Genetic variation among <Emphasis Type="Italic">Fusarium</Emphasis> isolates from onion,and resistance to <Emphasis Type="Italic">Fusarium</Emphasis> basal rot in related <Emphasis Type="Italic">Allium</Emphasis> species

The aim of this research was to study levels of resistance to Fusarium basal rot in onion cultivars and related Allium species, by using genetically different Fusarium isolates. In order to select genetically different isolates for disease testing, a collection of 61 Fusarium isolates, 43 of them from onion (Allium cepa), was analysed using amplified fragment length polymorphism (AFLP) markers. Onion isolates were collected in The Netherlands (15 isolates) and Uruguay (9 isolates), and received from other countries and fungal collections (19 isolates). From these isolates, 29 were identified as F. oxysporum, 10 as F. proliferatum, whereas the remaining four isolates belonged to F. avenaceum and F. culmorum. The taxonomic status of the species was confirmed by morphological examination, by DNA sequencing of the elongation factor 1-α gene, and by the use of species-specific primers for Fusarium oxysporum, F. proliferatum, and F. culmorum. Within F. oxysporum, isolates clustered in two clades suggesting different origins of F. oxysporum forms pathogenic to onion. These clades were present in each sampled region. Onion and six related Allium species were screened for resistance to Fusarium basal rot using one F. oxysporum isolate from each clade, and one F. proliferatum isolate. High levels of resistance to each isolate were found in Allium fistulosum and A. schoenoprasum accessions, whereas A. pskemense, A. roylei and A. galanthum showed intermediate levels of resistance. Among five A. cepa cultivars, ‘Rossa Savonese’ was also intermediately resistant. Regarding the current feasibility for introgression, A. fistulosum, A. roylei and A. galanthum were identified as potential sources for the transfer of resistance to Fusarium into onion.     

Accumulation of toxigenic <Emphasis Type="Italic">Fusarium</Emphasis> species and <Emphasis Type="Italic">Stenocarpella maydis</Emphasis> in maize grain grown under different cropping systems

Entomopathogenic nematodes in the genus Steinernema are associated with Xenorhabdus spp. bacteria. When steinernematid colonise an insect host the nematode-bacterium association overcomes the insect immune system and kills the host within 48 h. Xenorhabdus spp. produce secondary metabolites that are antifungal to protect nematode-infected cadavers from fungal colonization. The concentrated, or cell-free metabolites of X. szentirmaii exhibit high toxicity against various fungal plant pathogens and show potential as natural bio-fungicides. In the current study, we determined 1) thermo-stability, 2) dose-response, and 3) shelf-life of antifungal metabolites of X. szentirmaii against Monilinia fructicola (cause of brown rot of peach and other stone fruit) and Glomerella cingulata (cause of antharacnose). Thermo-stability was determined by autoclaving bacterial culture broths (121 °C and 15 psi for 15 min) and measuring fungal growth on in potato dextrose agar (PDA) containing 10% of the supernatants. Autoclaving had no impact on the antifungal activity of the secondary metabolites. Over a test period of 9 months, the activity of both extract types did not decline when stored at 4 or 20 °C. A dose-response study (10, 20, 40, 60, 80 and 100% supernatant-containing metabolite) using both phytopathogens demonstrated that a greater dose of supernatant increased antifungal activity. The antifungal-metabolite containing supernatant of X. szentirmaii has potential as a bio-fungicide. These results demonstrate the metabolite(s) are thermo-stable, they have a long shelf-life and require no stabilizing formulation, even at room temperature.     

Hyperspectral quantification of wheat resistance to <Emphasis Type="Italic">Fusarium</Emphasis> head blight: comparison of two <Emphasis Type="Italic">Fusarium</Emphasis> species

Experiments were conducted to determine the extent of Fusarium langsethiae infection in wheat, barley and oats grown under identical experimental conditions. In total, four experiments were conducted with both winter and spring sown experiments at two locations. The amount of F. langsethiae infection was determined by quantifying F. langsethiae DNA and quantifying the combined concentration of the trichothecene mycotoxins HT-2 and T-2 (HT-2 + T-2) in cereal head fractions (grain and rest of the head) after threshing at harvest. Results of the study showed that under identical experimental conditions, oats had the highest F. langsethiae DNA and HT-2 + T-2 concentrations compared to wheat and barley. This indicates that the high levels detected on UK oats compared to wheat and barley from surveys of commercial crops is a consequence of genetic differences rather than differences in agronomy applied to the cereal species. The concentration of HT-2 and T-2 per unit of F. langsethiae DNA in oats compared to wheat and barley was also significantly higher indicating host differences in either the stimulation of HT-2 and T-2 production or in the metabolism of HT-2 and T-2. The study also showed that the proportion of F. langsethiae DNA in threshed grains was significantly lower than that in the rest of the cereal head.     

Natural occurrence of <Emphasis Type="Italic">Fusarium</Emphasis> species and fumonisin on maize grains in Ethiopia

Fusarium species causing maize kernel rot are major threats to maize production, due to reduction in yield as well as contamination of kernels by mycotoxins that poses a health risk to humans and animals. Two-hundred maize kernel samples, collected from 20 major maize growing areas in Ethiopia were analyzed for the identity, species composition and prevalence of Fusarium species and fumonisin contamination. On average, 38 % (range: 16 to 68 %) of maize kernels were found to be contaminated by different fungal species. Total of eleven Fusarium spp. were identified based on morphological characteristics and by sequencing the partial region of translation elongation factor 1-alpha (EF-1α) gene. Fusarium verticillioides was the dominant species associated with maize kernels (42 %), followed by F. graminearum species complex (22.5 %) and F. pseudoanthophilium (13.4 %). The species composition and prevalence of Fusarium species differed among the areas investigated. Fusarium species composition was as many as eight and as few as four in some growing area. The majority of the maize samples (77 %) were found positive for fumonisin, with concentrations ranging from 25 μg kg^?1 to 4500 μg kg^?1 (mean: 348 μg kg^?1 and median: 258 μg kg^?1). Slight variation in fumonisin concentration was also observed among areas. Overall results indicate widespread occurrence of several Fusarium species and contamination by fumonisin mycotoxins. These findings are useful for intervention measures to reduce the impact of the main fungal species and their associated mycotoxins, by creating awareness and implementation of good agricultural practices.     

<Emphasis Type="Italic">Fusarium</Emphasis> species complex on maize in Switzerland: occurrence,prevalence, impact and mycotoxins in commercial hybrids under natural infection

The Fusarium species complex of maize kernels and stem pieces as well as mycotoxin contamination of commercial grain maize hybrids for animal feed were evaluated in Switzerland. Throughout 2 years, natural Fusarium infection varied significantly between the years and the locations and it ranged from 0.4% to 49.7% for kernels and from 24.2% to 83.8% for stem pieces. Using the agar plate method, 16 different Fusarium species were isolated from kernels and 15 from stem pieces. The Fusarium species composition, prevalence and impact differed between the north and the south and between kernel and stem piece samples. The dominant species on kernels in the north were F. verticillioides (32.9%), F. graminearum (31.3%), F. proliferatum (7.3%) and F. crookwellense (7.1%), in the south F. verticillioides (57.1%), F. subglutinans (24.6%), F. proliferatum (14.8%) and F. graminearum (1.5%) and on stem pieces F. equiseti (36.0%), F. verticillioides (20.1%), F. graminearum (9.5%), F. crookwellense (6.2%) and F. subglutinans (6.2%). In the south, fumonisin concentration of most hybrids exceeded guidance values for animal feed. Other Fusarium species isolated were F. avenaceum, F. culmorum, F. oxysporum, F. poae, F. sambucinum, F. semitectum, F. sporotrichioides, F. solani, F. tricinctum and F. venenatum. Maize hybrids varied in their susceptibility to Fusarium infection. Because of the high diversity of Fusarium species encountered in Switzerland representing a high toxigenic potential, we propose to screen maize hybrids for resistance against various Fusarium species and examine maize produce for several mycotoxins in order to ensure feed safety.     

<Emphasis Type="Italic">Fusarium</Emphasis> head blight incidence and detection of <Emphasis Type="Italic">Fusarium</Emphasis> toxins in wheat in relation to agronomic factors

We investigated incidences of Fusarium head blight (FHB) and concentrations of six mycotoxins (deoxynivalenol, nivalenol, 3-acetyldeoxynivalenol, T-2 toxin, HT-2 toxin and zearalenone) in wheat from 2010 to 2013. Field trials were conducted at the Experimental Station of Cultivar Testing in Chrz?stowo, Poland (53^o11’N, 17^o35’E). We examined the effects of four agronomic factors, including pre-crop type (corn, sugar beets and wheat), date of sowing (late autumn: November 8–December 9 or spring: March 29–April 19), fungicidal application (untreated or treated with two applications) and cultivar (Monsun, Cytra), on FHB index (FHBi) and mycotoxin levels in order to minimize the risk of wheat grain contamination by mycotoxins via integrated pest management methods. The dominant Fusarium species observed on wheat heads were F. culmorum, F. avenaceum (Gibberella avenacea) and F. graminearum (Gibberella zeae), at 21.1%, 17.2% and 7.1%, respectively. A monthly rainfall sum of 113.9 mm and a relatively low air temperature (monthly average 15.5 °C) resulted in the highest FHBi in untreated wheat (25.1%). Agronomic factors crucial for the FHB incidence were the pre-crop, fungicidal treatments and cultivar selection. In wheat planted after wheat or corn, the FHBi was higher compared with a pre-crop of sugar beet. A double application of fungicides at BBCH 30–32 with prothioconazole and spiroxamine and at a BBCH 65 with fluoxastrobin and prothioconazole effectively reduced the FHBi and mycotoxin concentrations, respectively, in grain. The cultivar ‘Cytra’ had a greater FHBi (10.4%) than ‘Monsun’ (4.6%), and grain infestations by Fusarium species were also greater in ‘Cytra’, at 16.5%, than in ‘Monsun’, at 11.2%. Untreated cv. Cytra grown after corn in spring produced grains with the highest amounts of the mycotoxins, deoxynivalenol, 3-acetyldeoxynivalenol, zearalenone and HT-2 (605, 103, 17.5 and 5.53 μg/kg, respectively). Total mycotoxin levels in wheat were correlated with five determinants: duration of the period between the end of flowering and the beginning of kernel abscission, FHBi, F. culmorum isolation, G. zeae isolation and Fusarium ratio (FR) as a % of total mould isolations. Although, the mean concentration of mycotoxins in grain did not exceed the maximum permissible values for unprocessed wheat our study suggests necessity to monitor and mitigate FHB risk for susceptible cultivars, when wheat spring sowing follows corn or wheat.     

Fusarium rot of hyacinth caused by <Emphasis Type="Italic">Gibberella zeae</Emphasis> (anamorph: <Emphasis Type="Italic">Fusarium</Emphasis><Emphasis Type="Italic"> graminearum</Emphasis>)

Severe rot of leaves, peduncles and flowers caused by Gibberella zeae (anamorph: Fusarium graminearum) was found on potted plants of hyacinth (Hyacinthus orientalis), a liliaceous ornamental, in greenhouses in Kagawa Prefecture, Japan, in January 2001. This disease was named “Fusarium rot of hyacinth” as a new disease because only the anamorph, F. graminearum, was identified on the diseased host plant. The authors contributed equally to this work. The fungal isolate and its nucleotide sequence data obtained in this study were deposited in the Genebank, National Institute of Agrobiological Sciences and the DDBJ/EMBL/GenBank databases under the accession numbers MAFF239499 and AB366161, respectively.     

Characterisation of novel <Emphasis Type="Italic">Fusarium graminearum</Emphasis> microsatellite markers in different <Emphasis Type="Italic">Fusarium</Emphasis> species from various countries

Fifteen novel microsatellite markers were isolated from Fusarium graminearum. The level of polymorphism at these novel and 13 previously published microsatellite markers was analysed in 33 F. graminearum strains from Europe, North America, and Nepal. The number of alleles for each of the novel markers ranged from 4 to 20 and gene diversity from 0.417 to 0.962. In comparison with the previously published markers, the resolution for distinguishing among different strains was slightly increased. Twenty-seven markers were also detectable in three F. culmorum strains and one F. crookwellense strain. None of the markers was detected in three F. avenaceum and four F. poae strains, underlining the potential use of these microsatellite markers for species differentiation.     

<Emphasis Type="Italic">Mycosphaerella</Emphasis> species occurring on <Emphasis Type="Italic">Eucalyptus globulus</Emphasis> in Portugal

Mycosphaerella leaf disease (MLD) is caused by species of Mycosphaerella and several anamorphic form genera that have been connected to Mycosphaerella. Until recently, MLD of eucalypts was largely ignored in Portugal. However, serious damage to Eucalyptus globulus has been reported since 1999 when frequent and severe defoliation of young trees was observed. The severity of this disease prompted a preliminary study of the Mycosphaerella species associated with major symptoms of a leaf blotch disease in commercial plantations of E. globulus in Portugal, which is presented here. The species were identified by molecular methods based on the ITS1-5.8S-ITS2 cluster, together with morphological characters. In addition to confirming the species previously recorded, Mycosphaerella vespa is reported for the first time from Portugal, while the status of Mycosphaerella grandis remains to be resolved.     

<Emphasis Type="Italic">Odontoglossum ringspot virus</Emphasis> causing flower crinkle in <Emphasis Type="Italic">Phalaenopsis</Emphasis> hybrids

A new disorder exhibiting flower crinkle on Phalaenopsis orchids bearing white flowers has been observed in Taiwan, China and Japan for several years. This disorder decreased the flower longevity and was considered as a physiological syndrome. The objective of this study was to identify and characterize the real causal agent of this new Phalaenopsis disorder. Five plants of Phalaenopsis hybrids “V3” (Phal. Yukimai × Phal. Taisuco Kochdian) with flower crinkle symptoms were collected and tested by enzyme-linked immunosorbent assay with antisera against 18 viruses. The extract of leaves and flowers from one diseased plant (96-Ph-16) reacted positively only to antiserum against Odontoglossum ringspot virus (ORSV), while those from the other four plants (96-Ph-7, 96-Ph-17, 96-Ph-18 and 96-Ph-19) reacted positively to the antisera against ORSV and Cymbidium mosaic virus (CymMV). Five ORSV isolates, one each from flowers of those five diseased Phalaenopsis orchids, were established in Chenopodium quinoa. A CymMV culture was isolated from the flowers of one of the ORSV/CymMV mix-infected Phalaenopsis orchids (96-Ph-19). To determine the causal agent of the flower crinkle disease, healthy Phalaenopsis seedlings were singly or doubly inoculated with the isolated ORSV and/or CymMV. Results of back inoculation indicated that ORSV is the sole causal agent of the crinkle symptom on petals of Phalaenopsis orchid. The CP gene of the ORSV isolates from this study shared 97.3–100% nucleotide identity and 96.2–100% amino acid identity with those of 41 ORSV isolates available in GenBank. This is the first report demonstrating ORSV as the sole virus causing flower crinkle disease on Phalaenopsis orchids.     

Nonhost resistance of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> against <Emphasis Type="Italic">Colletotrichum</Emphasis> species

Arabidopsis thaliana exhibits a durable resistance called nonhost resistance against nonadapted fungal pathogens. A. thaliana activates preinvasive resistance and terminates entry attempts by nonadapted fungi belonging to the genus Colletotrichum, which cause anthracnose disease in many plants. In the interaction between A. thaliana and nonadapted C. tropicale, the preinvasive resistance involves the PENETRATION 2-related antifungal secondary metabolite pathway and the ENHANCED DISEASE RESISTANCE 1-dependent antifungal peptide pathway. The development of invasive hyphae by C. tropicale owing to the reduction of preinvasive resistance then triggers the blockage of further hyphal expansion via the activation of the second layer of resistance, i.e., postinvasive resistance, which guarantees the robustness of the nonhost resistance of A. thaliana against Colletotrichum pathogens. Both the tryptophan-derived metabolic pathway and glutathione synthesis play critical roles in the postinvasive resistance against C. tropicale, although the molecular mechanism of postinvasive resistance remains to be elucidated. In this review, we describe the current understanding of the molecular background of the Arabidopsis nonhost resistance against Colletotrichum fungi and discuss perspectives for future research on this durable resistance.     

First report of blueberry red ringspot disease caused by <Emphasis Type="Italic">Blueberry</Emphasis><Emphasis Type="Italic">red</Emphasis><Emphasis Type="Italic">ringspot</Emphasis><Emphasis Type="Italic">virus</Emphasis> in Japan

Virus-like symptoms—red ringspots on stems and leaves, circular blotches or pale spots on fruit—were found on commercial highbush blueberry (Vaccinium corymbosum) cultivars Blueray, Weymouth, Duke and Sierra in Japan. In PCR testing, single DNA fragments were amplified from total nucleic acid samples of the diseased blueberry bushes using primers specific to Blueberry red ringspot virus (BRRV). Sequencing analysis of the amplified products revealed 95.7–97.7% nucleotide sequence identity with the BRRV genome. This paper is the first report of blueberry red ringspot disease caused by BRRV in Japan. The nucleotide sequence data reported in this paper are available in the GenBank/EMBL/DDBJ database as accessions AB469884 to AB469893 for BRRV isolates from Japan.     

Identity and pathogenicity of <Emphasis Type="Italic">Fusarium</Emphasis> species associated with crown rot on wheat (<Emphasis Type="Italic">Triticum</Emphasis> spp.) in Turkey

An extensive survey was carried out to collect Fusarium species colonizing the lower stems (crowns) of bread wheat (Triticum aestivum L.) and durum wheat (T. durum Desf.) from different wheat growing regions of Turkey in summer 2013. Samples were collected from 200 fields representing the major wheat cultivation areas in Turkey, and fungi were isolated from symptomatic crowns. The isolates were identified to species level by sequencing the translation elongation factor 1-alpha (TEF1-α) gene region using primers ef1 and ef2. A total of 339 isolates representing 17 Fusarium species were isolated. The isolates were identified as F. culmorum, F. pseudograminearum, F. graminearum, F. equiseti, F. acuminatum, F. brachygibbosum, F. hostae, F. redolens, F. avenaceum, F. oxysporum, F. torulosum, F. proliferatum, F. flocciferum, F. solani, F. incarnatum, F. tricinctum and F. reticulatum. Fusarium equiseti was the most commonly isolated species, accounting for 36% of the total Fusarium species isolated. Among the damaging species, F. culmorum was the predominant species being isolated from 13.6% of sites surveyed while F. pseudograminearum and F. graminearum were isolated only from 1% and 0.5% of surveyed sites, respectively. Six out of the 17 Fusarium species tested for pathogenicity caused crown rot with different levels of severity. Fusarium culmorum, F. pseudograminearum and F. graminearum caused severe crown rot disease on durum wheat. Fusarium avenaceum and F. hostae were weakly to moderately virulent. Fusarium redolens was weakly virulent. However, F. oxysporum, F. equiseti, F. solani, F. incarnatum, F. reticulatum, F. flocciferum, F. tricinctum, F. brachygibbosum, F. torulosum, F. acuminatum and F. proliferatum were non-pathogenic. The result of this study reveal the existence of a wide range of Fusarium species associated with crown rot of wheat in Turkey.     

Characterization of <Emphasis Type="Italic">Inago1</Emphasis> and <Emphasis Type="Italic">Inago2</Emphasis> retrotransposons in <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis>

From the genome of a Japanese field isolate of the rice blast fungus, Magnaporthe oryzae, we newly identified Inago1 and Inago2 LTR retrotransposons. Both elements were found to be Ty3/gypsy-like elements whose copies were dispersed within the genome of Magnaporthe spp. isolates infecting rice and other monocot plants. Southern hybridization patterns of nine re-isolates derived from conidia of the strain Ina168 produced after a methyl viologen treatment were not changed, indicating that the insertion pattern of Inago elements is relatively stable.     

Genetic diversity of <Emphasis Type="Italic">Rhizobium vitis</Emphasis> strains in Japan based on multilocus sequence analysis of <Emphasis Type="Italic">pyrG</Emphasis>, <Emphasis Type="Italic">recA</Emphasis> and <Emphasis Type="Italic">rpoD</Emphasis>

Forty-one strains of Rhizobium vitis, either tumorigenic (Ti) or nonpathogenic, were characterized using multilocus sequence analysis (MLSA) of the partial nucleotide sequences of pyrG, recA, and rpoD. The strains separated into seven clades. Rhizobium vitis (Ti) strains isolated from Japan were divided into five genetic groups (A to E), and nonpathogenic R. vitis strains were divided into two genetic groups (F and G). This result suggests that there are new genetic groups of R. vitis in Japan. Among these groups, members of A and B groups are widely distributed throughout Japan.     

Resistance of <Emphasis Type="Italic">Solanum</Emphasis> species to <Emphasis Type="Italic">Cucumber mosaic virus</Emphasis> subgroup IA and its vector <Emphasis Type="Italic">Myzus persicae</Emphasis>

Sixty-nine tomato genotypes representing nine Solanum species were evaluated for resistance to Cucumber mosaic virus (CMV) subgroup IA and its aphid vector Myzus persicae. Resistance was assessed by visual scoring of symptoms in the field under natural conditions, and in the greenhouse by artificial inoculations through aphid M. persicae and mechanical transmissions in the year 2007 and 2009. Considerable variation in responses was observed among the evaluation methods used. Field evaluations were found liable to errors as different levels were observed for the same genotypes in the different years, however mechanical inoculation was found to be the most useful in identifying CMV subgroup IA resistance, in contrast aphid transmission was most useful in identifying insect transmission resistance. All genotypes observed as highly resistant to CMV subgroup IA in the field or through vector transmission became systemically infected through mechanical inoculations. Using mechanical inoculation, six genotypes (TMS-1 of S. lycopersicum, LA1963 and L06049 of S. chilense, LA1353, L06145 and L06223 of S. habrochaites) were found resistant and another six (L06188 and L06238 of S. neorickii, L06219 of S. habrochaites, L05763, L05776 and L06240 of S. pennellii) were found tolerant showing mild symptoms with severity index (SI) ranging 1-2 and with delayed disease development after a latent period (LP) of 18–30 days. However, these genotypes were found to be resistant to highly resistant in the field and through inoculation by M. persicae; and they also supported low population levels of M. persicae except TMS-1. Another nine genotypes (LA2184 of S. pimpinellifolium L., LA2727 of S. neorickii, LA0111, L06221, L06127 and L06231 of S. peruvianum L., LA1306, L06057 and L06208 of S. chmielewskii) showing a susceptible response after mechanical inoculation were highly resistant, resistant and tolerant after M. persicae transmission. The resistant genotypes, identified in the present study can be exploited in the breeding programmes aimed at developing tomato varieties resistant to CMV subgroup IA and broadening the genetic base of CMV-resistant germplasm. The differences observed between mechanical and aphid transmission suggests that one should consider both evaluation methods for tomato germplasm screening against CMV subgroup IA.     

Pathogenicity of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> and <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> against <Emphasis Type="Italic">Galleria mellonella</Emphasis>

The greater wax moth Galleria mellonella L. (Lepidoptera: Pyralidae) is occasionally found in beehives and is a major pest of stored wax. Entomopathogenic fungi have recently received attention as possible biocontrol elements for certain insect pests. In this study, 90 isolates of Beauveria bassiana and 15 isolates of Metarhizium anisopliae were screened for proteases and lipases production. The results showed significant variations in the enzymatic action between the isolates. In the bioassay, the selected isolates evinced high virulence against the 4th instar of the G. mellonella larvae. The isolates BbaAUMC3076, BbaAUMC3263 and ManAUMC3085 realized 100% mortality at concentrations of 5.5 × 10⁶ conidia ml⁻¹, 5.86 × 10⁵ conidia ml⁻¹, and 4.8 × 10⁶ conidia ml⁻¹, respectively. Strong enzymatic activities in vitro did not necessarily indicate high virulence against the tested insect pest. The cuticle of the infected larvae became dark and black-spotted, indicating direct attack of fungus on the defense system of the insects. The LC₅₀ values were 1.43 × 10³, 1.04 × 10⁵ and 5.06 × 10⁴ for Bba3263AUMC, Bba3076AUMC and Man3085AUMC, respectively, and their slopes were determined by computerized probit analysis program as 0.738 ± 0.008, 0.635 ± 0.007 and 1.120 ± 0.024, respectively.     

Characterization of <Emphasis Type="Italic">Fusarium</Emphasis> species associated with tobacco diseases in Northwestern Argentina

Phenotypic traits are regularly used to diagnose the development of Fusarium head blight (FHB) in the field, whereas mycotoxin accumulation in wheat grains can only be accurately evaluated through costly methods, such as high–performance liquid chromatography (HPLC). The aim of this study was to determine whether: (i) the results provided by existing commercial decision support tools could be anticipated using phenotypic measurements, including a novel technique of computer–assisted image analysis of spikes; and (ii) these measurements could avoid using HPLC. We monitored the FHB development during two consecutive years in highly contaminated plots in the Burgundy region (France). Contamination by crop residues was simulated through a field inoculation with barley grains artificially colonized by Fusarium graminearum. The development of the disease on spikes and harvested grains was assessed on one tolerant and two susceptible wheat varieties. The accumulated amounts of mycotoxins were measured in harvested grains using HPLC. As expected, the measured traits revealed that the inoculum responsible for infection on spikes mainly came from residues left on the soil surface, and the susceptible varieties were more diseased than the tolerant variety. Weather conditions had a strong effect on disease development. The novel computer–assisted image analysis technique had a better prediction power of deoxynivalenol accumulation, was more objective and time–saving than classical visual symptom assessments. This assessment method could be suitable to supplement the use of existing prediction tools and might avoid systematic and costly mycotoxin measurements in likely infected plots.     

<Emphasis Type="Italic">Eremothecium coryli</Emphasis> and <Emphasis Type="Italic">E.</Emphasis><Emphasis Type="Italic">ashbyi</Emphasis> cause yeast spot of azuki bean

Yeast-like fungi were isolated from lesions on azuki bean (cv. Shin-Kyotodainagon) seeds that had been sucked by bean bugs in Kyoto Prefecture, Japan. On the basis of morphological and physiological characteristics and sequence data of the internal transcribed spacer (ITS) regions including the 5.8S rDNA, these yeasts were identified as Eremothecium coryli and E. ashbyi. Pathogenicity of those yeasts was confirmed by a reinoculation test. To our knowledge, this is the first report of the occurrence of yeast spot in azuki bean in Japan. The nucleotide sequence data reported are available in the GeneBank/EMBL/DDBJ database as accessions AB478291–AB478309 for E. coryli AZC1–19 and AB478310–AB478317 for E. ashbyi AZA1–8.     

<Emphasis Type="Italic">In vitro</Emphasis> bacterization of banana (<Emphasis Type="Italic">Musa</Emphasis> spp.) with native endophytic and rhizospheric bacterial isolates: Novel ways to combat <Emphasis Type="Italic">Fusarium</Emphasis> wilt

Compared to conventional planting material, micropropagated plantlets are highly susceptible to Fusarium wilt because they are free from beneficial root inhabitants. We aimed to introduce mixtures of beneficial microbes in the plantlets in the rooting medium under in vitro conditions rather than by field applications. Endophytes and rhizobacteria from different banana cultivars and plantation areas were screened and characterized. Under in vitro conditions, banana tissue culture plantlets were bacterized with the prospective endophytes, Bacillus subtilis strain EPB56 and EPB10 and the rhizobacteria, Pseudomonas fluorescens strain Pf1 and effects of in vitro bacterization were investigated against Fusarium oxysporum f. sp. cubense race 1 under glasshouse and field conditions. Inoculation of bananas during micropropagation allowed for the omission of minerals and salts as well as vitamins from the growing media while resulting in plantlets close to double size compared to the controls with full strength media. All endophyte and rhizobacteria strains tested resulted in significant reductions in Fusarium infection in the glasshouse and field and in significantly better plant growth. The three-way combination of bacteria resulted in 78% disease reduction and more than doubled the yields compared to the untreated controls across two field experiments. Three-way inoculation led to yields of 23 and 24 kg/ bunch compared to chemical disease control (13; 15 kg/bunch) and untreated controls (10; 13 kg/bunch) in the two field experiments. Under glasshouse conditions, activity of defence enzymes was significantly increased by all inoculation treatments. Inoculation in vitro led to the establishment of the microorganisms in the plant system before delivering to the farming community. Micropropagation combined with the establishment of a beneficial microbial consortium should complement the micropropagated plants for easier adaptation under field conditions.