Interactions Between Fusarium verticillioides, Aspergillus flavus, and Insect Infestation in Four Maize Genotypes in Lowland Africa

ABSTRACT An experiment was designed to compare cycles of selection of four maize genotypes for ear- and grain-quality characteristics, interactions with Aspergillus flavus and Fusarium verticillioides infection, and insect ear infestation in two seasons. Mean infection levels by A. flavus and F. verticillioides were significantly higher in inoculated rows than in the controls. The F. verticillioides-inoculated rows had significantly more coleopteran beetles and lepidopteran borers per ear than the controls and A. flavus-inoculated rows. Genotypes and cycles of selection within genotype were not different with respect to number of insects or percent fungal incidence in the ear, but they were different for husk extension, field weight, 100-grain weight, and grain density. Inoculation with either fungus resulted in significantly higher percentage of floaters (i.e., loss of grain density) and lower grain weight than the controls. Aflatoxin (B1 and B2) in A. flavus-inoculated rows averaged 327 ppb in the first season and 589 ppb in the second (dryer) season. Fumonisin levels in F. verticillioides-inoculated rows did not differ between seasons, with an average of 6.2 ppm across seasons. In the noninoculated control rows, fumonisin was significantly higher in the first (5.3 ppm) than in the second (3.1 ppm) season. For all genotypes, husk extension and yield parameters decreased in the fungal-inoculated treatments. General ear-rot scoring was significantly correlated with incidence of F. verticillioides in kernels and grain-weight loss but not with A. flavus in the grain.     

Infection and Fumonisin Production by Fusarium verticillioides in Developing Maize Kernels

ABSTRACT Fusarium ear rot and fumonisin contamination are serious problems for maize growers, particularly in the southeastern United States. The lack of maize genotypes highly resistant to infection by Fusarium verticillioides or to fumonisin contamination emphasizes the need for management strategies to prevent contamination by this mycotoxin. Information on the initial appearance of infection and fumonisin contamination of kernels and their increase over time is needed to determine if early harvest may be an appropriate control strategy. Maize ears from replicated studies at two locations in eastern North Carolina were harvested weekly, starting 2 weeks after pollination and continuing for 14 weeks. The percentage of kernels infected with F. verticillioides and the fumonisin contamination in the harvested samples were determined. Kernel infection by F. verticillioides and fumonisin contamination appeared as kernels neared physiological maturity and increased up to the average harvest date for maize in North Carolina. Beyond this date, the concentrations of fumonisin fluctuated. Under years conducive for fumonisin contamination, early harvest (greater than 25% grain moisture) may help reduce the level of contamination.     

亚洲玉米螟为害对玉米镰孢穗腐病发生程度的影响

为了明确亚洲玉米螟[Ostrinia furnacalis(Guenée)]在玉米雌穗不同发育期为害对玉米镰孢穗腐病发生程度的影响,分别在春播和夏播玉米吐丝、灌浆和乳熟期,以亚洲玉米螟为试虫,以轮枝镰孢菌为供试菌,单独或复合接种侵染。结果表明,无论春播还是夏播玉米,镰孢穗腐病病情指数均以复合处理最高,为31.57～88.80,单独接虫处理为16.27～55.00,单独接菌处理为19.02～59.47;玉米镰孢穗腐病的发生程度受玉米雌穗发育期和亚洲玉米螟为害程度的影响,其玉米螟为害级别为吐丝期>灌浆期>乳熟期;受螟害的介导作用,春播玉米雌穗被害率、螟害级别和镰孢穗腐病的发病率均显著重于夏播玉米,但夏播玉米镰孢穗腐病的病情指数高于春播玉米。螟害的介入明显加重了玉米镰孢穗腐病的发生,严重程度与雌穗发育时期密切相关。     

亚洲玉米螟与串珠镰孢菌复合侵染对玉米产量损失的影响

在玉米吐丝、灌浆和乳熟期,以亚洲玉米螟Ostrinia furnacalis(Guenée)为试虫,以串珠镰孢菌Fusarium verticillioides为供试菌种,研究玉米雌穗不同发育期亚洲玉米螟与串珠镰孢菌复合侵染对玉米产量的影响.结果表明:串珠镰孢菌与玉米螟复合侵染危害后玉米雌穗穗长缩短、籽粒数减少、穗粒腐病加重,使得玉米的产量性状发生改变,导致玉米产量和品质下降;接虫处理和复合处理均以玉米吐丝期对玉米产量构成因素影响最大,吐丝期复合处理单穗损失率为40.84%,在灌浆和乳熟期分别为27.92%和11.10%;在单独接菌条件下以灌浆期危害最重,单穗损失率为6.62%,而在吐丝和乳熟期分别为4.13%和0.65%.     

叶菌唑对轮枝镰刀菌的活性及作用机制

玉米穗腐病是严重的世界性真菌病害,而轮枝镰刀菌Fusarium verticillioide是我国玉米穗腐病的主要致病菌。为明确叶菌唑在我国玉米穗腐病防治中的应用潜力,采用菌丝生长速率法和孢子萌发法,分别测定了叶菌唑对2019至2021年采自我国山东、河南和江苏3个省份的35株轮枝镰刀菌菌丝生长以及分生孢子形成、萌发和芽管伸长的影响,并测定了该药剂对轮枝镰刀菌产毒(B族伏马毒素,type B fumonisins,FBs)能力及产毒基因表达的影响;通过测定叶菌唑处理后轮枝镰刀菌菌丝麦角甾醇、胞内甘油和丙二醛(MDA)含量以及电导率的变化,探究了其作用机制;同时评价了叶菌唑对田间玉米穗腐病的防治效果。结果显示：叶菌唑对35株轮枝镰刀菌均表现出较强的抑制活性,其对轮枝镰刀菌的菌丝生长以及分生孢子形成、萌发及芽管伸长具有显著的剂量依赖性抑制作用。其中：叶菌唑抑制菌丝生长的EC₅₀值在0.005～0.029μg/mL之间,平均EC₅₀值为(0.012±0.006)μg/mL;EC₅₀浓度的叶菌唑对轮枝镰刀菌分生孢子形成、萌发及芽...     

A mathematical simulation of growth of fusarium in maize ears after artificial inoculation

ABSTRACT Fusarium spp. in maize can contaminate the grain with mycotoxins if environmental conditions are favorable for fungal growth. To quantify the relationship between growth of Fusarium spp. and environmental conditions, a mathematical model was developed to simulate growth of F. graminearum and F. verticillioides on maize ears following silk inoculation in field experiments from 1992 to 1995. Each species was inoculated separately and as a mixture of the two for 3 of the 4 years on one maize hybrid. Disease progress in ears was measured by a visual rating scale that was converted to percent visual infection. Measurements were made at regular time intervals after silks were inoculated 5 days after silk emergence. Differential equations were used to relate growth rates of Fusarium spp. in maize ears to hourly air temperature and relative humidity and to daily precipitation. Integration of these equations over time produced quantitative estimates of fungal growth. Model calculations compared well with measurements (R(2) = 0.931, standard error of estimate [SEE] = 2.11%) of percent visual disease infection of maize ears over 3 years. The model was tested against a second set of data (R(2) = 0.89, SEE = 5.9%) in which silks were inoculated at nine different times after first silk emergence for each of 2 years (1994 and 1995) with the two species of fungi on the same maize hybrid. At this time, a silk function was developed to account for changes in the susceptibility of silks to disease. F. graminearum responded to wet conditions more than F. verticillioides, and for the conditions of this experiment, grew much faster than F. verticillioides when inoculated separately. When they were inoculated together, F. graminearum growth rates were much lower, indicating some interference by F. verticillioides. During 1993, weather conditions before inoculation reduced the growth of both species in silks.     

Relationships between seed germination, fumonisin content, and Fusarium verticillioides infection in selected maize samples from different regions of Costa Rica

Thirty-five maize seed samples from three geographic regions of Costa Rica were analysed for fumonisin content, germinability, and Fusarium verticillioides infection with and without surface disinfection. The concentration of fumonisins in the maize samples varied from 4 ng g⁻¹ to 16 000 ng g⁻¹ with an all over-all average of 2500 ng g⁻¹. There was a significant difference in fumonisin content between samples from Alajuela and Guanacaste. Germination of the seed samples ranged from 12% to 98% with significant differences between regions. F. verticillioides infection was 12–98% and 13–97% for surface-disinfected and nondisinfected seeds, respectively. There was a significant negative correlation ( r  = -0.52) between fumonisin content and seed germination, but no significant correlation was found between fumonisin content and F. verticillioides infection, or between F. verticillioides infection and seed germination. Most of the high fumonisin seed samples had some mechanical or insect damage. Whether or not the fumonisins had a direct effect on germination was not further established. It is concluded that the large differences in fumonisin content of maize seeds within and between regions are primarily caused by differences in seed quality, genetic diversity of F. verticillioides strains in natural populations, climatic differences between regions, and varietal differences. Some of the fumonisin levels found in this study coincide with levels associated with risks to humans and animals in other countries.     

Impact of primary infection site of <Emphasis Type="Italic">Fusarium</Emphasis> species on head blight development in wheat ears evaluated by IR-thermography

The effect of the primary infection site by Fusarium graminearum and F. culmorum within wheat ears on Fusarium head blight (FHB) was investigated under controlled conditions. FHB development was assessed visually and thermographically following inoculation by: (i) spraying ears, or injecting inoculum into spikelets on (ii) tip, (iii) centre and (iv) base of the ears, separately. Fusarium infection significantly increased the temperature span within ears 6 days post inoculation (dpi), especially infections starting at the ear tip. The temperature difference between air and ear was negatively correlated to FHB severity and enabled disease detection even 29 dpi. F. culmorum caused significant higher disease severity neither reflected in the frequency of infected kernels nor in thousand kernel weight (TKW). Spray inoculations had the strongest effect on TKW, whereas tip inoculations had no effect. Centre and base inoculations had intermediate effects on TKW, although FHB levels did not differ with the same trend among inoculation scenarios. The overall low correlations among FHB severity, infected kernels and TKW are explained by the pathogen spread within ears – downwards more than upwards – and the effect on yield formation which is lower for infections of the upper parts of ears. An exponential model showed high goodness of fit for gradients of infected kernels within ears (R ²  ≥ 70) except tip infection with F. culmorum. This study confirmed that FHB is a function of the primary infection site within ears. Thermography was useful to differentiate among infection scenarios and may be applied in breeding for FHB resistance.     

河北省玉米穗腐病病原菌鉴定及潜在产伏马毒素镰孢菌系统发育分析

 为明确河北省夏玉米区玉米穗腐病的发生情况、病原菌组成及地区对病原菌种类的影响,本团队于2016年和2017年在玉米收获前期对玉米穗腐病的发生情况进行调查,并通过生物学方法对随机采回的样本进行分离鉴定。田间调查结果表明,与2016年相比,2017年河北省玉米种植面积有所减少,玉米穗腐病的发病率有所下降。分离鉴定结果表明,引起河北省夏玉米区穗腐病的优势病原菌为拟轮枝镰孢菌(Fusarium verticillioides),分离频率为63.49%,其他病原菌如层出镰孢菌(F. proliferatum)、禾谷镰孢菌(F. graminearum)、变红镰孢菌(F. incarnatum)、黑曲霉(Aspergillus niger)、黄曲霉(A. flavus)、草酸青霉(Penicillium oxalicum)和哈茨木霉(Trichoderma harzianum)的分离频率分别为19.05%、6.35%、1.59%、14.29%、3.17%、9.52%和1.59%。为明确伏马毒素基因在潜在产伏马毒素镰孢菌鉴定中的作用,拟轮枝镰孢菌、层出镰孢菌和藤仓镰孢菌(F. fujikuroi)分别以EF-1α和FUM1基因序列为基础构建系统发育树。结果表明,两个系统发育树拓扑结构相似,伏马毒素基因可用于潜在产伏马毒素镰孢菌种的鉴定,基于FUM1基因的种间遗传距离大于基于EF-1α基因的种间遗传距离,而种内遗传距离的结论则与之相反。     

Relationships among resistances to fusarium and Aspergillus ear rots and contamination by fumonisin and aflatoxin in maize

ABSTRACT Fusarium verticillioides, F. proliferatum, and Aspergillus flavus cause ear rots of maize and contaminate the grain with mycotoxins (fumonisin or aflatoxin). The objective of this study was to investigate the relationships between resistance to Fusarium and Aspergillus ear rots and fumonisin and aflatoxin contamination. Based on a previous study of 143 recombinant inbred lines from the cross NC300 x B104, 24 lines with the highest and 24 lines with the lowest mean fumonisin concentration were selected for further evaluation. Paired plots of each line were inoculated with F. verticillioides and F. proliferatum or with A. flavus in replicated trials in 2004 and 2005 in Clayton, NC, and College Station, TX. The low-fumonisin group had significantly lower levels of fumonisin, aflatoxin, and Fusarium and Aspergillus ear rots. Across year-location environments, all four traits were significantly correlated; the genotypic correlation (r(G)) ranged from r(G) = 0.88 (aflatoxin and Aspergillus ear rot) to r(G) = 0.99 (Fusarium and Aspergillus ear rots). Quantitative trait loci (QTLs) were identified and their effects estimated. Two QTLs affected both toxin concentrations, one QTL affected both ear rots, and one QTL affected Aspergillus and Fusarium rots and fumonisin. These results suggest that at least some of the genes involved in resistance to ear rots and mycotoxin contamination are identical or genetically linked.     

The discovery of pyridalyl: a novel insecticidal agent for controlling lepidopterous pests

Synthesis of analogues of two compounds with known insecticidal activity, both of which contain a 3,3-dichloro-2-propenyloxy group, produced 2-(trifluoromethyl)-4-phenoxyphenyl 3,3-dichloro-2-propenyl ether, which had weak activity against lepidopterous larvae. Structural modifications around this lead compound led to the development of pyridalyl [Pleo, S-1812; 2,6-dichloro-4-(3,3-dichloroallyloxy)phenyl 3-[5-(trifluoromethyl)-2-pyridyloxy]propyl ether], which belongs to a new class of insecticides. Pyridalyl gives very good control of various lepidopterous and thysanopterous pests on cotton and vegetables, without phytotoxicity. It controls populations of Heliothis virescens F and Plutella xylostella (L) which are resistant to various currently used insecticides. It also produces unique insecticidal symptoms, so it may have a different mode of action from other existing insecticides. Pyridalyl is also less harmful than existing insecticides to various beneficial arthropods, so it should provide an important tool in IPM and insecticidal management programmes for the control of lepidopterous and thysanopterous pests. The first market introduction is expected in Japan and some Asian countries in the years between 2004 and 2005.     

安徽省玉米穗腐病主要致病镰孢菌的分离与鉴定

为明确安徽省玉米穗腐病主要致病镰孢菌的种类,采用单孢分离、形态学鉴定和分子生物学鉴定的方法,对2017年采集自安徽省6个玉米主产区——合肥市、蚌埠市、淮北市、阜阳市、亳州市和宿州市的玉米穗腐病病样进行了分离鉴定。在获得的455株镰孢菌中,拟轮枝镰孢菌、禾谷镰孢复合种、层出镰孢菌、尖孢镰孢菌和黄色镰孢菌的分离频率分别为59.13%、21.28%、13.68%、5.12%和0.79%。其中拟轮枝镰孢菌在各地的分布最广,属于安徽省的优势致病镰孢菌。     

甘肃玉米穗腐病样品中轮枝镰孢菌的分离鉴定及生物学特性

 为了明确甘肃玉米穗腐病的病原种类, 于2009年9月在甘肃省四大生态区采集玉米穗腐病样品, 以组织分离法进行病原物的分离培养, 对分离得到的镰孢菌菌落进行纯化和单孢分离后, 以形态学为基础, 参照Leisle分类系统进行鉴定。结果表明:共分离到271株镰孢菌菌株, 其中以轮枝镰孢菌(Fusarium verticillioides)和黄色镰孢菌(F. culmorum)出现频率高, 属优势种。按照柯赫氏法则对沈单16和金穗96832进行致病性测定, 证实了轮枝镰孢菌对玉米果穗的致病性。选取2株轮枝镰孢菌菌株进行EF-1α(tef) 基因序列分析, 将PCR产物回收测序后在GenBank上比对, 菌株GSLT4-3-2与GenBank中登记的轮枝镰孢菌FN179339、FN179345和FN179338;GSTS24-2-1与轮枝镰孢菌FN179343、FN179346、FN179340、FN179344和 FN252384亲缘关系最近, 序列同源性达100%。轮枝镰孢菌GSYC17-2-1的生长温度范围为15～35℃, 最适温度为28℃, pH 值为8;菌落在pH值为4～10的培养基上能够迅速扩展, pH值为7时产孢量最大;碳源对轮枝镰孢菌菌丝生长影响相对稳定, 而氮源对其营养生长影响的变幅较大;完全黑暗条件下, 菌丝扩展最快, 12 h光暗交替对产孢量有明显的促进作用;病原菌菌丝致死温度为70℃, 10 min。     

Control of loose smut (Ustilago nuda and U. tritici) infections in barley and wheat plants by foliar application of triadimefon

Foliar application of the broad-spectrum systemic fungicide triadimefon to loose smut-infected plants of wheat and barley produced a marked curative effect. Triadimefon was particularly effective when applied at the beginning of and during stem extension. In addition to reducing percentage plant infection, foliar application of this fungicide (particularly prior to flag leaf emergence) resulted in an increased proportion of infected ears which were only partially smutted. The curative effect of these early fungicide applications resulted in significant yield increases, as a consequence of increased grain number per m². Application of triadimefon to crops at ear emergence had no significant effect on percentage plant infection but resulted in lower levels of re-infection following artificial or natural inoculation of healthy ears, apparently as a result of a protectant effect of fungicide residues on or in the ovaries.     

Efficacy of Bacillus thuringiensis Cry3Aa protoxin and protease inhibitors against coleopteran storage pests

BACKGROUND: Environmental impacts and resistance to insecticides pose serious challenges to stored‐product insect and other types of pest control. Insect‐resistant transgenic grain is a potential alternative to fumigants, but candidate control proteins are needed, especially for coleopterans. Therefore, we evaluated the efficacy of a coleopteran‐active toxin, Bacillus thuringiensis Cry3Aa, with or without protease inhibitors, in laboratory feeding assays against coleopteran storage pests. RESULTS: In a comparison of the toxicity of Cry3Aa protoxin towards three species of coleopteran storage pests, Tenebrio molitor L. was found to be most sensitive, Tribolium castaneum (Herbst.) was most refractory and Rhyzopertha dominica F. displayed an intermediate response. For R. dominica, Cry3Aa combined with 3500 mg potato carboxypeptidase inhibitor or 5000 mg aprotinin kg^?1 diet resulted in both delayed development and increased mortality. Potato carboxypeptidase inhibitor and bovine aprotinin reduced the LC₅₀ of Cry3Aa for R. dominica two‐ and threefold respectively. Cry3Aa treatment resulted in fewer progeny from R. dominica, and progeny was further reduced when the protoxin was combined with potato carboxypeptidase inhibitor. CONCLUSIONS: These data support the hypothesis that a combination of Cry3Aa protoxin and protease inhibitors, particularly a potato carboxypeptidase inhibitor, may have applications in control strategies for preventing damage to stored products and grains by coleopteran pests. Published 2011 by John Wiley & Sons, Ltd.     

拟轮枝镰孢对多菌灵的敏感性及抗性菌株生物学性状和交互抗性

为评价玉米穗腐病主要致病菌拟轮枝镰孢Fusarium verticillioides对多菌灵的敏感性,采用菌丝生长速率法测定了采自山东、河北、河南、四川、甘肃、辽宁及宁夏7省(自治区)的168株玉米穗腐病拟轮枝镰孢对多菌灵的敏感性,并对经药剂驯化获得的拟轮枝镰孢抗多菌灵菌株主要生物学性状和交互抗性进行了研究。结果表明:多菌灵抑制拟轮枝镰孢菌丝生长的EC₅₀值在0.013 2~0.774 0 mg/L之间,平均EC₅₀值为(0.220 8 ± 0.143 7) mg/L。敏感性频率分布显示,供试病原菌群体中已出现对多菌灵敏感性下降的亚群体,但其中仍有35.71%的菌株对多菌灵的敏感性频率呈正态分布,因此可将此部分菌株的平均EC₅₀值 (0.081 4 ± 0.028 9) mg/L作为拟轮枝镰孢对多菌灵的相对敏感基线。药剂驯化共获得6株抗性菌株,抗性倍数在5.05~12.22之间。抗性菌株的菌丝生长速率及菌丝干重均低于亲本菌株,表明其抗性菌株的生物适合度有所降低,同时发现其抗药性均不能稳定遗传。室内交互抗性测定结果表明,拟轮枝镰孢对多菌灵的抗性与咪鲜胺、戊唑醇、三唑酮及吡唑醚菌酯之间均不存在交互抗性关系。     

MB和CMC液体培养基对禾谷镰孢产孢水平的影响

禾谷镰孢分生孢子定量接种是研究作物抗禾谷镰孢的必要手段?本试验利用绿豆(mung bean, MB)与羧甲基纤维素(carboxylmethyl cellulose, CMC)液体培养基在相同条件下培养禾谷镰孢, 比较两种培养基对禾谷镰孢产孢效率的影响; 并用两种培养基诱导分生孢子制成相同浓度孢子悬浮液, 对75个玉米家系进行人工接种鉴定, 检测不同培养基所产分生孢子致病性差异?结果表明, CMC培养基诱导分生孢子增长率(k2=1.125)大于MB培养基诱导增长率(k1=0.844)?在培养第14天, MB培养基诱导获得的分生孢子平均浓度为2.51×105个/mL, CMC培养基诱导的为3.62×105个/mL, 比MB培养基诱导多44.22%?用两种培养基诱生的孢子进行田间接种, 玉米穗腐病发病程度无明显差异?CMC培养基具有产孢快?孢子浓度高的优点, 是一种适宜禾谷镰孢分生孢子诱生的高效液体培养基?     

The contribution of ground-level inoculum of Fusarium culmorum to ear blight of winter wheat

In a series of field experiments in eastern England over 5 years, severe ear blight developed only in plots of winter wheat that were inoculated by spraying with conidial suspensions of Fusarium culmorum during anthesis, and in which infection was encouraged by rainfall or mist irrigation. In the absence of artificial inoculation of the ears, F. culmorum caused less extensive ear blight, and only where soil-surface inoculum was available after its application on infested plant material (colonized oat grains) up to 3–4 weeks before anthesis; it then developed most where significant rainfall occurred close to the time of anthesis. A warm, dry period following application of inoculum to the ground in late March contributed to increased infection of grain by F. culmorum , although ear blight was not increased. Ear infection therefore depended on adequate viable inoculum on infested plant debris within the crop, and conditions tending to favour brown foot rot development as well as, subsequently, rainfall and moist conditions during anthesis. These conditions did not occur together naturally during this period. Seedling infection by F. culmorum or Microdochium nivale made no significant contribution to ear blight. Inoculation of ears at anthesis with M. nivale or a locally obtained isolate of F. langsethiae did not produce ear blight symptoms. Possibilities for minimizing the availability of inoculum of F. culmorum and the implications for various options for ear-blight control are discussed.     

The infestation of tomato seed by Fusarium oxysporum f.sp. radicis-lycopersici

The infestation of seed by Fusarium oxysporum f.sp. radicis-lycopersici occurred at rates of 0-1 to 0-01 % in fruit on stem-infected plants. Direct infection of fruit in the flower or young developing fruit stage resulted in a grayish-brown lesion on the stylar end of the fruit or mummification. F. oxysporum f.sp. radicis-lycopersici was isolated from all seeds in such fruit. Picked fruit inoculated on the stem scar also became infected but 96 h after inoculation of the fruit, the seed was not infested or infected. The contact of clean seed with hands that had previously handled F. oxysporum f.sp. radicis-lycoperisid-infesied sawdust resulted in a high level of seed infestation. The fungus survived on seed sent across Canada and stored for up to 12 weeks. Treatment with NaOCl or 0·1 N HCl did not completely disinfest infested seed.     

Evaluating commercial maize hybrids for resistance to gibberella ear rot

An integral component of breeding maize for resistance to Fusarium graminearum ear rot is the identification of resistant genotypes. Since natural infection is not consistent from year to year, maize researchers must use manual techniques to inoculate the plant material with fungal spores. Information is presented here on site resistance of commercial maize hybrids to F. graminearum over three years and at two locations. Additionally, results of an investigation on the two predominant techniques of inoculating maize, the silk channel and kernel inoculation methods, are reported. Of 61 commercial hybrids tested, only two were ranked as moderately resistant to the fungus by both inoculation methods. These two hybrids also had a stable response to the F. graminearum infection across seven environments when the silk channel inoculation method was used. The majority of the hybrids were ranked as either susceptible or highly susceptible and less than 10% of the hybrids had a stable response to fungal infection. In the investigation of methodology, it was concluded that silk browning would be the least laborious way to identify the ideal time to complete silk channel inoculations. It was found that kernel inoculations using the pin inoculation method should take place between 11 and 15 days after 50% silking to achieve proper hybrid discrimination. Mist irrigation increased mold severity ratings and resulted in greater discrimination between hybrids with varying levels of resistance to F. graminearum infection.