Changing fitness of a necrotrophic plant pathogen under increasing temperature.

Warmer temperatures associated with climate change are expected to have a direct impact on plant pathogens, challenging crops and altering plant disease profiles in the future. In this study, we have investigated the effect of increasing temperature on the pathogenic fitness of Fusarium pseudograminearum, an important necrotrophic plant pathogen associated with crown rot disease of wheat in Australia. Eleven wheat lines with different levels of crown rot resistance were artificially inoculated with F. pseudograminearum and maintained at four diurnal temperatures 15/15°C, 20/15°C, 25/15°C and 28/15°C in a controlled glasshouse. To quantify the success of F. pseudograminearum three fitness measures, these being disease severity, pathogen biomass in stem base and flag leaf node, and deoxynivalenol (DON) in stem base and flag leaf node of mature plants were used. F. pseudograminearum showed superior overall fitness at 15/15°C, and this was reduced with increasing temperature. Pathogen fitness was significantly influenced by the level of crown rot resistance of wheat lines, but the influence of line declined with increasing temperature. Lines that exhibited superior crown rot resistance in the field were generally associated with reduced overall pathogen fitness. However, the relative performance of the wheat lines was dependent on the measure of pathogen fitness, and lines that were associated with one reduced measure of pathogen fitness did not always reduce another. There was a strong correlation between DON in stem base tissue and disease severity, but length of browning was not a good predictor of Fusarium biomass in the stem base. We report that a combination of host resistance and rising temperature will reduce pathogen fitness under increasing temperature, but further studies combining the effect of rising CO2 are essential for more realistic assessments.

Migrate or evolve: options for plant pathogens under climate change.

Findings on climate change influence on plant pathogens are often inconsistent and context dependent. Knowledge of pathogens affecting agricultural crops and natural plant communities remains fragmented along disciplinary lines. By broadening the perspective beyond agriculture, this review integrates cross-disciplinary knowledge to show that at scales relevant to climate change, accelerated evolution and changing geographic distribution will be the main implications for pathogens. New races may evolve rapidly under elevated temperature and CO2 , as evolutionary forces act on massive pathogen populations boosted by a combination of increased fecundity and infection cycles under favourable microclimate within enlarged canopy. Changing geographic distribution will bring together diverse lineages/genotypes that do not share common ecological niche, potentially increasing pathogen diversity. However, the uncertainty of model predictions and a lack of synthesis of fragmented knowledge remain as major deficiencies in knowledge. The review contends that the failure to consider scale and human intervention through new technology are major sources of uncertainty. Recognizing that improved biophysical models alone will not reduce uncertainty, it proposes a generic framework to increase focus and outlines ways to integrate biophysical elements and technology change with human intervention scenarios to minimize uncertainty. To synthesize knowledge of pathogen biology and life history, the review borrows the concept of 'fitness' from population biology as a comprehensive measure of pathogen strengths and vulnerabilities, and explores the implications of pathogen mode of nutrition to fitness and its interactions with plants suffering chronic abiotic stress under climate change. Current and future disease management options can then be judged for their ability to impair pathogenic and saprophytic fitness. The review pinpoints improving confidence in model prediction by minimizing uncertainty, developing management strategies to reduce overall pathogen fitness, and finding new sources of data to trawl for climate signatures on pathogens as important challenges for future research.

Comparative pathogenomics reveals horizontally acquired novel virulence genes in fungi infecting cereal hosts.

Comparative analyses of pathogen genomes provide new insights into how pathogens have evolved common and divergent virulence strategies to invade related plant species. Fusarium crown and root rots are important diseases of wheat and barley world-wide. In Australia, these diseases are primarily caused by the fungal pathogen Fusarium pseudograminearum. Comparative genomic analyses showed that the F. pseudograminearum genome encodes proteins that are present in other fungal pathogens of cereals but absent in non-cereal pathogens. In some cases, these cereal pathogen specific genes were also found in bacteria associated with plants. Phylogenetic analysis of selected F. pseudograminearum genes supported the hypothesis of horizontal gene transfer into diverse cereal pathogens. Two horizontally acquired genes with no previously known role in fungal pathogenesis were studied functionally via gene knockout methods and shown to significantly affect virulence of F. pseudograminearum on the cereal hosts wheat and barley. Our results indicate using comparative genomics to identify genes specific to pathogens of related hosts reveals novel virulence genes and illustrates the importance of horizontal gene transfer in the evolution of plant infecting fungal pathogens.

Fitness of three Fusarium pathogens of wheat.

Crown rot and head blight of wheat are caused by the same Fusarium species. To better understand their biology, this study has compared 30 isolates of the three dominant species using 13 pathogenic and saprophytic fitness measures including aggressiveness for the two diseases, saprophytic growth and fecundity and deoxynivalenol (DON) production from saprophytic colonization of grain and straw. Pathogenic fitness was generally linked to DON production in infected tissue. The superior crown rot fitness of Fusarium pseudograminearum was linked to high DON production in the stem base tissue, while Fusarium culmorum and Fusarium graminearum had superior head blight fitness with high DON production in grains. Within each species, some isolates had similar aggressiveness for both diseases but differed in DON production in infected tissue to indicate that more than one mechanism controlled aggressiveness. All three species produced more DON when infecting living host tissue compared with saprophytic colonization of grain or straw, but there were significant links between these saprophytic fitness components and aggressiveness. As necrotrophic pathogens spend a part of their life cycle on dead organic matter, saprophytic fitness is an important component of their overall fitness. Any management strategy must target weaknesses in both pathogenic fitness and saprophytic fitness.

Mycotoxins produced by Fusarium spp. associated with Fusarium head blight of wheat in Western Australia.

An isolated occurrence of Fusarium head blight (FHB) of wheat was detected in the south-west region of Western Australia during the 2003 harvest season. The molecular identity of 23 isolates of Fusarium spp. collected from this region during the FHB outbreak confirmed the associated pathogens to be F. graminearum, F. acuminatum or F. tricinctum. Moreover, the toxicity of their crude extracts from Czapek-Dox liquid broth and millet seed cultures to brine shrimp (Artemia franciscana) was associated with high mortality levels. The main mycotoxins detected were type B trichothecenes (deoxynivalenol and 3-acetyldeoxynivalenol), enniatins, chlamydosporol and zearalenone. This study is the first report on the mycotoxin profiles of Fusarium spp. associated with FHB of wheat in Western Australia. This study highlights the need for monitoring not just for the presence of the specific Fusarium spp. present in any affected grain but also for their potential mycotoxin and other toxic secondary metabolites.

Mycotoxins produced by Fusarium species associated with annual legume pastures and 'sheep feed refusal disorders' in Western Australia.

Sheep grazing in Western Australia can partially or completely refuse to consume annual Medicago pods contaminated with a number of different Fusarium species. Many Fusarium species are known to produce trichothecenes as part of their array of toxigenic secondary metabolites, which are known to cause feed refusal in animals. This study reports the identity of Fusarium species using species-specific PCR primers and a characterization of the toxigenic secondary metabolites produced by 24 Fusarium isolates associated with annual legume-based pastures and particularly those associated with sheep feed refusal disorders in Western Australia. Purification of the fungal extracts was facilitated by a bioassay-guided fractionation using brine shrimp. A number of trichothecenes (3-acetyldeoxynivalenol, deoxynivalenol, fusarenon-X, monoacetoxyscirpenols, diacetoxyscirpenol, scirpentriol, HT-2 toxin and T-2 toxin), enniatins (A, A1, B, and B1), chlamydosporol and zearalenone were identified using GC/MS and/or NMR spectroscopy. Some of the crude extracts and fractions showed significant activity against brine shrimp at concentrations as low as 5 µg ml(-1), and are likely to be involved in the sheep feed refusal disorders. This is the first report of chlamydosporol production by confirmed Fusarium spp.; of the incidence of F. brachygibbosum and F. venenatum in Australia and of F. tricinctum in Western Australia; and of mycotoxin production by Fusarium species from Western Australia.

Wheat crown rot pathogens Fusarium graminearum and F. pseudograminearum lack specialization.

This article reports a lack of pathogenic specialization among Australian Fusarium graminearum and F. pseudograminearum causing crown rot (CR) of wheat using analysis of variance (ANOVA), principal component and biplot analysis, Kendall's coefficient of concordance (W), and κ statistics. Overall, F. pseudograminearum was more aggressive than F. graminearum, supporting earlier delineation of the crown-infecting group as a new species. Although significant wheat line-pathogen isolate interaction in ANOVA suggested putative specialization when seedlings of 60 wheat lines were inoculated with 4 pathogen isolates or 26 wheat lines were inoculated with 10 isolates, significant W and κ showed agreement in rank order of wheat lines, indicating a lack of specialization. The first principal component representing nondifferential aggressiveness explained a large part (up to 65%) of the variation in CR severity. The differential components were small and more pronounced in seedlings than in adult plants. By maximizing variance on the first two principal components, biplots were useful for highlighting the association between isolates and wheat lines. A key finding of this work is that a range of analytical tools are needed to explore pathogenic specialization, and a statistically significant interaction in an ANOVA cannot be taken as conclusive evidence of specialization. With no highly resistant wheat cultivars, Fusarium isolates mostly differ in aggressiveness; however, specialization may appear as more resistant cultivars become widespread.

Genetic relationships between resistances to Fusarium head blight and crown rot in bread wheat (Triticum aestivum L.).

Fusarium head blight (FHB) and crown rot (CR) are two wheat diseases caused by the same Fusarium pathogens. Progress towards CR resistance could benefit from FHB-resistant germplasm if the same genes are involved in resistance to these two different diseases. Two independent studies were conducted to investigate the relationship between host resistances to these two diseases. In the first study 32 genotypes were assessed and no significant correlation between their reactions to FHB and CR was detected. The second study was based on a QTL analysis of a doubled haploid population derived from a variety with resistance to both diseases. Results from this study showed that loci conferring resistance to FHB and CR are located on different chromosomes. Together, these results suggest that, despite a common aetiology, different host genes are involved in the resistance against FHB and CR in wheat. Thus, although it is possible that genes affecting both diseases may exist in other germplasm or under different conditions, separate screening seems to be needed in identifying sources of CR resistance.

Passage through alternative hosts changes the fitness of Fusarium graminearum and Fusarium pseudograminearum.

Species of the necrotrophic fungal pathogen Fusarium that cause head blight and crown rot of cereals including wheat also infect a number of alternative host plants. This raises the prospect of more damaging pathogen strains originating and persisting as highly successful saprophytes on hosts other than wheat. The immediate impact on pathogenic (aggressiveness) and saprophytic (growth rate and fecundity) behaviour of six isolates with low, moderate or high initial aggressiveness was examined in two species of Fusarium after their passage through 10 alternative plant hosts. One passage through alternative hosts significantly reduced the pathogenic fitness of most isolates, but this change was not associated with a concomitant change in their overall saprophytic behaviour. The overall weak association between aggressiveness, fecundity and growth rate both before and after passage through the alternative hosts indicate that pathogenic and saprophytic fitness traits may be independently controlled in both Fusarium species. Thus, there was no trade-off between pathogenic and saprophytic fitness in these necrotrophic plant pathogens.

Multilocus sequence analysis of Fusarium pseudograminearum reveals a single phylogenetic species.

Fusarium pseudograminearum causes crown rot of wheat in Australia and most other wheat growing regions, but its evolutionary history is largely unknown. We demonstrate for the first time that F. pseudograminearum is a single phylogenetic species without consistent lineage development across genes. Isolates of F. pseudograminearum, F. graminearum sensu lato, and F. cerealis, were collected from four countries and four single copy, nuclear genes were partially sequenced, aligned with previously published sequences of these and related species, and analysed by maximum parsimony and Bayesian inference. Evolutionary divergence varied between genes, with high phylogenetic incongruence occurring between the gene genealogies. The absence of geographic differentiation between isolates indicates that the introduction of new fungal strains to a region has the potential to introduce new pathogenic and toxigenic genes into the native population through sexual recombination.

How will plant pathogens adapt to host plant resistance at elevated CO2 under a changing climate?

• To better understand evolution we have studied aggressiveness of the anthracnose pathogen, Colletotrichum gloeosporioides, collected from Stylosanthes scabra pastures between 1978 and 2000 and by inoculating two isolates onto two cultivars over 25 sequential infection cycles at ambient (350 ppm) and twice-ambient atmospheric CO2 in controlled environments. • Regression analysis of the field population showed that aggressiveness increased towards a resistant cultivar, but not towards a susceptible cultivar, that is no longer grown commercially. • Here we report for the first time that aggressiveness increased on both cultivars after a few initial infection cycles at twice-ambient CO2 as isolates adapted to combat enhanced host resistance, while at ambient CO2 this increased steadily for most cycles as both cultivars selected for increased aggressiveness. Genetic fingerprint and karyotype of isolates changed for some CO2 -cultivar combinations, but these were not related to changed aggressiveness. • At 700 ppm fecundity increased for both isolates, and this increased population size, in combination with a conducive microclimate for anthracnose from an enlarged plant canopy under elevated CO2 , could accelerate pathogen evolution.

A neuro-fuzzy framework for inferencing.

Earlier we proposed a connectionist implementation of compositional rule of inference (COI) for rules with antecedents having a single clause. We first review this net, then generalize it so that it can deal with rules with antecedent having multiple clauses. We call it COIN, the compositional rule of inferencing network. Given a relational representation of a set of rules, the proposed architecture can realize the COI. The outcome of COI depends on the choice of both the implication function and the inferencing scheme. The problem of choosing an appropriate implication function is avoided through neural learning. COIN can automatically find a 'good' relation to represent a set of fuzzy rules. We model the connection weights so as to ensure learned weights lie in [0,1]. We demonstrate through extensive numerical examples that the proposed neural realization can find a much better representation of the rules than that by usual implication and hence results in much better conclusions than the usual COI.