Impact of cycling temperatures on Fusarium verticillioides and Fusarium graminearum growth and mycotoxins production in soybean

BACKGROUND: Fusarium graminearum and F. verticillioides are two very important mycotoxigenic species as they cause diverse diseases in crops. The effects of constant and cycling temperatures on growth and mycotoxin production of these species were studied on soybean based medium and on irradiated soya beans. RESULTS: F. graminearum grew better when was incubated at 15, 20 and 15–20 °C (isothermal or cycling temperature) during 21 days of incubation. Maximum levels of zearalenone and deoxynivalenol (39.25 and 1040.4 µg g^?1, respectively) were detected on soya beans after 15 days of incubation and the optimal temperature for mycotoxin production was 15 °C for zearalenone and 20 °C for deoxynivalenol. F. verticillioides grew better at 25 °C in culture medium and at 15/20 °C and 15/25 °C on soybean seeds. Fumonisin B₁ was produced only in culture medium, and the maximum level (7.38 µg g^?1) was found at 15 °C after 7 days of incubation. CONCLUSION: When growth and mycotoxin production under cycling temperatures were predicted from the results under constant conditions, observed values were different from calculated for both species and substrate medium. Therefore, care should be taken if data at constant temperature conditions are to be extrapolated to real field conditions. Copyright © 2012 Society of Chemical Industry     

Fusarium graminearum Infection Strategy in Wheat Involves a Highly Conserved Genetic Program That Controls the Expression of a Core Effectome

Fusarium graminearum, the main causal agent of Fusarium Head Blight (FHB), is one of the most damaging pathogens in wheat. Because of the complex organization of wheat resistance to FHB, this pathosystem represents a relevant model to elucidate the molecular mechanisms underlying plant susceptibility and to identify their main drivers, the pathogen’s effectors. Although the F. graminearum catalog of effectors has been well characterized at the genome scale, in planta studies are needed to confirm their effective accumulation in host tissues and to identify their role during the infection process. Taking advantage of the genetic variability from both species, a RNAseq-based profiling of gene expression was performed during an infection time course using an aggressive F. graminearum strain facing five wheat cultivars of contrasting susceptibility as well as using three strains of contrasting aggressiveness infecting a single susceptible host. Genes coding for secreted proteins and exhibiting significant expression changes along infection progress were selected to identify the effector gene candidates. During its interaction with the five wheat cultivars, 476 effector genes were expressed by the aggressive strain, among which 91% were found in all the infected hosts. Considering three different strains infecting a single susceptible host, 761 effector genes were identified, among which 90% were systematically expressed in the three strains. We revealed a robust F. graminearum core effectome of 357 genes expressed in all the hosts and by all the strains that exhibited conserved expression patterns over time. Several wheat compartments were predicted to be targeted by these putative effectors including apoplast, nucleus, chloroplast and mitochondria. Taken together, our results shed light on a highly conserved parasite strategy. They led to the identification of reliable key fungal genes putatively involved in wheat susceptibility to F. graminearum, and provided valuable information about their putative targets.     

粮油食品防霉微生物筛选及其活性物质初步研究

为开发新型防霉剂,降低粮油食品储藏、运输、流通等过程中霉变对其品质和食用安全的影响,研究筛选获得具有强防霉活性的微生物。采用抑菌圈法,以禾谷镰刀菌（Fusarium graminearum）为指示菌筛选具有防霉活性的菌株。利用形态学观察、生化特征鉴定和16S rDNA基因序列比对进行菌株鉴定;通过不同温度、pH和蛋白酶K处理对防霉活性物质进行初步分析。结果显示：通过初筛获得33株对禾谷镰刀菌具有强抑制活性的菌株,复筛后选择的7株菌归属为芽孢杆菌属（Bacillus）;活性物质分析发现7株菌的活性物质均不耐高温和强酸碱,同时蛋白酶K处理对其防霉效果没有明显影响,初步推断为肽类;通过抑菌谱测定发现菌株ASAG 62对常见霉菌（产黄青霉Penicillium flavum、黑曲霉Aspergillus Niger、赭曲霉Aspergillus ochre、黄曲霉Aspergillus flavus、禾谷镰刀菌F. graminearum）均具有良好的抑制效果。     

对虾肠道中产毒镰孢菌抑制菌的筛选与鉴定

为了获得对产毒镰孢菌具有抑制效应的海洋细菌,从对虾肠道中分离细菌,并将分离得到的优势细菌与产T-2毒素的禾谷镰孢菌(Fusarium graminearum)(FG1207)进行固体对峙培养,具有抑菌圈的细菌菌落即为产毒镰孢菌的抑制菌。然后选取对FG1207具有抑制作用的细菌与FG1207进行液体共同培养,用LC-MS/MS技术检测菌悬液中T-2毒素含量,最后对具有抑制FG1207的生长和降解T-2毒素效果的细菌进行16s r RNA序列鉴定和VITEK2细菌生化鉴定。实验结果分离得到8株优势细菌,其中一株对FG1207的生长具有明显的抑制作用;LC-MS/MS检测发现该细菌与FG1207共同培养菌悬液中未检测到T-2毒素,说明该细菌不仅能够抑制产毒镰孢菌的生长,还能降解T-2毒素。经16s r RNA鉴定该细菌为海洋尼泊尔葡萄球菌(Nepal Staphylococcus Aureus),相似度为99.93%,VITEK2细菌生化鉴定的相似度为96.86%。     

禾谷镰刀菌对多菌灵抗性的监测及其演变规律

调查了江苏省1994-2007年小麦赤霉病的危害程度,监测了江苏省1985-2007年禾谷镰刀菌对多菌灵的田间抗性菌株频率,分析了禾谷镰刀菌对多菌灵的抗药性与小麦赤霉病发生程度及气象因子的相关.结果表明:小麦赤霉病在江苏省的苏南江浦等县中发生以上的年份占30.8%,而在江苏北部赣榆、高邮等县,小麦赤霉病中发生以上的年份占7.7%～15.4%.抗药性菌株频率的增减率与小麦赤霉病的发病程度、病情指数和小麦抽穗期4月中下旬的雨日呈极显著的相关关系.研究表明在农业生产上多菌灵防治小麦赤霉病仍然具有使用价值.     

丁香酚、香芹酚和百里香酚对禾谷镰刀菌的抑菌活性及机制

以转录组技术为基础,研究丁香酚、香芹酚和百里香酚对禾谷镰刀菌的抑制作用及作用机理。方法：借助体外抑菌实验测定丁香酚、香芹酚和百里香酚对禾谷镰刀菌菌丝生长、孢子萌发和呕吐毒素生物合成的有效作用浓度;通过测定丁香酚、香芹酚和百里香酚处理后菌丝液的电导率、ATP含量和丙二醛含量进一步从细胞水平探讨其作用位点;通过转录组学阐明丁香酚、香芹酚和百里香酚抑制禾谷镰刀菌生长和毒素合成的分子机制。结果：3 种植物精油活性成分中,百里香酚的抗真菌活性最强。丁香酚、香芹酚和百里香酚可破坏禾谷镰刀菌细胞膜完整性,引起电解质渗漏,能量代谢失衡。此外,3 种植物精油活性成分通过核糖体、线粒体结构和转录组水平上的酶合成发挥对禾谷镰刀菌的抑制作用。     

玉米穗,茎腐病禾谷镰孢菌和串珠镰孢菌同工酶分析

对沈阳、河北、吉林、北京等地区玉米穗、茎腐病禾谷镰孢菌和串珠镰孢菌进行了可溶性蛋白和同工酶聚丙烯酰胺凝胶电泳，以研究玉米穗、茎腐病同种镰孢菌是否存在寄生专化性或生理分化现象。结果表明：各地区的玉米穗、茎腐病串珠镰孢菌同源性高，亲缘关系近，相似性系数在0．86～0．98之间，菌株间差异仅表现为个体间多样性。而两种病害禾谷镰孢菌亲缘关系困地区不同而有差异，河北、吉林两地区穗、茎腐禾谷镰孢菌间均具有很近的亲缘关系，相似性系数在0．93以上；但沈阳地区禾谷镰孢菌亲缘关系相对较远，相似性系数仅为0．65～0．67。     

Impact of post-anthesis rainfall,fungicide and harvesting time on the concentration of deoxynivalenol and zearalenone in wheat

Field experiments were conducted to identify the impact of post-anthesis rainfall on the concentration of deoxynivalenol (DON) and zearalenone (ZON) in harvested wheat grain. Winter wheat plots were inoculated with Fusarium graminearum at stem extension (GS31) and prothioconazole was applied at mid-anthesis (GS65) to split plots and plots were subsequently mist irrigated for 5 days. Plots were either covered by polytunnels, irrigated by sprinklers or left as non-irrigated uncovered control plots after medium-milk (GS75). Plots were harvested either when ripe (GS92; early harvest) or three weeks later (late harvest). Fusarium head blight (FHB) was assessed each week from inoculation. At harvest, yield and grain quality was measured and grains were analysed for DON and ZON. Differences in rainfall resulted in contrasting disease pressure in the two experiments, with low FHB in the first experiment and high FHB in the second. Difference in FHB resulted in large differences in grain yield, quality and mycotoxin content. DON concentration was significantly (P < 0.05) higher in irrigated compared to covered and control plots in the first experiment, whereas in the second experiment, DON was significantly (P < 0.05) higher in the covered plots compared to the control and irrigated plots. ZON concentration was significantly (P < 0.05) higher in irrigated plots in both experiments. Later harvesting resulted in an approximate fivefold increase in ZON in the first experiment, but was not significantly different in the second experiment. Prothioconazole significantly (P < 0.05) reduced DON in both experiments, but gave inconsistent reductions to ZON. This is the first report to show that the post-anthesis rainfall can significantly increase ZON in wheat, which can increase further with a delayed harvest but may be significantly reduced with the application of prothioconazole. Importantly, in the absence of moisture late season, ZON remains at very low concentrations even when wheat is severely affected by FHB.     

Contamination of barley seeds with Fusarium species and their toxins in Spain: an integrated approach

Fusarium is a globally distributed fungal genus that includes different species pathogenic to cereals among others crops. Some of these Fusarium species can also produce toxic compounds towards animals and humans. In this work, the presence of the most important Fusarium toxins was determined in barley seeds from Spain, sampled according to European Union requirements. The results obtained were compared with the presence of mycotoxigenic species considered responsible for their synthesis by using species-specific polymerase chain reaction protocols. Fumonisins B₁ and B₂, zearalenone, trichothecenes type A (T-2 and HT-2) and trichothecenes type B (deoxynivalenol and nivalenol) were analysed by using high-performance liquid chromatography. Deoxynivalenol and zearalenone were detected in 72% and 38% of the barley samples, respectively, at levels below European Union limits in all cases. However, the co-occurrence of both toxins in 34% of the samples suggested that synergistic activity of these two mycotoxins should be evaluated. Nivalenol and HT-2/T-2 were detected at low levels in 17% and 10% of the samples, respectively. Fumonisins occurred in 34% of the samples at levels up to 300?µg/kg. This suggested that they might represent a risk in Spanish barley, and to our knowledge, this is the first report on the presence of fumonisins in barley in this country. The species-specific polymerase chain reaction assays to detect mycotoxin-producing Fusarium species showed a very consistent correlation between F. verticillioides detection and fumonisin contamination as well as F. graminearum presence and zearalenone, deoxynivalenol and nivalenol contamination in barley samples. The approach used in this study provided information of mycotoxin contamination of barley together with the identification of the fungal species responsible for their production. Detection of the species with the current polymerase chain reaction assay strategy may be considered predictive of the potential mycotoxin risk in this matrix.