Nivale Var Research Articles

RESEARCH ON MICROMYCETES IN OATS (AVENA SATIVA L.) UNDER THE CONDITIONS OF STUPINO DISTRICT, MOSCOW PROVINCE

Background. Fungal diseases are the main cause of the deterioration of grain quality and lower yield of cereals crops. Reduction of acreage, and, as a consequence, violation of crop rotation rules, oversaturation of fields with cereals, spring and summer droughts contribute to the mass emergence and widespreading of new obligate and facultative diseases in all regions of the country. Therefore, the search for a variety of donors and sources of sustainability is a vital task.Materials and methods. Materials engaged in the research were kernels of 41 oat accessions from the VIR collection of plant genetic resources, obtained in 2016–2018 in field crop rotations by the Genetic Diversity Research Department of ARHIBAN (Mikhnevo, Stupino District, Moscow Province). The specific diversity of micromycetes in grain was identified using Litvinov’s “wet chambers” technique and light microscopy.Results. The dominant pathocomplex of micromycetes in oat grain in Moscow Province was found to belong to the genera Alternaria (A. infectoria, A. tennuissima), Cladosporium (Cl. cladosporioides, Cl. herbarum) and Fusarium (F. avenaceum, F. culmorum, F. heterosporum, F. nivale var. nivale, F. oxysporum, F. poae, F. proliferatum, F. solani, F. sporotrichioides var. minus, and F. tricintum).Conclusion. The hulled cultivars ‘Bulanyi’ (VIR-15277, Russia, Moscow Prov.) and ‘Zakat’ (VIR-15384, Ukraine), and the naked cultivar Mestnyi (VIR-15290, UK) demonstrated the least contamination with pathogens: Fusarium fungi in these accessions were represented by one species – F. avenaceum.

Chemical composition, weed killer and antifungal activities of Tunisian thyme (Thymus capitatusHoff. et Link.) essential oils

The chemical composition, phytotoxic and antifungal activities of the essential oils isolated by hydrodistillation from the aerial parts of Tunisian thyme were evaluated. In order to find best results, different variables (i.e. provenance and phase of plant development) were considered. The chemical composition analysed by gas chromatography/mass spectrometry (GC/MS) revealed variability among provenances displaying interesting chemotypes and that carvacrol (63–84%), limonene (5–10%), δ-terpinene (0–7%), and β-myrcene (0–7%) were the main components. Antifungal ability of Thymus capitatus essential oils was tested by disc agar diffusion against five phytopathogenic fungi: Fusarium avenaceum, F. culmorum, Microdochium nivale var. nivale, Alternaria sp., and Bipolaris sorokiniana. High antifungal activity was observed for the essential oil isolated at the post-flowering development phase for the Bekalta provenance. There was no difference in the antifungal activities of oils isolated at the flowering phase ...

The effect of potential resistance inducers on development of Microdochium majus and Fusarium culmorum in winter wheat

The effect of potential resistance inducing chemicals on disease development of Fusarium head blight was studied in winter wheat (Triticum aestivum L.). As a pre-screening test, the effect of different treatments on development of Microdochium majus (syn. Microdochium nivale var. majus) was studied in detached leaves. Based on these tests, DL-3-aminobutyric acid, Bion (benzo-(1,2,3) thiadiazole-7-carbothioic acid S-methyl ester), and a foliar fertilizer containing potassium phosphite were selected for further studies. Greenhouse-grown winter wheat was sprayed with aqueous solutions of the potential resistance inducers 7 days prior to Fusarium culmorum point inoculation of the heads. Disease development was registered as number of bleached spikelets per inoculated spike. Spraying plants with the foliar fertilizer reduced the disease severity of F. culmorum by up to 40%. A reduced disease development of M. majus was also observed in detached leaves pre-treated with the foliar fertilizer. When the foliar fertilizer was added to the growth medium, a reduced in vitro growth of M. majus and F. culmorum was observed, indicating that the effect on disease development is at least partly due to a fungistatic effect. No significant reduction in disease development was observed in wheat pre-treated with DL-3-aminobutyric acid or Bion, although these compounds tended to reduce disease development, especially when applied in combination with other potential resistance inducers. We conclude that spraying winter wheat with a solution containing potassium phosphite can reduce development of M. majus and F. culmorum.

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

AbstractAn 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.

The effect of a mixture of seed-borne Microdochium nivale var. majus and Microdochium nivale var. nivale infection on Fusarium seedling blight severity and subsequent stem colonisation and growth of winter wheat in pot experiments

Greenhouse experiments were conducted in order to determine the impact of seed-borne Microdochium nivale var. nivale and var. majus inoculum, and seed treatment with a carboxin+thiram mixture, on the development of seedling blight, and on subsequent stem colonisation and growth of winter wheat (cv. Cadenza). Experiments were conducted at temperatures favourable (3°C) and unfavourable (22°C) to M. nivale. Seed-borne inoculum resulted in seedling blight symptom development when plants were grown at 3°C, but not when plants were grown at 22°C. For seedlings grown at 3°C, plants arising from heavily blighted seedlings developed more severe symptoms of stem colonisation, when compared with those arising from seedlings from carboxin+thiram treated seeds. In addition, the vigour of such plants (assessed by determining the number of tillers and ears per plant, stem length, green leaf area, dry weight and yield) was also significantly lower than for plants arising from carboxin+thiram treated seeds. Microdochium nivale var. majus and var. nivale appeared to have little effect on plant vigour from seedlings grown at 22°C. This is the first recorded incidence of seedling blight affecting subsequent plant growth. Microdochium nivale var. majus and var. nivale stem colonisation increased from growth stage (GS) 40–49 to harvest in plants raised from seedlings grown at both temperatures. Microdochium nivale var. majus and var. nivale were isolated from the second node at GS 40–49 and the third node at harvest of plants from seedlings grown at 3°C. For plants from seedlings raised at 22°C, M. nivale var. majus and var. nivale were isolated from the first node at GS 40–49 and the second node at harvest. Carboxin+thiram seed treatment decreased the extent and severity of stem colonisation on plants from seedlings grown at 22°C.

Colonization of winter wheat stem bases by communities of pathogenic fungi

The current study was focused on fungi colonizing the stem bases of wheat. Particularly, fungal species with significant harmful effect on wheat were studied. The dynamics of the stem base colonization by these fungi over the growing season were observed, and the association between individual pathogenic fungi was also evaluated. In addition, the fungal leaf sheaths penetration, which can cause physical disturbance, was assessed. The asymptomatic presence of fungal DNA in wheat tissue was confirmed. Microdochium nivale was found on the stem bases most frequently. Other fungi, such as Oculimacula spp., Rhizoctonia cerealis and Fusarium spp., were presented considerable less frequently. The statistical significant association was confirmed only between M. nivale var. nivale and M. nivale var. majus . Out of these, M. nivale tended to decrease in summer season, the frequency of occurrence of Oculimacula spp. and R. cerealis increased at later growth stages. Along with this, these species infected only plants...

The role of organic vs. conventional farming practice, soil management and preceding crop on the incidence of stem-base pathogens on wheat

A stem-base disease complex of winter wheat includes several causal agents, such as eyespot (Oculimacula yallundae and O. acuformis), sharp eyespot (Rhizoctonia cerealis) and brown foot rots (Fusarium graminearum, F. culmorum, F. avenaceum, F. po ae, Microdochium nivale vars. nivale and majus). Farming practices (organic and conventional), soil management (ploughing compared to min- and no-tillage) and the preceding crop (cereal, maize and oilseed rape) were studied as factors affecting the incidence of stem-base pathogens. The stem-base diseases were assessed visually and by molecular methods (PCR) on four winter wheat cultivars at four growth stages. The comparison of organic and conventional farming practices did not result in large differences in the relative incidence of stem-base pest species over the period of this study; nevertheless, a higher frequency of positive detection was recorded for three organisms (O. acuformis, O. yallundae and M. nivale var. nivale) under conventional farming. Other factors had none or only partial effects. Soil management didn’t affect the stem-base pathogens incidence significantly. The preceding crop maize reduced the incidence of M. nivale var. majus.

Components of Partial Disease Resistance in Wheat Detected in a Detached Leaf Assay Inoculated with Microdochium majus using First, Second and Third Expanding Seedling Leaves

AbstractThe use of first, second and third expanding seedling leaves of wheat (L1, L2 and L3 respectively), inoculated with conidial suspensions of Microdochium majus (syn. Microdochium nivale var. majus) in a detached leaf assay, for detecting components of partial disease resistance (PDR) was investigated across a range of wheat cultivars. Incubation periods (period from inoculation to first appearance of symptoms; a dull grey–green water‐soaked lesion) and latent periods (period from inoculation to the first appearance of sporodochia) were longest and lesions smallest on L3. The expression of PDR components on L2 was intermediate to those on L1 and L3. The longer latent periods on L3 typically occurred after leaf senescence contrasting with latent periods on L1 and L2 where sporulation most frequently occurred on relatively green leaf tissue. Cultivar differences in the first appearance of symptoms, incubation period, which occurred before any leaf senescence was observed, correlated significantly across all leaf positions. Similarly cultivar differences in latent period were correlated for L1 and L2. However, latent periods on L3, which were the least consistent between cultivars across experiments, were not correlated with those of L1 or L2 in any experiment. The results indicate that due to the delay in sporulation until after leaf senescence, observations on latent period in L3 are less representative of what occurs in the whole plant where infection of living tissue is of greatest interest. This work indicates that the selection of the first or second expanding leaf of wheat is optimal for the use in the detached leaf assay using M. majus for studying components of PDR.

Phylogenetic analysis of EF-1 alpha gene sequences from isolates of Microdochium nivale leads to elevation of varieties majus and nivale to species status

Degenerate PCR primers were designed based on EF-1 alpha (EF-1alpha) gene sequences of several filamentous fungi retrieved from sequence databases. These primers were used to isolate a partial sequence, approximately 830 bp in length of the EF-1alpha from isolates of Microdochium nivale obtained from various geographic locations across the world. Two distinct groups of isolates were evident among those isolates examined. Sequence homology for comparisons within group was 99.7% for group A and 99.8% for group B. Primers specific to either group A or group B sequences were designed and tested on isolates from around the world. Comparisons were made with primers previously reported for the two varieties of M. nivale and revealed that Group A type isolates correlated with M. nivale var. majus and group B isolates with M. nivale var. nivale. The primers from this study and those previously reported were in agreement for all isolates with the exception of one isolate (NRRL 3289) which failed to amplify with previously published M. nivale primers. Sequence analysis of NRRL 3289 suggested that it was an isolate of M. nivale var. nivale as indicated by the EF-1alpha based primers developed in this study. This study provides sequence based phylogenetic evidence of two species and when taken together with biological differences reported, leads to the recognition of M. majus comb. nov. (syn. Fusarium nivale var. majus). Descriptions of the two species are provided.

Evaluation of Components of Fusarium Head Blight Resistance in Soft Red Winter Wheat Germ Plasm Using a Detached Leaf Assay

A large environmental influence on phenotypic estimates of disease resistance and the complex polygenic nature of Fusarium head blight (FHB) resistance in wheat (Triticum aestivum) are impediments to developing resistant cultivars. The objective of this research was to investigate the utility of a detached leaf assay, inoculated using inoculum from isolates of Microdochium nivale var. majus, to identify components of FHB resistance among 30 entries of U.S. soft red winter wheat in the 2002 Uniform Southern FHB Nursery (USFHBN). Whole plant FHB resistance of the USFHBN entries was evaluated in replicated, mist-irrigated field trials at 10 locations in eight states during the 2001-2002 season. Incubation period (days from inoculation to the first appearance of a dull gray-green water-soaked lesion) was the only detached leaf variable significantly correlated across all FHB resistance parameters accounting for 45% of the variation in FHB incidence, 27% of FHB severity, 30% of Fusarium damaged kernels, and 26% of the variation in grain deoxynivalenol (DON) concentration. The results for incubation period contrasted with previous studies of moderately resistant European cultivars, in that longer incubation period was correlated with greater FHB susceptibility, but agreed with previous findings for the Chinese cultivar Sumai 3 and CIMMYT germ plasm containing diverse sources of FHB resistance. The results support the view that the detached leaf assay method has potential for use to distinguish between specific sources of FHB resistance when combined with data on FHB reaction and pedigree information. For example, entry 28, a di-haploid line from the cross between the moderately resistant U.S. cultivar Roane and the resistant Chinese line W14, exhibited detached leaf parameters that suggested a combination of both sources of FHB resistance. The USFHBN represents the combination of adapted and exotic germ plasm, but four moderately resistant U.S. commercial cultivars (Roane, McCormick, NC-Neuse, and Pat) had long incubation and latent periods and short lesion lengths in the detached leaf assay as observed in moderately FHB resistant European cultivars. The dichotomy in the relationship between incubation period and FHB resistance indicates that this may need to be considered to effectively combine exotic and existing/adapted sources of FHB resistance.

Resistance of wheat to Fusarium spp. in an in vitro seed germination assay and preliminary investigations into the relationship with Fusarium head blight resistance

Resistance of wheat cultivars from the UK, 2003 recommended list were evaluated against Fusarium culmorum, F. graminearum, Microdochium nivale var. majus and var. nivale, F. avenaceum and F. poae using a seed germination assay and related to previously published data on detached leaf assays, using M. nivale, and UK recommended list Fusarium head blight (FHB) resistance ratings. Significant cultivar differences were found after inoculation with all fungal species with the exception of F. poae where the decline in the percentage of seed germinating relative to the controls was small. Correlations of the percentage seed germinating inoculated with the remaining Fusarium spp. and M. nivale were high (r = 0.68; P < 0.01 to r = 0.94; P < 0.001). Overall, F. graminearum caused the greatest reduction with a mean seed germination of 61.7% relative to the controls followed by F. avenaceum (65.5%), M. nivale var. majus (67.2%), F. culmorum (76.6%), M. nivale var. nivale (89.2%) and was least for F. poae (92.5%). The resistance detected in the germination assay was significantly correlated to whole plant FHB resistance ratings (r s = 0.45; P < 0.05) but was not correlated to partial disease resistance (PDR) components detected using the detached leaf assay, namely, incubation period, latent period and lesion length. The results suggest that while resistances detected in the seed germination and detached leaf assays may in part share a common genetic basis to FHB resistance, resistances detected in both in vitro assays are independent of each other. Resistances detected in the seed germination assay had a lower correlation with FHB resistance ratings against F. culmorum, than those detected by latent period in the detached leaf assay.

Quantitative Detection of Fusarium Species in Wheat Using TaqMan

Fusarium head blight (FHB) of wheat and other small-grain cereals is a disease complex caused by several fungal species. To monitor and quantify the major species in the FHB complex during the growing season, real-time PCR was developed. TaqMan primers and probes were designed that showed high specificity for Fusarium avenaceum, F. culmorum, F. graminearum, F. poae and Microdochium nivale var. majus. Inclusion of an internal PCR control and serial dilutions of pure genomic DNAs allowed accurate determination of the concentration of fungal DNA for each of these species in leaves, ears as well as harvested grains of winter wheat. The DNA concentration of F. graminearum in grain samples correlated (r 2 = 0.7917) with the incidence of this species on the grain as determined by isolation from individual kernels. Application of the TaqMan technology to field samples collected in 40 wheat crops in the Netherlands during the growing season of 2001 revealed that M. nivale var. majus predominated on leaves early in the season (GS 45–65). Ears and harvested grains from the same fields, however, showed F. graminearum as the major species. In 2002, grain samples from 40 Dutch fields showed a much wider range of species, whereas in ears from 29 wheat crops in France, F. graminearum was the predominant species. The concentration of DON correlated equally well with the incidence of the DON-producing species F. culmorum and F. graminearum in the grain samples (r 2 = 0.8232) as well as with total DNA of both these species (r 2 = 0.8259). The Fusarium TaqMan technology is an important tool to quantify and monitor the dynamics of individual species of the complex causing FHB in cereals during the growing season. This versatile tool has been applied in a comparison of different genotypes, but can also be applied to other disease management systems, e.g. fungicide treatments.

Development and Evaluation of an in vitro Detached Leaf Assay forc Pre-Screening Resistance to Fusarium Head Blight in Wheat

An in vitro detached leaf assay, involving the inoculation of detached leaves with Microdochium nivale, was further developed and used to compare with whole plant resistance ratings to Fusarium head blight (FHB) of 22 commercial cultivars and published information on 21 wheat genotypes, identified as potential sources for FHB resistance. An incubation temperature of 10 °C and isolates of M. nivale var. majus of intermediate pathogenicity were found to be the most suitable for the differential expression of several components of partial disease resistance (PDR), namely incubation period, latent period and lesion length, in wheat genotypes used in the detached leaf assay. There were highly significant differences (P < 0.001) for each component of PDR within commercial cultivars and CIMMYT genotypes. Positive correlations were found between incubation period and latent period (r = 0.606; P < 0.001 and r = 0.498; P < 0.001, respectively, for commercial cultivars and CIMMYT genotypes), inverse correlations between incubation period and lesion length (r = -0.466; P < 0.01 and r = −0.685; P < 0.001, respectively) and latent period and lesion length (r = −0.825; P < 0.001 and r = −0.848; P < 0.001, respectively). Spearman rank correlations between individual PDR components and UK 2003 recommended list ratings were significant for incubation period (rs = 0.53; P < 0.05) and latent period (rs = 0.70; P < 0.01) but not for lesion length (r s = −0.26). Commercial cultivars identified with high resistances across all three PDR components in the detached leaf assay also had high whole plant FHB resistance ratings, with the exception of cv. Tanker which is more susceptible than the results of the detached leaf assay suggested, indicating an additional susceptibility factor could be present. Agreement between resistances found in the detached leaf assay and resistance to FHB suggests resistances detected in detached leaves are under the same genetic control as much of the resistances expressed in the wheat head of the commercial cultivars evaluated. In contrast, high resistances in each of the PDR components were associated with higher susceptibility across 19 CIMMYT genotypes previously evaluated as potential breeding sources of FHB resistance (incubation period: r = 0.52; P < 0.01, latent period: r = 0.53; P < 0.01, lesion length: r = −0.49; P < 0.01). In particular, the CIMMYT genotypes E2 and E12 together with Summai #3, known to have high levels of whole plant FHB resistance, showed low levels of resistance in each PDR component in the detached leaf assay. Such whole plant resistances, which are highly effective and not detected by the detached leaf assay, do not appear to be present in Irish and UK commercial cultivars. The most resistant Irish and UK commercial cultivars were comparable to the genotype Frontana and the most resistant CIMMYT germplasm evaluated in the leaf assay.

Competitive interactions between Microdochium nivale var. majus, M. nivale var. nivale and Fusarium culmorum in planta and in vitro.

Microdochium nivale var. majus and var. nivale are economically important fungal pathogens of cereal seedlings, stem bases and ears, as is the toxigenic species Fusarium culmorum. Competition experiments on seedlings support an earlier report of differential host preference between the varieties of M. nivale on wheat and rye seedlings at 15 degrees C, but showed that it does not extend across a broad range of temperatures. The studies showed that, although interaction is disadvantageous to the less virulent pathogen, it does not confer an advantage to the more virulent pathogen. In mixed inoculum experiments on wheat seedlings at 15 degrees C and 20 degrees C, F. culmorum suppressed the growth of both varieties of M. nivale. However, if M. nivale var. majus became established on the seedlings, it was able to co-suppress colonization of wheat seedlings by F. culmorum. In contrast M. nivale var. nivale did not suppress F. culmorum significantly. The growth of M. nivale var. majus and F. culmorum was also co-suppressed in liquid culture. Significantly, the accumulation of deoxynivalenol mycotoxin was also reduced in the mixed in vitro culture compared with axenic culture of F. culmorum. However, in vitro interaction studies on solidified media were of only limited use in predicting the outcome of competitions in planta.

Major changes in Fusarium spp. in wheat in the Netherlands

The re-emergence of fusarium head blight throughout the world and especially in Western Europe prompted a survey of the situation in the Netherlands. To allow for a high throughput screening of large numbers of samples, a diagnostic PCR method was developed to detect the most common species of Fusarium occurring on wheat. Seven primer pairs were tested for their ability to identify isolates of Fusarium avenaceum, F. culmorum, F. graminearum, F. poae, F. proliferatum and Microdochium nivale var. majus and M. nivale var. nivale. Each primer pair only generated a PCR product with the corresponding Fusarium species and all PCR fragments had different molecular sizes. This allowed the generation of these amplicons using a mixture of all seven primer pairs. The robustness of this multiplex PCR encouraged us to screen a large series of isolates collected in 2000 and 2001. In both years 40 fields were sampled leading to a collection of 209 isolates from 2000 and 145 isolates from 2001. The results of the multiplex PCR demonstrated that F. graminearum was the most abundant species in the Fusarium complex on wheat in both years. This is in sharp contrast to reports from the 1980s and early 1990s, which found F. culmorum as the predominant species. Primers derived from the tri7 and tri13 genes, which are implicated in the acetylation and oxygenation of the C-4 atom of the backbone of the trichothecene molecule, were used to discriminate between deoxynivalenol and nivalenol (NIV) producers. The populations of F. culmorum and F. graminearum both showed a slight increase in NIV-producers in 2001.

Evidence for differential host preference in Microdochium nivale var. majus and Microdochiumnivale var. nivale

The pathogenicity of Microdochium nivale var. majus and var. nivale was tested on wheat, rye and oat seedlings using both visual disease scoring and quantitative PCR measurements. In an individual inoculation trial at 10°C var. majus and var. nivale were strongly pathogenic towards wheat and rye, with var. nivale causing slightly greater disease in rye. At this temperature only var. nivale caused significant disease of oats. In a further experiment M. nivale was inoculated as a series of mixtures of the two varieties and incubated at 15°C. The ratio of the varieties present in the inoculum and present at harvest was analysed by quantitative PCR and this enabled a coefficient of selection to be calculated for the varieties on each host. M. nivale var. majus showed a weak selective advantage over var. nivale on wheat (0.33 ± 0.08) and oat seedlings (0.35 ± 0.016) and M. nivale var. nivale showed a strong selective advantage over var. majus on rye seedlings (0.92 ± 0.26). The isolates were also compared for sensitivity to benzoxazolinone (BOA), a hydroxamic acid compound derived from rye leaves. M. nivale var majus was found to be significantly more sensitive to BOA than M. nivale var. nivale, indicating a possible mechanism for the selective advantage of var. nivale growing on rye. This is the first substantiated indication of a significant difference in host preference between Microdochium nivale var. majus and var. nivale.

Visual disease and PCR assessment of stem base diseases in winter wheat

Stem base disease (eyespot, sharp eyespot and brown foot rot) was assessed visually and by the polymerase chain reaction (PCR) technique on single plants sampled at four‐week intervals in two crops of winter wheat grown in the UK in 1992–3. PCR assays were conducted for Fusarium avenaceum, F. culmorum, both varieties of Microdochium nivale, both eyespot‐causing species of Tapesia and Rhizoctonia cerealis. PCR diagnoses were compared with visual diagnoses at each time point. Eyespot was caused principally by T. acuformis (R‐type) and developed rapidly late in the season. Visual diagnoses of eyespot were largely confirmed by PCR but T. acuformis was detected in many plants lacking eyespot symptoms. R. cerealis was detected at relatively low incidences on both sites, and sharp eyespot visual diagnoses did not correlate with the incidence of any of the pathogens assayed by PCR. Brown foot rot, caused principally by Microdochium nivale var. majus, accumulated earlier in the season than eyespot. Overall, visual diagnoses of stem base disease coincided poorly with PCR data until after growth stage (GS) 30.

Towards the Development of a Novel In Vitro Strategy for Early Screening of Fusarium Ear Blight Resistance in Adult Winter Wheat Plants

A novel in vitro bioassay is described for screening Fusarium ear blight (FEB) resistance in adult winter wheat plants. Seven winter wheat cultivars were assessed for components of partial disease resistance as 28 day-old detached leaf segments in the laboratory using isolates of Microdochium nivale var. nivale and M. nivale var. majus. Results were compared with disease data obtained at anthesis using the same cultivars as whole plants and the same isolates under glasshouse conditions. Significant cultivar differences were observed using detached leaves, with cv. Avalon (a Fusarium culmorum ear susceptible cultivar) having the shortest leaf incubation period, greatest leaf lesion development and shortest leaf latent period compared to cv. Spark (a Fusarium culmorum ear resistant cultivar), which had the longest leaf incubation period, least leaf lesion development and longest leaf latent period. Using whole plants, cv. Avalon had the shortest ear incubation period and greatest ear disease severity, whilst cv. Spark had the longest incubation period and least ear disease severity. Overall, cultivars of intermediate F. culmorum ear resistance expressed intermediate responses to M. nivale isolates, using both detached leaves and whole plants. Significant correlations were found with ear disease severity and ear incubation period in whole plants and components of partial disease resistance in detached leaves, with significant correlations obtained between leaf incubation period and ear disease parameters using the M. nivale var. nivale isolate. In addition, leaf latent period and leaf lesion size showed significant correlations with whole plant reactions using M. nivale var. nivale and var. majus isolates. The in vitro screening of cultivars as detached leaves using M. nivale isolates may offer a real possibility of a rapid bioassay for the early screening of FEB resistance in wheat and other cereals.

Morphology of sporodochia of Microdochium nivale var. nivale on cereal hosts using cryoscanning electron microscopy

This paper reports on the use of cryoscanning electron microscopy for the observation of sporodochia produced by the artificial inoculation of detached leaves of wheat, barley and oats with Microdochium nivale var. nivale . Morphological differences were apparent between sporodochia on different hosts. The use of high-speed and slow-speed freezing resulted in differences in the appearance of sporodochia; slow-speed freezing caused a loss in detail of macroconidia with damage and deformation of the specimens.

Genetic diversity of Microdochium nivale isolates from turfgrass

Conserved primers were used in a polymerase chain reaction to amplify the ITS region of the rDNA of 100 Microdochium nivale isolates collected from different turfgrasses in southern Ontario. The profile of the restriction digestion of the amplified ITS region revealed that all the M. nivale isolates analysed belonged to var. nivale. RAPD profiling and RFLP analyses of the IGS regions of rDNA revealed extensive genetic diversity within var. nivale. With RAPD markers, the average similarity coefficient was 66% and the estimate of genotypic diversity was 0.179. Population subdivision analysis showed that 92.2% of the total genetic diversity was found among individuals within populations compared to 7.8% among populations. In dendrograms derived from genetic distances using RAPD and IGS-RFLP markers, there was some evidence for host specialization. Most RAPD markers were shared by individuals from different turfgrasses, and the populations were not highly differentiated. The high level of genotypic diversity detected within populations and the low level of genetic differentiation among populations show that recombination and migration are likely playing important roles in the population biology of M. nivale var. nivale.