小麦条锈菌吸器母细胞超微结构的研究

本文利用电镜技术研究了小麦条锈菌吸器母细胞的超微特征。观察发现,小麦条锈菌吸器母细胞大都位于胞间菌丝的顶部,但也可形成于两菌丝细胞间,同一菌丝细胞上可着生多个吸器母细胞,吸器母细胞的多核现象较为普遍。吸器母细胞壁由６层组成,而其隔膜可分辩为４层。细胞化学染色结果表明隔膜突中含有多糖类物质,吸器母细胞隔膜突的出现和消失与吸器的发育密切相关。本文结果表明小麦条锈菌吸器母细胞的一些超微特征明显不同于其它     

种衣剂17号包衣对小麦条锈菌发育影响的超微结构研究

本研究采用电镜种衣剂１７号泽小麦条锈菌发育的影响。观察结果表明,该种衣剂引病菌和寄主细胞内发生了一系列变化。病菌菌丝和吸器内脂肪粒和液泡明显增加;菌丝壁和吸器壁呈不规则加厚;菌丝分枝处玩隔膜产生或隔膜畸形;有的吸器母细胞产生的畸形人侵栓,大都不能穿管寄主细胞壁,初生吸器外音质同内沉积有染色较深的物质,次生吸器可产生多个不规则分枝,但不能扩张膨大,菌丝外渗的物质可能引起寄主细胞的坏死;大多数受侵寄主     

三裂叶豚草锈菌侵染寄主叶片的显微和超微观察

【背景】三裂叶豚草是我国重要的外来入侵植物之一,其传播速度快,已给我国造成巨大的经济损失。近年来发现的三裂叶豚草锈菌是一种对其具有生物防治潜力的病原菌。【方法】本文利用显微技术研究了三裂叶豚草锈菌的侵染过程及其对寄主结构的影响。【结果】三裂叶豚草锈菌菌丝可从多处侵入同一个叶肉细胞,胞间菌丝与叶肉细胞相接触可使部分细胞壁增厚。锈菌侵染使三裂叶豚草叶脉末梢导管分枝增多,造成三裂叶豚草水分代谢失调;叶片细胞内膜系统破碎化,细胞器结构受到不同程度的破坏,导致细胞内膜系统紊乱,细胞器结构稳定性降低。【结论与意义】豚草锈菌侵染破坏了三裂叶豚草叶片的细胞结构。本研究为深入研究豚草锈菌的致病机理奠定了基础。     

小麦条锈菌吸器超微结构和细胞化学的研究

本文应用电镜技术和细胞化学方法,对小麦条锈菌吸器和入侵点的超微结构进行了研究。小麦条锈菌吸器由呈管状的颈部和顶端膨大的吸器体组成,颈部壁和吸器体壁相互连贯,均为两层,并且含有多糖物质。在颈部中段存在有一染色较深的颈环结构。观察发现吸器中的多核现象极为普遍。细胞化学染色结果表明:在吸器外间质内分布有多糖物质;经蛋白消酶解处理后,吸器外间质中可观察到染色较深的纤丝状物质。在入侵点部位,吸器母细胞壁因局部增厚而呈凸镜状,入侵栓壁由内、外两层构成,这两层分别与吸器母细胞壁的第六层和第五层相连接。本研究还观察到同一入侵点产生两个入侵栓的现象。     

小麦条锈菌吸器母细胞超微结构的研究

本文利唱电镜技术研究了小麦条锈菌吸器母细胞的超微特征。观察发现,小麦条锈菌吸器母细胞大部位于胞间菌丝的顶部,但也可形成于两菌丝细胞间,同一菌丝细胞上可着生多个吸器母细胞,吸器母细胞的多核现象较为普遍。吸器母细胞壁由6层组成,而其隔膜可分辩为4层。细泡化学染色结果表明隔膜突中含有多糖类物质,吸器母细胞隔膜突的出现和消失与吸器的发育密切相关。本文结果表明小麦条锈菌吸器母细胞的一些超微特征明显不同于其它锈菌。     

种衣剂17号包衣对小麦条锈菌发育影响的超微结构研究

本研究采用电镜技术研究了种衣剂17号对小麦条锈菌发育的影响。观察结果表明,该种衣剂引起病菌和寄主细胞内发生了一系列变化。病菌菌丝和吸器内脂肪粒和液泡明显增加;菌丝壁和吸器壁呈不规则加厚;菌丝分枝处无隔膜产生或隔膜畸形;有的吸器母细胞产生的畸形入侵栓,大都不能穿透寄主细胞壁,初生吸器外间质内沉积有染色较深的物质,次生吸器可产生多个不规则分枝,但不能扩张膨大;菌丝外渗的物质可能引起寄主细胞的坏死;大多数受侵寄主细胞可分泌形成较大的胼胝质,有时寄主细胞分泌的物质可将吸器体完全包围起来。上述结果表明,种衣剂17号不仅可直接作用于条锈菌,而且也可通过影响寄主而间接地影响病菌。     

光强对人参叶片显微和超微结构的影响

在５％、２０％和３５％３种透光率棚下,人参叶片的比叶重、叶厚度,单位叶面积的叶肉细胞数目,叶肉细胞表面积和细胞层数随着光强的增加而增加,５０％透光率棚下叶片开始减少,５％和２０％透光率棚下叶片叶肉细胞主要呈横向排列,胞间隙较大,３５％透光率棚下叶片叶肉细胞排列致密,胞间隙变小,靠近上一有皮形成一层呈纵向排列的栅栏状细胞;５０５透光率棚下叶片叶肉细胞为大而圆形,胞间隙变大,排列也不规则。５％透光率棚     

白粉菌侵染对小麦叶片显微及超微结构的影响白粉菌侵染对小麦叶片显微及超微结构的影响

通过半薄及超薄切片,比较了正常和受白粉菌感染的小麦叶片细胞的显微及超微结构的差异。观察结果发现(1)受感染小麦叶肉细胞的细胞壁上局部沉积大量团状电子致密颗粒;(2)叶绿体形状由原来的椭圆形转变成圆形,叶绿体膜破裂,类囊体膨大,基粒片层排列疏松,同时,叶绿体内嗜锇性颗粒数量增加;(3)线粒体膜解体,内含物分散到了细胞质中     

小麦条锈菌胞间菌丝的超微结构和细胞化学研究

本文应用电镜技术和细胞化学方法,对小麦条锈菌寄主胞间菌丝的超微结构进行了研究。观察发现:小麦条锈菌胞间菌丝有两种类型,即具隔膜菌丝和无隔膜菌丝。在胞间菌丝中,多核现象极为普遍。常规染色和细胞化学染色结果表明:胞间菌丝的细胞壁由四层组成,隔膜由三层构成,细胞壁的内层与隔膜的外层相连,细胞壁和隔膜中含有蛋白质和多糖物质。隔膜的发育可分三个阶段,即隔膜突的形成,隔膜壁的延伸和隔膜孔结构的形成。本研究中还观察到胞间菌丝间的融合现象。本文的研究结果表明:小麦条锈菌胞间菌丝的一些特征显然不同于其它锈菌。     

条锈菌与慢锈小麦品种互作的超微结构*

条锈菌与慢锈小麦品种互作的超微结构研究表明,随着慢锈性的表达,条锈菌的胞间菌丝发育受抑,细胞器泡囊化解体。吸器母细胞和吸器发育受阻,最终解体、坏死。吸器的受抑、坏死主要表现在吸器体形成后期。在锈菌与寄主互作的交界面,吸器外质膜皱褶、电子致密度增高,吸器外间质加宽,并有大量电子致密度加深的物质沉积。与抗病品种相比, 慢锈品种中病菌受抑、坏死程度轻,寄主细胞的过敏性坏死机率也较低。     

Histopathology of Leaf Rust Infection and Development in Wheat Genotypes containing Lr12 and Lr13

Evidence exists that certain genes for resistance to leaf rust in wheat, e.g. Lr13 and Lr34 , may interact with other genes to condition higher levels of resistance than that conferred by each gene individually. In this study, the hypothesis that Lr12 and Lr13 , both genes for adult plant resistance to Puccinia recondita Roberge ex. Desmaz f. sp. tritici Eriks. and Henn., interact to confer an improved level of resistance, was investigated using fluorescence and phase-contrast microscopy. Flag leaf segments of monogenic and digenic Thatcher lines, sampled 64 and 240 h post-inoculation, were stained with Uvitex 2B and screened, using fluorescence microscopy, for development of infection structures or host response. To study cell wall appositions, specimens were stained with trypan blue and a solution of picric acid in methyl salicylate. Aborted penetration, consisting of nonpenetrating appressoria and aborted substomatal vesicles, showed that inhibition of fungal growth in wheat lines containing Lr12 and/or Lr13 was activated, to a certain degree, before haustoria were formed. At 240 h after inoculation colony size indicated that fungal colonies in the Lr gene combination lines were generally smaller than in the parents, but not necessarily smaller than those in a line with Lr13 only. Host cell necrosis was more frequently associated with infection sites, specifically of pathotype UVPrt2, in the combination lines than in the parents. The morphology of cell wall appositions varied considerably from a narrow, luminous zone slightly wider in the centre, to a thick central part opposite the haustorium mother cell, sharply decreasing towards both ends. Histological assessments could, however, not conclusively prove pronounced resistance enhancement or unconventional resistance mechanisms due to combining the genes Lr12 and Lr13 .     

Proteome of monoclonal antibody‐purified haustoria from Puccinia triticina Race‐1

Puccinia triticina causes leaf rust, a disease that causes annual yield losses in wheat. It is an obligate parasite that invades the host leaf and forms intracellular structures called haustoria, which obtain nutrients and suppress host immunity using secreted proteins called effectors. Since effector proteins act at the frontier between plant and pathogen and help determine the outcome of the interaction, it is critical to understand their functions. Here, we used a direct proteomics approach to identify effector candidates from P. triticina Race 1 haustoria isolated with a specific monoclonal antibody. Haustoria were >95% pure and free of host contaminants. Using high resolution MS we have identified 1192 haustoria proteins. These were quantified using normalized spectral counts and spanned a dynamic range of three orders of magnitude, with unknown proteins and metabolic enzymes as the most highly represented. The dataset contained 140 candidate effector proteins, based on the presence of a signal peptide and the absence of a known function for the protein. Some of these candidates were significantly enriched with cysteine, with up to 13 residues per protein and up to 6.8% cysteine in composition.     

Methyl jasmonate stimulates polyamines biosynthesisand resistance against leaf rust in wheat plants

Abstract

Leaf rust of wheat (Triticum aestivum L.), incited by Puccinia recondita ex Desm. f. sp. tritici Eriks, is one of the most important wheat diseases in Egypt. Methyl jasmonate (MJ) is a potential plant elicitor which induces a wide range of chemical and anatomical defense reactions in conifers and might be used to increase systemic resistance against biotic damage. In the greenhouse, different concentrations of MJ (10, 20 and 30 mM) were applied as seed soaking plus foliar spray or only as foliar spray to control leaf rust and induction of secondary compound production in leaves of wheat plants. Foliar spray was applied after 30 and 50 days of sowing. Results indicated that all concentrations and treatments reduced the severity of rust disease caused by P. recondita f. sp. tritici in wheat leaves during 45 days of inoculations. Disease incidence was decreased significantly in MJ-treated plants as seed soaking plus foliar spray with 20 and 30 mM when compared to 10 mM MJ or control plants. The study revealed that, with increasing concentrations of MJ, the secondary metabolites were greatly increased. Endogenous levels of both free and conjugated putrescine, spermidine and spermine increased in response to the elicitor. Activities of polyamine biosynthetic enzymes of ornithine decarboxylase (ODC) and polyamine oxidase (PAO) displayed up to threefold increases relative to untreated control. Moreover, significant increases in activities of plant defense-related protein, enzymes as peroxidase and chitinase as well as free and conjugated phenols contents were recorded in treated plants compared with untreated and infected plants. Furthermore, MJ treatment increased the chlorophyll-a, chlorophyll-b and carotenoids pigments contents, the higher increase was obtained with combined treatment between seeds soaking plus foliar spray at 20 and 30 mM of MJ. Under field conditions, three concentrations of MJ, i.e. 10, 20 and 30 mM as combined treatment between seeds soaking plus foliar spray or only as foliar spray were applied to study their effect against rust disease. Foliar spray was applied after 30 and 80 days of sowing. Results showed that the high reduction in disease severity was obtained with combined treatments between seeds soaking plus foliar spray with MJ at 20 and 30 mM compared with other treatments and control. At the same time, all treatments increased the growth and grain yield of wheat plants. It could be suggested that combination treatment between seeds soaking plus foliar spray with methyl jasmonate might be used commercially for controlling rust disease of wheat plants under field conditions.     

Microscopic Observation on the Development of Puccinia striiformis in the Spike and Stem of Wheat

The majority of germ tubes of the pathotype CYR32 of Puccinia striiformis f.sp. tritici formed on the surface of spike organs of the susceptible wheat cv. Suwon 11 penetrated through the stomatal pore, only a few germ tubes formed small appressoria over the stomata. In the lemma, palea and glume, the stripe rust fungus spread between the parenchyma cells close to the inner epidermal layer, but the fungus did not develop between the thick‐walled cells near the outer epidermal layer of these organs. In the awn and stem, spread of the stripe rust was confined to the intercellular spaces of the chlorophyll parenchyma, beneath the invaded stomatal pore of the epidermis and the urediniospores to be released disrupted the epidermis. In the caryopsis, the spread of hyphae was restricted to the intercellular spaces of the pericarp cells.     

Early molecular diagnosis and detection of Puccinia striiformis f. sp. tritici in China

Aims: Wheat stripe (yellow) rust, caused by Puccinia striiformis f. sp. tritici (Pst), is the most important foliar disease on wheat in China. Early molecular diagnosis and detection of stripe rust will provide a useful aid to the accurate forecast and seasonal control of this destructive disease. Our objective was to develop PCR assays for the rapid identification and detection of P. striiformis. Methods and Results: The genomic DNA of P. striiformis and P. triticina were amplified by a pair of primers derived from conserved β‐tubulin gene sequence. A 235‐bp specific DNA fragment of P. striiformis was isolated and purified. Based on its sequence, another two primer sets were designed successfully to obtain new sequence‐characterized amplified region (SCAR) markers of P. striiformis, which could be amplified in all test isolates of P. striiformis, whereas no DNA fragment was obtained in other nontarget wheat pathogens. The detection limit of the primer set YR (f)/YR (r1) was 2·20 pg μl^?1. The new SCAR markers of P. striiformis can also be detected in Pst‐infected wheat leaves postinoculated for 2 days. Conclusions: Our assays are significantly faster than the conventional methods used in the identification of P. striiformis. Significance and Impact of the Study: Development of a simple, high‐throughput assay kit for the rapid diagnosis and detection of wheat stripe rust would be anticipated in a further study.     

感染叶锈菌的小麦细胞间隙液中激发子的定性及初步分离

感染叶锈菌后小麦叶片细胞间隙液(1wF)中有激发子存在,它(们)能诱导寄主PAL、PO活性的增强及细胞过敏性坏死的产生。这种有诱导活性的物质分别用NaIO4和蛋白酶处理证明含有糖基和蛋白质成分,可能是糖蛋白。IWF经凝胶过滤分离后,各部分诱导不同的防卫反应的活性强度不等。因此IWF中可能含有几种不同成分的激发子,或是同种成分而聚合度不同的激发子。     

温室条件下条形柄锈菌体细胞重组的分子确证

为了获得温室条件下条形柄锈菌发生体细胞重组而导致毒性变异的直接证据,本研究选取7个美国条形柄锈菌小麦专化型菌系和2个美国条形柄锈菌大麦专化型菌系按照夏孢子颜色和专化型与毒性差异组成9对菌系组合,对于室内混合接种产生的子代菌系用具有不同抗性的小麦或大麦品种进行筛选,采用毒性分析及SSR分子标记技术对条形柄锈菌体细胞重组现象进行了研究。对获取的413个单孢子代菌系进行的毒性分析结果显示,有84个单孢子代菌系的毒性谱表现与亲本菌系不同,初步证明体细胞重组过程的存在。SSR标记分析结果显示,11对SSR引物中有6对引物在5对菌系组合的28个毒性谱不同的单孢子代菌系中,检测发现3个单孢菌系的扩增条带与其亲本菌系不同,且表现为亲本菌系扩增条带的重组,为体细胞重组菌系。这一结果从分子水平上证明了条形柄锈菌在室内接种条件下可以通过体细胞重组产生新小种而导致毒性变异。     

小麦抗条锈病近等基因系感染条锈病后丁布含量变化

选用2套遗传背景不同的抗条锈病近等基因系作为供试寄主材料,研究了不同抗条锈病近等基因系丁布的含量及其在感病过程中丁布含量的动态变化.结果表明,在未受病菌侵染情况下,2套分别含有Yr2、Yr9和YrSpP基因的近等基因系（抗病系）与其轮回亲本Taichung 29、铭贤169(感病系)间丁布含量没有显著差异（P＞0.05）.接种条锈病菌后,感病系在病菌侵染初期丁布含量下降,而抗病系在病菌侵染初期丁布含量迅速大幅度上升.感染条锈病最终导致感病植株丁布含量比未接种的植株明显减少, 感病系的减少幅度明显高于抗病系.在整个病程中,抗病系丁布的含量始终高于感病系,表明接种条件下小麦植株体内丁布含量变化与小麦抗条锈近等基因系的抗性有关.