminearum能够产生一系列细胞壁降解酶,即果胶甲基酯酶(PE)、多聚半乳糖醛酸酶(PG)、果胶甲基半乳糖醛酸酶(PMG)、多聚半乳糖醛酸反式消除酶(PGTE)、果胶甲基反式消除酶(PMTE)和纤维素酶(Cx)。Fusarium graminearum产生的细胞壁降解酶活性明显高于Pythium aphanidermatum。病菌在活体内和活体外产生的细胞壁降解酶活性明显不同。酶动力学研究表明:Fusarium graminearum产生的各种细胞壁降解酶均有特定的最适反应条件。水解酶类的PG、PMG、Cx最大酶活的pH分别为6.0、5.0、6.0,温度分别为50、40、50℃;裂解酶类的PGTE和PMTE最大酶活的pH均为9.0,温度分别为30、20℃,与其它病菌产生的细胞壁降解酶的特性基本相同。 相似文献
7.
番茄芝麻斑病原菌产生的细胞壁降解酶种类及其活性变化 总被引：3，自引：0，他引：3
番茄芝麻斑点病原菌能够产生一系列细胞壁降解酶,即果胶甲基酯酶(PE)、多聚半乳糖醛酸酶(PG)、果胶甲基半乳糖醛酸酶(PMG)、多聚半乳糖醛酸反式消除酶(PGTE)、果胶甲基反式消除酶(PMTE)和纤维素酶(Cx)。酶动力学研究表明:产生的各种细胞壁降解酶均有特定的最适反应条件。水解酶类的PG、PMG、Cx最大酶活的pH均为5.0,温度均为50℃;裂解酶类的PGTE和PMTE最大酶活的pH均为9.0,温度均为30℃,与其他病菌产生的细胞壁降解酶的特性基本相同。在活体外,随着培养天数的增加PG、PMG、PGTE、PMGE、Cx的活性都大幅度增加,但所有酶活性都明显低于活体内的酶活性。 相似文献
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为了明确芒果炭疽病病菌(Colletotrichum gloeosporioides)漆酶与该菌侵染相关致病因子之间的关系,本文以愈创木酚为底物筛选获得的高产漆酶菌株A2为材料,测定了漆酶粗提液对芒果炭疽病病菌致病力的影响,并通过半定量RT-PCR法,分析了在不同侵染时段漆酶基因lac1和其他酶基因的表达。结果表明,漆酶粗提液单独不能致病,但能促进该病原菌在寄主中的扩展;在侵染过程中,lac1表达与黑色素合成酶(thd和scd)基因表达相关性更高,与细胞壁降解酶(ecg和pel)基因表达也有一定相关性,但不及前者高。芒果胶孢炭疽菌分泌的漆酶有助于该菌在侵染芒果过程中的扩展,且对侵染相关致病因子黑色素合成酶和细胞壁降解酶基因的表达有一定影响。初步推测,漆酶lac1除了能通过降解芒果组织中的酚类物质、促进病原菌的扩展之外,也可以通过干扰与致病力相关的黑色素合成和细胞壁降解酶的产生来影响芒果胶孢炭疽菌致病力。 相似文献
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两种季铵盐阳离子表面活性剂对番茄灰霉病菌的毒理效应 总被引：2，自引：1，他引：1
为探讨季铵盐阳离子表面活性剂1227和C8-10对番茄灰霉病菌Botrytis cinerea的毒理效应,运用电子显微镜、氧电极、紫外-可见分光光度计等测定了1227、C8-10对番茄灰霉病菌分生孢子萌发、细胞壁结构、呼吸作用和三种细胞壁降解酶的影响,并在番茄叶片活体上接种经药剂处理的番茄灰霉病菌,观察药剂对其侵染活性的影响.结果显示,两种季铵盐阳离子表面活性剂5μg/mL处理可使分生孢子芽管变短变粗、基部明显膨大,显著抑制聚甲基半乳糖醛酸酶、多聚半乳糖醛酸酶和羧甲基纤维素酶的活性,并可显著影响番茄灰霉病菌在番茄活体叶片上的侵染活性;40μg/mL处理可导致菌体细胞壁表面出现多处破损、细胞内含物聚集等现象;100μg/mL处理显著抑制分生孢子的呼吸作用;250μg/mL和1 000μg/mL处理对病原菌菌丝呼吸产生破坏作用.说明两种季铵盐杀菌剂高剂量能够破坏菌体细胞壁结构,导致番茄灰霉病菌呼吸完全停止和死亡,而低剂量可以抑制细胞胞壁降解酶的活性,降低其致病力. 相似文献
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细胞壁降解酶在油茶炭疽病菌致病过程中的作用研究 总被引：1，自引：0，他引：1
为明确细胞壁降解酶在油茶炭疽病菌致病过程中的作用,本文研究了活体内外炭疽病菌产生的细胞壁降解酶活性及其对叶片的降解情况。结果表明,活体外以羧甲基纤维素钠(CMCNa)为诱导底物,羧甲基纤维素酶(Cx酶)和漆酶活性最高;以柑橘果胶为诱导底物,果胶酶活性最高;以油茶叶为诱导底物,纤维素酶、果胶酶和漆酶可产生较高活力;并且经5种诱导物诱导的酶液对叶片均有降解作用。发病叶片的各部位,以病健交界处细胞壁降解酶活性最高。接种4d后开始发病,其细胞壁降解酶活性迅速增强;6d后滤纸酶(FPA)、β-葡萄糖苷酶和漆酶活性达最大值,分别为4.53、7.44、1.21U/mg;而Cx酶和果胶酶在第8天时酶活性最高,分别为15.79和25.49U/mg;接种10~16d,酶活性比较稳定。上述结果表明,纤维素酶、果胶酶和漆酶在油茶炭疽病菌致病过程中起重要作用。 相似文献
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光镜和电镜观察表明,禾顶囊壳小麦变种(Gaeumannomyces graminis var.tritici,小麦全蚀病菌)对小麦种子根的侵染过程可分为侵入前、侵入表皮层、进入皮层和进入中柱等4个连续阶段。麦根接菌后在15℃下培养,48 h后侵入表皮层细胞,60 h后进入皮层,120 h后进入中柱。病原菌主要以侵染菌丝直接侵入表皮层,表皮细胞间隙和根毛基细胞是主要侵入部位,少数由附着枝侵入。菌丝穿透细胞壁有明显的酶解作用特征,菌丝先端前方胞壁上还产生电子密物质。皮层细胞是病原菌定殖和发展的主要场所,病原菌还能离解胞间层,形成胞外空间,特别有利于菌丝和菌丝束的扩展。在侵入位点的寄主细胞壁和质膜之间,形成多种形状的木质管,其数量与侵入菌丝的数目相对应,但木质管不能阻止菌丝进入细胞。菌丝进入中柱后,可阻塞导管和筛管。小麦细胞发生退行性病变,尤以细胞壁膨大崩坏和早期质壁分离最明显,细胞间隙还产生性质不明的黄色物质。 相似文献
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The infection process of Fusarium avenaceum on wheat spikes and the alteration of cell wall components in the infected host tissue were examined by means of electron microscopy and cytochemical labelling techniques following spray inoculation at growth stage (GS) 65 (mid-flowering). Macroconidia of the pathogen germinated with one to several germ-tubes 6–12 h after inoculation (hai) on host surfaces. The germ-tubes did not penetrate host tissues immediately, but extended and branched on the host surfaces. Hyphal growth on abaxial surfaces of the glume, lemma and palea was scanty 3–4 days after inoculation (dai) and no direct penetration of the outer surfaces of the spikelet was observed. Dense mycelial networks formed on the inner surfaces of the glume, lemma, palea and ovary 36–48 hai. Penetration of the host tissue occurred 36 hai by infection hyphae only on the adaxial surfaces of the glume, lemma, palea and upper part of ovary. The fungus penetrated the cuticle and hyphae extended subcuticularly or between the epidermal wall layers. The subcuticular growth phase was followed by penetration of the epidermal wall, and hyphae spread rapidly inter- and intracellularly in the glume, lemma, palea and ovary. During this necrotrophic colonization phase of the wheat spike, a series of alterations occurred in the host tissues, such as degeneration of cytoplasm and cell organelles, collapse of host cells and disintegration of host cell walls. Immunogold labelling techniques showed that cell walls of spike tissues contained reduced amounts of cellulose, xylan and pectin near intercellular hyphae or infection pegs compared to walls of healthy host tissues. These studies suggest that cell wall degrading enzymes produced by F. avenaceum facilitated rapid colonization of wheat spikes. The different penetration properties of abaxial and adaxial surfaces of the spikelet tissues as well as the two distinct colonization strategies of host tissues by F. avenaceum are discussed. The penetration and colonization behaviour of F. avenaceum in wheat spikelets resembled that of F. culmorum and F. graminearum, although mycotoxins produced by F. avenaceum differed from those of the latter two Fusarium species. 相似文献
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小麦穗组织中脱氧镰刀菌烯醇毒素的免疫细胞化学定位 总被引：5，自引：0，他引：5
采用免疫细胞化学技术对禾谷镰刀菌(Fusarium graminearum)在侵染小麦穗部过程中产生的脱氧镰刀菌烯醇毒素(deoxynivalenol,DON)进行了定位分析。在接种后24h,当菌丝在外稃、内稃的内侧表面扩展而尚未侵入寄主细胞前,病菌已分泌DON,并且DON已扩散到寄主组织内。在菌丝细胞内,DON主要被定位于细胞质、线粒体及细胞壁上;在寄主细胞中DON主要分布于细胞壁、叶绿体、细胞质和内质网上。在侵染初期(接种后2 d),菌丝仅能在寄主细胞间隙扩展,随寄主组织中DON浓度的升高,寄主细胞相应发生了一系列病理变化。随寄主细胞坏死(接种后3~4d),病菌进入坏死的寄主细胞。上述结果表明,DON在禾谷镰刀菌的侵染、致病和定殖过程中起着重要的作用。毒素标记结果表明病菌产生的毒素可通过穗轴微管束组织从侵染部位向上、向下转输,毒素向上的转输量明显高于向下转输 相似文献
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The fungal infection of emmer grain (Triticum dicoccum) with Fusarium graminearum and Fusarium culmorum was investigated at the level of the proteome. High‐resolution two‐dimensional gel electrophoresis and mass spectrometry were used to identify proteins that were differentially expressed in response to fungal infection of emmer. Moreover, the effects of natural field conditions at two locations on the carbon and nitrogen contents and the mycotoxin concentration of emmer grains were evaluated. Inoculation of emmer with a mixture of the two Fusarium species led to infection of the ears, with deoxynivalenol concentrations up to 10 mg kg?1 in the grain. Carbon concentration and crude protein content were not significantly changed, but 10 distinct proteins changed in abundance. Stress‐related proteins, such as a serine protease inhibitor, a thaumatin‐like protein that reduced fungal growth and the starch hydrolysis β‐amylase increased upon infection, whereas the stress‐related proteins peroxidase, peroxiredoxin, a starch‐synthesis protein (a glycosyltransferase) and a fungal cell wall degrading protein (a chitinase) decreased. Furthermore, levels of three storage proteins in emmer grains were affected by Fusarium infection: α‐gliadin decreased and two globulins increased upon infection. 相似文献
15.
Histopathological assessment of infection by the crown rot pathogen Fusarium pseudograminearum in wheat seedling tissues was performed using fluorescence microscopy. The coleoptiles and leaf sheaths of four host cultivars of differing susceptibility were examined. Leaf sheaths were most frequently penetrated via stomata, indicated by initial lesions forming at the guard cells. Internally, cell wall penetration was facilitated by penetration structures which appeared as hyphal swellings or septate foot‐shaped appressoria. Colonization of leaf sheaths resulted in the re‐emergence of hyphae from stomata on both surfaces of the sheath. These hyphae are hypothesized to have two major roles; first as exploratory hyphae for colonization of new tissues, and secondly as sites of profuse conidial production. The formation of conidia on the leaf sheath surface was only recorded on the most susceptible bread wheat genotype. No other major differences in host–pathogen interactions were observed among these cultivars. Almost all cell types in the leaf sheath tissues were extensively colonized, except for the vascular bundles and silica cells. This investigation provides the first comprehensive assessment of F. pseudograminearum infection structures and growth patterns during the infection of wheat seedlings. 相似文献
16.
为探讨小菜蛾Plutella xylostella对病原真菌入侵的免疫防御机制,利用抑制消减杂交技术构建蝉拟青霉Paecilomyces cicadae侵染的小菜蛾幼虫的抑制性差减文库,并对文库进行鉴定。ESTs序列聚类分析共获得412个独立基因。通过同源比对,28.24%ESTs为未知功能基因,71.76%ESTs与已知功能基因序列相似,包括免疫相关、金属离子结合和加工、核酸和蛋白代谢及加工、细胞信号、细胞结构、形状和流动、能量代谢、胁迫和解毒以及其它基因。鉴定出39个可能参与小菜蛾免疫蝉拟青霉侵染的免疫基因,包括：识别分子、蛋白酶及蛋白酶抑制剂、效应因子及其它免疫基因。qRT-PCR结果表明肽聚糖识别蛋白、器官芽生长因子、葛佬素、溶菌酶、酚氧化酶原激活蛋白酶3以及转铁蛋白基因均可诱导表达。免疫相关基因在不同微生物诱导下的表达模式存在不同。结果表明当病原真菌蝉拟青霉侵染寄主小菜蛾后,小菜蛾体内发生了一系列的复杂反应,小菜蛾抵御病原真菌侵染是一个多途径共同作用的复杂过程。该研究为进一步研究小菜蛾抵御病原真菌入侵的分子机制提供基础信息。 相似文献
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赤霉病是我国小麦上的重要病害,品种抗病性利用是控制病害发生的重要措施,明确小麦抗赤霉病资源的抗性类型,有利于小麦抗赤霉病育种。2003年和2004年对9个常用抗源在穗期进行单花滴注和喷雾接种,研究其抗侵染和抗扩展性,并对病穗中的脱氧雪腐镰刀菌烯醇(DON)的含量进行分析。结果表明,望水白和苏麦3号具较好的抗侵染和抗扩展能力,其中望水白的抗扩展性最好;感染赤霉病后,DON在5个抗源穗组织中的含量差异显著,DON在望水白和繁60096穗组织中积累量明显比在苏麦3号、延岗坊主和翻山小麦低。通过对望水白/安农8455遗传群体两年的病小穗率和病穗中DON毒素含量的比较,发现二者具有一定的相关性,且受环境影响比较大。 相似文献
18.
Wanyoike Mary Wanjiru Kang Zhensheng Heinrich Buchenauer 《European journal of plant pathology / European Foundation for Plant Pathology》2002,108(8):803-810
Cytological studies were carried out to elucidate the importance of cell wall degrading enzymes (CWDE) during infection of wheat spikes by Fusarium graminearum. Scanning electron micrographs revealed that at 6–24 hours after inoculation (hai) of single spikelets with macroconidia of F. graminearum, the fungus germinated by forming several germ tubes and developed a dense hyphal network in the cavity of the spikelet. At 24–36hai, the fungus formed infection hyphae which invaded the ovary and inner surface of the lemma and palea. Transmission electron microscopical studies revealed that the fungus extended inter- and intracellularly in the ovary, lemma and rachis and caused considerable damage and alterations to the host cell walls. In different tissues of healthy and F. graminearum-infected wheat spikes the cell wall components cellulose, xylan and pectin were localized by means of enzyme-gold and immuno-gold labelling techniques. Localization of cellulose, xylan and pectin showed that host cell walls which were in direct contact with the pathogen surface had reduced gold labelling compared to considerable higher labelling densities of walls distant from the pathogen–host interface or in non-colonized tissues. The reduced gold labelling densities in the infected host cell walls indicate that these polysaccharide degrading enzymes might be important pathogenicity factors of F. graminearum during infection of wheat spikes. The results revealed that, infection and colonization of wheat spikes by F. graminearum and reactions of infected host tissue were similar to those reported for F. culmorum. 相似文献