Impact of competitive fungi on Trichothecene production by Fusarium graminearum

Bioassays were used to determine the production of the trichothecene mycotoxin, deoxynivalenol (DON), by two isolates of Fusarium graminearum when grown in association with potentially competitive fungi and an antifungal chemical, 6-pentyl-alpha-pyrone (6PAP). The presence of 6PAP in the culture medium reduced DON production by as much as 80%, but this effect was reduced for the F. graminearum isolate that most efficiently metabolized the added 6PAP. A 6PAP-producing Trichoderma isolate grown in a competition assay system with the F. graminearum isolates was also able to substantially reduce DON production. When Fusarium isolates (F. crookwellense, F. culmorum, F. subglutinans, F. poae, F. equiseti, F. avenaceum, and F. sambucinum), which co-occur with F. graminearum in New Zealand maize plants (Zea mays), were grown in competition assays, the effect on DON production was variable. However, all isolates of F. subglutinans tested were shown to cause reductions in DON production (by 13-76%, mean = 62%). F. subglutinans frequently co-occurs with F. graminearum, but its presence can vary with location and time of the season. When the competitive fungus tested was also a trichothecene producer (e.g., of nivalenol), both toxins were produced in the assay medium. The results indicate that mycotoxin production by F. graminearum can be affected by the presence of particular competitive fungi. These results have implications for an ecological understanding of pathogenicity and of mycotoxin accumulation in plants. Early establishment of F. subglutinans, for example, may act as a biological control mechanism providing a temporary protection against invasion by more commonly toxigenic fusaria such as F. graminearum.     

Enhancement of trichothecene production in Fusarium graminearum by cobalt chloride

The effects of cobalt chloride on the production of trichothecene and ergosterol in Fusarium graminearum were examined. Incorporation experiments with (13)C-labeled acetate and leucine confirmed that both 3-acetyldeoxynivalenol and ergosterol were biosynthesized via a mevalonate pathway by the fungus, although hydroxymethyl-glutaryl CoA (HMG-CoA) from intact leucine was able to be partially used for ergosterol production. Addition of cobalt chloride at concentrations of 3-30 μM into liquid culture strongly enhanced 3-acetyldeoxynivalenol production by the fungus, whereas the amount of ergosterol and the mycelial weight of the fungus did not change. The mRNA levels of genes encoding trichothecene biosynthetic proteins (TRI4 and TRI6), ergosterol biosynthetic enzymes (ERG3 and ERG25), and enzymes involved in the mevalonate pathway (HMG-CoA synthase (HMGS) and HMG-CoA reductase (HMGR)) were all strongly up-regulated in the presence of cobalt chloride. Precocene II, a specific trichothecene production inhibitor, suppressed the effects of cobalt chloride on Tri4, Tri6, HMGS, and HMGR, but did not affect erg3 and erg25. These results indicate that cobalt chloride is useful for investigating regulatory mechanisms of trichothecene and ergosterol production in F. graminearum.     

Long chain alkanes in silk extracts of maize genotypes with varying resistance to Fusarium graminearum

The alkane content of the silks of nine maize genotypes was analyzed to investigate the role of silk wax in resistance to Fusarium graminearum. Silk samples were collected 2, 4, 6, and 8 days after silk emergence and divided into three sections: exposed silk, silk channel silk, and silk that is under the husk and overlying the kernels. Four major unbranched alkanes (C(25), C(27), C(29), and C(31)) and three isoalkanes (C(27i), C(29i), and C(31i)) were identified. Total alkane contents were highest in the exposed silk followed by the silk channel silk, with the lowest in the youngest silk closest to the kernels. In the silk channel and overlying kernel silks, the moderately resistant inbred CO272 consistently had the highest alkane content. None of the other inbreds with improved resistance had as high a level of alkanes as CO272, indicating that alkane content is not a major mechanism of resistance.     

Fumonisin production by Fusarium verticillioides strains isolated from maize in Mexico and development of a polymerase chain reaction to detect potential toxigenic strains in grains

Fumonisins are mycotoxins produced by Fusarium verticillioides (Sacc. Nirenberg) in maize (Zea mays L.), a staple crop in Mexico. In this study, we report the isolation and identification of 67 Fusarium strains isolated from maize kernels collected in Northwest and Central Mexico. The strains were characterized regarding fumonisin B(1) production and the presence of the FUM1 gene. F. verticillioides was the predominant species isolated in both geographic regions, but the isolates from Northwest Mexico produced higher levels of fumonisin. A polymerase chain reaction (PCR)-based method, to detect a region of the FUM1 gene involved in fumonisin biosynthesis, was developed and employed to detect mycotoxigenic fungi in pure culture and in contaminated maize. The presence of the FUM1 gene was associated with fumonisin production in most isolates, except seven that did not synthesize fumonisin but contained the gene in their genome. The PCR method allowed the direct detection of fungal contamination in ground corn and could be employed to screen for the presence of potential mycotoxigenic fusaria.     

玉米籽粒冲击破碎特性试验研究

农业物料的冲击破碎特性对运输、排种和粉碎等机械设备的工作参数设定具有重要参考意义。该研究以玉米籽粒为研究对象,设计并搭建了单颗粒物料冲击破碎试验装置,开展了5种不同含水率（10.86%±0.13%,13.87%±0.18%,17.24%±0.08%、20.23%±0.19%和23.46%±0.23%）玉米籽粒的冲击破碎试验,并以籽粒破碎模式、临界速度和破碎程度表征籽粒的破碎特性。试验结果表明,玉米籽粒的破碎模式可分为未破碎、破裂和破碎3种形式,其中破裂形式包括整体破裂和局部破裂;以碎粒中大颗粒质量与玉米籽粒质量间的比值W界定玉米籽粒的破碎模式,并建立了不同含水率玉米籽粒的W值与冲击速度间的回归模型,模型的决定系数均大于0.92,确定了5种不同含水率（含水率由低到高）玉米籽粒的临界破裂速度分别为11.45、15.54、19.84、30.49和34.28 m/s,临界破碎速度分别为19.55、23.75、28.68、42.07和46.79 m/s,表明玉米籽粒的临界破裂速度和破碎速度均随含水率的增加而增加;采用响应曲面法建立了破碎颗粒平均粒径与冲击速度和含水率之间的回归模型,模型的决定系数为0.96,方差分析结果表明冲击速度、含水率及其交互作用对玉米籽粒的破碎程度影响极显著（P<0.01）,且破碎程度随冲击速度的提高而增加,随含水率的提高而降低。该研究可为玉米籽粒处理设备的工作参数设定提供参考,并为农业物料颗粒的冲击破碎特性研究提供理论依据和试验方法参考。     

Analysis of the Fusarium mycotoxins fusaproliferin and trichothecenes in grains using gas chromatography-mass spectrometry

A method is described using gas chromatography-mass spectrometry (GC-MS) for the simultaneous detection of the Fusarium mycotoxins fusaproliferin and seven trichothecenes from grains. Sample purification of the raw extract was carried out with commercial solid phase extraction columns, and the recovery of the more polar analytes was increased by rinsing the column with acetonitrile. A significant matrix effect was found for the analysis of fusaproliferin and trichothecenes; thus, the calibrants should be prepared in a blank matrix. The response was linear in the range used. The mean recovery for fusaproliferin was 60.4 or 62.9%, depending on the spiking level. With respect to the trichothecenes, the recovery was generally higher (70.2-125.3%). The method proved to be repeatable for the analysis of fusaproliferin and trichothecenes. The limit of detection for fusaproliferin in the blank matrix mixture was 50 microg/kg, and that for trichothecenes was 5-15 microg/kg. Thirty-eight Finnish grain samples were analyzed for fusaproliferin and trichothecenes with the method developed. Fusaproliferin was not detected in any of the samples. The mean levels of deoxynivalenol, 3-acetyldeoxynivalenol, nivalenol, HT-2 toxin, and T-2 toxin in Finnish grain samples were 272, 17, 150, 40, and <20 microg/kg, respectively.     

氮素供应对玉米子粒铁累积的影响

谷物子粒的铁营养是影响人及动物营养状况的重要因素之一,但对子粒铁营养的生理限制因素了解十分缺乏。氮素供应可能通过影响营养器官中的物质转移从而影响子粒铁的供应。本研究以玉米为材料,在田间两个氮水平(施氮、不施氮)下研究了吐丝至成熟期玉米从土壤中铁的吸收量、不同营养器官中铁的输出量、及其对子粒铁的表观贡献率,探讨了氮素供应对这些过程的影响。结果表明,氮素供应对子粒铁浓度没有显著影响,两个氮水平下,茎(含叶鞘)中铁的净输出量为1.25～1.71.mg/plant、输出率为40.4%～48.2%,对子粒铁的表观贡献率为50.9%～69.8%。中部叶片(穗位叶及其上、下各两片叶)中铁的含量表现为净增加。不施氮时营养器官中铁的净输出量和输出率低于施氮,而吸铁量对子粒铁的表观贡献率却高于施氮。两个氮水平下,吐丝至成熟期子粒铁的源(营养器官输出铁与从土壤中吸收的铁之和)的供应能力均大于子粒铁的累积能力(累积量),说明铁的吸收和转运可能不是决定子粒铁浓度的主要因素,子粒中铁的卸载及代谢可能是限制子粒铁营养的主要生理过程。     

两株高效溶磷菌的溶磷能力及其对玉米生长和红壤磷素形态的影响

[目的]磷在土壤中易于固定,且向有效态的转化能力弱.研究两株高效溶磷菌活化土壤中的磷素的能力,为提高红壤供磷能力提供指导.[方法]以溶磷菌株伯克霍尔德菌(Burkholderia)XQP35(P35)、拉乌尔菌(Raoultella)SQP80(P80)为研究对象,以磷酸铝、磷酸铁、植酸钙和卵磷脂替代液体NBRIP培养...     

Fumonisin disruption of ceramide biosynthesis in maize roots and the effects on plant development and Fusarium verticillioides-induced seedling disease

The fungus Fusarium verticillioides infects maize and produces fumonisins, inhibitors of ceramide synthase. Seeds of the cultivar Silver Queen were inoculated with fumonisin-producing or non-fumonisin-producing strains of F. verticillioides. Leaf lesion incidence and severity of effects on root and stalk growth were significantly correlated with fumonisin in roots and disruption of sphingolipid metabolism in roots. Uninoculated seeds grown in soil watered with solutions of fumonisin B1 exhibited above-ground symptoms indicative of F. verticillioides-induced seedling disease and dose-dependent reduction in root mass that was inversely correlated with fumonisin B1, sphingoid bases, and sphingoid base 1-phosphates in roots. There was also evidence of an adaptive response to disrupted sphingolipid metabolism in both the virulence and watering assays, suggesting induction of pathways responsible for metabolism of sphingoid base 1-phosphates after prolonged exposure. The results suggest that fumonisin, and its effects on sphingolipids, could contribute to all aspects of F. verticillioides maize seedling disease.     

Properties of the protein and carbohydrate fractions in immature wheat kernels

The time course of compositional changes in the polysaccharide and protein fractions, and of changes in protein structural properties, was investigated in grains of two Italian durum wheat cultivars, Ofanto and Duilio, and the pattern of changes was found to be specific to each cultivar. The fructo-oligosaccharide content in ripening kernels was different between cultivars, as was the pattern of starch accumulation. Evaluation of gluten proteins by Western-blotting using broad-specificity anti-gliadin antibodies showed the sudden appearance of immunoreactive proteins during ripening. Protein surface hydrophobicity and thiol content and accessibility were evaluated in conditions apt at assessing changes in protein interaction and/or in aggregative patterns. Proteins in the two cultivars showed differences in their surface hydrophobicity and in their content of accessible thiols. The simultaneous evaluation of changes in the individual components indicates that use of immature grains for the production of functional foods will require accurate monitoring of the rapidly evolving biopolymer patterns, and careful choice of the cultivar.     

Role of maize hybrids and their chemical composition in Fusarium infection and fumonisin production

This study was designed to investigate the role of hybrids in maize Fusarium section Liseola interaction and fumonisin production, with particular emphasis on the occurrence and accumulation of hidden fumonisins in maize (masking phenomenon). In this 2 year study, naturally infected field crops were chosen with 10 maize hybrids, six of them grown in both years. Maize samples collected in 2010 showed a higher incidence of fungal infection as well as higher fumonisin contamination than those obtained in 2009 but a very similar incidence of F. section Liseola. Fumonisin masking was confirmed in raw maize, with a lower amount of hidden forms as compared to free fumonisins detected in the year with higher contamination. The chemical composition of the different hybrids was determined and correlated with the contamination data: the results obtained highlight the main role of fatty acids, with a higher fumonisin contamination in hybrids showing a higher linoleic acid content and a higher masking action in hybrids with higher oleic to linoleic ratio. These results represent a good basis to explain maize hybrid susceptibility to F. section Liseola infection, fumonisin contamination, and masking not related to a specific commercial hybrid but extendable to all hybrids.     

Fumonisin production in the maize pathogen Fusarium verticillioides: genetic basis of naturally occurring chemical variation

Fumonisins are polyketide-derived mycotoxins produced by the maize pathogen Fusarium verticillioides. Previous analyses identified naturally occurring variants of the fungus that are deficient in fumonisin C-10 hydroxylation or that do not produce any fumonisins. In the current study, gene deletion and genetic complementation analyses localized the C-10 hydroxylation deficiency to a cytochrome P450 monooxygenase gene in the fumonisin biosynthetic gene (FUM) cluster. Sequence analysis indicated that the hydroxylation deficiency resulted from a single nucleotide insertion that caused a frame shift in the coding region of the gene. Genetic complementation localized the fumonisin-nonproduction phenotype to the polyketide synthase gene in the FUM cluster, and sequence analysis indicated that the nonproduction phenotype resulted from a nucleotide substitution, which introduced a premature stop codon in the coding region. These results provide the first direct evidence that altered fumonisin production phenotypes of naturally occurring F. verticillioides variants can result from single point mutations in the FUM cluster.     

Fusarium graminearum TRI14 is required for high virulence and DON production on wheat but not for DON synthesis in vitro

Fusarium head blight (FHB) of wheat (Triticum aestivum L.), caused by the fungus Fusarium graminearum, is a major concern worldwide. FHB grain is reduced in yield, may fail to germinate, and is often contaminated with deoxynivalenol, a trichothecene mycotoxin linked to a variety of animal diseases and feed refusals. Annual losses in the tens of millions of dollars due to FHB underscore the need to develop improved methods of disease control and prevention. Previous research has identified deoxynivalenol biosynthesis as a virulence factor on wheat. Recently, we found that the TRI14 gene of F. sporotrichioides, closely related to F. graminearum, was not required for synthesis of a related trichothecene, T-2 toxin. TRI14 does not share similarity with any previously described genes in the databases. In this study, we examined the role that F. graminearum TRI14 may play in both deoxynivalenol synthesis and in virulence on wheat. TRI14 deletion mutants synthesize deoxynivalenol on cracked maize kernel medium and exhibit wild-type colony morphology and growth rate on complex and minimal agar media. However, FHB assays on greenhouse-grown wheat indicate that FgDeltaTri14 mutants cause 50-80% less disease than wild type and do not produce a detectable quantity of deoxynivalenol on plants. We discuss a number of possible roles that TRI14 may play in the disease process.     

Patterns of trichothecene production, genetic variability, and virulence to wheat of Fusarium graminearum from smallholder farms in Nepal

Fusarium graminearum causes wheat head blight and contaminates grain with the trichothecenes 4-deoxynivalenol and nivalenol. Sequence analysis of trichothecene genes indicates that nivalenol production is the ancestral trait; however, deoxynivalenol producers occur worldwide and predominate in North and South America and in Europe. Analysis of a large field population (>500 strains) from Nepal identified three groups that were both genetically distinct and polymorphic for trichothecene production: SCAR1 comprising 95% deoxynivalenol producers, SCAR2 comprising 94% nivalenol producers, and SCAR3/5 comprising 34% deoxynivalenol producers/63% nivalenol producers. The ability to cause wheat head blight differed between SCAR groups and trichothecene chemotypes: deoxynivalenol producers were more virulent than nivalenol producers across all three SCAR groups and within the SCAR3/5 genetic background. These data support the hypothesis that production of deoxynivalenol rather than nivalenol confers a selective advantage to this important wheat pathogen.     

磷对不同玉米品种生长、体内磷循环和分配的影响

以杂交玉米蠡玉16和冀单28为供试作物,采用供高磷（250 mol/L）和低磷（5 mol/L）营养液的石英砂培养方法,研究2个玉米品种的各器官干重和磷积累与分配、体内磷在木质部和韧皮部中的循环、流动及磷的吸收和利用效率。结果表明,与供高磷处理相比,低磷处理的2个玉米品种各营养器官的干重、磷含量和木质部中运输的磷量显著降低; 而磷在体内韧皮部的再循环显著增加,并且玉米各部位叶片活化出的磷主要是通过韧皮部循环至根中后,再经过木质部向上部新生叶运输的; 体内光合产物与磷向上部叶的运输是不同步的过程。低磷时,与冀单28相比,蠡玉16的根冠比高010,整株干重和磷含量增加269%和120%,磷吸收和利用效率提高121%和133%,木质部总磷向上部叶运输的比例高306%。说明低磷条件下,磷高效玉米品种生物量大是由于具有较大的根冠比,木质部中更大比例的磷被分配到上部新生叶以及其具有较高的磷吸收和利用效率。     

Production of trichothecenes and other secondary metabolites by Fusarium culmorum and Fusarium equiseti on common laboratory media and a soil organic matter agar: an ecological interpretation

Fusarium culmorum and F. equiseti were characterized with regard to production of trichothecenes and other secondary metabolites. Results following growth on laboratory media are interpreted with the aim of increasing the understanding of fungal metabolism in the field environment. While trichothecene production was detected for 94 of 102 F. culmorum isolates, only 8 of 57 F. equiseti isolates were positive. Profiles of secondary metabolites were compared by following growth on yeast extract sucrose agar (YES), potato sucrose agar (PSA), and an agar medium, prepared from soil organic matter (SOM), which was included to simulate growth conditions in soil. SOM supported the production of chrysogine by F. culmorum. The two species utilized the media differently. F. culmorumproduced zearalenone (ZEA) on YES, whereas some F. equiseti isolates produced ZEA on PSA. Other F. equiseti isolates produced equisetin. These differences may reflect that F. culmorum depends on a pathogenic life style while F. equiseti has a more saprotrophic mode of existence.     

利用玉米秸秆制备生物杀菌剂的研究

通过拮抗性筛选分离出对蔬菜枯萎病原菌具有很强抑制作用的木霉菌株“T41”,并利用玉米秸秆粉为原料,采用液固两相发酵工艺制成木霉菌剂。该菌剂对多种蔬菜枯萎病防治效果显著且稳定,并高于化学杀菌剂防治效果     

Fast and sensitive screening method for detection of trichothecenes in maize by using protein synthesis inhibition in cultured fibroblasts

A fast and sensitive bioassay with hamster (BHK-21 C13) fibroblasts for the detection of toxic trichothecenes in maize is described. Cells are exposed to pure toxins or crude maize extracts for 30 min. The mixture is then incubated with [1-14C]-leucine for an additional 60 min and the radioactivity incorporated into the protein of the washed cells is determined. The sensitivity of the assay was in the range 1-10 ng/mL (or 50 ppb in maize) for T-2, HT-2, and diacetoxyscirpenol. At least 1000-fold higher concentrations of non-trichothecene mycotoxins and plant toxins were necessary to cause an inhibition of protein synthesis in the cells. Of 24 maize samples tested, 14 gave a positive response in this assay and the presence of trichothecenes could be confirmed chemically in 11 samples. Therefore, the described bioassay is proposed as a useful screening method for cytotoxic trichothecenes in maize.     

钾对不同基因型玉米生长、体内钾循环和分配的影响

钾素循环和再利用对维持植物生长和提高钾利用效率非常重要。本文以杂交玉米豫玉23和兴农998为供试作物,采用供应高钾（1.85 mmol/L）和低钾（0.1 mmol/L）营养液的石英砂培养方法,研究不同基因型玉米各器官干重和钾积累及分配、 体内钾在木质部和韧皮部中的循环、 流动及钾效率等。结果表明,低钾处理10 d后,与高钾处理相比,低钾降低两玉米品种各器官干重净增量、 钾浓度、 钾积累量、 钾吸收效率和木质部中钾的运输量,增加钾利用效率和源叶中韧皮部输出的钾,提高钾通过韧皮部的再循环量及占木质部运输总量的比例。低钾处理的豫玉23和兴农998上部叶均为钾库,钾积累量的38.5%和70.3%是由各自中、 下部叶韧皮部输出的钾供给,但体内光合产物和钾向上部叶的运输是不完全同步的过程。两品种比较,低钾处理下的豫玉23较兴农998干重净增量提高,在于前者具有较大根量、 较高钾吸收速率和较多钾素吸收量,木质部中有更多的钾通过叶片韧皮部输出进行循环运输进入上部叶;但豫玉23对介质中钾的吸收大于体内钾的再利用。钾营养高效基因型玉米应该具有较强的吸收和利用体内钾的能力。     

Fumonisins: probable role as effectors in the complex interaction of susceptible and resistant maize hybrids and Fusarium verticillioides

Fusarium verticillioides is best known for its worldwide occurrence on maize resulting in highly variable disease symptoms, ranging from asymptomatic to severe rotting and wilting and fumonisin production. The aim of this study was to investigate the influence of hybrid genotypes in the early stages of F. verticillioides infection, and the role of fumonisins as effectors in the outcome of this complex interaction. Disease symptoms, growth parameters, root morphology, and fungal colonization were evaluated at 7, 14, and 21 days after planting in seedlings from maize seeds of resistant (RH) and susceptible (SH) hybrids inoculated with F. verticillioides or watered with solutions of fumonisins. F. verticillioides induced growth enhancement or retardation depending on the plant genetic background and the fungal colonization rate, while fumonisins caused severe reduction in biomass and fitness. Seedlings watered with high fumonisin concentrations displayed lesions similar to those seen in F. verticillioides maize seedling disease, and also elicited inhibitory effects on root growth and morphology and on functional properties. In summary, these data strongly suggest a dual role for fumonisins in the F. verticillioides-maize interaction, acting as pathogenic factors at high concentrations, or triggering the plant detoxification mechanisms at low levels.