Ear Rot Symptoms Research Articles

First Report of Fusarium verticillioides Causing Fusarium Ear Rot of Corn in Jordan.

Sweet corn (Zea mays L.) is one of the most popular crops grown in Jordan. Fusarium verticillioides (Sacc.) is a major pathogen of corn and a producer of mycotoxin fumonisins (Blacutt et al. 2018). During September and October 2019, ear rot symptoms were observed on ̴30% of the sweet corn variety Mis Dolce grown in the Jordan Valley. The disease caused substantial losses, including damage to greater than 50% of the kernels within 15-20 days after harvesting. A total of 350-corn kernels were randomly taken from 70 plants distributed in five fields with a total area of 2 ha. About 35% of the samples showed typical symptoms of the disease. Discolored corn kernels were surface sterilized with 5% NaOCl solution for 1 min, then rinsed three times with sterilized distilled water (SDW), plated on potato dextrose agar (PDA) at 25°C, and incubated in the dark for 7 days. Twelve putative isolates of the genus Fusarium were hyphal-tipped on new PDA plates. Isolates were cultured on synthetic low-nutrient agar (SNA) with a ca. 1 × 2-cm strip of sterile filter paper on the agar surface (Nirenberg 1976). Cultures were incubated for 10 to 14 days at 20°C in dark conditions. When sporulation was observed, agar blocks were mounted on a microscopic slide with a drop of lactophenol cotton blue and examined under the microscope at 400x. Colonies grew rapidly with abundant pink to violet aerial hyphae. Sporodochia formed on the agar, and the aerial conidiophores branched sparsely, often alternately or oppositely, terminating with up to three verticillate phialides. Macroconidia were abundant, falcate to straight, three- to five-septate, with a distinct foot cell, 27 to 73 × 3.1 to 5.6 μm. Microconidia produced on polyphialides and aggregating in heads, were unicellular, ovoidal, or ellipsoidal, 4.4 to 17 × 1.5 to 4.5 μm (Fig. 1A, B, C, D, E, and F). Based on morphological characteristics, isolates were tentatively identified as F. verticillioides (Al-Hatmi et al. 2016; Guarro 2013). Two representative isolates were DNA extracted and the translation elongation factor 1-α gene (TEF1) was amplified (O'Donnell et al. 1998), and sequenced from both directions at Macrogen Inc, South Korea. The consensus sequences of the two isolates Fvcorn2021JO-03 (OK040159) and Fvcorn2021JO-04 (OK040160) were used as BLASTn query on the NCBI website and were 100% and 99% similar with F. verticilloides JF740717 and JF740737 accessions, respectively. Similarly, the two isolates were 100% and 99.85% similar with F. verticilloides reference sequences MH582332 and MH582327 on the Fusarium MLST database, respectively. The pathogenicity of the two isolates was tested on 15 cobs by injecting 2 ml of a 2.5 × 105 conidia/ml suspension into the silk channel and into kernel wounds of the primary ear (three replicates) for each treatment (Reid and Hamilton 1996). Inoculated kernels were incubated at 25°C for 2 weeks in plastic boxes. The healthy kernels were injected with 2 ml of SDW as a negative control. Grains started to rot after 2 weeks, in the form of a thick, cottony, crimson-looking growth between the ear and its covers, with only some grains or a group of a adjacent grains rotting, and then white lines appear on the outer skin of the grain, yielding symptoms similar to those in the field (Fig. 2A, B, C, and D). The fungus was re-isolated from the inoculated kernels and was morphologically identified as F. verticilloides thus fulfilling Koch's postulates. The fumonisins-producing potential of the isolated F. verticillioides was confirmed using the AgroQuant Total Fumonisins Assay (Romer Labs, Singapore). To our knowledge, this is the first report of F. verticillioides causing Fusarium ear rot on corn in Jordan. Further investigation is needed to gain a better understanding of the spatio-temporal dynamics of this novel pathogen.

Association mapping and genomic prediction for ear rot disease caused by Fusarium verticillioides in a tropical maize germplasm

AbstractFusarium ear rot (FER), caused by Fusarium verticillioides (Sacc.) Nirenberg, is one of the major ear diseases that affect both yield and grain quality in maize (Zea mays L.), especially in tropical environments. Fusarium genetic resistance is a complex trait, controlled by several small‐effect genes and strongly influenced by the environment. We applied a comprehensive genome‐wide association study and genomic prediction for ear rot and starburst symptoms, using 291,633 high‐quality single nucleotide polymorphism (SNPs) markers in 320 tropical maize inbred lines, in two distinct locations in Brazil's southern region. Three SNPs were significantly associated with starburst symptoms, each associated with 6–8% of the phenotypic variance, and with gene models that have expression levels in ears, pericarp, and kernels, corresponding to disease infection period and suggesting some role in FER resistance. No significant SNP was associated with FER, which is an indication that the genetic architecture for this trait is highly polygenic, with potentially many variants having small effects that are not detectable in the association mapping analysis. We observed genomic prediction accuracies ranging from 0.34 to 0.4 for FER and starburst, respectively. Further research is required to validate these significant SNPs and their relationship to genes affecting FER resistance, and also to improve genomic prediction accuracies across different genetic backgrounds.

Sexual reproduction parameters in Fusarium verticillioides populations from maize in Brazil

Fusarium verticillioides is an important plant pathogen causing infections on ears and/or stalks of maize. This study aimed to compare sexual reproduction parameters in populations of F. verticillioides from distinct regions in the different Brazilian climatic zones where maize is grown. Fertility and effective population size were calculated from data generated by crossing field isolates with female fertile testers. Three hundred fungi isolates were obtained from maize kernels with Fusarium ear rot symptoms in different geographic regions of Brazil in the years 2012 and 2013. For the entire population, 231 out of 300 isolates were cross-fertile with tester isolates. MAT1–1 and MAT1–2 idiomorphs of the fertile isolates segregated in a 105:126 ratio. Female isolates (hermaphrodites) accounted for 96 out of 231 fertile isolates, while 135 were male-only isolates. The ratio of mating types within the population [Ne(mt)] was 99% of the count for the Brazilian population when the mating type idiomorphs were used as predictors to estimate the population size. However, when this calculation was made on the basis of the female fertile isolates, the Ne(f) was 83%. When the total population was divided by geographical/climatic regions, the frequencies of female fertile isolates were 33 out of 80 fertile isolates from the Northeast (equatorial tropical climate); 49 out of 73 fertile isolates from the Central-West (tropical climate); and only 14 out of 78 fertile isolates from the South (temperate climate), indicating that the sexual reproduction in F. verticillioides is likely favored by a tropical climate in Brazil. Brazilian populations of F. verticillioides show high fertility and high effective population size, suggesting the possibility of frequent sexual reproduction in the field, especially in populations from Tropical Climate regions of Brazil.

Pathogenicity of Fumonisin-producing and Nonproducing Strains of Aspergillus Species in Section Nigri to Maize Ears and Seedlings

Species of Aspergillus section Nigri are commonly associated with maize kernels, and some strains can produce fumonisin mycotoxins. However, there is little information about the extent to which these fungi contribute to fumonisin contamination in grain, the damage they cause to maize ears, or their effects on maize seed germination and seedling health. We compared fumonisin-producing and nonproducing strains of A. niger, A. welwitschiae, A. phoenicis, A. tubingensis, and A. carbonarius from the United States and Italy in laboratory and field studies to assess their ability to contribute to fumonisin contamination, to cause maize ear rot, and to affect seed germination and seedling growth. In laboratory experiments, some strains of each Aspergillus species reduced germination or seedling growth, but there was high variability among strains within species. There were no consistent differences between fumonisin-producing and nonproducing strains. In field studies in Iowa and Illinois, strains were variable in their ability to cause ear rot symptoms, but this was independent of the ability of the Aspergillus strains to produce fumonisins. Contamination of grain with fumonisins was not consistently increased by inoculation with Aspergillus strains compared with the control, and was much greater in F. verticillioides-inoculated treatments than in Aspergillus-inoculated treatments. However, the ratio of the FB analogs FB2 and FB1 was altered by inoculation with some Aspergillus strains, indicating that FB2 production by Aspergillus strains occurred in the field. These results demonstrate the pathogenic capabilities of strains of Aspergillus in section Nigri, but suggest that their effects on maize ears and seedlings are not related to their ability to produce fumonisins, and that fumonisin contamination of grain caused by Aspergillus spp. is not as significant as that caused by Fusarium spp.

The influence of flavonoids in maize pericarp on fusarium ear rot symptoms and fumonisin accumulation under field conditions

Breeding efforts have been undertaken to increase resistance of maize to fusarium ear rot (FER) and to fumonisin accumulation. Flavonoids in the pericarp of the kernels are considered particularly able to reduce the fumonisin accumulation. The aim of this 2‐year field study was to assess the effect of flavonoids on FER symptoms and fumonisin contamination in maize kernels using two isogenic hybrids, one providing pigmentation in the pericarp (P1‐rr) and the other without it (P1‐wr). FER incidence (FERi), FER severity (FERs), the incidence of infections caused by Fusarium spp. in symptomless kernels (FF) and fumonisin contamination (FUM) were assessed in both hybrids. Significant differences between the two hybrids were detected mainly in 2012 trials where P1‐rr showed lower FERi (P < 0·01), FF (P < 0·05) and FUM (P < 0·1) than P1‐wr. Site, characterized by local temperature and precipitation, played a relevant role in modelling all the measured variables, as its effect was highly significant in both years, whether they were considered individually or altogether. The interaction of hybrid with location was a significant (P < 0·001) source of variation only for FF. FF, together with FERi, was also significantly (P < 0·001) influenced by the interaction of hybrid with year. In general, FUM was more influenced by year and location parameters, such as temperatures during late ripening, than by flavonoid presence in kernel pericarp. The results indicate that flavonoid pigments alone may not be an important component in the resistance of maize to fumonisin accumulation.

First Report of Fusarium Ear Rot of Maize Caused by Fusarium andiyazi in China.

Maize (Zea mays L.) is an important food crop worldwide. Some Fusarium species cause maize ear rot leading to significant yield losses and, for some Fusarium species, potential risk of mycotoxin contamination. In 2013, a survey was conducted to determine the population composition of Fusarium species on maize in Dongyang, Zhejiang Province, China, where about 5% of maize ears in each field were found with reddish-white mold. Symptomatic maize ears were collected from several cultivars including forage corn Zhedan724 and Zhengdan958, sweet corn Chaotian4 and Chaotian135, and waxy corn Heinuo181 and Zhenuoyu6; no association between the disease and maize cultivars was observed. Maize kernels showing a pink or white mold were surface-disinfested with 70% ethanol and 10% sodium hypochlorite, followed by three rinses with sterile distilled water and placed onto potato dextrose agar (PDA). After 3 days of incubation at 25°C in the dark, mycelia were transferred to fresh PDA and purified by the single-spore isolation method (4). Species were identified based on morphological characteristics (2), and sequence analysis of the translation elongation factor-1α (TEF) gene. The results indicated that Fusarium verticillioides Sacc. (84.6%) is the main causal agent of maize ear rot in this region. However, morphological characteristics of two strains (7.7%) from the same field were found to be identical to F. andiyazi Marasas, Rheeder, Lampr., K.A. Zeller & J.F. Leslie. Colonies on PDA showed floccose to powdery mycelium and pale-purple pigmentation. Hyaline and straight or slightly curved macroconidia were observed with 3- to 6-septate and a slightly curved apical cell. Chlamydospores were absent. In order to validate this result, partial translation elongation factor (TEF-1α, 646 bp) gene sequences of isolates were generated (GenBank Accession No. KJ137019) (1). BLASTn analysis of TEF-1α with the GenBank database revealed 99.7% sequence identity to F. andiyazi (JN408195 and JN408196), and much lower (94 to 98%) identity with other Fusarium spp. Thus, both morphological and molecular criteria supported identification of the strains as F. andiyazi. A pathogenicity test was performed on maize cv. Zhengdan958 in a greenhouse. Four days post-silk emergence, a 2-ml conidial suspension (105 macroconidia/ml) of each isolate was injected into each of 10 maize ears through the silk channel. An equal amount of sterile distilled water was injected into 10 ears as a control. Typical Fusarium ear rot symptoms (reddish-white mold), which were observed in the ears inoculated with these strains 20 days after inoculation, were similar to the original symptoms in the sampling sites, and no symptoms were observed on the water control ears. The same fungus was re-isolated from the infected kernels using the method described above. F. andiyazi are the major pathogens of sorghum (2) and also proved to attack maize kernels recently (3). To our knowledge, this is the first report of F. andiyazi causing Fusarium ear rot on maize in China. Further investigation is needed to gain a better understanding of the spatial and temporal dynamics of this new pathogen. Also, the new species must be considered in the development of maize cultivars with broad-based resistance to the pathogens.

First Report of Fusarium temperatum Causing Fusarium Ear Rot on Maize in Northern China.

In China, several diseases of maize (Zea mays L.) including ear rot are caused by Fusarium spp., leading to significant yield losses and potential risk of mycotoxin contamination (2,3). In 2013, a survey was conducted to determine the population composition of Fusarium species on maize ears in Jilin Province. Symptomatic maize ears with pink or white mold were collected and surface disinfested with 70% ethanol and 10% sodium hypochlorite, followed by three rinses with sterile distilled water and placed onto potato dextrose agar (PDA). After 3 days of incubation at 25°C in the dark, newly grown-out mycelia were transferred onto fresh medium and purified by the single-spore isolation method (4). Fusarium spp. were identified by morphological characteristics (2) and sequence analysis of translation elongation factor-1α (TEF) gene (1). A large number of Fusarium spp. were found including F. graminearum species complex and F. verticillioides. In addition, a new species, F. temperatum, recently described in Belgium (2), was also identified. F. temperatum was originally described as F. subglutinans, but a robust polyphasic approach proved it to be a new biological species closely related to F. subglutinans (2). Previous studies had reported ~15% of Fusarium maize ear rot in Jilin was F. subglutinans. In this study, we found both F. subglutinans s. str. and F. temperatum in the proportion of 16.3% and 9.2%, respectively. Similar to previous studies (2), colonies of our strains on PDA were initially white cottony mycelium that become pinkish white. Conidiophores formed abundantly on SNA that were erect, branched, and terminated in 1 to 3 phialides. Microconidia were abundant, hyaline, 0 to 2 septa, obovoid to oval, and not produced in chains. Chlamydospores were absent. Typically macroconidia were falcate, 3 to 5 septate (mostly 4 septate), hyaline with a curved and blunt apical cell and a distinct foot-shaped basal cell. In order to validate this result, partial translation elongation factor (TEF-1α, 629 bp) gene sequences of isolates were generated (GenBank Accession No. KJ137018) (1). BLASTn analysis revealed 100% sequence identity to F. temperatum (HM067690). A pathogenicity test was performed on maize cv. Zhengdan958. Four days after silk emergence, 2 ml conidial suspension (105 macroconidia/ml) of each isolate was injected into each of 10 maize ears through silk channel. Control plants were inoculated with sterile distilled water. Twenty days after inoculation, typical Fusarium ear rot symptoms (reddish-white mold) was observed on inoculated ears and no symptoms were observed on water controls. Koch's postulates were fulfilled by re-isolating the same fungus from the infected seeds. Although F. temperatum was reported to attack maize kernels in southern China where the annual average temperatures are moderately high (3), to our knowledge, this is the first report of F. temperatum causing Fusarium ear rot in northern China, where the winter is long and very cold, the annual average temperature is 4 to 5°C, and the lowest temperature is lower than -35°C. This indicated that F. temperatum was widely distributed in different ecological regions in China. Furthermore, the northeast spring corn region that includes Jinlin is the most important corn belt, with corn production of this region accounting for 42% of the total corn production in China. Therefore, we should pay more attention to the new species in this region and consider them in the development of maize cultivars with broad-based resistance to the pathogens.

Toxigenicity of Fusarium Species in Gibberella fujikuroi Species Complex (GFSC) Associated With Stalk and Ear Rot Disease of Corn

Fusarium stalk and ear rot disease did not only cause significant losses of yield but also produced mycotoxins that are harmful to animals and human. This study was conducted to elucidate three major mycotoxins i.e. fumonisin B1 (FUMB1), moniliformin (MON), and beauvericin (BEA) produced by the Fusarium spp. isolated from corn showing typical stalk and ear rot symptoms in Indonesia, Malaysia and Thailand. Twenty selected strains of Fusarium species in Gibberella fujikuroi species complex i.e. F.verticillioides, F. proliferatum, F. subglutinans, and F. konzum were analyzed for production of the three mycotoxins by using an Ultra Performance Liquid Chromatography (UPLC). All strains of F. verticillioides and F. proliferatum produced FUMB1 at high levelsand MON at low levels. Many strains of F. verticillioides (67%) and F. proliferatum (50%) did not produce BEA while the others produced BEA at low levels. Two strains of F. subglutinans did not produce FUMB1 but produced MON at low levels. One strain of F. subglutinans produced BEA and the other one did not produce the toxin. Two strains of F. konzum produced both MON and BEA at low levels but only one strain produced FUMB1 at a low level. These mycotoxins have not been reported from Fusarium spp. in Gibberella fujikuroi species complex isolated form stalk and ear rot diseases of corn in these areas. Therefore, concerted efforts must be made to educate all stake holders about the presence and health hazards of these mycotoxins.

Effects of planting date and environmental factors on fusarium ear rot symptoms and fumonisin B1accumulation in maize grown in six North American locations

Fusarium ear rot, primarily caused byFusarium verticillioides,causes losses in grain yield and quality and can result in contamination of grain by mycotoxins, primarily fumonisin B1. Disease severity and fumonisin B1contamination vary considerably among maize‐growing regions and from year to year. A 2 year field study was conducted in six locations in the USA, to evaluate the roles of planting date, maize hybrid, rainfall, temperature and insect pests in the variation in fusarium ear rot symptoms and fumonisin B1contamination. Grain samples were inspected to determine percentage of kernels with fusarium ear rot symptoms, categorized as ‘moulded’ or ‘starburst’; grain was also analysed by ELISA for fumonisin B1. Hybrid and planting date frequently had significant effects (P ≤ 0·05) on fusarium ear rot and fumonisin B1contamination. Earlier planting consistently resulted in lower ear rot severity, fumonisin B1levels and insect damage. Mould symptoms were highly correlated with thrips populations (Frankliniella occidentalis) (r = 0·78) and with fumonisin B1concentration (r = 0·89). The starburst symptom was not as closely correlated with thrips (r = 0·33) or fumonisin B1(r = 0·18). A multiple linear regression model identified highly significant effects on fumonisin B1for thrips, lower average daily precipitation after flowering, and location. These results strengthen the evidence that locations with high populations of ear‐infesting thrips and dry conditions after pollination have an increased risk of fumonisin contamination, and suggest that high fumonisin levels are much more likely in visibly moulded kernels compared to those with ‘starburst’ symptoms.

Relationships of immature and adult thrips with silk‐cut, fusarium ear rot and fumonisin B1 contamination of maize in California and Hawaii

Field experiments were conducted in California and Hawaii in order to investigate the relationships between thrips feeding in maize ears and fusarium ear rot and silk‐cut symptoms. Half the plots in each experiment were treated with insecticides following pollination. Thrips populations within ears were enumerated at six stages of ear development. Grain was examined microscopically and the percentages of kernels with silk‐cut and ear rot symptoms were quantified by weight. Fumonisin B1 contamination in grain was measured by ELISA. Immature stages of thrips predominated, and maximum thrips populations occurred 21 days after pollination. Insecticides reduced thrips numbers, as well as silk‐cut, ear rot symptoms and fumonisin B1 contamination. Immature thrips populations were more strongly correlated with silk‐cut/ear rot symptoms (R = 0·75) and fumonisin B1 accumulation (R = 0·53), than were adult thrips (R = 0·48 and 0·36, respectively). Silk‐cut kernels all had ear rot symptoms and the percentage of kernels with symptoms was highly correlated with fumonisin B1 contamination (R = 0·84). Results suggest that thrips are not occasional feeders, but can complete a substantial portion of their life cycle on maize ears. The results also indicate that thrips activity may be a cause of silk‐cut symptoms, and this may be the mechanism that connects thrips activity with fusarium ear rot and fumonisin contamination of grain.

Associations of planting date, drought stress, and insects with Fusarium ear rot and fumonisin B1 contamination in California maize

Fusarium ear rot, caused by Fusarium verticillioides, is one of the most common diseases of maize, causing yield and quality reductions and contamination of grain by fumonisins and other mycotoxins. Drought stress and various insects have been implicated as factors affecting disease severity. Field studies were conducted to evaluate the interactions and relative influences of drought stress, insect infestation, and planting date upon Fusarium ear rot severity and fumonisin B1 contamination. Three hybrids varying in partial resistance to Fusarium ear rot were sown on three planting dates and subjected to four irrigation regimes to induce differing levels of drought stress. A foliar-spray insecticide treatment was imposed to induce differing levels of insect injury. Populations of thrips (Frankliniella spp.), damage by corn earworm (Helicoverpa zeae), Fusarium ear rot symptoms, and fumonisin B1 levels were assessed. There were significant effects of hybrid, planting date, insecticide treatment, and drought stress on Fusarium ear rot symptoms and fumonisin B1 contamination, and these factors also had significant interacting effects. The most influential factors were hybrid and insecticide treatment, but their effects were influenced by planting date and drought stress. The more resistant hybrids and the insecticide-treated plots consistently had lower Fusarium ear rot severity and fumonisin B1 contamination. Later planting dates typically had higher thrips populations, more Fusarium ear rot, and higher levels of fumonisin B1. Insect activity was significantly correlated with disease severity and fumonisin contamination, and the correlations were strongest for thrips. The results of this study confirm the influence of thrips on Fusarium ear rot severity in California, USA, and also establish a strong association between thrips and fumonisin B1 levels.

Pyrethroid application timing to control European corn borer (Lepidoptera: Crambidae) and minimize fumonisin contamination in maize kernels

Ostrinia nubilalis (Hubner) (European corn borer) is the main maize pest in Central and South Europe and it promotes Fusarium verticillioides infection on maize grains, which is able to produce fumonisins. The objective of this study was to determine the effect of the timing of pyrethroid treatments on European corn borer damage, fungal ear rot and fumonisin contamination. The field experiments were performed from 2005 to 2007 in NW Italy. Four application timings were compared to an untreated control. The insecticide treatments were applied at approximately 10 days intervals, starting from the end of flowering. The last treatment was performed approximately 15 days after the ECB flight peak. At harvest, the ears were rated for the incidence and severity of European corn borer damage and fungal ear rot symptoms, and the harvested kernels were analyzed for fumonisins B 1 +B 2 . In all the years, the treatments applied 7–10 days before the European corn borer adult flight peak showed the best efficacy to co...

옥수수 이삭썩음병에 관여하는 Fusarium속균의 동정

In 2007, a total of 77 isolates of Fusarium spp. were obtained from ear rot symptoms of corns collected from 5 locations in Gangwon Province, Korea. The fungal isolates were identified based on their morphological features. Out of the isolates, fifteen isolates were identified as Fusarium verticillioides which formed microconidia in long chains on monophialides. Four isolates were identified as F. subglutinans which formed microconida only on false heads. Six isolates were identified as F. graminearum which produced red pigment in PDA culture. Besides these Fusarium species, F. napiform, F. nygamai, and F. oxysporum were identified from the rest isolates. To assess for genetic diversity of the isolates, a random amplified polymorphic DNA(RAPD) technique was carried out using URP primers. The results from the RAPD analysis showed that the isolates from corn were divided into 6 groups. These RAPD groups of the Fusarium species corresponded to morphological characters of the Fusarium species. The phylogenetic analysis of most isolates by DNA sequencing of EF-1<TEX>$\alpha$</TEX> gene corresponded to morphological characters of the Fusarium species. The results of pathogenicity tests by two inoculation methods revealed that F. verticillioides, F. graminearum and F. subglutinans are strongly pathogenic to corn stalks.

Management of fumonisin contamination in maize kernels through the timing of insecticide application against the European corn borer Ostrinia nubilalis Hübner

The European corn borer (ECB), Ostrinia nubilalis, is the principal pest of maize in Central and South Europe. It is known to promote Fusarium verticillioides infection in maize grain, a recognized producer of fumonisins. Field experiments were performed in 2006 and 2007 in two sites in NW Italy to determine the effects of the timing of insecticide application on ECB damage, fungal ear rot and fumonisin contamination under natural conditions. Different insecticide application timings were compared, from maize flowering to approximately 15 days after the flight peak of adult ECB. At harvest, the ears were rated for incidence and severity of ECB damage, fungal ear rot symptoms and fumonisin (FB1 + FB2) contamination. In all years/sites, treatments applied at the beginning of consistent ECB flight activity were most effective in controlling insect damage on ears. Fungal ear rot and fumonisin contamination were significantly affected by ECB control. The efficacy of the best timing of insecticide application in controlling fumonisin contamination was, on average, 93% compared to the untreated control. Contamination levels of these mycotoxins increased with either an earlier or later treatment. Furthermore, an earlier insecticide application showed lower fumonisin contamination than a treatment applied after the adult flight peak. Production of maize kernels and maize-based foods that do not exceed the maximum international and EU permitted levels for fumonisins could be enhanced by appropriate insecticide treatment against second generation ECB. The optimum time for insecticide application is between the beginning of consistent adult flight activity and the flight peak.

Impact of insecticide treatments on Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) and their influence on the mycotoxin contamination of maize kernels

European corn borer (ECB), Ostrinia nubilalis (Hübner), is the main maize pest in Central and Southern Europe and promotes the infection of maize with Fusarium spp., which produce mycotoxins. The objective of this study was to determine the effect of insecticide treatments on ECB damage, Fusarium infection and mycotoxin contamination. The field experiments were performed from 2000 to 2006 in three locations in north-western Italy. At harvest, ears were rated for the incidence and severity of ECB damage and Fusarium ear rot symptoms, and the harvested kernels were analysed for fumonisins and zearalenone. In all the years except 2003, the chemical treatment had a significant effect on ECB incidence and severity. The ear damage was reduced, on average, by 44.1%. Fusarium ear rot and fumonisin contamination were affected by ECB control. The occurrence of the mycotoxin was significantly reduced, on average by 68%. The differences between the treatments were less clear in the year with highest ECB pressure. No significant difference was observed between the different insecticide classes. No relations were detected between the ECB activity and the occurrence of zearalenone. This research indicates that the production of kernels with low fumonisin content may be enhanced by an insecticide treatment against the second ECB generation.

First Report of Ear Soft Rot of Corn (Zea mays) Caused by Burkholderia gladioli in the United States.

During the summer of 2005, an uncharacterized disease was observed on sweet corn 'Mirai 301BC' commercially grown in Sunflower County, Mississippi. Initial symptoms developing at the base of the ear on interior husk leaves were brown, water-soaked, irregular lesions. These gradually enlarged up to 10 cm in diameter. Market value was significantly affected when the corn ears had visible symptoms of this disease. Bacterial cell streaming was observed at a magnification of ×675 from the diseased husk. A bacterium was consistently isolated from lesions on nutrient broth yeast (NBY) agar. Colonies on NBY were yellowish white, slightly convex, shiny, and circular with entire margins. Isolates MS102 and MS103, which were chosen for further characterization, were gram negative, lacked arginine dihydrolase, did not produce fluorescent pigment on Pseudomonas F medium, accumulated poly-β-hydroxybutyrate, and grew aerobically. The isolates were able to utilize l-arabinose, d-mannitol, N-acetylglucosamine, capric acid, malic acid, adipic acid, and phenylacetic acid, but not d-maltose. These characteristics are the same as those described previously for Burkholderia gladioli (3). Analysis of fatty acid methyl ester profiles (Sherlock version TSBA 4.10; Microbial Identification System, Newark, DE) characterized the isolates as B. gladioli (similarity indices: 0.23 to 0.38) and revealed that they have C16:0 3OH, the most characteristic fatty acid for the genus Burkholderia. Confirmation was made by PCR amplification of the nearly complete16S rRNA gene (1,471 bp; GenBank Accession No. EU053154) using universal primers (forward: 5'-AGAGTTTGATCCTGGCTCAG and reverse: 5'-GGCTACCTTGTTACGACTTC). DNA sequence analysis demonstrated that the 16S rRNA gene of the bacterium shared highest identities (99.4 to 99.6%) with that of B. gladioli strains 321gr-6, 223gr-1, and S10 (4). A PCR product (approximately 300 bp) characteristic of B. gladioli also was obtained from both isolates using species-specific primers GLA-f and GLA-r (2). To confirm pathogenicity, cell suspensions (108 CFU/ml in phosphate buffer) of isolates MS102 and MS103 were injected into interior husk leaves of field-grown sweet corn with a 20-gauge needle and syringe (2 ml per ear). Control corn ear husks were injected with phosphate buffer. After 3 days, ear rot symptoms were observed on all plants inoculated with the isolates but not those injected with phosphate buffer. Cell suspension of isolates dropped on nonwounded husks also incited the same symptoms as those inoculated with the syringe. Koch's postulates were fulfilled with reisolation from the inoculated tissues. The identity of the reisolated pathogen was proved by sequencing the 16S rRNA gene. This disease was previously reported in Brazil (1). To our knowledge, this is the first report of B. gladioli causing a disease of corn in the United States. Although the impact of this disease was not observed from 2005 to 2006 because of dry weather and rotation to other crops in the affected field, there is a potential that the bacterium could become established in corn-producing areas as a member of the corn ear rot complex if environmental conditions are favorable. Reference: (1) I. M. G. Almeida et al. Arq. Inst. Biol. Sao Paulo 66:141, 1999. (2) N. Furuya et al. J. Gen. Plant Pathol. 68:220, 2002. (3) M. Gillis et al. Int. J. Syst. Bacteriol. 45:274, 1995. (4) R. Nandakumar et al. Phytopathology (Abstr.) 95(suppl.):S73, 2005.

Molecular mapping of QTLs for resistance toGibberellaear rot, in corn, caused byFusarium graminearum

Gibberella ear rot, caused by the fungus Fusarium graminearum Schwabe, is a serious disease of corn (Zea mays) grown in northern climates. Infected corn is lower yielding and contains toxins that are dangerous to livestock and humans. Resistance to ear rot in corn is quantitative, specific to the mode of fungal entry (silk channels or kernel wounds), and highly influenced by the environment. Evaluations of ear rot resistance are complex and subjective; and they need to be repeated over several years. All of these factors have hampered attempts to develop F. graminearum resistant corn varieties. The aim of this study was to identify molecular markers linked to the genes for resistance to Gibberella ear rot. A recombinant inbred (RI) population, produced from a cross between a Gibberella ear rot resistant line (CO387) and a susceptible line (CG62), was field-inoculated and scored for Gibberella ear rot symptoms in the F4, F6, and F7 generations. The distributions of disease scores were continuous, indicating that resistance is probably conditioned by multiple loci. A molecular linkage map, based on segregation in the F5 RI population, contained 162 markers distributed over 10 linkage groups and had a total length of 2237 cM with an average distance between markers of 13.8 cM. Composite interval mapping identified 11 quantitative trait loci (QTLs) for Gibberella ear rot resistance following silk inoculation and 18 QTLs following kernel inoculation in 4 environments that accounted for 6.7%-35% of the total phenotypic variation. Only 2 QTLs (on linkage group 7) were detected in more than 1 test for silk resistance, and only 1 QTL (on linkage group 5) was detected in more than 1 test for kernel resistance, confirming the strong influence of the environment on these traits. The majority of the favorable alleles were derived from the resistant parent (CO387). The germplasm and markers for QTLs with significant phenotypic effects may be useful for marker-assisted selection to incorporate Gibberella ear rot resistance into commercial corn cultivars.

Pedigree selection for Gibberella ear rot resistance in maize

The pedigree method is often used for developing inbred lines in maize (Zea mays L.). This study was conducted to assess the effectiveness of pedigree selection for improving resistance to Gibberella ear rot in four maize populations. Selection was based on the severity of ear rot symptoms after inoculation with macroconidial suspensions of Fusarium graminearum (Schwabe) into the silk channel (for two populations) and into the developing kernels (for two other populations). Samples of the selfed families (S1 to S5), recovered from remnant seed from the selection programs, were evaluated for disease resistance during three years, using inoculation and evaluation protocols similar to those used during selection. Among-family selection was effective in both of the populations selected after silk inoculation and in one of the populations selected after kernel inoculation. Responses to selection were more evident in later than in earlier generations for both types of inoculation. Changes in the estimated genetic gain over generations were consistent with changes in the variances among families, which tended to increase in early generations and to decrease in later generations. Selection after kernel inoculation seemed to have been more effective than selection after silk inoculation in developing families with more stable resistance. Based on the results obtained here, it seems that responses to family selection could be accelerated without increasing operational costs by increasing selection intensity in later generations and inoculating fewer plants per family.

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.

Fumonisins in Maize: Can We Reduce Their Occurrence?

Fumonisins in Maize: Can We Reduce Their Occurrence?