Potential of fungal antagonists for biocontrol of Fusarium spp. in wheat and maize through competition in crop debris

Pathogenic Fusarium spp. cause head blight in wheat or ear rot in maize leading to yield losses and also a reduction in quality due to mycotoxin contamination of the grain. Infected crop residues are the main inoculum source for epidemics. Saprophytic fungi, obtained from cereal tissues or necrotic tissues of other crops, were screened for their ability to colonise wheat straw and maize stalks and to suppress sporulation of pathogenic Fusarium spp. Results of bio-assays conducted under controlled conditions were variable among Fusarium spp. and host substrates for most antagonists tested, such as yeasts, Trichoderma spp. and non-pathogenic Fusarium spp. Isolates of Clonostachys rosea consistently suppressed sporulation of F. culmorum and F. graminearum on wheat straw, and of F. culmorum, F. graminearum, F. proliferatum and F. verticillioides on maize stalks. Isolates of C. rosea, C. cladosporioides and F. equiseti were applied to pieces of maize stalks or flowering ears in preliminary experiments conducted under field conditions. The colonisation of stalk pieces by pathogenic Fusarium spp. was assessed after 9 months. Colonisation of stalk pieces by pathogenic Fusarium spp. was significantly reduced at several sampling dates. However, results obtained with the antagonists were not consistent for all sampling dates and between experiments.     

防风根际真菌的分离鉴定及其生防潜力评价

作为世界性分布的镰刀属常见病原真菌,尖孢镰刀菌（Fusarium oxysporum）和木贼镰刀菌（F. equiseti）对包括经济作物及药用植物等的生长均有较大危害。利用源自于植物根际土壤的有益微生物防控尖孢镰刀菌和木贼镰刀菌引起的真菌性病害,是目前较为理想的植物病害管理策略。为获得可有效抑制尖孢镰刀菌和木贼镰刀菌的生防菌源,于防风根际土壤中分离纯化真菌104株,基于平板对峙法筛选获得1株对尖孢镰刀菌和木贼镰刀菌具有显著抑制效果的菌株MR-43,结合形态特征、ITS序列分析,将其确定为Sirastachys castanedae（GenBank登录号：OK287148.1）,隶属于Sirastachys组进化分支,发现其可宿生于植物根际土壤。基于盆栽试验法,探究MR-43在防风栽培土壤中的定殖能力,评价MR-43对由尖孢镰刀菌引起的防风枯萎病和由木贼镰刀菌引起的防风根腐病的防病能力,及其对防风植株的促生能力。结果显示,Sirastachys castanedae MR-43对尖孢镰刀菌、木贼镰刀菌的抑菌率为57%以上,且经多次验证抑菌效果稳定;菌株MR-43可稳定定殖于防风栽培土壤并实现有效扩繁;菌株MR-43的孢子悬液可有效控制防风枯萎病和防风根腐病,平均防效达68.52%,与多菌灵、代森锰锌的防病效果无显著性差异（P>0.05）;此外,MR-43对防风植株生长具有明显促进作用。因此,Sirastachys castanedae MR-43在防风枯萎病、根腐病等真菌性病害管理方面具有较好的开发价值及应用潜力。     

Contamination of malt barley and wheat by Fusarium graminearum and Fusarium culmorum from the crop years 2001-2003 in eastern Croatia

This study investigated infection levels with Fusarium graminearum and Fusarium culmorum in malt barley and wheat in eastern Croatia. The contamination was surveyed over three consecutive crop years (2001–2003) on five locations for barley and three wheat cultivating locations. F. graminearum loads reached levels of potentially serious threat for the commercial production of malting raw materials in both cereals (up to 29.1%). On the other hand, the mean percentage of kernels infected with F. culmorum was low to medium (up to 6.1%). The fungal invasions for years and locations were affected by meteorologic and other environmental factors and the pattern seemed to be consistent with species-specific optimal conditions reported by other authors.     

镉胁迫对绢毛委陵菜-丛枝菌根真菌共生体根系结构和生理的影响

为从生理水平上揭示绢毛委陵菜（Potentilla sericea L.）-丛枝菌根真菌共生体对镉胁迫的应答机制,以绢毛委陵菜多年生植株为试验材料,分别接种根内根生囊霉（Rhizophagus intraradices）和摩西球囊霉（Funneliformis mosseae）,80 d后以不同浓度的镉胁迫处理并测定菌根侵染率,研究侵染率较高菌种的共生体植株的生理指标变化和结构变化。结果表明：摩西球囊霉较根内根生囊霉对绢毛委陵菜菌根侵染率高,达100%,而根内根生囊霉侵染率为89.33%;接种摩西球囊霉真菌的植物,当镉质量浓度达10 mg?L^-1时,细胞壁小范围破裂、液泡出现变小的趋势,高浓度镉（20 mg?L^-1）处理时,根系细胞壁破裂,细胞无法维持原有形态;镉胁迫下,Cd（5 mg?L^-1）~ Cd（10 mg?L^-1）MDA含量显著上升（P<0.05）,Cd（0 mg?L^-1）~Cd（5 mg?L^-1）SOD活性和脯氨酸含量变化显著（P<0.05）,可溶性蛋白含量和叶绿素含量显著减少（P<0.05）,Cd（10 mg?L^-1）~Cd（15 mg?L^-1）可溶性蛋白含量和叶绿素含量变化不显著（P>0.05）,植物可以抵抗一定浓度范围的镉胁迫。综合来看,接种摩西球囊菌可以增强绢毛委陵菜耐镉胁迫能力,为利用绢毛委陵菜修复镉污染土壤提供了理论依据。     

Potential for control of seedling blight of wheat caused by Fusarium graminearum and related species using the bacterial endophyte Bacillus mojavensis

Fusarium-infected wheat seed decreases germination, seedling emergence, and causes post emergence seedling death, and can contribute to wheat scab and ear rot of maize, with consequent production of mycotoxins such as deoxynivalenol and zearalenone. Current seed treatments have proved ineffective in controlling seedling blight and scab. A patented endophytic bacterial strain, Bacillus mojavensis RRC 101, and several other strains of this species were studied to determine in vitro antagonism to some Fusarium species and to assess the potential of this bacterium to serve as an endophytic biocontrol for seedling blight of wheat produced by species within the F. graminearum complex, as well as other species of Fusarium. Seedling emergence and seed germination were two tests used as indicators of seedling blight. These tests were conducted in growth rooms with two wheat cultivars highly susceptible to scab, Norm and Pioneer 2552, and other cultivars with varying resistance to scab. The results indicated that all strains of this bacterium were antagonistic in vitro to the strains of F. graminearum and its seven related species, as well as four strains of F. pseudograminearum and the two strains of F. verticillioides. Germination of the highly scab susceptible cultivar 2552 was increased from 77 to 97% when planted in soil containing a mixed inoculum of F. graminearum and related species. Seedling emergence in the very susceptible wheat cultivar Norm increased from 20 to 82% when treated with the bacterium. The data indicated that inoculating wheat kernels with B. mojavensis reduced seedling blight of wheat produced by F. graminearum and related Fusarium species indicating the potential for this bacterium as a biocontrol under field condition.     

Trichothecene Content of Rye and Wheat Genotypes Inoculated with a Deoxynivalenol- and a Nivalenol-producing Isolate of Fusarium culmorum

Head blight caused by Fusarium culmorum may lead to yield reduction and the contamination of cereal grain with the mycotoxins deoxynivalenol (DON), 3-acetyl deoxynivalenol (3-ADON), nivalenol (NIV), fusarenone-X (FUS), and others. In this study, the covariation between DON and NIV accumulation of 12 rye and eight wheat genotypes that differed in resistance were analysed by inoculating them with a DON-and a NIV-producing isolate, respectively, in three locations. The resistance traits head blight rating and plot yield relative to the uninoculated plots of the same genotype were assessed and the contents of DON, 3-ADON, NIV, and FUS in the grain were analysed by gas chromatography with mass spectrometry. The NIV-producing isolate was significantly (P=0.05) less aggressive and led to a considerably lower mean NIV content in the grain compared with the aggressiveness and mean DON content of the DON-producing isolate (19.5 mg NIV/kg grain versus 48.4 mg DON/kg). Wheat and rye genotypes significantly differed in their DON and NIV accumulation. All genotypes reacted in a similar manner to both chemotypes of F. culmorum for the resistance traits and the respective mycotoxin contents with the exception of one wheat variety, that caused a change in rank order for mycotoxin content. In conclusion, resistance to head blight and tolerance to mycotoxin accumulation seems to be most likely the same for DON- and NIV-producing isolates of F. culmorum .     

Production of a Mixed Inoculum of Fusarium oxysporum Fo47 and Pseudomonas fluorescens C7 to Control Fusarium Diseases

Strains of non-pathogenic Fusarium oxysporum and Pseudomonas fluorescens are effective biocontrol agents against Fusarium diseases. Use of strain Fo47 of F. oxysporum in combination with strain C7 of P. fluorescens improves disease control in comparison to application of Fo47 alone. To develop a product based on a combination of these two strains, it would be beneficial to produce and formulate both organisms together. When an irradiated peat was inoculated with these strains either alone or in combination, they grew actively and reached similar maximum population densities irrespective of the initial inoculum concentration (1×10³ and 1×10⁶ g^-1 of peat) and mode of inoculation. These populations survived well with only a slight decrease in population densities after 11 months storage at 25°C. A bioassay was performed to evaluate peat and microbial suspension inocula of C7 and Fo47 alone or in combination against the flax pathogen, F. oxysporum f. sp. lini. Biocontrol activity of the peat-produced inocula was as effective as the suspensions. There was no dose-response relationship for C7, and when C7 was used as a mixed inoculum with Fo47, the biocontrol activity was always greater than that provided by Fo47 alone. This process could be used to produce an effective mixed inoculum with a shelf life of one year or more.     

Isolation of 4-acetyl-benzoxazolin-2-one (4-ABOA) and diferuloylputrescine from an extract of gibberella ear rot-resistant corn that blocks mycotoxin biosynthesis, and the insect toxicity of 4-ABOA and related compounds

Bioassay-directed fractionation of an extract of a gibberella ear rot-resistant corn hybrid resulted in the isolation of a fraction that inhibited mycotoxin production, but not growth, of Fusarium culmorum. The fraction contained diferuloylputrescine and a new allelochemical 4-acetyl-benzoxazolin-2-one (4-ABOA). This report describes the isolation of the bioactive fraction and tests of the effect of 4-ABOA on 3-acetyl deoxynivalenol (3ADON) and aflatoxin production. In addition, data are reported on the effects of 4-ABOA and related compounds on a number of insect species and some preliminary studies on the mechanism of insect toxicity. The effect of 4-ABOA on F. culmorum and Apergillus flavus was similar. Production of toxin was reduced without a concomitant reduction of growth. A 50% inhibition of 3ADON production occurred at ca. 4 μM and 0.12 mM for alfatoxin. 4-ABOA was slightly toxic to Spodoptera frugiperda.     

茄病镰刀菌单克隆抗体的制备及胶体金免疫层析试纸条的研发*

Objective: The cell lines secreting specific monoclonal antibodies (McAbs) were prepared by using Fusarium solani, one of the pathogenic fungi causing root rot of Fritillaria thunbergii, and the colloidal gold immunochromatographic test strip based on McAbs was developed to provide scientific basis for detecting root rot of F. thunbergii. Methods: Hybridoma technology was used to obtain cell lines that could secrete specific McAbs against F. solani using the whole protein extract of F. solani as the antigen. The specificity, titer, sensitivity and binding protein of McAbs were detected by indirect ELISA and Western blot. Colloidal gold particles were prepared by trisodium citrate reduction method and McAbs were labeled to prepare colloidal gold immunochromatographic strip. Results: Three cell lines secreting specific McAbs against F. solani were obtained, which were named as FsA3, FsG6 and FsD4. The detection sensitivity of FsA3 was 24.41 ng / mL, and that of both FsG6 and FsD4 was 12.21 ng / mL. FsA3, FsG6 and FsD4 had strong reactions to F. solani, and had no cross-reaction to Alternaria tenuissima, A. alternata, Botrytis cinerea, F. equiseti, F. incarnatum, F. oxysporum, Phoma sp., and Phomopsis oblonga. The colloidal gold immunochromatographic strip based on FsG6 showed only a quality control line when detecting the tissue culture seedlings of F. thunbergii. When 100 ng F. solani antigen or the samples of F. thunbergii infested with root rot disease were detected, there were visible quality control lines and test lines. Conclusion: The specificity and sensitivity of the McAbs and test strip are sufficient to detect F. solani isolated from diseased strains of F. thunbergii, which provides the technical support for the rapid detection of root rot of F. thunbergii in the field.     

Influence of Growth Stage on Fusarium Head Blight and Deoxynivalenol Production in Wheat

Fusarium head blight is a major concern for wheat production worldwide. The fungi that cause the disease may infect head tissues from flowering to late stages of kernel development, but a better understanding of the influence of the time of infection on grain weight reduction and mycotoxin accumulation resulting from the infection process is needed. We investigated the influence of wheat reproductive stage at the time of inoculation on disease and grain quality parameters, especially production of deoxynivalenol (DON) in mature grains. Heads of Norm wheat were spray inoculated with a macroconidial suspension of a DON‐producing isolate of Fusarium graminearum at each of six reproductive stages from flowering to hard dough. Plants were incubated in a mist chamber for 48 h and then moved to the greenhouse until maturity. Norm wheat was susceptible at all stages inoculated but the highest grain weight reduction and DON accumulation occurred in plants inoculated past flowering to late milk stages. However, high incidences of kernel infection and significant levels of DON accumulation resulted from inoculations as late as the hard dough stage, even though there was no corresponding reduction in grain weight compared to non‐inoculated plants. The occurrence of commercially significant levels of DON in plump, high‐yielding wheat may result from infections that occur during favourable environments well after the flowering stages. Late infection and DON production should therefore be a future research focus for wheat breeding and integrated management of FHB and an important consideration for grading systems that employ the presence of visibly damaged kernels as a means of estimating DON content of wheat.     

Occurrence and Distribution of Microdochium and Fusarium Species Isolated from Durum Wheat in Northern Tunisia and Detection of Mycotoxins in Naturally Infested Grain

An outbreak of Fusarium Head Blight of durum wheat occurred in 2004 being localized in sub-humid and higher semi-arid region of Northern Tunisia. A mycological survey carried out throughout these regions, revealed that 78% of the prospected fields were infested. Results of the morphological and molecular identification, showed that the most common species isolated from diseased wheat spikes was Microdochium nivale var. nivale (63.5%), followed by Fusarium culmorum (26%), F. pseudograminearum (9%) and F. avenaceum (1.5%). To evaluate mycotoxin content of naturally infected grain, the amounts of trichothecene mycotoxin deoxynivalenol (DON) in harvested grain from 45 fields were quantified by RIDASCREEN DON Enzyme Immunoassay Kit (ELISA) . This study showed that the infection levels in freshly harvested grain were very low and the maximum deoxynivalenol (DON) level of the positive samples was 53 ppb. This is the first report on the natural occurrence of DON in naturally infected wheat grain sampled from Northern Tunisia.     

Growth inhibition of the cereal root pathogens Rhizoctonia solani AG8, R. oryzae and Gaeumannomyces graminis var. tritici by a recombinant 42-kDa endochitinase from Trichoderma harzianum

The objective of this study was to determine the effectiveness of a 42-kDa endochitinase coded by the ThEn42 gene from Trichoderma harzianum as a potential source of transgenic resistance to Rhizoctonia root rot of barley caused by Rhizoctonia solani AG8 and/or R. oryzae. The gene cThEn42 was codon optimized (GC content increased from 53.3 to 65.1%) and then synthesized to produce the modified cThEn42GC in Pichia pastoris for in vitro tests. Two expression vectors were constructed: one with the fungal signal peptide and the fungal activation peptide [FSP-FAP-cThEn(GC)] and the other with barley chitinase 26 signal peptide followed by the fungal signal and activation peptides [SP(HVChi26)-FSP-FAP-cThEn(GC)]. N-terminal sequencing showed that, of two proteins secreted into liquid medium, FSP was cleaved off faithfully in one protein and both FSP and FAP were cleaved from the other protein. Purified endochitinase provided strong in vitro inhibition of both R. solani AG8 and R. oryzae. The enzyme had an intermediate inhibitory activity against Gaeumannomyces graminis var. tritici, and no inhibitory activity against Fusarium graminearum, F. pseudograminearum, and F. culmorum.     

三七连作土壤浸提液对其根腐病菌的化感效应

采用甲醇、乙酸乙酯和水分别按液土比3∶1、6∶1和9∶1对三七连作土壤进行浸提,研究其浸提液对三七根腐病菌生长和种群数量的影响。结果表明: 平板培养72 h后,甲醇、乙酸乙酯和水浸提液对尖镰孢菌和腐皮镰孢菌的菌丝生长均表现为化感促进,其中,甲醇和乙酸乙酯浸提液对尖镰孢菌的化感效应指数为14.0%～19.8%和16.2%～20.2%,高于水浸提液的8.9%～14.2%,且不同浸提比例之间差异不显著;而甲醇浸提液对链格孢菌菌丝生长表现为化感抑制,且抑制效应在浸提比例为3∶1时最强,达到-33.2%～-38.5%,乙酸乙酯和水浸提液对链格孢菌菌丝生长无显著影响。土壤培养4周后,甲醇、乙酸乙酯和水浸提液均能增加土壤中尖镰孢菌的数量,其中,水浸提液的增加效应最强,达到每克干土3.49×10⁶～9.56×10⁶拷贝数,高于甲醇(每克干土1.68×10⁴～6.73×10⁴拷贝数)和乙酸乙酯浸提液(每克干土1.77×10⁴～3.72×10⁴拷贝数),且这种增加效应随浸提比例的增加逐渐减弱;水浸提液和低浸提比例的甲醇提取液均能增加土壤中腐皮镰孢菌的数量,而重茬土壤浸提液对链格孢菌的数量影响不显著。因此,三七连作土壤浸提液对根腐病菌如尖镰孢菌和腐皮镰孢菌均表现出明显的化感促进效应,这可能是再植三七易发生根腐病等土传病害的原因之一。     

Deoxynivalenol-nonproducing Fusarium graminearum Causes Initial Infection,but does not Cause DiseaseSpread in Wheat Spikes

Fusarium graminearum is a major pathogen that causes fusarium head blight (FHB) in wheat and produces deoxynivalenol (DON) in infected grain. In previous studies, the trichodiene synthase gene (Tri5) in the fungal strain GZ3639 was disrupted to produce the DON-nonproducing strain GZT40.In this report, the virulence of strains GZ3639 and GZT40 was tested on wheat cultivars with various resistance levels by using methods of spray inoculation and injection inoculation with fungal conidia. Under field and greenhouse conditions, strain GZ3639 produced significantly more disease symptoms and reduced more yield than strain GZT40 in all wheat cultivars tested. Conidia of strain GZT40 germinated and infected inoculated spikelets, but disease symptoms were limited to inoculated spikelets without spread to uninoculated spikelets. When strain GZT40 was inoculated using the spray method, multiple initial infection sites in a spike resulted in higher levels of disease symptoms than in spikes inoculated by a single injection. Greenhouse tests confirmed that strain GZT40 did not produce DON in the infected kernels following either inoculation method. The results confirm that DON production plays a significant role in the spread of FHB within a spike, and are the first report that DON production is not necessary for initial infection by the fungus. This revised version was published online in June 2006 with corrections to the Cover Date.     

东北红豆杉—无花果复合种植对两种植物生长和土壤酶活性影响

设计了室内控制条件下的东北红豆杉和无花果的复合种植实验，并与单一种植东北红豆杉和单一种植无花果的模式进行了比较。分别考察了上述3种模式下，东北红豆杉和无花果的生长指标、光合作用以及土壤酶活性差异。结果表明：①在生长量方面，复合种植3个月的东北红豆杉的株高增长率为152.6%，是单一种植的东北红豆杉株高增长率的1.56倍；复合种植3个月的无花果，其当年生枝的生长量为68.4 cm³，比单一种植的无花果高36.3%。②在光合作用指标方面，在800 μmol·m^-2·s^-1光照强度下，复合种植3个月的东北红豆杉和无花果净光合速率与相应的单一种植相比，分别提高了19.0%和5.3%。③在土壤酶活性方面，在7月份，复合种植模式下的土壤蔗糖酶、脲酶和酸性磷酸酶活性比单一种植东北红豆杉土壤相应酶活性分别提高51.6%、58.5%和50.8%，比单一种植物无花果土壤相应酶活性分别提高85.5%、47.5%和71.9%；土壤多酚氧化酶活性在不同种植模式之间无显著差异（P>0.01）。综上，东北红豆杉-无花果复合种植可以提高土壤蔗糖酶、脲酶和酸性磷酸酶活性，对两种植物生长均具有促进作用。上述结果可为在生产实践上进行东北红豆杉-无花果复合经营提供参考。     

The Effect of Nitrogen Fertilization on Fusarium Head Blight Development and Deoxynivalenol Contamination in Wheat

The impact of nitrogen (N) fertilization on the development of Fusarium head blight (FHB) in wheat and the resulting deoxynivalenol (DON) contamination in the kernels was studied. In a first experiment, the disease was assessed on two locations under natural infection pressure. Five different types of nitrogen fertilizer (both organic and mineral) were investigated, each applied at five input rates from 0 to 160 kg N/ha. With all fertilizers, a significant increase of disease intensity was observed with increasing N input, while the type of N fertilizer had poor or no effects on FHB. Depending on the fertilizer used, the percentage of diseased spikelets increased from 2.2% at zero N rate up to 6.6% at 160 kg N input per hectare. In a second series of trials, three spring wheat varieties including one Durum wheat line were artificially inoculated with a Fusarium graminearum and a F. culmorum strain, known producers of DON. A mineral N fertilizer was applied at five input levels from 0 to 160 kg N/ha. A significant increase in FHB intensity and DON contamination in the grain was observed with increasing N from 0 to 80 kg/ha. At higher input rates, relevant in contemporary crop husbandry, disease intensity and toxin contamination remained at constant levels. It is concluded that adaptation of N fertilization represents no relevant tool in managing FHB in practical wheat cultivation.     

On-farm experiments over 5 years in a grain maize/winter wheat rotation: effect of maize residue treatments on <Emphasis Type="Italic">Fusarium graminearum</Emphasis> infection and deoxynivalenol contamination in wheat

Over the course of 5 years, different maize residue treatments were conducted on 14 zero tillage on-farm sites in Switzerland to evaluate their effect on the development of Fusarium head blight (FHB) and the contamination with the mycotoxin deoxynivalenol (DON) in winter wheat grains and wheat straw following grain maize. Two experimental series with three and five different treatments were carried out, respectively. Fusarium graminearum (Schwabe) was the predominant FHB-causing species with an overall incidence of 15% infected wheat grains. A significant correlation between symptoms in the field, F. graminearum incidence and DON content in wheat grains and wheat straw was observed. The average DON content in both wheat grains and wheat straw was approximately 5,000 μg/kg and thus several times higher than the European maximum limit of 1,250 μg/kg for unprocessed small-grain cereals for human consumption. Of all grain samples, 74% were above the maximum limit. Pooled over both experimental series, the average reduction of DON in grains through treatments of the maize residue compared with a control treatment ranged between 21 and 38%. The effect of various other factors, including the year, the wheat variety, the site, the maize hybrid and the production system was evaluated as well. The year and the wheat variety were the most important FHB influencing factors. Over all treatments, the variety Levis showed a fivefold higher average DON content compared with the variety Titlis. From different categories of maize residue particles, intact pieces of 5–15 cm length were strongly correlated with F. graminearum incidence and DON content in grains. During the time course of this study, the recommendation from a preliminary version of the internet-based DON forecasting system FusaProg to apply or to omit a fungicide treatment was correct in 32 out of 42 cases. The results are currently being used to optimise the FusaProg models. This study has shown that in a grain maize/winter wheat rotation, the DON content in wheat grains frequently exceeded the European maximum limit, even with a thorough treatment of maize residues and less susceptible wheat varieties. Hence, in order to reduce the contamination risk in a zero tillage system, the crop rotation needs to be modified.     

Characterisation of<Emphasis Type="Italic">fusarium graminearum</Emphasis> and<Emphasis Type="Italic">F. culmorum</Emphasis> isolates by mycotoxin production and aggressiveness to wheat

A total of 27Fusarium culmorum isolates from Germany and 41F. graminearum isolates from Kenya were investigated for aggressiveness and mycotoxin production on wheat ears. In addition, ergosterol content of the kernels from ears inoculated withF. graminearum was determined and theF. culmorum isolates were tested for mycotoxin productionin vitro. For both pathogens, isolates markedly differed in aggressiveness. 59% and 37% of theF. culmorum isolates produced NIV and DON, respectively,in vivo andin vitro. The DON-producing isolates also produced 3-acDONin vitro. The more aggressive isolates produced mainly DON while the less aggressive isolates produced mainly NIV. 12% and 85% of theF. graminearum isolates produced NIV and DON, respectively. The highly aggressive isolates produced higher amounts of DON, aggressiveness being highly correlated to DON content in the kernels. NIV-producing isolates were less aggressive. Ergosterol content of kernels was moderately correlated to aggressiveness but highly correlated to DON content. Disease severity was associated with kernel weight reduction.     

Cytotoxicity induced by nivalenol, deoxynivalenol, and fumonisin B1 in the SF-9 insect cell line

The toxicity of the mycotoxins nivalenol (NIV), deoxynivalenol (DON), and fumonisin B1 (FB1) was studied in the lepidopteran Spodoptera frugiperda (SF-9) cells, by the trypan blue dye-exclusion and 3-(4,5-dimethylthiozole-2-yl)-2,5-biphenyl tetrazolium bromide (MTT) tests, uptake analyses of cytotoxicity, and cell metabolism, respectively. Deoxyribonucleic acid analysis by flow cytometry was used to identify apoptosis and cell cycle distribution. After 48 h of exposure, the MTT and trypan blue dye-exclusion tests indicated that NIV was significantly more toxic than DON, and both were significantly more toxic than FB1. The IC50 (mycotoxin concentration resulting in 50% inhibition of proliferation) values for NIV and DON were 4.5 and 41 microM, and the CC50 (mycotoxin concentration that caused 50% cytotoxicity) values were 9.5 and 45 microM, respectively. At the highest concentration of FB1 (100 microM), there was 80% viability. With the same incubation time, cell cycle distribution showed an arrest of cells in the G0/G1 phase in the presence of NIV (up to 0.3 microM), DON (up to 3 microM), and FB1 (up to 10 microM). Morphological evidence of apoptosis was related to the toxicity of the substances in that the more toxic NIV induced late apoptosis, whereas DON and FB1 produced less-severe morphological changes characteristic of early apoptosis. This study suggests that NIV is more toxic than DON, which in turn is more toxic than FB1. These mycotoxins can modify the normal progression of the cell cycle and induce an apoptotic process.     

Induced resistance by the mutualistic endophyte, Fusarium oxysporum strain 162, toward Meloidogyne incognita on tomato

The non-pathogenic endophytic fungus, Fusarium oxysporum strain 162, originally isolated from the endorhiza of tomato roots, reduces damage caused by Meloidogyne incognita, by inhibiting juvenile penetration of and development in the root. However, little is known about the mode of action of this endophyte fungus against the nematode. This study aimed at investigating how the endophyte affects nematode motility and survival and if induced resistance plays a role in the relationship. In a previous study, F. oxysporum strain 162 decreased nematode penetration of tomato up to 60%. In experiments using a split-root chamber to test for induced resistance, nematode penetration, number of galls, and number of egg masses were investigated 2 and 5 weeks after nematode inoculation. Split-root plants treated with F. oxysporum strain 162 showed 26-45% less nematode penetration, 21-36% less galls and a 22-26% reduction in the number of egg masses in the roots not directly inoculated with the fungus when compared to untreated control plants in repeated tests. In conclusion, inoculation of tomato plants with the non-pathogenic fungal endophyte F. oxysporum strain 162 resulted in a signficant reduction of nematode infection, which was in part due to induced resistance in the first 2-3 weeks after fungal inoculation.