Thomas Miedaner

A European Database of Fusarium graminearum and F. culmorum Trichothecene Genotypes

Fusarium species, particularly Fusarium graminearum and F. culmorum, are the main cause of trichothecene type B contamination in cereals. Data on the distribution of Fusarium trichothecene genotypes in cereals in Europe are scattered in time and space. Furthermore, a common core set of related variables (sampling method, host cultivar, previous crop, etc.) that would allow more effective analysis of factors influencing the spatial and temporal population distribution, is lacking. Consequently, based on the available data, it is difficult to identify factors influencing chemotype distribution and spread at the European level. Here we describe the results of a collaborative integrated work which aims (1) to characterize the trichothecene genotypes of strains from three Fusarium species, collected over the period 2000–2013 and (2) to enhance the standardization of epidemiological data collection. Information on host plant, country of origin, sampling location, year of sampling and previous crop of 1147 F. graminearum, 479 F. culmorum, and 3 F. cortaderiae strains obtained from 17 European countries was compiled and a map of trichothecene type B genotype distribution was plotted for each species. All information on the strains was collected in a freely accessible and updatable database (www.catalogueeu.luxmcc.lu), which will serve as a starting point for epidemiological analysis of potential spatial and temporal trichothecene genotype shifts in Europe. The analysis of the currently available European dataset showed that in F. graminearum, the predominant genotype was 15-acetyldeoxynivalenol (15-ADON) (82.9%), followed by 3-acetyldeoxynivalenol (3-ADON) (13.6%), and nivalenol (NIV) (3.5%). In F. culmorum, the prevalent genotype was 3-ADON (59.9%), while the NIV genotype accounted for the remaining 40.1%. Both, geographical and temporal patterns of trichothecene genotypes distribution were identified.

Developments in breeding cereals for organic agriculture

The need for increased sustainability of performance in cereal varieties, particularly in organic agriculture (OA), is limited by the lack of varieties adapted to organic conditions. Here, the needs for breeding are reviewed in the context of three major marketing types, global, regional, local, in European OA. Currently, the effort is determined, partly, by the outcomes from trials that compare varieties under OA and CA (conventional agriculture) conditions. The differences are sufficiently large and important to warrant an increase in appropriate breeding. The wide range of environments within OA and between years, underlines the need to try to select for specific adaptation in target environments. The difficulty of doing so can be helped by decentralised breeding with farmer participation and the use of crops buffered by variety mixtures or populations. Varieties for OA need efficient nutrient uptake and use and weed competition. These and other characters need to be considered in relation to the OA cropping system over the whole rotation. Positive interactions are needed, such as early crop vigour for nutrient uptake, weed competition and disease resistance. Incorporation of all characteristics into the crop can be helped by diversification within the crop, allowing complementation and compensation among plants. Although the problems of breeding cereals for organic farming systems are large, there is encouraging progress. This lies in applications of ecology to organic crop production, innovations in plant sciences, and the realisation that such progress is central to both OA and CA, because of climate change and the increasing costs of fossil fuels.

Involvement of trichothecenes in fusarioses of wheat, barley and maize evaluated by gene disruption of the trichodiene synthase (Tri5) gene in three field isolates of different chemotype and virulence

SUMMARY Fusarium graminearum is the main causative agent of Fusarium head blight on small grain cereals and of ear rot on maize. The disease leads to dramatic yield losses and to an accumulation of mycotoxins. The most dominant F. graminearum mycotoxins are the trichothecenes, with deoxynivalenol and nivalenol being the most prevalent derivatives. To investigate the involvement of trichothecenes in the virulence of the pathogen, the gene coding for the initial enzyme of the trichothecene pathway was disrupted in three field isolates, differing in chemotype and in virulence. From each isolate three individual disruption mutants were tested for their virulence on wheat, barley and maize. Despite the different initial virulence of the three wild-type progenitor strains on wheat, all disruption mutants caused disease symptoms on the inoculated spikelet, but the symptoms did not spread into other spikelets. On barley, the trichothecene deficient mutants showed no significant difference compared to the wild-type strains: all were equally aggressive. On maize, mutants derived from the NIV-producing strain caused less disease than their wild-type progenitor strain, while mutants derived from DON-producing strains caused the same level of disease as their progenitor strains. These data demonstrate that trichothecenes influence the virulence of F. graminearum in a highly complex manner, which is strongly host as well as moderately chemotype specific.

Revealing the genetic architecture of FHB resistance in hexaploid wheat (Triticum aestivum L.) by QTL meta-analysis

Fusarium head blight (FHB) in wheat results in reduced yield and quality and in accumulation of mycotoxins. The objective of this study was to identify genomic regions in wheat involved in the control of FHB resistance applying a QTL meta-analysis approach by combining QTL of 30 mapping populations to propose independent meta-QTL (MQTL). A consensus map was created on which initial QTL were projected. Nineteen MQTL comprising 2–13 initial QTL with widely varying confidence intervals were found on 12 chromosomes. Some of them coincided with genomic regions previously identified (e.g. chromosomes 3BS, 6B), however, some MQTL were newly detected by this study. Separate analysis of populations with the same resistant parent showed a rather high consistency for the Chinese spring wheat donor ‘Sumai 3’, but little consistency for the Chinese donor ‘Wangshuibai’ and the Swiss donor ‘Arina’. According to our results breeders can in future (1) choose parents for crossing not comprising the same resistance loci or QTL intervals, (2) exploit new MQTL, and (3) select markers of some of these MQTL to be used in marker-assisted selection.

Stacking quantitative trait loci (QTL) for Fusarium head blight resistance from non-adapted sources in an European elite spring wheat background and assessing their effects on deoxynivalenol (DON) content and disease severity.

Fusarium head blight (FHB) is a devastating disease in wheat that reduces grain yield, grain quality and contaminates the harvest with deoxynivalenol (DON). As potent resistance sources Sumai 3 and its descendants from China and Frontana from Brazil had been analysed by quantitative trait loci (QTL) mapping. We introgressed and stacked two donor QTL from CM82036 (Sumai 3/Thornbird) located on chromosomes 3B and 5A and one donor QTL from Frontana on chromosome 3A in elite European spring wheat and estimated the effects of the three individual donor QTL and their four combinations on DON, Fusarium exoantigen content, and FHB rating adjusted to heading date. One class with the susceptible QTL alleles served as control. Each of the eight QTL classes was represented by 12–15 F3-derived lines tested in F5 generation as bulked progeny possessing the respective marker alleles homozygously. Traits were evaluated in a field experiment across four locations with spray inoculation of Fusarium culmorum. All three individual donor-QTL alleles significantly reduced DON content and FHB severity compared to the marker class with no donor QTL. The only exception was the donor-QTL allele 3A that had a low, but non-significant effect on FHB severity. The highest effect had the stacked donor-QTL alleles 3B and 5A for both traits. They jointly reduced DON content by 78% and FHB rating by 55% compared to the susceptible QTL class. Analysis of Fusarium exoantigen content illustrates that lower disease severity is associated with less mycelium content in the grain. In conclusion, QTL from non-adapted sources could be verified in a genetic background of German elite spring wheat. Within the QTL classes significant (P<0.05) genotypic differences were found among the individual genotypes. An additional phenotypic selection would, therefore, be advantageous after performing a marker-based selection.

Deoxynivalenol and Nivalenol Production by Fusarium culmorum Isolates Differing in Aggressiveness Toward Winter Rye.

ABSTRACT A susceptible synthetic winter rye population was inoculated with 42 isolates of Fusarium culmorum, originating from nine European countries and Australia, at two field locations in Germany. Significant (P = 0.01) genetic variation in aggressiveness of isolates of F. culmorum was observed across both field locations. Field samples were used to determine deoxynivalenol (DON), nivalenol (NIV), and ergosterol (ERG) contents. The 42 isolates also were incubated on rye grain in vitro, and DON and NIV contents were analyzed. Thirty-four isolates produced DON, and seven isolates produced NIV at both field locations and in vitro. Mean DON contents ranged from 0.5 to 64.6 mg/kg in grain from field trials and from 0.3 to 376.3 mg/kg in grain incubated in vitro; mean NIV contents ranged from 17.6 to 30.4 mg/kg in grain from field trials and from 0.8 to 381.0 mg/kg in grain incubated in vitro. No correlation was found between the DON content of field-grown grain and grain incubated in vitro. NIV-producing isolates originated from the Netherlands, Germany, Italy, and Australia. More aggressive isolates produced higher mean DON contents in grain in field trials (r = 0.69; P = 0.01). However, DON production rate per unit of fungal biomass, estimated as the DON/ERG ratio at harvest, was not correlated with aggressiveness. Toxin production seemed to be a common feature in F. culmorum. In vitro assays reliably distinguished DON- and NIV-producing types of F. culmorum; however, these assays could not predict production of DON by these isolates in the field.

Detection of segregation distortion loci in triticale (x Triticosecale Wittmack) based on a high-density DArT marker consensus genetic linkage map

BackgroundTriticale is adapted to a wide range of abiotic stress conditions, is an important high-quality feed stock and produces similar grain yield but more biomass compared to other crops. Modern genomic approaches aimed at enhancing breeding progress in cereals require high-quality genetic linkage maps. Consensus maps are genetic maps that are created by a joint analysis of the data from several segregating populations and different approaches are available for their construction. The phenomenon that alleles at a locus deviate from the Mendelian expectation has been defined as segregation distortion. The study of segregation distortion is of particular interest in doubled haploid (DH) populations due to the selection pressure exerted on the plants during the process of their establishment.ResultsThe final consensus map, constructed out of six segregating populations derived from nine parental lines, incorporated 2555 DArT markers mapped to 2602 loci (1929 unique). The map spanned 2309.9 cM with an average number of 123.9 loci per chromosome and an average marker density of one unique locus every 1.2 cM. The R genome showed the highest marker coverage followed by the B genome and the A genome. In general, locus order was well maintained between the consensus linkage map and the component maps. However, we observed several groups of loci for which the colinearity was slightly uneven. Among the 2602 loci mapped on the consensus map, 886 showed distorted segregation in at least one of the individual mapping populations. In several DH populations derived by androgenesis, we found chromosomes (2B, 3B, 1R, 2R, 4R and 7R) containing regions where markers exhibited a distorted segregation pattern. In addition, we observed evidence for segregation distortion between pairs of loci caused either by a predominance of parental or recombinant genotypes.ConclusionsWe have constructed a reliable, high-density DArT marker consensus genetic linkage map as a basis for genomic approaches in triticale research and breeding, for example for multiple-line cross QTL mapping experiments. The results of our study exemplify the tremendous impact of different DH production techniques on allele frequencies and segregation distortion covering whole chromosomes.

Genetic Mapping of Pathogenicity and Aggressiveness of Gibberella zeae (Fusarium graminearum) Toward Wheat

ABSTRACT Gibberella zeae is the major fungal pathogen of Fusarium head blight of wheat and produces several mycotoxins that are harmful to humans and domesticated animals. We identified loci associated with pathogenicity and aggressiveness on an amplified fragment length polymorphism based genetic map of G. zeae in a cross between a lineage 6 nivalenol producer from Japan and a lineage 7 deoxynivalenol producer from Kansas. Ninety-nine progeny and the parents were tested in the greenhouse for 2 years. Progeny segregated qualitatively (61:38) for pathogenicity:nonpathogenicity, respectively. The trait maps to linkage group IV, which is adjacent to loci that affect colony pigmentation, perithecium production, and trichothecene toxin amount. Among the 61 pathogenic progeny, the amount of disease induced (aggressiveness) varied quantitatively. Two reproducible quantitative trait loci (QTL) for aggressiveness were detected on linkage group I using simple interval analysis. A QTL linked to the TRI5 locus (trichodiene synthase in the trichothecene pathway gene cluster) explained 51% of the variation observed, and a second QTL that was 50 centimorgans away explained 29% of the phenotypic variation. TRI5 is tightly linked to the locus controlling trichothecene toxin type. The two QTLs, however, were likely part of the same QTL using composite interval analysis. Progeny that produced deoxynivalenol were, on average, approximately twice as aggressive as those that produced nivalenol. No transgressive segregation for aggressiveness was detected. The rather simple inheritance of both traits in this interlineage cross suggests that relatively few loci for pathogenicity or aggressiveness differ between lineage 6 and 7.

Marker-Assisted Selection for Disease Resistance in Wheat and Barley Breeding

Marker-assisted selection (MAS) provides opportunities for enhancing the response from selection because molecular markers can be applied at the seedling stage, with high precision and reductions in cost. About each of 50 genes conferring monogenic resistances and hundreds of quantitative trait loci (QTL) for quantitative disease resistances have been reported in wheat and barley. For detecting single-major gene resistance, MAS could be easily applied, but is often not necessary because the resistances are selected phenotypically. In quantitative disease resistances, MAS would be very useful, but the individual QTL often have small effects. Additionally, only a few monogenic resistances are durable and only a few QTL with high effects have been successfully transferred into elite breeding material. Further economic and biological constraints, e.g., a low return of investment in small-grain cereal breeding, lack of diagnostic markers, and the prevalence of QTL-background effects, hinder the broad implementation of MAS. Examples in which MAS has been successfully applied to practical breeding are the wheat rust resistance genes Lr34 and Yr36, the eyespot resistance gene Pch1, the recessive resistance genes rym4/rym5 to barley yellow mosaic viruses, mlo to barley powdery mildew, and two QTL for resistance to Fusarium head blight in wheat (Fhb1 and Qfhs.ifa-5A). Newly identified broad-spectrum resistance genes/QTL conferring resistance to multiple taxa of pathogens offer additional perspectives for MAS. In the future, chip-based, high-throughput genotyping platforms and the introduction of genomic selection will reduce the current problems of integrating MAS in practical breeding programs and open new avenues for a molecular-based resistance breeding.

Quantitative-genetic basis of aggressiveness of 42 isolates of fusarium culmorum for winter rye head blight

Forty-two isolates of Fusarium culmorum obtained from diseased plant parts collected from fields in nine European countries and Australia were tested on a synthetic winter rye population. A spore suspension of each isolate was sprayed during midanthesis in five environments (location-year combinations) onto the heads. All isolates were pathogenic as judged by head blight rating scored on a 1 to 9 scale and grain weight relative to the noninoculated control. Isolates differed, however, in their ability to cause disease (aggressiveness). Quantitative geno-typic variation for aggressiveness occurred, while isolate-environment interaction variance, although significant, accounted for only 14% of total variance averaged over both traits. Correlations for aggressiveness across environments ranged from 0.6 to 0.8 (P = 0.01). Estimates of heritability on an entry-mean basis were high (h 2 = 0.9) for both traits, indicating that a substantial proportion of the phenotypic variation was caused by genetic effects. It is concluded that aggressiveness of F. culmorum is inherited as a complex trait.