Host Range ofVerticillium dahliae in cultivated species in Crete

To determine the host range ofVerticillium dahliae among the cultivated species in Crete, Greece, studies were carried out during 1992–2000. Based on disease symptoms observed on 28 vegetable and forage species grown in a field naturally infected byV. dahliae, and an extensive survey of the most common cultivated species grown under natural conditions, seven hosts belonging to four botanical families not previously reported as susceptible to Verticilliun wilt, and 12 hosts belonging to seven families new for Greece, were recorded. The worldwide new hosts are: anise (Anethum graveolens), chard (Beta vulgaris ssp.cicla), chickpea (Cicer arietinum), wild sweet pea (Lathyrus ochrus), lentil (Lens culinaris), marigold (Tagetes erecta) and vetch (Vicia sativa). These species could be infected by their hosts-of-originV. dahliae isolates during pathogenicity tests. http://www.phytoparasitica.org posting Jan. 13, 2002.     

Plant Host Range of Verticillium longisporum and Microsclerotia Density in Swedish Soils

Verticillium longisporum is a soil-borne fungal pathogen causing vascular wilt of Brassica crops. This study was conducted to enhance our knowledge on the host range of V. longisporum. Seven crop species (barley, oat, oilseed rape, pea, red clover, sugar beet and wheat) and five weed species (barren brome, black-grass, charlock, cleavers and scentless mayweed) all common in southern Sweden were evaluated for infection by response to V. longisporum. Oat, spring wheat, oilseed rape, scentless mayweed and charlock inoculated with V. longisporum in a greenhouse showed stunting to various degrees close to the fully ripe stage. Based on the extent of microsclerotia formation, explants were separated into four groups: for pea and wheat, <5% of the samples had formed microsclerotia; for scentless mayweed, 5–10%; for oat, 10–20%; and for charlock and oilseed rape >80%. The results suggest that plant species outside the Brassicaceae can act as reservoirs of V. longisporum inoculum. Soil inoculum densities in nine fields were monitored over a period of 12 months, which ranged from 1 to 48 cfu g⁻¹ soil. Density of microsclerotia was lowest just after harvest, reaching its maximum six months later. No significant correlation between inoculum density in soil and disease incidence on oilseed rape plants was found. However, the data suggest that a threshold of 1 cfu g⁻¹ soil is needed to cause disease on oilseed rape. Species identification based on microsclerotia morphology and PCR analysis showed that V. longisporum dominated in soil of seven, and V. dahliae in two of the nine fields studied.     

棉花黄萎病防治策略

黄萎病是棉花生产中的最主要病害之一,广泛分布于世界各产棉国.由于该病是土壤传播的维管束病害,防治难度较大,至今尚无特效的防治药剂,只能依靠种植抗病品种为主的综合防治措施.但是,我国目前棉花品种的抗病性只能达到抗至耐病水平,致使该病在环境条件适宜时连续流行为害.控制该病的猖獗为害,已成为棉花生产可持续发展的主要问题之一.     

Vegetative-compatibility groups in Verticillium dahliae from Israel

Nitrate-nonutilizing (nit) mutants were used to determine vegetative compatibility among 34 isolates of Verticillium dahliae from cotton, potato, olive, eggplant, chrysanthemum and tomato from 12 sites in Israel. Based on the formation of complementary heterokaryons, 33 isolates were assigned to two vegetative- compatibility groups (VCGs): one VCG contained 15 isolates from cotton, eggplant, chrysanthemum and olive; and the other VCG contained 18 isolates from potato, olive and cotton. The status of an additional isolate from tomato, which was compatible with both VCGs, remained unclear. In a limited pathogenicity test with 10 isolates, two (from tomato and eggplant) were pathogenic on tomato, eggplant and cotton; most isolates from cotton were pathogenic on cotton and eggplant only; and one from cotton was non-pathogenic. Fewer isolates were pathogenic on tomato than on cotton or eggplant. The diversity of vegetative compatibility found in our V. dahliae collection is comparable to that found in studies of American populations.     

Sources of Verticillium dahliae Affecting Lettuce

ABSTRACT Since 1995, lettuce in coastal California, where more than half of the crop in North America is grown, has consistently suffered from severe outbreaks of Verticillium wilt. The disease is confined to this region, although the pathogen (Verticillium dahliae) and the host are present in other crop production regions in California. Migration of the pathogen with infested spinach seed was previously documented, but the geographic sources of the pathogen, as well as the impact of lettuce seed sparsely infested with V. dahliae produced outside coastal California on the pathogen population in coastal California remain unclear. Population analyses of V. dahliae were completed using 16 microsatellite markers on isolates from lettuce plants in coastal California, infested lettuce seed produced in the neighboring Santa Clara Valley of California, and spinach seed produced in four major spinach seed production regions: Chile, Denmark, the Netherlands, and the United States (Washington State). California produces 80% of spinach in the United States and all seed planted with the majority infested by V. dahliae comes from the above four sources. Three globally distributed genetic populations were identified, indicating sustained migration among these distinct geographic regions with multiple spinach crops produced each year and repeated every year in coastal California. The population structure of V. dahliae from coastal California lettuce plants was heavily influenced by migration from spinach seed imported from Denmark and Washington. Conversely, the sparsely infested lettuce seed had limited or no contribution to the Verticillium wilt epidemic in coastal California. The global trade in plant and seed material is likely contributing to sustained shifts in the population structure of V. dahliae, affecting the equilibrium of native populations, and likely affecting disease epidemiology.     

Host specialization of Verticillium dahliae, with emphasis on isolates from cocoa (Theobroma cacao)

Isolates of Verticillium dahliae from cocoa (four Brazilian and one Ugandan) were screened against cocoa, aubergine, tomato, cotton, and pepper. Isolates originating from hosts other than cocoa were also inoculated. In general, all inoculated crops were systemically invaded by isolates from cocoa. Isolates from each host tended to be more aggressive on their original host. All isolates from cocoa were pathogenic to cocoa, but exhibited various degrees of aggressiveness to other crops. They induced severe symptoms on aubergine, but few symptoms on tomato, although colonization occurred up to the stem apex in both cases. Likewise, symptomless invasion of pepper was found with some Brazilian isolates. The Ugandan isolate was significantly more aggressive on cotton and pepper than the Brazilian isolates.
A Brazilian isolate from cocoa from the State of Bahia was also used to inoculate 12 common weed species from the same geographical area. Four species showed wilt symptoms, while V. dahliae was readily recovered from the stems of a further four species. The role of alternative hosts on disease spread in cocoa growing areas is discussed.     

番茄黄萎病病原菌研究

近年在黑龙江省保护地番茄上发现一种番茄植株黄萎病害,从其病茎上分离得到10个菌株。将其接种番茄幼苗,发病症状与自然发病症状完全一致,并从接种病株上重新分离到该菌。经鉴定,病原菌为大丽轮枝菌(Ver-tillium dahliae Kleb)。对其生物学特性研究表明:该菌生长最低温度为5℃,最适范围20-25℃,以22.5℃生长最适。孢子萌发温度为5-30℃,最适温度为25℃。病菌菌丝在pH4-8都能生长,以pH 5生长最好。     

Genetic variability of tolerance to Verticillium albo-atrum and Verticillium dahliae in Medicago truncatula

Verticillium wilt caused by Verticillium albo-atrum and Verticillium dahliae is responsible for yield losses in many economically important crops. The capacity of pathogenic fungi to adapt to new hosts is a well-known threat to the durability of resistant crop varieties. In the present work, 25 Medicago truncatula genotypes from a core collection and six V. albo-atrum and V. dahliae strains were used to study the potential of non-host Verticillium strains isolated from different plant species to infect this legume plant and the plant’s susceptibility to the pathogens. The experiment was designed as factorial with randomised complete blocks and with three repetitions. The wilt symptoms caused by V. albo-atrum and V. dahliae were scored on a disease index scale from 0 to 4, during 30 days after inoculation of 10-day-old plantlets. Disease severity was quantified by maximum symptom scores (MSS) and areas under the disease progress curves (AUDPC), which integrate the time course of symptom development. Highly significant differences were observed among plant genotypes and fungal strains, and their interaction was also significant. The correlation between MSS and AUDPC was 0.86. The most severe symptoms were caused by the alfalfa strain V. albo-atrum V31-2 and the least severe by V. dahliae JR2, as shown by mean values obtained on the 25 genotypes. M. truncatula genotype TN8.3 was the most susceptible genotype by mean values obtained with the six fungal strains, whereas F11013-3, F83005.9 and DZA45.6 were highly resistant to all strains studied. The results can be used to choose parents for studying the genetics of resistance in Medicago truncatula to Verticillium strains with different levels of aggressiveness.     

Sulphur supply impairs spread of Verticillium dahliae in tomato

Vascular wilt caused by the soil-borne fungus Verticillium dahliae is a major yield and quality-limiting disease across a broad spectrum of crop plants worldwide. Sulphur-enhanced plant defence mechanisms provide an opportunity to effectively and environmentally safely constrain the wilt disease levels in planta. To evaluate the influence of sulphur nutrition on the protective potential of these mechanisms, two near-isogenic tomato genotypes differing in fungal susceptibility, were treated with low or supra-optimal sulphur supply. Microscopic analysis revealed a significant sulphur-induced decrease in the amount of infected vascular cells in both genotypes. However, plant shoot and severely pathogen-affected root growth did not display this distinct alleviating influence of sulphur nutrition. Rates of leaf photosynthesis were impeded by Verticillium dahliae infection in both genotypes especially under low sulphur nutrition. However, assimilate transport rates in the phloem sap were enhanced by fungal infection more in the resistant genotype and under high sulphur nutrition suggesting a stronger sink for assimilates in infected plant tissues possibly involved in sugar-induced defence. A SYBR Green-based absolute quantitative Real-Time assay using a species-specific primer was developed which sensitively reflected sulphur nutrition-dependent changes in fungal colonization patterns. High sulphur nutrition significantly reduced fungal spread in the stem in both tomato genotypes. Concentrations of selected sulphur-containing metabolites revealed an increase of the major anti-oxidative redox buffer glutathione under high sulphur nutrition in response to fungal colonization. Our study demonstrates the existence of sulphur nutrition-enhanced resistance of tomato against Verticillium dahliae mediated by sulphur-containing defence compounds.     

Hosts of Verticillium dahliae in Kriti (Greece)*

Naturally wilted cultivated and weed species were collected from several fields in Kriti (GR) over the years 1992/1997, and tested by isolation for the presence of Verticillium dahliae. The fungus was most frequently recovered from cultivated Solanaceae and Cucurbitaceae. Infection of many known Verticillium hosts was confirmed, and the fungus was found for the first time in Kriti in pistachio and grapevine and in several common weeds.     

Spatial Patterns of Microsclerotia of Verticillium dahliae in Soil and Verticillium Wilt of Cauliflower

ABSTRACT The spatial patterns of microsclerotia of Verticillium dahliae in soil and wilt symptoms on cauliflower were determined at three sites in each of two fields in 1994 and 1995. Each site was an 8 x 8 grid divided into 64 contiguous quadrats (2 by 2 m each). Soil samples were collected to a depth of 15 cm with a probe (2.5 cm in diameter), and samples from four sites in each quadrat were bulked. Plants in each quadrat were cut transversely, and the number of plants with vascular discoloration and the number without discoloration were recorded. The soil was assayed for microsclerotia by the modified Anderson sampler technique. Lloyd's index of patchiness (LIP) was used as an indicator to evaluate the aggregation of microsclerotia in the field. Spatial autocorrelation and geostatistical analyses were also used to assess the autocorrelation of microsclerotia among quadrats. The LIP for microsclerotia was greater than 1, indicating aggregation of propagules; however, the degree of aggregation at most sites was not high. Significant autocorrelation within or across rows was detected in some spatial autocorrelograms of propagules, and anisotropic patterns were also detected in some oriented semivariograms from geostatistical analyses for microsclerotia, indicating the influence of bed preparation in the fields on pathogen distribution. The parameter estimates p and theta in the beta-binomial distribution and the index of dispersion (D) associated with the distribution were used to assess the aggregation of diseased plants at each site. A random pattern of wilt incidence was detected at 7 of 12 sites, and an aggregated pattern was detected at 5 of 12 sites. The degree of aggregation was not high. A regular pattern of wilt severity was detected at all sites. The high disease incidence (77 to 98%) observed at 11 of the 12 sites could be explained by high inoculum density.     

Phylogenetic analysis of Verticillium dahliae vegetative compatibility groups

The evolutionary relationships among Verticillium dahliae vegetative compatibility (VCG) subgroups VCG1A, VCG1B, VCG2A, VCG2B, VCG4A, VCG4B, and VCG6 were investigated by parsimony analysis of amplified fragment length polymorphism (AFLP) fingerprints and sequences of six DNA regions (actin, beta-tubulin, calmodulin, and histone 3 genes, the ITS 1 and 2 regions of the rDNA, and a V. dahliae-specific sequence), using 101 isolates of diverse host and geographic origin. Polymorphisms in gene sequences among isolates of different VCGs were very low and individual gene genealogies provided very little resolution at the VCG level. The combined analysis of all DNA regions differentiated all VCG subgroups except for isolates in VCG1A and VCG1B. VCG clonal lineages in V. dahliae and evolutionary relationships among them were resolved independently by analyses of AFLP fingerprints, multiple gene genealogies, and the combined data set of AFLP fingerprinting and multiple gene genealogies. Two main lineages (I and II) were identified with lineage II comprising two closely related subgroups of VCGs. Lineage I included VCG1A, VCG1B, and VCG2B334; and lineage II included, VCG2A and VCG4B (subclade 1); and VCG2B824, VCG4A, and VCG6 (subclade 2). VCG subgroups were monophyletic except for VCG2B that appeared polyphyletic. Limiting the parsimony analysis either to AFLP fingerprints or DNA sequences would have obscured intra-VCG differentiation. Therefore, the dual approach represented by the independent and combined analyses of AFLP fingerprints and DNA sequences was a highly valuable method for the identification of phylogenetic relationships at the intraspecific level in V. dahliae.     

绿色荧光蛋白基因标记棉花黄萎病菌

以携带有潮霉素抗性筛选标记的pCTHyg载体为骨架,构建了含有增强型绿色荧光蛋白基因sGFP的载体pCH-sGFP,并通过农杆菌介导的遗传转化法导入引起棉花黄萎病的高致病力大丽轮枝菌Vd991,获得了sGFP整合到大丽轮枝菌基因组的转化株。通过转化子荧光信号、生长表型和致病力筛选鉴定,获得了1株与Vd991生长和致病力无显著差异且荧光信号强烈的转化株Vd gfp77。侵染棉花根部试验表明,Vd-gfp77侵染棉花后快速扩展繁殖,子代仍然能发出强烈的荧光信号。本试验绿色荧光蛋白标记大丽轮枝菌的成功构建,为后续大丽轮枝菌侵染棉花过程的组织学和致病机理研究提供了良好的研究材料。     

大丽轮枝菌核糖体基因ITS区段的特异扩增

 根据棉花黄萎病菌(Verticillium dahliae)核糖体基因ITS区段的碱基编码序列,设计合成了1对为26 bp的PCR特异扩增引物(引物1:5'CATCAGTCTCTCTGTTTATACCAACG,和引物2:3'CGATGCGAGCTGTAACTACTACGCAA),进行了大丽轮枝病菌PCR特异扩增试验。试验结果表明:本试验设计合成的这对引物,能对大丽轮枝菌全基因组DNA和人工接种棉花黄萎病病株组织特异地扩增到大丽轮枝菌核糖体基因ITS区段的324 bp分子片段,该对引物可用于棉花黄萎病的分子鉴定和分子监测。本试验结果对棉花黄萎病的早期诊断和病原菌的检测和监测有潜在的应用价值。     

Biofumigation potential of Brassicaceae cultivars to Verticillium dahliae

The biofumigation potential of brassicaceous green manures to Verticillium dahliae was systematically assessed. In a toxicological study, five different isothiocyanates (ITCs) were tested using a bioassay with sterile quartz sand artificially infested with microsclerotia. For 2-propenyl ITC, a LD₉₀ value of 88.7 nmol g^?1 was determined. Furthermore, 2-propenyl ITC with a dose of 150 nmol g^?1 soil was tested in 22 naturally infested soils. The efficiency varied from 9 % to 92 % and was negatively correlated with the organic carbon content of the soils, indicating that in many soils much higher concentrations will be necessary to achieve sufficient control. To evaluate the biofumigation potential of different Brassicaceae, 19 cultivars of Brassica juncea, Rhaphanus sativus and Sinapis alba were grown in the field. Biomass production was measured and glucosinolate concentrations were analyzed. Simulating the field situation, the biofumigation effect of cultivars was assessed in a standardized laboratory bioassay with microsclerotia-infested sterile quartz sand amended with freeze-dried ground plant tissues. Amendments of B. juncea shoot tissue reduced the number of viable microsclerotia significantly with efficiencies from 69.3 to 81.3 %. Total potentially released amounts of 2-propenyl ITC ranged between 50.6–78.1 nmol g^?1sand and indicate a clear ITC-related suppression. However these are considered too low for effective control in practice as low release efficiencies mean that effective levels fall well short of the estimated 150 nmol g^?1 of soil required. In comparison with B. juncea, the R. sativus and S. alba were less effective due to lower concentrations and/or toxicity of the ITC released. In summary, the biofumigation potential of the cultivars tested appears insufficient alone for effective control, especially on soils with higher (>1.0 %) organic carbon content.     

芽胞杆菌BvR001对大丽轮枝菌抑制效果研究

为研发绿色、安全、高效的作物黄萎病生物防治产品,本研究以生防菌株BvR001为研究对象,通过对黄萎病菌的抑菌效果测定、可湿性粉剂研制、在寄主根际的定殖能力与防效测定,明确该菌株对黄萎病具有防控效果.gyrB序列检测和系统发育树构建分析表明,该菌株属于芽胞杆菌属,并且与贝莱斯芽胞杆菌Bacillus velezensis...     

棉花黄萎病菌提取物对棉花的抗病效果

近年来棉花黄萎病发生严重 ,重病田病株率高达 95 %以上。目前生产上缺少抗病品种 ,防治方法不健全 ,田间防治效果不理想 ,生产上急需新的防治措施。随着环境和生态问题的日益突出 ,诱导抗病性研究日益被人们重视。本研究从诱导抗病性原理出发 ,旨在探索能够有效激发、诱导棉花抗黄萎病的因子 ,并取得一些结果 ,现报道如下。1　材料与方法1 .1　供试材料　棉种为中棉 1 6 ;棉花黄萎病菌 (Verticillium dahliae Kleb)由本室分离鉴定。1 .2　激发子的制备　将棉花黄萎病菌在马铃薯葡萄糖液体培养基中 ,5℃下恒温振荡培养两周后 ,滤纸过滤 ,滤…