小麦穗组织中脱氧镰刀菌烯醇毒素的免疫细胞化学定位

 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     

不同小麦品种(系)对赤霉病的抗性和麦穗组织中DON毒素积累分析

 为明确不同小麦品种(系)对赤霉病的抗性和麦穗组织中DON毒素积累水平,培育和利用抗赤霉病和DON毒素积累的品种提供资源和依据,本研究采用单小花滴注接种法对河南省的106个小麦品种(系)抗赤霉病性进行鉴定分析,并用ELISA测定了病穗组织中DON毒素水平。结果表明不同小麦品种(系)对赤霉病的抗性有显著差异,106个小麦品种(系)中未发现抗病和中抗材料,中感品种(系)有华育198、郑麦103和春丰0021等14个,占13.2%;感病的有曌式2010-06、百农898和中麦63等92个,占86.8%。不同小麦品种(系)籽粒、颖壳和穗轴中DON毒素积累水平有显著差异,籽粒中DON毒素水平在(0.70~287.63)mg/kg之间,其中郑03876、豫保1号和中麦63 的DON毒素水平在2 mg/kg 以下,为抗毒素材料;其他的103个品种DON毒素水平大于2 mg/kg;颖壳和穗轴中的DON毒素水平在(51.03~392.87)mg/kg之间,普遍比籽粒中DON毒素含量高。籽粒中DON毒素水平与小麦品种(系)的平均病害严重度间呈极显著正相关。     

长江中下游地区亚洲镰刀菌NX-2毒素群体的检测

由禾谷镰刀菌复合种(Fusarium graminearum species complex,FGSC)引起的麦类赤霉病,是农业生产上的重要病害。为明确中国长江中下游冬小麦主产区小麦赤霉病菌种的构成及其地理分布,对2008年从江苏、浙江和湖北3省采集的656株小麦赤霉病菌株进行了分类鉴定。结果显示,其中558个菌株为亚洲镰刀菌(Fusarium asiaticum),98个菌株为禾谷镰刀菌(Fusarium graminearum sensu stricto),表明中国长江中下游冬小麦主产区小麦赤霉病的主要致病菌是亚洲镰刀菌。选择亚洲镰刀菌(F.asiaticum)为研究对象,通过PCR-RFLP的方法对其进行产NX-2毒素菌株的检测。结果没有检测到产NX-2毒素菌株,表明中国长江中下游冬小麦主产区并未出现NX-2毒素群体。本研究旨在了解NX-2毒素群体在中国长江中下游地区的地理分布,为进一步研究麦类赤霉病菌群体遗传多样性和演化趋势奠定基础,为麦类赤霉病的防治和毒素污染的控制提供理论依据。     

禾谷镰刀菌组蛋白乙酰转移酶FgHat1在DON毒素生物合成中的功能

 由禾谷镰刀菌引起的小麦赤霉病是小麦最主要的真菌病害之一。禾谷镰刀菌侵染小麦时分泌脱氧雪腐镰刀菌烯醇(DON)真菌毒素,威胁人畜健康。本研究通过对组蛋白乙酰转移酶基因FgHAT1的功能研究,发现该基因编码的蛋白定位于细胞核并调控组蛋白H4的乙酰化。Fghat1敲除突变体在生长发育和致病过程中表现正常,但毒素合成存在显著缺陷。敲除FgHATI导致参与DON毒素合成的TRI基因转录水平降低,突变体产毒相关的细胞分化也表现异常。外源添加cAMP可以有效回复突变体的产毒缺陷,表明FgHat1对毒素的调控与cAMP信号通路有关。研究结果表明组蛋白表观修饰和胞内信号通路之间存在联系,这两者的交叉互作对DON毒素合成的精确调控至关重要。     

河南省首次发现梨孢镰刀菌引起的小麦赤霉病

2015年从河南省田间小麦赤霉病病穗上分离得到一种生长速度较慢的镰刀菌, 通过形态学和分子鉴定明确其分类地位, 通过田间单小花滴注法和喷雾法接种测定其致病力, 并通过高效液相色谱串联质谱分析对麦穗中的毒素种类进行测定, 明确其产毒特征?结果表明:分离得到的8个菌株均为梨孢镰刀菌, 在马铃薯葡萄糖琼脂(PDA)培养基上为白色菌落, 菌落底部产生少量红色色素, 平均生长速度为13.3 mm/d; 小型分生孢子为椭球形葡萄状, 平均大小为7.1 μm×5.8 μm, 未见大型分生孢子和厚垣孢子; 致病力弱, 且不侵染穗轴, 单小花滴注法接种条件下平均病级为0.1, 喷雾法接种条件下平均病小穗率为6.5%; 供试的8个镰刀菌菌株均不产生T-2和HT-2毒素, 均产生雪腐镰刀菌烯醇(NIV)毒素, NIV毒素含量水平为371.74~5 282.80 μg/kg, 其中3个菌株产生少量脱氧雪腐镰刀菌烯醇(DON)毒素(86.13~227.22 μg/kg)?     

小麦赤霉病穗组织中黄色镰刀菌和DON毒素的定量分析

为阐明小麦赤霉病穗组织中黄色镰刀菌Fusarium culmorum(简称Fc)与毒素产生之间的关系.分别用TaqMan探针实时定量PCR和ELISA法,对接种4天后的17个小麦品系穗组织中的FcDNA和脱氧雪腐镰刀菌烯醇(DON)含量进行了测定.在Fc DNA定量检测中,TaqMan探针实时PCR技术具有高度的稳定性和可重复性.不同小麦品系的Fc DNA含量差异显著,感病品种比抗病品种高100倍左右;DON含量也不相同,Fc生物量的增加伴随着DON产量的提高,两者之间存在对数关系(y=19758 lnx-111230).菌量较低时,DON含量相对较大;而菌量较高时,DON含量相对偏低.     

不同小麦品种抗赤霉病性类型和抗毒素积累能力分析

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。     

小麦四种镰刀菌毒素的ELISA检测及毒素污染分析

为明确镰刀菌毒素在小麦中的污染与分布状况,选取以四种镰刀菌毒素的特异性抗体制备的ELISA试剂盒,检测了来自全国范围内的183个冬小麦、46个春小麦品种(系)样品中的镰刀菌毒素含量,并对不同麦区小麦样品中脱氧雪腐镰刀菌烯醇(DON)、伏马(fumonisin)、单端孢霉烯醇(T-2)和玉米赤霉烯酮(ZON)四种毒素含量及污染率进行比较分析.结果表明,我国小麦品种(系)中镰刀菌毒素污染比较普遍,但污染水平较低,均符合国家规定标准.四种镰刀菌毒素中以DON毒素为主,其平均含量为73.87μg/kg.不同麦区间DON、fumonisin和T-2毒素的含量存在显著差异(P<0.01),而ZON毒素的含量在麦区间差异不显著(P>0.05),东北春麦晚熟组、长江中下游冬麦组和黄淮冬麦区南片冬水组镰刀菌毒素总体含量较高.DON与ZON毒素污染率在麦区间存在显著差异(P<0.01),而fumonisin与T-2毒素污染率差异不显著(P>0.05).在不同麦区间,东北春麦晚熟组的DON毒素的含量和污染率均为最高.     

不同杀菌剂对小麦赤霉病及籽粒DON毒素的控制效果

为探明不同杀菌剂对小麦赤霉病和小麦籽粒DON毒素(包括DON、3-ADON和15-ADON)的控制效果, 采用菌丝生长速率法测定了12种药剂对禾谷镰刀菌野生型菌株PH-1的室内活性, 同时采用液相色谱-串联质谱法(LC-MS)测定了这些药剂对DON毒素的抑制效果, 并开展了小麦赤霉病及籽粒DON毒素的田间防治试验。结果表明, 12种原药对菌丝生长抑制活性强弱依次为氟唑菌酰羟胺>咪鲜胺>戊唑醇>丙硫菌唑>叶菌唑>氰烯菌酯>氟环唑>多菌灵>甲基硫菌灵>吡唑醚菌酯>嘧菌酯>井冈霉素A。氟环唑EC50和EC90离体胁迫均刺激DON毒素产生, 其他杀菌剂EC50和EC90胁迫均抑制DON毒素产生。田间试验结果表明, 200 g/L氟唑菌酰羟胺SC、30%丙硫菌唑OD和20%叶菌唑WP病指防效和DON防效为87.68%~94.77%; 430 g/L戊唑醇SC、25%氰烯菌酯SC、45%咪鲜胺EW、25%氟环唑SC、50%多菌灵WP和70%甲基硫菌灵WP病指防效和DON防效为57.63%%~85.49%; 250 g/L吡唑醚菌酯EC和250 g/L嘧菌酯SC病指防效分别为72.18%和51.98%, DON防效分别为43.06%和-7.96%; 24%井冈霉素A AS病指防效和DON防效分别为42.37%和62.87%。药剂离体和田间控毒效果不完全一致, 赤霉病有效防控是DON防控的前提, 病害防效与DON防效不完全一致, 本研究为小麦赤霉病及籽粒DON毒素防控提供了科学依据。     

我国不同麦区小麦镰刀菌毒素检测及毒素污染分析

为明确镰刀菌毒素在我国不同小麦产区的污染与分布,利用超高效液相色谱-串联质谱仪对全国214个小麦品种籽粒中的7种镰刀菌毒素含量进行了测定和比较分析。结果表明:7种镰刀菌毒素的平均回收率在95.02%～116.52%之间,相对标准差为6.32%～21.50%。所有测试小麦样品均被镰刀菌毒素污染,7种毒素中以雪腐镰刀菌烯醇(nivalenol,NIV)含量最高,即使在NIV含量最低的长江中下游冬麦区,平均含量也高达182.56μg/kg,虽然目前并没有关于NIV的相关限量标准,但也应该予以足够重视;脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)含量较高,在8.07～337.23μg/kg之间,低于国家限量标准1 000μg/kg;T-2含量在4.05～78.05μg/kg之间,低于现有国际限量标准100μg/kg;玉米赤霉烯酮(zearalmone,ZEA)含量在5.30～12.50μg/kg之间,低于国家限量标准60μg/kg;其它3种毒素含量较低,均低于17.73μg/kg。不同麦区间毒素比较发现,DON和3-乙酰脱氧雪腐镰刀菌烯醇的含量在长江中下游冬麦区最高,分别为43.09μg/kg和6.39μg/kg,且显著高于其它麦区;来自长江中下游冬麦组和黄淮冬麦区南片水地组的小麦品种籽粒中毒素T-2和HT-2的含量显著高于其它麦区;其余毒素在不同麦区间均无显著差异。     

Real-time PCR Quantification and Mycotoxin Production of Fusarium graminearum in Wheat Inoculated with Isolates Collected from Potato, Sugar Beet, and Wheat

ABSTRACT Fusarium graminearum causes Fusarium head blight (FHB) in small grains worldwide. Although primarily a pathogen of cereals, it also can infect noncereal crops such as potato and sugar beet in the United States. We used a real-time polymerase chain reaction (PCR) method based on intergenic sequences specific to the trichodiene synthase gene (Tri5) from F. graminearum. TaqMan probe and primers were designed and used to estimate DNA content of the pathogen (FgDNA) in the susceptible wheat cv. Grandin after inoculation with the 21 isolates of F. graminearum collected from potato, sugar beet, and wheat. The presence of nine mycotoxins was analyzed in the inoculated wheat heads by gas chromatography and mass spectrometry. All isolates contained the Tri5 gene and were virulent to cv. Grandin. Isolates of F. graminearum differed significantly in virulence (expressed as disease severity), FgDNA content, and mycotoxin accumulation. Potato isolates showed greater variability in producing different mycotoxins than sugar beet and wheat isolates. Correlation analysis showed a significant (P < 0.001) positive relationship between FgDNA content and FHB severity or deoxynivalenol (DON) production. Moreover, a significant (P < 0.001) positive correlation between FHB severity and DON content was observed. Our findings revealed that F. graminearum causing potato dry rot and sugar beet decay could be potential sources of inoculum for FHB epidemics in wheat. Real-time PCR assay provides sensitive and accurate quantification of F. graminearum in wheat and can be useful for monitoring the colonization of wheat grains by F. graminearum in controlled environments, and evaluating wheat germplasms for resistance to FHB.     

Effect of infection timing on fusarium head blight and mycotoxin accumulation in open- and closed-flowering barley

ABSTRACT Barley has two flowering types, chasmogamous (open-flowering) and cleistogamous (closed-flowering). We examined the effect of the timing of Fusarium graminearum infection on Fusarium head blight (FHB) and mycotoxin accumulation in barley cultivars with different flowering types using greenhouse experiments. In the first experiment, 13 cultivars were spray inoculated at two different developmental stages, and the severity of FHB was evaluated. The effect of the timing of infection differed among cultivars. Cleistogamous cultivars were resistant at anthesis but susceptible at 10 days after anthesis, whereas chasmogamous cultivars were already susceptible at anthesis. In the second experiment, five cultivars were inoculated at three different developmental stages and the concentrations of deoxynivalenol (DON) and nivalenol (NIV) in mature grain were analyzed. Cleistogamous cultivars accumulated more mycotoxins (DON and NIV) when inoculated 10 or 20 days after anthesis than when inoculated at anthesis, whereas chasmogamous cultivars accumulated more mycotoxins when inoculated at anthesis. Thus, the most critical time for F. graminearum infection and mycotoxin accumulation in barley differs with cultivar, and likely is associated with the flowering type. Late infection, even without accompanied FHB symptoms, was also significant in terms of the risk of mycotoxin contamination.     

Pathogenicity and In Planta Mycotoxin Accumulation Among Members of the Fusarium graminearum Species Complex on Wheat and Rice

ABSTRACT Fusarium head blight (FHB), or scab, is a destructive disease of small grains caused by members of the Fusarium graminearum species complex, comprised of at least nine distinct, cryptic species. Members of this complex are known to produce mycotoxins including the trichothecenes deoxynivalenol (DON) along with its acetylated derivatives and nivalenol (NIV). In this study, 31 strains, belonging to eight species of this complex and originating from diverse hosts or substrates, were tested for differences in aggressiveness and mycotoxin production. Large variation among strains, both in terms of their aggressiveness and the ability to produce trichothecenes on a susceptible cultivar of wheat was found; variation appears to be a strain-specific rather than species-specific characteristic. While pathogenicity was not influenced by the type of mycotoxin produced, a significant correlation was observed between the amount of the dominant trichothecene (DON and its acetylated forms or NIV) produced by each strain and its level of aggressiveness on wheat. Some isolates also were tested for their ability to infect rice cv. M201, commonly grown in the United States. While tested strains were capable of infecting rice under greenhouse conditions and causing significant amount of disease, no trichothecenes could be detected from the infected rice florets.     

Trichothecene mycotoxin genotypes of Fusarium graminearum sensu stricto and Fusarium meridionale in wheat from southern Brazil

A total of 82 fungal isolates was obtained from wheat kernel samples affected by fusarium head blight collected from 20 locations in southern Brazil. Polymerase chain reaction (PCR) assays were used to characterize trichothecene mycotoxin genotypes [deoxynivalenol (DON), nivalenol (NIV) and two acetylated derivatives of DON]. To identify isolates that producing DON and NIV, portions of the Tri13 gene were amplified. To identify 3-acetyl-deoxynivalenol (3-ADON) and 15-acetyl-deoxynivalenol (15-ADON) genotypes, portions of Tri3 and Tri12 were amplified. Nearly all of the isolates studied (76/82) were of the DON/15-ADON genotype. Six of the isolates were of the NIV genotype. The DON/3-ADON genotype was not observed. Portions of three genes were sequenced from representative isolates of the NIV and DON/15-ADON genotypes and compared with sequences from curated reference isolates of Fusarium in GenBank. blast queries for individual gene sequences and pairwise comparisons of percentage identity and percentage divergence based on 1676 bp of concatenated DNA sequence suggested that the isolates representing the DON/15-ADON genotype were Fusarium graminearum sensu stricto and the isolates representing the NIV genotype were Fusarium meridionale . This is the first detailed report of trichothecene mycotoxin genotypes of F. graminearum and F. meridionale in Brazil.     

Within-field variation of Fusarium graminearum isolates for aggressiveness and deoxynivalenol production in wheat head blight

Fusarium head blight (FHB), caused by Fusarium graminearum sensu stricto (s.s.), causes tremendous annual yield losses in wheat worldwide. Variation of aggressiveness of isolates from individual field populations in terms of FHB infection and deoxynivalenol (DON) concentration in the host are important population parameters reflecting parasitic ability. Our main objective was to estimate the variation of both traits within three populations of F. graminearum s.s., each consisting of 30 single-spore isolates collected from small wheat fields in Germany, and to compare it with 11 isolates of a collection (F. graminearum collection) from four countries. The same isolates were characterized using 19 single-sequence repeat markers. All isolates were spray inoculated on a moderately resistant spring wheat cultivar at two field locations over 2 years (i.e., in four environments). The genotypic proportion of phenotypic variance (σ(2)(G)) within populations was significant (P < 0.01) for both traits, and the σ(2)(G) × environment interaction was even more important for mean FHB severity. Ranges in mean FHB severity and DON concentration in the host were only slightly smaller for the field populations than for the F. graminearum collection. Both traits were significantly (P < 0.05) correlated within and across populations. A further partitioning of σ(2)(G) revealed 72% of σ(2)(G) within and 28% of σ(2)(G) across populations for both traits. Molecular variance of the three populations was similarly distributed (73.6% within versus 26.4% between populations). In view of this high within-field variation for traits of parasitic ability and selection, neutral molecular markers, multiple resistance genes of different origin should be employed in wheat breeding programs to obtain a long-term stable FHB resistance.     

Evaluation of type I fusarium head blight resistance of wheat using non-deoxynivalenol-producing fungi

A series of experiments was conducted to determine whether type I resistance (resistance to initial infection) to fusarium head blight (FHB) in wheat could be assessed using fungal species/isolates that do not produce deoxynivalenol (DON), a mycotoxin critical to the spread of Fusarium graminearum in the wheat spike. It was shown that, while the non-toxin-producing species Microdochium nivale and M. majus could infect following spray inoculation of wheat spikes, they were unable to spread within the spike following point inoculation. However, although these species might reveal type I resistance, they are not highly pathogenic towards wheat. A nivalenol (NIV)-producing isolate of F. graminearum caused high levels of disease following spray inoculation, but spread only very slowly within the spike and rarely induced bleaching above the point of inoculation. It is proposed that spray inoculation with an appropriate, aggressive, non-DON-producing FHB pathogen may be used to characterize type I resistance to complement point inoculation with a DON-producing isolate to assess type II resistance (resistance to spread within the spike).     

Fusarium head blight of cereals in Denmark: species complex and related mycotoxins

Quantitative real-time polymerase chain reaction differentiating 10 Fusarium spp. and Microdochium nivale or M. majus was applied to a total of 396 grain samples of wheat, barley, triticale, oat, and rye sampled across Denmark from 2003 to 2007, along with selected samples of wheat and barley from 1957 to 2000, to determine incidence and abundance of individual Fusarium spp. The mycotoxins deoxynivalenol (DON), nivalenol, zearalenone, T-2, and HT-2 were quantified using liquid chromatography-double mass spectrometry. Major differences in the Fusarium species complex among the five cereals as well as great yearly variation were seen. Fusarium graminearum, F. culmorum, and F. avenaceum were dominant in wheat, with DON as the dominant mycotoxin. F. langsethiae, F. culmorum, and F. avenaceum were dominant in barley and oat, leading to relatively high levels of the mycotoxins T-2 and HT-2. F. graminearum, F. culmorum, and F. avenaceum dominated in triticale and rye. The nontoxigenic M. nivale/majus were present in significant amounts in all cereal species. Wheat and barley samples from 1957 to 1996 exhibited no or very low amounts of F. graminearum, indicating a recent increase of this pathogen. Biomass and mycotoxin data exhibited good correlations between Fusarium spp. and their corresponding mycotoxins under field conditions.     

Effect of nitrogen application at anthesis on Fusarium head blight and mycotoxin accumulation in breadmaking wheat in the western part of Japan

In the western part of Japan, two wheat cultivars, Nishinokaori and Minaminokaori, are currently cultivated for breadmaking. Breadmaking wheat requires a higher protein content compared to the Japanese noodle wheat (the major type of wheat in Japan). This high protein level in the grain is obtained by top-dressing with nitrogen (N) near anthesis. Because such N applications may increase levels of Fusarium head blight (FHB) and consequent mycotoxin [deoxynivalenol (DON) and nivalenol (NIV)] accumulation in the grain, the effect of N application (0, 4, and 8 g/m²) at anthesis on FHB and mycotoxin accumulation in Nishinokaori and Minaminokaori was tested in the greenhouse in 2004 and 2005 and in two fields in 2006. In the greenhouse, plants were spray inoculated at 3, 10, and 20 days after N treatment. In field experiments, colonized maize kernels, which generate ascospores during the testing season, served as inoculum. In all experiments for both cultivars, N application at anthesis significantly increased grain protein as expected, but had no significant effect on FHB and DON and NIV levels in grain. These results suggest that, at least in these cultivars, N can be applied close to anthesis without increasing the risk of FHB and mycotoxin (DON and NIV) accumulation.     

Resistance against <Emphasis Type="Italic">Fusarium graminearum</Emphasis> and the relationship to β-glucan content in barley grains

Fusarium head blight (FHB) caused by Fusarium graminearum (FG) is a destructive disease impacting barley worldwide. The disease reduces the grain yield and contaminates grains with mycotoxins, such as the trichothecene deoxynivalenol (DON). Although the infection mainly affects the grain yield, little is known about its impact on grain structural and biochemical properties. Yet, such information is instrumental to characterize the facets of resistance in the grains. After artificial inoculation of six barley cultivars with FG in a 2 years field test, different levels of symptoms on spikes, of colonisation of grains and of DON content were observed. The infections caused a reduction in grain weight and an average decrease of 10% of the β-glucan content in grains, indicating alterations of grain filling, composition and structure. According to our results, we postulate the presence of two distinct resistance mechanisms in the grain, tolerance to grain filling despite infection as well as the inhibition of mycotoxin accumulation. Differently to wheat, in barley, type IV resistance (tolerance of the grain to infection) is directly linked with type III resistance (resistance against kernel infection). The resistance against toxin accumulation (named type V resistance in wheat) appeared to be independent to all other resistance types. Generally, the resistance was significantly influenced by the environment and by genotype x environment interactions explaining the generally weak stability of resistance in barley. Interestingly, a significant and inverse relationship between DON contamination and β-glucan content in grains suggests that high β-glucan content in grains contributes to type V resistance.     

Maize maturity and the development of gibberella ear rot symptoms and deoxynivalenol after inoculation

Development of gibberella ear rot disease symptoms and the accumulation of the mycotoxin deoxynivalenol (DON) in maize ears inoculated via the silk with Fusarium graminearum was determined at various times after inoculation. Ten hybrids ranging in maturity from early to late, were inoculated with a conidial suspension in 1993 and 1994 and harvested every 2 weeks for 14 weeks after inoculation. Disease symptom evaluations were conducted on all 10 hybrids; five of these hybrids were further analysed for DON concentrations. Disease symptoms reached a maximum and stabilized by 6 weeks after inoculation, approximately at physiological maturity (35% kernel moisture) for the early hybrids and the late dent stage of maturity for later hybrids. Deoxynivalenol accumulation was correlated with symptom development but did not stabilize at 6 weeks for all genotypes. Hybrid maturity did not influence symptom development or DON accumulation, but environment did. For the evaluation of hybrids, assessments of resistance to fungal invasion and mycotoxin accumulation based on symptom development could be made much earlier than the current 12-14 week harvest time commonly used in inoculated experiments.