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Infection of onion by Alternaria porri and Stemphylium vesicarium was investigated under a range of controlled temperatures (4–25°C) and leaf wetness periods (0–24 h). Conidia of A. porri and S. vesicarium germinated within 2 h when incubated at 4°C. Terminal and intercalary appressoria were produced at similar frequencies at or above 10°C. The maximum number of appressoria was produced after 24 h at 25°C. Penetration of leaves by both pathogens was via the epidermis and stomata, but the frequency of stomatal penetration exceeded that of epidermal penetration. There was a strong correlation ( R 2 > 90%) between appressorium formation and total penetrations at all temperatures. Infection of onion leaves occurred after 16 h of leaf wetness at 15°C and 8 h of leaf wetness at 10–25°C, and infection increased with increasing leaf wetness duration to 24 h at all temperatures. Interruption of a single or double leaf wetness period by a dry period of 4–24 h had little effect on lesion numbers. Conidia of A. porri and S. vesicarium separately or in mixtures caused similar numbers of lesions. Alternaria porri and S. vesicarium are both potentially important pathogens in winter-grown Allium crops and purple leaf blotch symptoms were considered to be a complex caused by both pathogens. 相似文献
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ABSTRACT Field observations in four pear orchards during 5 years from April to October indicated that days with uninterrupted wetness of variable length represented 83.9% of the total days studied. However, days with surface wetness interruptions and with high relative humidity (RH) (>/=90%) without wetness occurred with a frequency of 7.1 and 6.2%, respectively. Accordingly, the effect of interruption of 24-h wetness periods by dry periods of high or low RH on infections caused by Stemphylium vesicarium on pear was determined. Pear plants inoculated with conidia of S. vesicarium were exposed to a 12-h wet period followed by a dry period of variable length (0, 3, 6, 12, 18, or 24 h) and a second wet period of 12 h. The dry period consisted either of low (60%) or high (96%) RH. The infection process was irreversibly stopped under low RH during dry periods between wetness, but continued at high RH. The effect of high RH on disease severity in the absence of wetness was also determined. Pear plants inoculated with S. vesicarium were exposed to periods of variable length (3 to 24 h) either at high RH (96%) in the presence of wetness or at high RH (96%) without wetness. No infections were observed on plants incubated under high RH without wetness, indicating that conidia of S. vesicarium require the presence of a water film in the plant surface to develop infections on pear. 相似文献
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新疆大蒜叶疫病的研究——Ⅰ症状及病原菌研究鉴定 总被引：3，自引：0，他引：3
大蒜叶疫病在新疆发生于6~8月,主要为害大蒜叶片。根据发病症状、病原菌形态,分离物培养性状、产孢习性及人工接种等研究结果,确定本病系由真菌Stemphylium vesicarium (Wallr)Simmons的侵染所致。该病在我国尚属首次报道。 相似文献
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Surveys between 1989 and 1993 in the major garlic production areas of Spain identified a new leaf spot disease, characterized by white and purple lesions followed by extensive necrosis. Isolation and pathogenicity tests with fungal isolates taken from these spots indicated that Stemphylium vesicarium was the causal agent. Pseudothecia of the teleomorph stage, Pleospora sp., were found on leaf debris from affected plants. Inoculation of garlic and onion plants with residues carrying mature pseudothecia, or with ascospore suspensions obtained from the pseudothecia, resulted in the development of white and purple leaf spots. Wetness periods longer than 24 h were required for symptom development under controlled conditions. Isolates of S. vesicarium from garlic, onion and asparagus caused disease in all three hosts. In garlic, cv. Blanco de Vallelado was most susceptible, while lines B4P17 and B6P1, and cvs Iberose and Golourose were less susceptible to the disease. 相似文献
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Singh P Bugiani R Cavanni P Nakajima H Kodama M Otani H Kohmoto K 《Phytopathology》1999,89(10):947-953
ABSTRACT Culture filtrates of a pathogenic isolate (IT37) of Stemphylium vesicarium, causing brown spot of European pear, induced veinal necrosis only on pear leaves susceptible to the pathogen. Two host-specific toxins, SV-toxins I and II, were purified from culture filtrates of IT37 by successively using Amberlite XAD-2 resin adsorption, cellulose thin-layer chromatography, and high-performance liquid chromatography under three different sets of conditions. Susceptible cultivars showed veinal necrosis at a SV-toxin I concentration of 0.01 to 0.1 mug/ml, whereas resistant cultivars were insensitive to the toxin at 1,000 mug/ml. SV-toxins I and II caused a dose-dependent increase in electrolyte loss from susceptible leaf tissues. No increase in electrolyte loss was detected in leaf tissues from resistant cultivars. The results of physiological studies indicated that SV-toxins appear to have an early effect on plasma membranes of susceptible leaves. Spores of a nonpathogenic isolate induced necrotic lesions on susceptible leaves in the presence of a small amount of toxin. SV-toxins were detected in intercellular fluids obtained from diseased leaves after inoculation with the pathogen. The results indicate that SV-toxins I and II produced by S. vesicarium can be characterized as host-specific toxins. 相似文献
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将制备并提纯的抗Xcc单克隆抗体、多克隆抗体及重组抗体,采用间接ELISA方法对比其效价和特异性,筛选用于胶体金速测卡的最优抗体;采用柠檬酸三钠还原法制备胶体金颗粒,标记抗Xcc单抗Xcc-2D8,固定于金标垫上;利用双抗夹心原理,将单抗Xcc-2D6和羊抗鼠抗体分别固定于硝酸纤维素膜上,作为检测线(T线)和质控线 (C线),经试验条件优化,组装胶体金速测卡密封干燥保存.对Xcc菌悬液及柑橘样本浸泡液进行检查.结果表明ELISA方法筛选出2株特异性强、稳定性好的抗体(Xcc-2D8效价为1∶128 000倍、Xcc-2D6效价为1∶256 000倍),应用于胶体金速测卡的研发;所研制速测卡对Xcc菌悬液的最低检测下限为1×103～1×104 cfu/mL,对多个地区的206份柑橘样本进行实际检测所得结果与qPCR方法所得结果具有较高一致性(kappa值=95.15％),混杂的柑橘组织液对检测结果无影响,检测时间控制在10 min以内,检测常见细菌及植物病菌显示特异性良好,各批次速测卡无批间差异,速测卡保存方便稳定性良好. 相似文献
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柑橘溃疡病是重要的柑橘细菌性病害,为探明壳聚糖和枯草芽孢杆菌对柑橘溃疡病的防治潜力,本文用平板菌落计数法评价了3种粘度壳聚糖对柑橘溃疡病菌的杀菌效果,以改良的纸碟法测定2株枯草芽孢杆菌(Bacillus subtilis)发酵液对柑橘溃疡病菌的抑菌效果。综合处理0.5 h、3 h和6 h的结果显示,3种粘度壳聚糖对柑橘溃疡病的杀菌强弱依次为:粘度12粘度100粘度20。枯草芽孢杆菌菌株BS 101和54-6在YPG培养基中有氧发酵3 d产生发酵液原液的抑菌圈直径分别为3.45 cm和3.21 cm,比0.1 g/mL氨苄青霉素的抑菌圈分别大45%和35%。可见,发酵液中存在对溃疡病菌高度有效的成分。因此,认为粘度12的壳聚糖和芽孢杆菌菌株BS 101和54-6具有防治柑橘溃疡病的潜力。 相似文献
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Xanthomonas citri subsp. citri (Xac) is the causal agent of citrus bacterial canker, an important disease for the citrus industry. Studies of Xac survival in environments outside of the lesion performed in the past may have underestimated the viable population because the recovery was based on the ability of the bacterium to grow on culture media. This study monitored survival of Xac that express green fluorescent protein (GFP) in two different forms: the native protein, and a protein that is unstable due to a specific oligopeptide tail targeted by proteases within the bacterium. Transformed strains of Xac were verified to be stable in their expression of GFP and to show no differences in virulence and fitness compared to wild type strains. Evaluation of protein stability confirmed that strains with unstable GFP only expressed and fluoresced in metabolically active cells, and not in dead bacteria. Fluorescence of unstable GFP strains under confocal microscopy was used to track bacterial survival and biofilm formation on leaf and fruit surfaces. After spray inoculation, aggregates of fluorescing cells of unstable GFP strains formed biofilms on leaves and fruit. Bacterial cells that aggregated on the surfaces only survived when protected from desiccation. Aggregation of viable bacteria in biofilms confirms their role in pathogen survival outside of lesions and protection from bactericide treatments in the field or in the fruit disinfection process. 相似文献
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U. Melcher J. Fletcher 《European journal of plant pathology / European Foundation for Plant Pathology》1999,105(6):519-533
Spiroplasmas are members of the Class Mollicutes, wall-less prokaryotes having a high adenosine–thymidine content in their small genomes. Spiroplasma citri is a plant pathogen that inhabits phloem. Like other phytopathogenic spiroplasmas and the related phytoplasmas, it is transmitted from plant to plant by phloem-feeding leafhoppers that serve as alternate hosts for the spiroplasma as well as vectors. Genetic information in spiroplasmas is carried on a circular chromosome, on plasmids and/or in virus genomes. A picture emerging from recent research on the S. citri genome is one of frequent and often extensive variation, resulting from a number of different mechanisms. Expansion and contraction events must continually be occurring in about equal proportions so that the net genome size varies within defined boundaries. Particularly impressive are large changes in genome size that can occur in only a few generations. As with most organisms, genetic variation in S. citri results from variation in extrachromosomal DNA content, changes due to DNA replication and repair processes and changes due to recombination. The implied flux of genetic information into and out of the S. citri genome should be beneficial to the bacterium, allowing it, with its small genome size, to adapt to new environments. 相似文献
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传统PCR方法不能诊断柑橘溃疡病菌(Xanthomonas citri subsp.citri Gabriel)的死活状态,往往导致假阳性检测结果.本研究将特异性核酸染料叠氮溴化乙锭(ethidium monoazide bromide,EMA)与PCR技术结合,旨在建立柑橘溃疡病活菌的快速检测技术.根据柑橘溃疡病菌独有的保守蛋白基因设计特异性引物扩增出278 bp的靶带,PCR反应的检测下限为25个细胞/25 μL或2.75 pg/25 μL.EMA-PCR结果表明:当卤钨灯曝光时间1 min,EMA终浓度为1.0 mg/L时,能有效抑制1.0×108 cfu/mL死菌的扩增;当EMA的浓度小于30 mg/L时,EMA对上述相同浓度活菌靶基因的扩增没有明显的抑制.EMA-PCR对死活混合菌的扩增表明,活菌数在6.875×101～6.875×105 cfu/PCR范围时,荧光强度与混合体系中活菌的对数值有线性关系.基于以上建立的EMA-PCR活体检测技术,对疑似带病柑橘材料进行检测,结果发现能降低柑橘溃疡病菌检测过程中的假阳性,有望为柑橘溃疡病的检疫检验提供更科学的技术手段. 相似文献
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试验结果表明,本菌(Stemphylium vesicarium(Wallr.) Simons)以病残体上分生孢子和菌丝体在田间和冷藏室越冬,9月至翌年6月检查,分生孢子萌发率为16.4~41.3%。人工接种试验指出,不论有无伤口,病菌均可侵染大蒜,但洋葱与大葱在伤口接种条件下病情较重。在自然条件下,大蒜的斑点型病状出现于6月下旬,7月为病情高峰期。影响病情的天气因素,主要是7月份温度和7月中旬的降水量(r=0.69)。如果7月雨次多、雨时长,病情即极为严重,可导致蒜田绝收。化学防治试验结果,以腐霉利防效最佳。从7月上旬开始,每间隔10~13天喷药1次。喷药2~3次即可取得较好的防治效果。 相似文献
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以柑橘溃疡病菌DNA为模板,对抗铜相关基因copA和copB进行了PCR扩增和克隆,获得了大小分别为1 782 bp 和1 095 bp的目标片段。构建了这2个基因的原核表达载体 (pET-copA和pET-copB),并在大肠杆菌 [Escherichia coli BL21(DE3)] 中成功诱导表达。利用原核表达的融合蛋白免疫大耳白兔,制备了抗copA和copB原核表达蛋白的多克隆抗体。用间接ELISA法测定了所制备的多克隆抗体的效价均为1∶6 400。用所制备的抗体分别对copA和copB的原核表达产物进行Western blot分析的结果显示,在相应位置产生了较强的免疫反应条带,表明所制备抗体能特异性地与相应的抗原发生免疫反应。 相似文献
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N. Kositcharoenkul O. Chatchawankanphanich A. Bhunchoth W. Kositratana 《Plant pathology》2011,60(3):436-442
A single‐tube nested PCR was developed for detection of Xanthomonas citri subsp. citri (Xcc), the causal agent of citrus canker disease. The assay targets the pthA gene of Xcc and utilizes different annealing temperatures for the two primer pairs. It reliably detected as few as 1·0 × 102 Xcc cells, and was unaffected by the presence of PCR inhibitors. It was 10‐fold and 8500‐fold more sensitive than standard PCR and ELISA, respectively. Increased sensitivity was also achieved via the use of a washing method for DNA extraction, as opposed to direct extraction from leaf tissue. When evaluated for Xcc detection in 90 samples collected from affected pomelo orchards, the single‐tube nested PCR was superior to standard PCR, detecting the pathogen in 67 vs. 54 samples. It was also able to detect Xcc from samples with and without symptoms. This assay can be used as a rapid and sensitive technique for routine Xcc detection in field samples for surveillance of citrus canker. 相似文献
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The copper-based products widely used for control of citrus canker may lead to the development of Xanthomonas citri subsp. citri (X. citri) resistant to copper (CuR). However, the study of copper sensitivity of X. citri strains from Paraná state, Brazil, did not reveal the existence of CuR, but copper tolerant (CuT) strains. The aim of this study was to describe for the first time the existence of CuT X. citri and compare the genetic determinants that differentiate the CuT strains from the sensitive (CuS) and CuR strains. CuT strains supported intermediate concentrations of copper in comparison to CuS and CuR. CuT strains lack the gene clusters copLAB or copABCD responsible for copper resistance in CuR strains and the large plasmids (c. ≥200 kb) that normally carry these genes. The nucleotide sequences of chromosomal homologous genes cohLAB, involved in copper homeostasis, were 100% similar in strains of all phenotypes. CuT strains differed from CuS strains by the higher expression of the homologous chromosomal genes cohA and cohB in the presence of copper. CuT X. citri strains are not precursors of CuR strains and do not pose a threat to the efficient use of copper-based bactericides for management of citrus canker in citrus orchards. Copper resistance and tolerance are distinct phenotypes and should not be used as synonyms. The proper characterization of the sensitivity to copper leads to a more confident monitoring of the distribution of copper resistant populations of X. citri and adoption of containment measures only when necessary. 相似文献
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