Genetic Analysis of the Role of Trichothecene and Fumonisin Mycotoxins in the Virulence of Fusarium

The phytotoxicity of the Fusarium trichothecene and fumonisin mycotoxins has led to speculation that both toxins are involved in plant pathogenesis. This subject has been addressed by examining virulence of trichothecene and fumonisin-nonproducing mutants of Fusarium in field tests. Mutants were generated by transformation-mediated disruption of genes encoding enzymes that catalyze early steps in the biosynthesis of each toxin. Two economically important species of Fusarium were selected for these studies: the trichothecene-producing species Fusarium graminearum, which causes wheat head blight and maize ear rot, and the fumonisin-producing species F. verticillioides, which causes maize ear rot. Trichothecene-non-producing mutants of F. graminearum caused less disease than the wild-type strain from which they were derived on both wheat and maize, although differences in virulence on maize were not observed under hot and dry environmental conditions. Genetic analyses of the mutants demonstrated that the reduced virulence on wheat was caused by the loss of trichothecene production rather than by a non-target mutation induced by the gene disruption procedure. Although the analyses of virulence of fumonisin-non-producing mutants of F. verticillioides are not complete, to date, the mutants have been as virulent on maize ears as their wild-type progenitor strains. The finding that trichothecene production contributes to the virulence of F. graminearum suggests that it may be possible to generate plants that are resistant to this fungus by increasing their resistance to trichothecenes. As a result, several researchers are trying to identify trichothecene resistance genes and transfer them to crop species.     

Epidemiology of Fusarium Diseases and their Mycotoxins in Maize Ears

Fusarium species cause two distinct diseases on ears of maize, Fusarium ear rot (or pink ear rot) and Gibberella ear rot (or red ear rot), both of which can result in mycotoxin contamination of maize grain. The primary causal agent for Fusarium ear rot is Fusarium verticillioides, but F. subglutinans and F. proliferatum are also important. Gibberella ear rot is caused primarily by F. graminearum, but F. culmorum can also be important, especially in Europe. Aspects of the epidemiology of both diseases have been studied for decades, but only recently have efforts been made to synthesize this information into comprehensive models of disease development. Much of the work on F. graminearum has focused on Fusarium head blight of small-grain crops, but some of the results obtained are also relevant to maize. The primary mycotoxins produced by these fungi, fumonisins and deoxynivalenol, have differing roles in the disease-cycle, and these roles are not completely understood, especially in the case of fumonisins. Progress is being made toward accurate models for risk assessment of both diseases, but key challenges remain in terms of integrating models of pre- and post-infection events, quantifying the roles of insects in these diseases, and characterizing interactions among competing fungi and the environment.     

小麦赤霉病菌多菌灵抗性群体的扩散路径分析——基于致病菌种类及所产毒素化学型鉴定和抗药性检出的时序性

通过对江苏、安徽、山东、河南、湖北、河北和四川7省小麦赤霉病菌对多菌灵抗性及敏感菌株Fusarium asiaticum和F.graminearum的鉴定、所产生毒素的化学型及多菌灵抗性菌株检出时序性的分析,初步推测了小麦赤霉病菌对多菌灵抗性群体在中国麦区的扩散路径。结果表明:江淮流域的江苏、安徽、湖北3省和四川省小麦赤霉病菌对多菌灵的抗性或敏感菌株优势群体均是F.asiaticum,而黄淮流域的山东、河南2省及河北省小麦赤霉病菌对多菌灵的敏感菌株优势群体为F.graminearum,抗性菌株优势群体则为F.asiaticum。江苏、安徽、山东和河南抗多菌灵菌株F.asiaticum产生毒素的化学型为3-AcDON和NIV,并以3-AcDON为主。江苏省连续使用多菌灵防治小麦赤霉病长达20多年后才检测到田间抗性菌株,而近年来检测到田间抗性菌株的山东、河南2省用多菌灵防治赤霉病的历史较短,且为偶尔使用,药剂的选择压力相对较小,因此推测山东和河南麦区出现的小麦赤霉病菌抗多菌灵菌株可能是通过种子调运及联合收割机跨区作业等方式从抗药性发生较早的江淮麦区流入的。     

Genetic variation among <Emphasis Type="Italic">Fusarium</Emphasis> isolates from onion,and resistance to <Emphasis Type="Italic">Fusarium</Emphasis> basal rot in related <Emphasis Type="Italic">Allium</Emphasis> species

The aim of this research was to study levels of resistance to Fusarium basal rot in onion cultivars and related Allium species, by using genetically different Fusarium isolates. In order to select genetically different isolates for disease testing, a collection of 61 Fusarium isolates, 43 of them from onion (Allium cepa), was analysed using amplified fragment length polymorphism (AFLP) markers. Onion isolates were collected in The Netherlands (15 isolates) and Uruguay (9 isolates), and received from other countries and fungal collections (19 isolates). From these isolates, 29 were identified as F. oxysporum, 10 as F. proliferatum, whereas the remaining four isolates belonged to F. avenaceum and F. culmorum. The taxonomic status of the species was confirmed by morphological examination, by DNA sequencing of the elongation factor 1-α gene, and by the use of species-specific primers for Fusarium oxysporum, F. proliferatum, and F. culmorum. Within F. oxysporum, isolates clustered in two clades suggesting different origins of F. oxysporum forms pathogenic to onion. These clades were present in each sampled region. Onion and six related Allium species were screened for resistance to Fusarium basal rot using one F. oxysporum isolate from each clade, and one F. proliferatum isolate. High levels of resistance to each isolate were found in Allium fistulosum and A. schoenoprasum accessions, whereas A. pskemense, A. roylei and A. galanthum showed intermediate levels of resistance. Among five A. cepa cultivars, ‘Rossa Savonese’ was also intermediately resistant. Regarding the current feasibility for introgression, A. fistulosum, A. roylei and A. galanthum were identified as potential sources for the transfer of resistance to Fusarium into onion.     

Screening of cultivated and wild adzuki bean for resistance to race 3 of <Emphasis Type="Italic">Cadophora gregata</Emphasis> f. sp. <Emphasis Type="Italic">adzukicola</Emphasis>, cause of brown stem rot

Two diseases of adzuki bean, brown stem rot (BSR, caused by Cadophora gregata f. sp. adzukicola) and adzuki bean Fusarium wilt (AFW, caused by Fusarium oxysporum f. sp. adzukicola), are serious problems in Hokkaido and have been controlled using cultivars with multiple resistance. However, because a new race of BSR, designated race 3, was identified, sources of parental adzuki bean for resistance to race 3 were needed. Therefore, we examined 67 cultivars and lines of cultivated and wild adzuki bean maintained at the Tokachi Agricultural Experiment Station using a root-dip inoculation method. Consequently, nine adzuki bean cultivars, one wild adzuki bean accession and 30 lines (including two lines resistant to all the three races of BSR and AFW) were confirmed to be resistant or tolerant to race 3 of BSR, and we found a cultivar Akamame as well as a wild adzuki bean Acc2515 to be a new source for a resistance gene to the race 3. This cultivar also holds promise as a source of resistance against other races of BSR and AFW.     

Fusarium wilt of basil in Greece: Foliar infection and cultivar evaluation for resistance

Fusarium wilt of basil (Ocimum basilicum), caused byFusarium oxysporum f.sp.basilici, is reported for the first time in Greece. Foliage inoculation of young plants resulted in a downward movement of the pathogen to the crown and roots and 20–30% plant mortality. Of 14 commercial basil cultivars evaluated for possible disease resistance using young plants, six out of eight large-leaved cultivars were found resistant, while all six small-leaved ones were susceptible. http://www.phytoparasitica.org posting Feb. 23, 2004.     

Fungi on roots and stem bases of asparagus in the Netherlands: species and pathogenicity

A survey was made to identify the most important soilborne fungal pathogens of asparagus crops in the Netherlands. Ten plants were selected from each of five fields with a young (1–4 y) first planting, five fields with an old (6–13 y) first planting and five fields with a young replanting. The analysis included fungi present in the stem base and the roots of plants with symptoms of foot and root rot or showing growth decline without specific disease symptoms. Isolates of each species were tested for pathogenicity to asparagus on aseptically grown plantlets on Knop's agar. Symptoms were caused byFusarium oxysporum, F. culmorum, Botrytis cinerea, Penicillium verrucosum var.cyclopium, Cylindrocarpon didymum, Phialophora malorum, Phoma terrestris andAcremonium strictum. F. oxysporum was by far the most common species and was isolated from 80% of the plants. Not all of its isolates were pathogenic to asparagus. Symptoms were caused by 67%, 78% and 93% of the isolates obtained from young first plantings, old first plantings and replantings, respectively.F. culmorum was isolated from 31% of the plants. Two other notorious pathogens of asparagus,F. moniliforme andF. proliferatum, did not occur in our samples.Species causing symptoms in the vitro test that were found on more than 5% of the plants were additionally tested for their pathogenicity in pot experiments.F. oxysporum f.sp.asparagi caused severe foot and root rot, significantly reduced root weights and killed most of the plants.F. culmorum caused lesions on the stem base often resulting in death of the plant.P. terrestris, a fungus only once reported as a pathogen of asparagus, caused an extensive root rot, mainly of secondary roots that became reddish. The fungus was isolated in only a few samples and is not to be regarded as an important pathogen in Dutch asparagus crops.P. malorum caused many small brown lesions on the stem base and incidentally also on the upper part of small main roots. This is the first report of its pathogenicity to asparagus. The fungus is one of the organisms inciting spear rust and it reduced crop quality rather than crop yield.P. verrucosum var.cyclopium andC. didymum did not cause symptoms in pot experiments.Because of its predominance on plants with foot and root rot and its high virulence,F. oxysporum f.sp.asparagi was considered to be the main soilborne pathogen of asparagus in the Netherlands.     

Development of PCR assays to Tri7 and Tri13 trichothecene biosynthetic genes, and characterisation of chemotypes of Fusarium graminearum, Fusarium culmorum and Fusarium cerealis

Fusarium graminearum, Fusarium culmorum and Fusarium cerealis are major causal agents of Fusarium Head Blight (scab) which is a disease of global significance in all cereal growing areas. These fungi produce trichothecene mycotoxins, principally nivalenol (NIV) and deoxynivalenol (DON). Genes Tri13 and Tri7 from the trichothecene biosynthetic gene cluster convert DON to NIV (Tri13) and NIV to 4-acetyl-NIV (Tri7). We have developed positive–negative PCR assays based on these two genes, which accurately indicate a DON or NIV chemotype in F. graminearum, F. culmorum and F. cerealis. These assays are useful in assessing the risk of trichothecene contamination, and can be informative in epidemiological studies. All NIV chemotype isolates studied have functional copies of both Tri13 and Tri7, and all DON-producing isolates have both genes disrupted or deleted. We have identified several mutations in these genes, which are conserved across F. graminearum lineage, RAPD and SCAR groupings and between the three species. There appears to be evidence of inter-species hybridisation within the trichothecene biosynthetic gene cluster.     

Molecular,biological and physiological characterizations of resistance to phenamacril in Fusarium graminearum

Fusarium head blight (FHB), caused by Fusarium graminearum, is one of the most devastating wheat diseases in China. Phenamacril is a novel cyanoacrylate fungicide with a unique chemical structure and specific mode of action against Fusarium spp. In this study, the molecular, biological and physiological characteristics of laboratory‐induced mutants of F. graminearum with resistance to phenamacril were investigated. Compared to the wildtype strains, the phenamacril‐resistant mutants showed obvious defects in various biological and physiological characteristics, including vegetative growth, carbon source utilization, response to oxidative and osmotic stresses, sensitivity to cell wall and cell membrane integrity inhibitors, cell membrane permeability, glycerol accumulation and pathogenicity. The phenotypes of the phenamacril‐resistant mutants exhibited many variations. Sequencing indicated that the three parental strains studied were identical, and the mutants TXR1, TXR2, BMR1, BMR2, SYR1 and SYR2 each had a single point mutation in the amino acid sequence encoded by the myosin‐5 gene (FGSG_01410). These results provide new reference information for future investigations concerning the resistance mechanism of F. graminearum to phenamacril and could offer important relevant data for the management of FHB caused by F. graminearum.     

Effect of benomyl on soil fungi associated with rye. 1. Effect on the incidence of sharp eyespot caused by Rhizoctonia cerealis

Effects of benomyl on incidence of pathogens affecting the culm base of rye were studied in field trials and growth chamber experiments. Spraying of the crop with the fungicide at a high dosage (2.4 kg.ha^–1) resulted in a tenfold increase of sharp eyespot caused byRhizoctonia cerealis and reduced foot rot symptoms caused by fusaria by 50%. In a field trial at a low dosage (0.24 kg.ha^–1) a slight increase of sharp eyespot was observed. In one year, probably because of wet conditions during the infection period, sharp eyespot did not occur in either benomyl-treated or untreated plots, but eyespot caused byPseudocercosporella herpotrichoides was abundant. Its occurrence was reduced from 74% affected culm bases in untreated plots to 8% and 1% in plots that received 0.24 and 2.4 kg.ha^–1 of the fungicide, respectively.In growth chambers seedlings were grown in two sandy soils inoculated withR. cerealis. The soil was kept dry at about 35% of the moisture holding capacity. In plots with benomyl (1 mg.kg^–1; moisture content 11% of fresh weight), fewer seedlings emerged than in plots without the fungicide. This result was highly significant (P<0.01) for one soil but not for the other. The number of seedlings that remained free of disease symptoms was higher (P<0.01) in untreated than in fungicide-treated plots of both soils.Isolates of pathogens obtained from diseased culms were tested for their sensitivity to benomyl. Growth of all of them includingR. cerealis was inhibited, although not always completely suppressed, at 10 g.ml^–1 on potato-dextrose agar. ED₅₀ values of most isolates ofR. cerealis were between 2.2 and 3.1 g.ml^–1. The fungus was slightly but consistently less sensitive thanF. culmorum. Mycelial growth ofF. nivale was appreciably more sensitive than that of the otherFusarium spp. from cereals.P. herpotrichoides andF. nivale were the most sensitive pathogens tested with ED₅₀ values of <1 g.ml^–1. Accordingly,F. nivale was absent on culms from treated plots. In a growth chamber experiment, seedlings were protected from infection by supplying the fungicide (1 mg.kg^–1) to previously inoculated soil.In a laboratory assay the effect of benomyl on microbial antagonism toR. cerealis was estimated for rhizosphere soil. Enhanced incidence of sharp eyespot in treated crops was associated with adverese effects of the fungicide on microbial antagonism. There is presumptive evidence thatR. cerealis is suppressed by bacteria after wet periods during the vegetation period of the crop and by fungi after dry periods. Only fungal antagonism, which may be less effective, is affected by benomyl. The response to benomyl of the microflora in different soils varied. Reasons for this inconsistency are suggested.Samenvatting In veldproeven en in een klimaatkamer werd de invloed van benomyl op het optreden van voetziekten in rogge onderzocht. In veldjes die bespoten waren met een hoge dosis van het fungicide (in totaal 2.4 kg.ha^–1) bleken tienmaal zoveel halmen met scherpe oogvlekken, veroorzaakt doorRhizoctonia cerealis, voor te komen dan in onbespoten veldjes. Daarentegen was voetrot veroorzaakt doorFusarium-soorten met 50% verminderd. In een volgende veldproef, waarbij een voor de praktijk geadviseerde dosis (0.24 kg.ha^–1) was toegepast, werd een lichte toename van scherpe oogvlekken waargenomen.In een ander jaar trad scherpe oogvlekkenziekte in het geheel niet op, ook niet in met benomyl behandelde veldjes. De vochtige omstandigheden tijdens de infectieperiode zijn daarvan waarschijnlijk de oorzaak. Daarentegen werd de oogvlekkenziekte, welke doorPseudocercosporella herpotrichoides werd veroorzaakt, veel aangetroffen. In de onbehandelde veldjes waren 74% van de halmen aangetast tegen 8 en 1% in de veldjes die met het fungicide waren behandeld in doseringen van 0.24 en 2.4 kg.ha^–1.De invloed van het fungicide op de aantasting van kiemplanten werd in klimaatkamerproeven onderzocht. Daartoe werden twee zandgronden metR. cerealis geënt. De grond werd droog gehouden (op 35% van het waterhoudend vermogen). In grond met fungicide (1 mg.kg^–1) was de opkomst minder dan in grond zonder fungicide. Dit was zeer significant (P<0.01) voor één van de beide zandgronden, maar niet voor de andere. Het aantal gezonde kiemplanten was in beide gevallen duidelijk hoger (P<0.01) voor de onbehandelde grond.De isolaten van ziekteverwekkers uit aangetaste halmen werden op hun gevoeligheid voor het fungicide getoetst. Op aardappel-glucoseagar werden alle isolaten in hun groei geremd bij een benomyl-concentratie van 10 g.ml^–1.R. cerealis was iets minder gevoelig danF. culmorum. Voor het overgrote deel van de isolaten vanR. cerealis lag de ED₅₀ waarde tussen 2,2 en 3,1 g.ml^–1. De myceliumgroei vanF. nivale werd meer geremd dan die van de andereFusarium-soorten.P. herpotrichoides enF. nivale waren met een ED₅₀ waarde van <1 g.m.^–1 de gevoeligste pathogenen die uit de halmvoeten werden geïsoleerd. Dat de populatie vanF. nivale in benomylhoudende grond wordt onderdrukt, blijkt uit (1) het feit dat de schimmel niet voorkwam op halmen uit behandelde veldjes en (2) de bescherming tegen infectie van kiemplanten als aan de besmette grond fungicide (1 mg.kg^–1) was toegevoegd.In laboratoriumproeven werd de invloed van benomyl op het microbiële antagonisme in rhizosfeergrond tegenR. cerealis bepaald. Een toename in het optreden van scherpe oogvlekkenziekte in behandelde gewassen bleek gepaard te gaan met een remming van het antagonisme tegen de ziekteverwekker. Er zijn sterke aanwijzingen datR. cerealis na vochtige perioden tijdens de vegetatieperiode door bacteriën wordt onderdrukt en na droge perioden door schimmels. Het antagonisme van de laatste groep lijkt minder effectief te zijn en alleen dit antagonisme wordt door benomyl verlaagd. Tenslotte wordt een mogelijke oorzaak aangegeven voor de ongelijke respons op het fungicide van het microbieel antagonisme in verschillende gronden.     

Allelism of theFcu-1 andFoc genes conferring resistance to fusarium wilt in cucumber

The inheritance of resistance toFusarium oxysporum f.sp.cucumerinum race 1 was determined in the cucumber cv. WIS-248 by analyzing segregation of F₁, F₂, and BC populations of crosses with the susceptible cv. Straight-8. Resistance was conferred by a single dominant gene. In an allelism test, it was proven that theFcu-1 gene, which confers resistance toF. oxysporum f.sp.cucumerinum races 1 and 2 in cucumber cv. SMR-18 and theFoc gene, which confers resistance toF. oxysporum f.sp.cucumerinum race 2 in cucumber cv. WIS-248, are indistinguishable.     

Resistance Reaction of Conifer Species (European Larch, Norway Spruce, Scots Pine) to Infection by Selected Necrotrophic Damping-off Pathogens

Three conifer species (European larch, Norway spruce, Scots pine) were investigated for their resistance to five damping-off pathogens (Rhizoctonia solani, Fusarium solani, F. oxysporum, F. culmorum, F. avenaceum). Inoculation of the primary roots of seedlings with these pathogens caused host cell death which did not prevent the invasive growth of these fungi; seedlings that had formed secondary and tertiary roots could overcome the infection to a significant degree. Infections with R. solani caused significant mortality to all the conifer species. In contrast, the tree species expressed different levels of resistance when challenged with the Fusarium isolates, with Norway spruce being the most resistant compared to uninoculated controls. Some of the Fusarium isolates were more pathogenic to certain hosts than others; F. oxysporum for European larch, F. avenaceum for Scots pine, F. solani for European larch; only F. culmorum was significantly pathogenic to Norway spruce. No significant differences in disease severity were observed at different soil pH (4.3–7.5). Disease progression was delayed at lower (10–15°C) rather than higher temperatures (20–25°C).     

玉米内生菌L10的分离、鉴定及拮抗活性

为获得对玉米茎腐病主要病原菌禾谷镰孢Fusarium graminearum有明显拮抗作用的玉米内生菌,采用平板对峙法从成熟健康玉米茎秆中筛选禾谷镰孢拮抗菌株,并分析其抗菌谱;通过形态特征、生理生化特性及16S rDNA序列分析进行菌种鉴定;利用盆栽生防试验检测其对玉米茎腐病的防治效果。结果表明,共分离获得了164株玉米内生细菌菌株,其中L10菌株对禾谷镰孢具有较好的抑制效果,抑菌圈半径达1.68 cm;该菌对玉米大斑病菌Setosphaeria turcica、层出镰孢F. proliferatum、禾谷镰孢F. graminearum、拟轮枝镰孢F. verticilliodes、玉米弯孢叶斑病菌Curvularia lunata、玉米小斑病菌Bipolaris maydis、立枯丝核菌Rhizoctonia solani、茄链格孢Alternaria solani共8种植物病原菌均有拮抗作用,尤其对禾谷镰孢抑制效果最佳;结合形态特征、生理生化性质及16S rDNA序列分析,将L10菌株鉴定为多粘类芽胞杆菌Paenibacillus polymyxa。L10菌株脂肽类物质对禾谷镰孢菌具有较好的抑制活性,且盆栽生防试验结果显示该菌株对玉米茎腐病具有一定的防治效果。表明菌株L10对玉米镰孢茎腐病的防治具有一定潜力。     

黄淮海夏玉米主产区穗腐病病原菌的分离鉴定

为明确我国黄淮海夏玉米主产区玉米穗腐病的病原菌种类、优势种群及虫害、年度、省份对病原菌的影响,以形态学为基础,结合分子生物学方法对2013、2015年随机采自河南、河北、山东3省的155份玉米穗腐病样品进行分离鉴定。结果表明,引起黄淮海夏玉米主产区玉米穗腐病的主要致病菌为镰孢菌Fusarium spp.,包括拟轮枝镰孢F.verticillioides、禾谷镰孢F.graminearum、层出镰孢F.proliferatum、木贼镰孢F.equiseti及藤仓镰孢F.fujikuroi,分离频率分别为49.7%、28.4%、12.3%、3.9%和1.3%;其次为木霉菌Trichoderma spp.,包括哈茨木霉T.harzianum、绿色木霉T.viride和棘孢木霉T.asperellum,分离频率分别为8.4%、3.2%和5.2%;青霉菌Penicillium spp.分离频率较低,为14.2%;曲霉菌Aspergillus spp.包括黑曲霉A.niger和黄曲霉A.flavus,分离频率分别为2.6%和1.9%。研究表明,黄淮海主产区玉米穗腐病优势病原菌为拟轮枝镰孢、禾谷镰孢和木霉菌,不同省份不同年度间病原菌种类及优势病原菌存在差异,虫害能加重玉米穗腐病的发生。     

甘草根腐病病原菌鉴定

 甘草(Glycyrrhiza uralensis Fish.)别名甜草、蜜草、甜根子, 为豆科多年生草本植物, 以根与根茎入药, 具有补脾益气、清热解毒、祛痰止咳、缓急止痛、调和诸药之功效, 是我国临床常用的中药材, 也可用作食品添加剂。甘草主要分布在我国的内蒙古、甘肃和宁夏, 在青海、陕西、新疆、黑龙江、辽宁、吉林、河北、山西等地局部地区也有分布。宁夏盐池县是我国乌拉尔甘草的重要产区, 面积大、贮量多、品质好, 1995年被誉为“中国甘草之乡” ^[1]。近年来, 野生甘草遭到了大规模采挖, 甘草蕴藏量急剧减少, 目前主要通过人工种植来满足市场需求。随着甘草种植面积的不断扩大, 甘草病虫害日趋严重, 根腐病危害尤为突出, 直接影响甘草的产量和品质, 造成巨大经济损失。     

Effect of solarization and fumigant applications on soilborne pathogens and root-knot nematodes in greenhouse-grown tomato in Turkey

A study was conducted in two greenhouses with a history of Fusarium crown and root rot (Fusarium oxysporum f.sp.radicis-lycopersici, Forl) and root-knot nematodes (Meloidogyne javanica andM. incognita). During the 2005–06 growing season, the effectiveness of soil disinfestation by solarization in combination with low doses of metham-sodium (500, 750, 1000 and 1250l ha⁻¹) or dazomet (400 g ha⁻¹), was tested against soilborne pathogens and nematodes in an attempt to find a suitable alternative to methyl bromide, which is soon to be phased out. Solarization alone was not effective in the greenhouse with a high incidence ofForl. In the greenhouse with a low level ofForl, all the treatments tested reduced disease incidence, and were therefore considered to be applicable for soil disinfestation. In addition, root-knot nematode density decreased with all the treatments tested in both of the greenhouses.     

Sensitivity ofFusarium graminearum to fungicide JS399-19:In vitro determination of baseline sensitivity and the risk of developing fungicide resistance

The baseline sensitivity ofFusarium graminearum Schwade [teleomorph =Gibberella zeae (Schweinitz) Petch] to the fungicide JS399-19 (development code no.) [2-cyano-3-amino-3-phenylacrylic acetate] and the assessment of risk to JS399-19 resistancein vitro are presented. The mean EC₅₀ values for JS399-19 inhibiting mycelial growth of three populations of wild-typeF. graminearum isolates were 0.102±0.048, 0.113±0.035 and 0.110±0.036 μg ml⁻¹, respectively. Through UV irradiation and selection for resistance to the fungicide, we obtained a total of 76 resistant mutants derived from five wild-type isolates ofF. graminearum with an average frequency of 1.71 × 10⁻⁷% and 3.5%, respectively. These mutants could be divided into three categories of resistant phenotypes with low (LR), moderate (MR) and high (HR) level of resistance, determined by the EC₅₀ values of 1.5–15.0 μg ml⁻¹, 15.1–75.0 μg ml⁻¹ and more than 75.0 μg ml⁻¹, respectively. There was no positive cross-resistance between JS399-19 and fungicides belonging to other chemical classes, such as benzimidazoles, ergosterol biosynthesis inhibitors and strobilurins, suggesting that JS399-19 presumably has a new biochemical mode of action. Although the resistant mutants appeared to have comparable pathogenicity to their wild-type parental isolates, they showed decreased mycelial growth on potato-sucrose-agar plates and decreased sporulation capacity in mung bean broth. Nevertheless, most of the resistant mutants possessed fitness levels comparable to their parents and had MR or HR levels of resistance. As these studies yielded a high frequency of laboratory resistance inF. graminearum, appropriate precautions against resistance development in natural populations should be taken into account. http://www.phytoparasitica.org posting August 7, 2008.     

Effect of Dose Rate of Azoxystrobin and Metconazole on the Development of Fusarium Head Blight and the Accumulation of Deoxynivalenol (DON) in Wheat Grain

Glasshouse studies were undertaken to determine if fungicides used for the control of Fusarium head blight (FHB) result in elevated concentrations of the trichothecene mycotoxin, deoxynivalenol (DON) in harvested wheat grain. Metconazole and azoxystrobin, at double, full, half or quarter the manufacturer's recommended dose rate, were applied to ears of wheat (cv. Cadenza), artificially inoculated with conidia of either Fusarium culmorum or F. graminearum. Metconazole demonstrated high activity against both pathogens, reducing significantly the severity of FHB and the DON concentrations at each of the four dose rates tested when compared to untreated controls. Applications of azoxystrobin significantly reduced FHB and DON compared to unsprayed controls. However, their effectiveness was significantly less than that of metconazole and no dose rate response was observed. Quantification of the amount of trichothecene-producing Fusarium present in harvested grain was determined using a competitive PCR assay based on primers derived from the trichodiene synthase gene (Tri5). Simple linear regression analyses revealed strong relationships between the amount of trichothecene-producing Fusarium present in grain and the DON concentrations (r ²=0.72–0.97). It is concluded that fungicides, applied for the control of FHB, affect DON concentrations indirectly by influencing the amount of trichothecene-producing Fusarium species present in wheat grain. There was no evidence that fungicide applications directly increase the concentration of DON in grain.     

水稻恶苗病菌田间抗咪鲜胺菌株的适合度及其交互抗性

恶苗病是水稻生产上较为严重的种传真菌病害,咪唑类广谱内吸性杀菌剂咪鲜胺是目前防治该病害的主要药剂。以对咪鲜胺抗性及敏感的田间水稻恶苗病菌为试材,研究了其适合度及对几种常用杀菌剂的交互抗性。结果显示:抗性菌株的抗药性可稳定遗传,其温度敏感性与敏感菌株无明显差异,部分抗性菌株在菌丝生长速率、产孢量、孢子萌发率和致病力方面显著高于田间敏感菌株;咪鲜胺与三唑类及2-氰基丙烯酸酯类杀菌剂之间均无交互抗性。研究表明,对咪鲜胺产生抗性的水稻恶苗病菌具有较强的适合度,在田间自然条件下有可能形成优势群体,因此需合理轮换使用不同作用机制的杀菌剂,以延缓其抗药性的发展。     

Fusarium rot of hyacinth caused by <Emphasis Type="Italic">Gibberella zeae</Emphasis> (anamorph: <Emphasis Type="Italic">Fusarium</Emphasis><Emphasis Type="Italic"> graminearum</Emphasis>)

Severe rot of leaves, peduncles and flowers caused by Gibberella zeae (anamorph: Fusarium graminearum) was found on potted plants of hyacinth (Hyacinthus orientalis), a liliaceous ornamental, in greenhouses in Kagawa Prefecture, Japan, in January 2001. This disease was named “Fusarium rot of hyacinth” as a new disease because only the anamorph, F. graminearum, was identified on the diseased host plant. The authors contributed equally to this work. The fungal isolate and its nucleotide sequence data obtained in this study were deposited in the Genebank, National Institute of Agrobiological Sciences and the DDBJ/EMBL/GenBank databases under the accession numbers MAFF239499 and AB366161, respectively.