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The SnTox1 effector is a virulence factor of the fungal pathogen Stagonospora nodorum (Berk.), which interacts with the host susceptibility gene Snn1 in a gene-for-gene manner and causes necrosis on the leaves of sensitive wheat genotypes. It is known that salicylic acid (SA), jasmonic acid (JA) and ethylene are the key phytohormones involved in plant immunity. To date, effectors of various pathogens have been discovered that can manipulate plant hormonal pathways and even use hormone crosstalk to promote disease development. However, the role of SnTox1 in manipulating hormonal pathways has not been studied in detail. We studied the redox status and the expression of twelve genes of hormonal pathways and two MAPK genes in six bread wheat cultivars sensitive and insensitive to SnTox1 with or without treatment by SA, JA and ethephon (ethylene-releasing agent) during infection with the SnTox1-producing isolate S. nodorum 1SP. The results showed that SnTox1 controls the antagonism between the SA and JA/ethylene signaling pathways. The SA pathway was involved in the development of susceptibility, and the JA/ethylene pathways were involved in the development of wheat plants resistance to the Sn1SP isolate in the presence of a SnTox1-Snn1 interaction. SnTox1 hijacked the SA pathway to suppress catalase activity, increase hydrogen peroxide content and induce necrosis formation; it simultaneously suppresses the JA and ethylene hormonal pathways by SA. To do this, SnTox1 reprogrammed the expression of the MAPK genes TaMRK3 and TaMRK6 and the TF genes TaWRKY13, TaEIN3 and TaWRKY53b. This study provides new data on the role of SnTox1 in manipulating hormonal pathways and on the role of SA, JA and ethylene in the pathosystem wheat S. nodorum. [ABSTRACT FROM AUTHOR]

Background: Currently, the role of microRNAs in plant immune responses is being actively studied. Thus, our aim was to research the effect of Stagonospora nodorum (Berk.) NEs SnToxA and SnTox3 on the expression of miRNAs involved in the wheat–S. nodorum interaction and to determine the role of phytohormones in this process. Methods: The expressions of nine conserved microRNAs were studied by quantitative real-time polymerase chain reaction in three different wheat genotypes of bread spring wheat (Triticum aestivum L.) infected with S. nodorum. Phytohormone treatments (trans-zeatin, 2-chloroethylphosphonic acid (etefone is the chemical precursor of ethylene), and salicylic acid) were applied. The results were compared with disease symptoms, the redox status of plants, and the expression of fungal necrotrophic effector (NE) genes of SnToxA and SnTox3 and genes of SnPf2, SnStuA, alongside SnCon7 transcription factors (TFs). Results: Salicylic acid (SA) and cytokinins (CK) are involved in the development of defense reactions in wheat plants against S. nodorum, by regulating the expression of fungal NEs and TFs genes, inducing an oxidative burst in all three wheat genotypes. Moreover, ethylene enhanced the virulence of the pathogen by increasing the expression of fungal NE and TF genes, thereby resulting in a decrease in the generation of reactive oxygen species in all three cultivars. The nine miRNAs played a role in the development of wheat resistance against S. nodorum. NE SnTox3 mainly suppressed the expression of three miRNAs: miR159, miR393, and miR408, while NE SnToxA suppressed miR166 expression. Conversely, treatment with CK and SA increased the expression of miR159 and miR408; treatment with CK increased the expression of miR393 and miR166. Ethylene inhibited the expression of miR159, miR408, miR393, and miR166. Suppression of miP159 expression by NE SnTox3 was most likely associated with the activation of the ethylene signaling pathway. NEs SnToxA and SnTox3 suppressed the expression of miR408, whose role most likely consisted of inhibiting the catalase activity, via SA and CK regulation. In addition, NE SnToxA hijacked the SA signaling pathway and manipulated it for fungal growth and development. Fungal TFs SnPf2 and SnStuA could be involved in the regulation of these processes indirectly through the regulation of the expression of NE genes. Conclusions: The results of this work show, for the first time, the role of microRNAs in the development of wheat resistance against S. nodorum and the effect of S. nodorum NEs SnToxA and SnTox3 on the activity of plant microRNAs.

The fungus Stagonospora nodorum Berk. is the causative agent of Septoria nodorum blotch (SNB) of wheat. The most important factors of Stagonospora nodorum virulence include numerous fungal necrotrophic effectors (NEs) encoded by SnTox genes. They interact with the matching products of host susceptibility genes (Snn). SnTox-Snn interactions are mirror images of classical gene-for-gene interactions and lead to the development of disease. We have studied the SnTox3-Snn3 interaction, resulting in the development of infection on leaves and formation of extensive lesions. The mechanism of SnTox3 action is likely to be linked to the regulation of redox metabolism and the influence on ethylene synthesis in the wheat plants, although the molecular mechanisms are not fully unveiled. To characterize the SnTox3-Snn3 interaction, we used S. nodorum isolates differing in the expression of the NEs genes SnTox3 (SnB (Tox3+), Sn4VD (Tox3–)) and two soft spring wheat (Triticum aestivum L.) cultivars, contrasting in resistance to the SNB agent and differing in the allelic composition of the susceptibility locus Snn3-B1: Kazakhstanskaya 10 (susceptible) and Omskaya 35 (resistant). We carried out a comparative assessment of the transcriptional activity patterns of genes responsible for ethylene biosynthesis (TaACS1, TaACО) and signaling pathway (TaEIL1, TaERF1) by real-time PCR and estimated the redox state of wheat plants infected with different isolates of S. nodorum by spectrometry. The induction of ethylene biosynthesis and signaling has been shown to result from gene-for-gene interaction between Snn3-B1 and SnTox3. The results of plant redox status estimation showed that ethylene inhibited accumulation of hydrogen peroxide in SnTox3-sensitive genotypes by regulating the operation of various pro-/antioxidant enzymes at the transcriptional and posttranslational levels. Our results suggest that NE SnTox3 influences ethylene biosynthesis and signaling, thereby regulating redox metabolism in infected wheat plants as necessary for successful host colonization at the initial phases of infection, which ultimately leads to extensive lesions due to fast pathogen reproduction.

Stagonospora nodorum blotch (SNB) is an economically important wheat disease caused by the necrotrophic fungus Parastagonospora nodorum. SNB resistance in wheat is controlled by several quantitative trait loci (QTLs). Thus, identifying novel resistance/susceptibility QTLs is crucial for continuous improvement of the SNB resistance. Here, the hard winter wheat association mapping panel (HWWAMP) comprising accessions from breeding programs in the Great Plains region of the US, was evaluated for SNB resistance and necrotrophic effectors (NEs) sensitivity at the seedling stage. A genome-wide association study (GWAS) was performed to identify single‐nucleotide polymorphism (SNP) markers associated with SNB resistance and effectors sensitivity. We found seven significant associations for SNB resistance/susceptibility distributed over chromosomes 1B, 2AL, 2DS, 4AL, 5BL, 6BS, and 7AL. Two new QTLs for SNB resistance/susceptibility at the seedling stage were identified on chromosomes 6BS and 7AL, whereas five QTLs previously reported in diverse germplasms were validated. Allele stacking analysis at seven QTLs explained the additive and complex nature of SNB resistance. We identified accessions ('Pioneer-2180' and 'Shocker') with favorable alleles at five of the seven identified loci, exhibiting a high level of resistance against SNB. Further, GWAS for sensitivity to NEs uncovered significant associations for SnToxA and SnTox3, co-locating with previously identified host sensitivity genes (Tsn1 and Snn3). Candidate region analysis for SNB resistance revealed 35 genes of putative interest with plant defense response-related functions. The QTLs identified and validated in this study could be easily employed in breeding programs using the associated markers to enhance the SNB resistance in hard winter wheat. [ABSTRACT FROM AUTHOR]

Use of chemical pesticides poses a threat for environment and human health, so green technologies of crop protection are of high demand. Some microbial proteins able to activate plant defense mechanisms and prevent the development of resistance in plant pathogens, may be good alternative to chemicals, but practical use of such elicitors is limited due to need to protect them against adverse environment prior their delivery to target receptors of plant cells. In this study we examined a possibility to encapsulate heat resistant FKBP-type peptidyl prolyl cis-trans isomerase (PPIase) from Pseudomonas fluorescens, which possesses a significant eliciting activity in relation to a range of plant pathogens, in sodium alginate microparticles and evaluated the stability of resulted complex under long-term UV irradiation and in the presence of proteinase K, as well as its eliciting activity in three different “plant-pathogen” models comparing to that of free PPIase. The obtained PPIase-containing microparticles consisted of 70% of sodium alginate, 20% of bovine serum albumin, and 10% of PPIase. In contrast to free PPIase, which lost its eliciting properties after 8-h UV treatment, encapsulated PPIase kept its eliciting ability unchanged; after being exposed to proteinase K, its eliciting ability twice exceeded that of free PPIase. Using “tobacco-TMV”, “tobacco-Alternaria longipes”, and “wheat-Stagonospora nodorum” model systems, we showed that encapsulation process did not influence on the eliciting activity of PPIase. In the case of the “wheat-S. nodorum” model system, we also observed a significant eliciting activity of alginate-albumin complex and almost doubled activity of encapsulated PPIase as compared to the free PPIase. As far as we know, this is the first observation of a synergistic interaction between alginate and other compound possessing any bioactive properties. The results of the study show some prospects for a PPIase use in agriculture.

The centre of origin of the globally distributed wheat pathogen Parastagonospora nodorum has remained uncertain because only a small number of isolates from the Fertile Crescent were included in earlier population genetic and phylogeographic studies. We isolated and genetically analysed 193 P. nodorum strains from three naturally infected wheat fields distributed across Iran using 11 neutral microsatellite loci. Compared to previous studies that included populations from North America, Europe, Africa, Australia, and China, the populations from Iran had the highest genetic diversity globally and also exhibited greater population structure over smaller spatial scales, patterns typically associated with the centre of origin of a species. Genes encoding the necrotrophic effectors SnToxA, SnTox1, and SnTox3 were found at a high frequency in the Iranian population. By sequencing 96 randomly chosen Iranian strains, we detected new alleles for all three effector genes. Analysis of allele diversity showed that all three effector genes had higher diversity in Iran than in any population included in previous studies, with Iran acting as a hub for the effector diversity that was found in other global populations. Taken together, these findings support the hypothesis that P. nodorum originated either within or nearby the Fertile Crescent with a genome that already encoded all three necrotrophic effectors during its emergence as a pathogen on wheat. Our findings also suggest that P. nodorum was the original source of the ToxA genes discovered in the wheat pathogens Phaeosphaeria avenaria f. sp. tritici 1, Pyrenophora tritici‐repentis, and Bipolaris sorokiniana. [ABSTRACT FROM AUTHOR]

Stagonospora leaf and glume blotch (SLGB) of wheat caused by Parastagonospora nodorum (formerly Stagonospora nodorum) has recently emerged as a major problem in changing climatic conditions of Himachal Pradesh (HP), especially during delayed winter rains. In the present studies symptomatology, morpho-cultural as well as molecular marker based identification showed the prevalence of disease in the state and conclusively proved that leaf and glume blotch of wheat is caused by P. nodorum. The test pathogen showed 100% homology with other reported P. nodorum isolates by rDNA (ribosomal DNA) analysis. In addition, the amplification of rDNA region of 36 P. nodorum isolates representing various agro-ecological areas of HP and one infected wheat leaf sample generated an amplicon of ~ 449-bp with JB433 (5′-ACACTCAGTAGTTTACTACT-3′) and JB434 (5′-TGTGCTGCGCTTCAATA-3′) P. nodorum specific primer pair whereas no amplification was observed with the genomic DNA of Septoria titici, Stemphylium vesicarium and healthy wheat leaf sample. This study on integration of morpho-cultural and microscopic methods along with PCR based technique could form basis for routine diagnosis of the SLGB in wheat samples during early growth stages of crop in the seed production fields. [ABSTRACT FROM AUTHOR]

Ethylene, salicylic acid (SA), and jasmonic acid are the key phytohormones involved in plant immunity, and other plant hormones have been demonstrated to interact with them. The classic phytohormone cytokinins are important participants of plant defense signaling. Crosstalk between ethylene and cytokinins has not been sufficiently studied as an aspect of plant immunity and is addressed in the present research. We compared expression of the genes responsible for hormonal metabolism and signaling in wheat cultivars differing in resistance to Stagonospora nodorum in response to their infection with fungal isolates, whose virulence depends on the presence of the necrotrophic effector SnTox3. Furthermore, we studied the action of the exogenous cytokinins, ethephon (2-chloroethylphosphonic acid, ethylene-releasing agent) and 1-methylcyclopropene (1-MCP, inhibitor of ethylene action) on infected plants. Wheat susceptibility was shown to develop due to suppression of reactive oxygen species production and decreased content of active cytokinins brought about by SnTox3-mediated activation of the ethylene signaling pathway. SnTox3 decreased cytokinin content most quickly by its activated glucosylation in an ethylene-dependent manner and, furthermore, by oxidative degradation and inhibition of biosynthesis in ethylene-dependent and ethylene-independent manners. Exogenous zeatin application enhanced wheat resistance against S. nodorum through inhibition of the ethylene signaling pathway and upregulation of SA-dependent genes. Thus, ethylene inhibited triggering of SA-dependent resistance mechanism, at least in part, by suppression of the cytokinin signaling pathway.

The genetic control of adult plant resistance to Stagonospora nodorum blotch (SNB) is complex, consisting of genes with minor effects interacting in an additive manner. Earlier studies detected quantitative trait loci (QTL) for flag leaf resistance in successive years on chromosomes 1B, 2A, 2D, and 5B using SSR- and DArT-based genetic maps of progeny from the crosses EGA Blanco/Millewa, 6HRWSN125/WAWHT2074, and P92201D5/P91193D1. Similarly, QTL for glume resistance detected in successive years and multiple environments were identified on chromosomes 2D and 4B from genetic maps of P92201D5/P91193D1 and 6HRWSN125/WAWHT2074, respectively. The SSR- and DArT-based genetic maps had an average distance of 6.5, 7.8, and 9.7 cM between marker loci for populations EGA/Millewa, P92201D5/P91193D1, and 6HRWSN125/WAWHT2074, respectively. This study used single nucleotide polymorphism (SNP) markers from the iSelect Infinium 90K genotyping array to fine-map genomic regions harbouring QTL for flag leaf and glume SNB resistance, reducing the average distance between markers to 2.9, 3.3, and 3.4 cM for populations P92201D5/P91193D1, EGA/Millewa, and 6HRWSN125/WAWHT2074, respectively. Increasing the marker density of the genetic maps with SNPs did not identify any new QTL for SNB resistance but discriminated previously identified co-located QTL into separate but closely linked QTL. [ABSTRACT FROM AUTHOR]

The paper describes various ways of storing strains of three septoriosis (leaf spot disease) pathogens (Stagonospora nodorum, Septoria tritici and Stagonospora avenae f. sp. triticea) included in the State Collection of Phytopathogenic Microorganisms of All-Russian Research Institute of Phytopathology. The advantages and disadvantages of these methods were noted, the results of long-term studies on the viability and pathogenicity of strains under various storage conditions were presented. The most reliable methods and optimal storage times for different types of septoriosis pathogens were determined. [ABSTRACT FROM AUTHOR]

Stagonospora nodorum blotch (SNB) caused by Parastagonospora nodorum is a serious disease of wheat worldwide. In the United States, the disease is prevalent on winter wheat in many eastern states, and its management relies mainly on fungicide application after flag leaf emergence. Although SNB can occur prior to flag leaf emergence, the relationship between the time of disease onset and yield has not been determined. Such a relationship is useful in identifying a threshold to facilitate prediction of disease onset in the field. Disease occurred in 390 of 435 disease cases that were recorded across 11 counties in North Carolina from 2012 to 2014. Using cases with disease occurrence, the effect of disease onset on yield was analyzed to identify a disease onset threshold that related time of disease onset to yield. Regression analysis showed that disease onset explained 32% of the variation in yield (P < 0.0001) and from this relationship, day of year (DOY) 102 was identified as the disease onset threshold. Below-average yield occurred in 87% of the disease cases when disease onset occurred before DOY 102 but in only 28% of those cases when onset occurred on or after DOY 102. Subsequently, binary logistic regression models were developed to predict the occurrence and onset of SNB using preplanting factors and cumulative daily infection values (cDIV) starting 1 to 3 weeks prior to DOY 102. Logistic regression showed that previous crop, latitude, and cDIV accumulated 2 weeks prior to DOY 102 (cDPV.2) were significant {P < 0.0001) predictors of disease occurrence, and wheat residue, latitude, longitude, and cDIV.2 were significant (P < 0.0001) predictors of disease onset. The disease onset model had a correct classification rate of 0.94 and specificity and sensitivity rates >0.90. Performance of the disease onset model based on the area under the receiver operating characteristic curve (AUC), k, and the true skill statistic (TSS) was excellent, with prediction accuracy values >0.88. Similarly, internal validation of the disease onset model based on AUC, k, and TSS indicated good performance, with accuracy values >0.88. This disease onset prediction model could serve as a useful decision support tool to guide fungicide applications to manage SNB in wheat. [ABSTRACT FROM AUTHOR]

Long-term research work was performed in the years 1996-2002 on the occurrence and significance of winter wheat glume blotch (Stagonospora nodorum) in the region of middle-southern Poland. It was stated that the disease occurred every year at different intensity and this was dependent on the amount of rainfall during the vegetative period, especially in June. Winter wheat was strongly infected in the years 1999 and 2002 when rainfall in June was the highest. The lowest level of infection was recorded in the year 2000 characterized by the lowest rainfall in this month. Full chemical protection with broad spectrum fungicides performed once at GS 49-55 and always significantly increased grain yield, but the increase was the highest in the years of the highest ear infection by S. nodorum. It was concluded that in experimental period glume blotch of winter wheat could be considered as a disease of the greatest importance in the region of middle - Southern Poland, and thus creation a serious threat to winter wheat cultivated in this area.

1879-2720 leaf samples from 8-13 stations of Hungary were collected in March, April, May and June 2000-2002. Drechslera tritici-repentis, Septoria tritici, Stagonospora nodorum and Bipolaris sorokiniana were found in the leaf samples. The occurrence of necrotrophic pathogens was highest (10,79%) in 2001 and lowest (2,63%) in 2002. The occurrence and rate of the necrotrophic pathogens changed significantly among years and locations. The resistance of cultivars based on natural infection could not be properly evaluated, as the level of the epidemic was mostly low and contradictory among locations. Additionally forty four cultivars were tested in Szeged after winter wheat in protected and unprotected environment. Among the necrotrophic pathogens, the dominant pathogens were D. tritici-repentis (2001 and 2002) and S. nodorum (2001). The differences in variety resistance were significant. The biotrophic (leaf rust, yellow rust and powdery mildew) as well as the mentioned necrotrophic pathogens caused significant yield decrease in both years. According to values of correlation coefficients, the influence of biotrophic pathogens was greater on the yield.

Parastagonospora nodorum is a necrotrophic fungal pathogen causing Septoria nodorum blotch on wheat. We have identified nine necrotrophic effector-host dominant sensitivity gene interactions, and we have cloned three of the necrotrophic effector genes, including SnToxA, SnToxl, and SnTox3. Because sexual populations of P. nodorum are difficult to develop under lab conditions, genome-wide association study (GWAS) is the best population genomic approach to identify genomic regions associated with traits using natural populations. In this article, we used a global collection of 191 P. nodorum isolates from which we identified 2,983 singlenucleotide polymorphism (SNP) markers and gene markers for SnToxA and SnTox3 to evaluate the power of GWAS on two popular wheat breeding lines that were sensitive to SnToxA and SnTox3. Strong marker trait associations (MTA) with P. nodorum virulence that mapped to SnTox3 and SnToxA were first identified using the marker set described above. A novel locus in the P. nodorum genome associated with virulence was also identified as a result of this analysis. To evaluate whether a sufficient level of marker saturation was available, we designed a set of primers every 1 kb in the genomic regions containing SnToxA and SnTox3. Polymerase chain reaction amplification was performed across the 191 isolates and the presence/absence polymorphism was scored and used as the genotype. The marker proximity necessary to identify MTA flanking SnToxA and SnTox3 ranged from 4 to 5 and 1 to 7 kb, respectively. Similar analysis was performed on the novel locus. Using a 45% missing data threshold, two more SNP were identified spanning a 4.6-kb genomic region at the novel locus. These results showed that the rate of linkage disequilibrium (LD) decay in P. nodorum and, likely, other fungi is high compared with plants and animals. The fast LD decay in P. nodorum is an advantage only if sufficient marker density is attained. Based on our results with the SnToxA and SnTox3 regions, markers are needed every 9 or 8 kb, respectively, or in every gene, to guarantee that genes associated with a quantitative trait such as virulence are not missed. [ABSTRACT FROM AUTHOR]

A simple and inexpensive method for extraction of genomic DNA from mycelia of filamentous fungi was developed and the standardization of the protocol is described. The protocol includes both mycelium culturing and DNA extraction. By incorporating a selected commercial sand product into the culture media, the collection and the grinding of mycelia was simple and quick. The protocol of DNA extraction was derived from the alkaline lysis method for plasmid DNA extraction and modified to facilitate efficient extraction of fungal genomic DNA. It has increased efficiency and fidelity compared to the other simple DNA extraction protocols. Relative to a commonly used commercial DNA extraction kit, this extraction protocol is faster, easier and can generate more DNA without compromising quality. From a single sample of mycelium, genomic DNA can be prepared within 20 min, including concentration measurement. The DNA can be used in restriction enzyme digestion and as a template in PCR to amplify DNA fragments larger than 3000 base pairs. This method is recommended for DNA extraction in fungal phylogenetic studies when hundreds or thousands of isolates from one or a few closely-related species are subjected to PCR-based genotyping. [ABSTRACT FROM PUBLISHER]

Agrobacterium tumefaciens-mediated random mutagenesis was used to generate insertional mutants of the fungus Leptosphaeria maculans. Of 91 transformants screened, only one (A3) produced lesions of reduced size on cotyledons of canola (Brassica napus). Genes flanking the T-DNA insertion had the best matches to an alcohol dehydrogenase class 4 (ADH4)-like gene (Adh4L) and a 3-ketoacyl-CoA thiolase gene (Thiol) and were expressed in mutant A3 in vitro and in planta at significantly higher levels than in the wild type. This is the first report of a T-DNA insertion in fungi causing increased gene expression. Transformants of the wild-type isolate expressing both Adh4L and Thiol under the control of a heterologous promoter had similar pathogenicity to mutant A3. Ectopic expression of only thiolase resulted in loss of pathogenicity, suggesting that thiolase overexpression was primarily responsible for the reduced pathogenicity of the A3 isolate. The thiolase gene encoded a functional protein, as shown by assays in which a nontoxic substrate (2, 4 dichlorophenoxybutyric acid) was converted to a toxic product. The use of a translational fusion with a reporter gene showed thiolase expressed in organelles that are most likely peroxisomes.

Background: Stagonospora nodorum blotch (SNB), Fusarium head blight (FHB) and stem rust (SR), caused by the fungi Parastagonospora (synonym Stagonospora) nodorum, Fusarium graminearum and Puccinia graminis, respectively, signiicantly reduce yield and quality of wheat. Three resistance factors, QSng.sfr-3BS, Fhb1 and Sr2, conferring resistance, respectively, to SNB, FHB and SR, each from a unique donor line, were mapped previously to the short arm of wheat chromosome 3B. Based on published reports, our hypothesis was that Sr2 is the most distal, Fhb1 the most proximal and QSng.sfr-3BS is in between Sr2 and Fhb1 on wheat chromosome arm 3BS. Results: To test this hypothesis, 1600 F2 plants from crosses between parental lines Arina, Alsen and Ocoroni86, conferring resistance genes QSng.sfr-3BS, Fhb1 and Sr2, respectively, were genotyped and phenotyped for SNB along with the parental lines. Five closely linked single-nucleotide polymorphism (SNP) markers were used to make the genetic map and determine the gene order. Conclusions: The results indicate that QSng.sfr-3BS is located between the other two resistance genes on chromosome 3BS. Knowing the positional order of these resistance genes will aid in developing a wheat line with all three genes in coupling, which has the potential to provide broad-spectrum resistance preventing grain yield and quality losses. [ABSTRACT FROM AUTHOR]

This review has a personal, plant pathologist's outlook on plant breeding. It touches upon some generalities, among which the "three stages" of plant breeding, participatory plant breeding and biotechnology in plant breeding. It delves deep into modern molecular studies on leaf blotch (anamorph Septoria tritici) and glume blotch (anamorph Septoria nodorum) of wheat. Epidemiological knowledge of the teleomorphs Mycosphaerella graminicola and Stagonospora nodorum has progressed with great strides. Consequences for applied plant breeding slowly become visible.

Stagonospora nodorum blotch (SNB), caused by the fungus Parastagonospora nodorum, is a major disease of wheat (Triticum aestivum). Residue from a previously infected wheat crop can be an important source of initial inoculum, but the effects of infected residue on disease severity and yield have not previously been quantified. Experiments were conducted in Raleigh and Salisbury. North Carolina, in 2012, 2013, and 2014 using the moderately susceptible winter wheat cultivar DG Shirley. In 2014, the highly susceptible cultivar DG 9012 was added to the experiment and the study was conducted at an additional site in Tyner, North Carolina. Four (2012) or six (2013 and 2014) wheat residue treatments were applied in the field in a randomized complete block design with five replicates. Treatments in 2012 were 0, 30, 60, and 90% residue coverage of the soil surface, while 10 and 20% residue treatments were added in 2013 and 2014. Across site-years, disease severity ranged from 0 to 50% and increased nonlinearly (P < 0.05) as residue level increased, with a rapid rise to an upper limit and showing little change in severity above 20 to 30% soil surface coverage. Residue coverage had a significant (P < 0.05) effect on disease severity in all site-years. The effect of residue coverage on yield was only significant (P < 0.05) for DG Shirley at Raleigh and Salisbury in 2012 and for DG 9012 at Salisbury in 2014. Similarly, residue coverage significantly (P < 0.05) affected thousand-kernel weight only of DG 9012 in 2014 at Raleigh and Salisbury. Our results showed that when wheat residue was sparse, small additions to residue density produced greater increases in SNB than when residue was abundant. SNB only led to effects on yield and test weight in the most disease-conducive environments, suggesting that the economic threshold for the disease may be higher than previously assumed and warrants review. [ABSTRACT FROM AUTHOR]

Tan spot and Stagonospora nodorum blotch (SNB), often occurring together, are two economically significant diseases of wheat in the Northern Great Plains of the United States. They are caused by the fungi Pyrenophora tritici-repentis and Parastagonospora nodorum, respectively, both of which produce multiple necrotrophic effectors (NE) to cause disease. In this work, 120 hard red winter wheat (HRWW) cultivars or elite lines, mostly from the United States, were evaluated in the greenhouse for their reactions to the two diseases as well as NE produced by the two pathogens. One P. nodorum isolate (Sn4) and four Pyrenophora tritici-repentis isolates (Pti2, 331-9, DW5, and AR CrossBIO) were used separately in the disease evaluations. NE sensitivity evaluation included ToxA, Ptr ToxB, SnToxl. and SnTox3. The numbers of lines that were rated highly resistant to individual isolates ranged from 11 (9%) to 30 (25%) but only six lines (5%) were highly resistant to all isolates, indicating limited sources of resistance to both diseases in the U.S. adapted HRWW germplasm. Sensitivity to ToxA was identified in 83 (69%) of the lines and significantly correlated with disease caused by Sn4 and Pti2, whereas sensitivity to other NE was present at much lower frequency and had no significant association with disease. As expected, association mapping located ToxA and SnTox3 sensitivity to chromosome arm 5BL and 5BS, respectively. A total of 24 potential quantitative trait loci was identified with -log (P value) > 3.0 on 12 chromosomes, some of which are novel. This work provides valuable information and tools for HRWW production and breeding in the Northern Great Plains. [ABSTRACT FROM AUTHOR]

Stagonospora nodorum blotch (SNB) is a fungal disease of wheat caused by Stagonospora nodorum. The fungus attacks epigeous parts of the plant causing direct damage to kernel with heavy yield losses. This study aimed at evaluating the effect of S. nodorum infection on the electrophoretic protein pattern of durum wheat kernels. The protein pattern of kernels harvested from plants artificially infected in the field and of kernels inoculated in vitro, was defined by SDS-PAGE. Starting from this experimental set, two sub-samples were analysed, the first without further manipulation, the second after an incubation at different time intervals on water agar plates. The results did not disclose any change in the protein pattern of the first sub-sample of kernels whereas a profound alteration of their protein profile occurred after incubation. This result would suggest that albeit the pathogen has the ability to degrade host proteins is unable to express the same ability during field infection. [ABSTRACT FROM AUTHOR]

Background The fungus Stagonospora nodorum is a necrotrophic pathogen of wheat. It causes disease by secreting proteinaceous effectors which interact with proteins encoded by dominant susceptibility genes in the host. The outcome of these interactions results in necrosis, allowing the fungus to thrive on dead plant material. The mechanisms of these effectors though are poorly understood. In this study, we undertake a comprehensive transcriptomics, proteomic and metabolomic approach to understand how a susceptible wheat cultivar responds to exposure to the Stagonospora nodorum effector protein SnTox3. pathogenicity-related proteins and the induction of cell death. Collapse of the photosynthetic machinery was also apparent at the transcriptional and translational level. SnTox3-infiltrated wheat leaves also showed a strong induction of enzymes involved in primary metabolism consistent with increases in hexoses, amino acids and organic acids as determined by primary metabolite profiling. Methionine and homocysteine metabolism was strongly induced upon exposure to SnTox3. Pathogenicity in the presence of homocysteine was inhibited confirming that the compound has a role in plant defence. Consistent with the strong defence responses observed, secondary metabolite profiling revealed the induction of several compounds associated with plant defence, including the phenylpropanoids chlorogenic acid and feruloylquinic acid, and the cyanogenic glucoside dhurrin. Serotonin did not accumulate subsequent to SnTox3 infiltration. Conclusions These data support the theory that the SnTox3 effector protein elicits a host cell death response to facilitate the pathogen's necrotrophic infection cycle. Our data also demonstrate that the mechanism of SnTox3 appears distinct from the previously characterised Stagonospora nodorum effector SnToxA. Collectively, this comprehensive analysis has advanced our understanding of necrotrophic effector biology and highlighted the complexity of effector-triggered susceptibility. [ABSTRACT FROM AUTHOR]

Stagonospora nodorum is a necrotrophic fungal pathogen that causes Stagonospora nodorum blotch (SNB), a yield- and quality-reducing disease of wheat. S. nodorum produces a set of necrotrophic effectors (NEs) that interact with the products of host sensitivity genes to cause cell death and increased susceptibility to disease. The focus of this study was determination of NE sensitivity among 25 winter wheat cultivars, many of them from the southeastern United States, that are susceptible to SNB, as well as the moderately resistant 'NC-Neuse'. Thirty-three isolates of S. nodorum previously collected from seven southeastern U.S. states were cultured for NE production, and the culture filtrates were used in an infiltration bioassay. Control strains of Pichia pastoris that expressed SnToxA, SnToxl, or SnTox3 were also used. All SNB-susceptible cultivars were sensitive to at least one NE, while NC-Neuse was insensitive to all NEs tested. Among the sensitive lines, 32% contained sensitivity gene Tsnl and 64% contained sensitivity gene Snn3. None were sensitive to SnToxl. Additionally, 10 molecular markers for sensitivity genes Tsnl, Snnl, Snn2, and Snn3 were evaluated for diagnostic potential. Only the marker Xfcp623 for Tsnl was diagnostic, and it was in perfect agreement with the results of the infiltration bioassays. The results illuminate which NE sensitivity genes may be of concern in breeding for resistance to SNB in the southeastern United States. [ABSTRACT FROM AUTHOR]

Stagonospora nodorum glume blotch (SNG), caused by the necrotrophic fungus Stagonospora nodorum, is one of the economically important diseases of bread wheat ( Triticum aestivum L.). Resistance to SNG is known to be quantitative and previous studies of a recombinant inbred line (RIL) population identified a major quantitative trait locus (QTL) for resistance to SNG on the short arm of chromosome 3B. To localize this QTL ( QSng.sfr- 3BS) with high resolution, we constructed a genetic map for the QTL target region using information from sequenced flow-sorted chromosomes 3B of the two parental cultivars 'Arina' and 'Forno', the physical map of chromosome 3B of cultivar 'Chinese Spring' and BAC-clone sequences. The mapping population of near-isogenic lines (NIL) was evaluated for SNG resistance in field infection tests. NILs segregated for disease resistance as well as for plant height; additionally, we observed a high environmental influence on the trait. Our analysis detected a strong negative correlation of SNG resistance and plant height. Further analysis of the target region identified two linked loci associated with SNG resistance. One of them was also associated with plant height, revealing an effect of QSng.sfr- 3BS on plant height that was hidden in the RIL population. This result demonstrates an unexpectedly high genetic complexity of resistance controlled by QSng.sfr- 3BS and shows the importance of the study of QTL in mendelized form in NILs. [ABSTRACT FROM AUTHOR]

The relative abundances of DNA of Mycosphaerella graminicola and Phaeosphaeria nodorum in archived wheat samples are closely correlated with UK anthropogenic emissions of oxidized sulphur over the last 160 years. To test whether this could be a causal relationship, possible modes of action of sulphur on the two fungi were examined. Mycelial growth of the two fungi in solutions of sulphurous acid was similar. Sulphurous acid at p H 4 reduced percentage germination of P. nodorum conidia more strongly than M. graminicola conidia. In spray inoculations of wheat cv. Squarehead's Master, Cappelle Desprez and Riband with water or sulphurous acid (p H 4), the ratio of leaves infected by P. nodorum to leaves infected by M. graminicola was increased by factors of 2·5, 2·1 and 0·6, respectively at pH 4. The same three cultivars of wheat were grown in sand and vermiculite and fertilized with nutrient solution containing 2·5 or 0·5 m m sulphate. Both pathogens infected less frequently at 2·5 m m sulphate, by a factor of about 2. The severity of infection by M. graminicola was reduced on all three cultivars by a factor of about 4·5 at 2·5 m m sulphate, but severity of P. nodorum was reduced only by a factor of about 2. Both elevated free sulphate concentrations in soil and sulphite in rainwater could therefore increase the prevalence of P. nodorum relative to M. graminicola, which is consistent with the historical changes in abundance. [ABSTRACT FROM AUTHOR]

This paper considers multiplicative models for the analysis of half-diallels with parents (selfcrosses) and graphical displays based on such models, including biplots. It is shown that models respecting the symmetry of the diallel two-way classification are inherently restrictive because multiplicative terms for specific combining ability (SCA) effects are forced to have the same sign for all self-crosses. The models are illustrated using an example from wheat (Triticum aestivum L). Limitations of other multiplicative models for diallel analysis such as the AMMI and GGE models are also discussed. It is concluded that multiplicative models are useful for the analysis of diallels when the pattern of SCA effects can be well represented by just one or two multiplicative terms. When SCA effects have a more complex structure, however, it may be preferable to resort to classical diallel analysis. [ABSTRACT FROM AUTHOR]

A reverse genetics approach was used to investigate the role of γ-aminobutyric acid metabolism in the wheat pathogenic fungus Stagonospora nodorum. The creation of mutants lacking Sdh1, the gene encoding succinic semialdehyde dehydrogenase, resulted in strains that grew poorly on γ-aminobutyric acid as a nitrogen source. The sdh1 mutants were more susceptible to reactive oxygen stress but were less affected by increased growth temperatures. Pathogenicity assays revealed that the metabolism of γ-aminobutyric acid is required for complete pathogenicity. Growth assays of the wild-type and mutant strains showed that the inclusion of γ-aminobutyric acid as a supplement in minimal media (i.e., not as a nitrogen or carbon source) resulted in restricted growth but increased sporulation. The addition of glutamate, the precursor to GABA, had no effect on either growth or sporulation. The γ-aminobutyric acid effect on sporulation was found to be dose dependent and not restricted to Stagonospora nodorum with a similar effect observed in the dothideomycete Botryosphaeria sp. The positive effect on sporulation was assayed using isomers of γ-aminobutyric acid and other metabolites known to influence asexual development in Stagonospora nodorum but no effect was observed. These data demonstrate that γ-aminobutyric acid plays an important role in Stagonospora nodorum in responding to environmental stresses while also having a positive effect on asexual development. [ABSTRACT FROM AUTHOR]

A reverse genetics approach was used to investigate the role of γ-aminobutyric acid metabolism in the wheat pathogenic fungus Stagonospora nodorum. The creation of mutants lacking Sdh1, the gene encoding succinic semialdehyde dehydrogenase, resulted in strains that grew poorly on γ-aminobutyric acid as a nitrogen source. The sdh1 mutants were more susceptible to reactive oxygen stress but were less affected by increased growth temperatures. Pathogenicity assays revealed that the metabolism of γ-aminobutyric acid is required for complete pathogenicity. Growth assays of the wild-type and mutant strains showed that the inclusion of γ-aminobutyric acid as a supplement in minimal media (i.e., not as a nitrogen or carbon source) resulted in restricted growth but increased sporulation. The addition of glutamate, the precursor to GABA, had no effect on either growth or sporulation. The γ-aminobutyric acid effect on sporulation was found to be dose dependent and not restricted to Stagonospora nodorum with a similar effect observed in the dothideomycete Botryosphaeria sp. The positive effect on sporulation was assayed using isomers of γ-aminobutyric acid and other metabolites known to influence asexual development in Stagonospora nodorum but no effect was observed. These data demonstrate that γ-aminobutyric acid plays an important role in Stagonospora nodorum in responding to environmental stresses while also having a positive effect on asexual development. [ABSTRACT FROM AUTHOR]

Stagonospora nodorum and Pyrenophora tritici-repentis produce the effector Tox A that interacts with the dominant susceptibility gene in wheat, Tsn1. However, the way in which Tox A induces cell death and causes disease is unclear., Here, we performed comprehensive metabolite profiling of Tox A-infiltrated wheat ( Triticum aestivum) to observe the secondary metabolite response to this effector., A strong induction of secondary metabolism subsequent to Sn Tox A infiltration was observed, including the monoamine serotonin. We established a novel role for serotonin as a phytoalexin in wheat and demonstrated that serotonin strongly inhibited sporulation of S. nodorum. Microscopy revealed that serotonin interferes with spore formation and maturation within pycnidial structures of the fungus. Subsequent analysis of S. nodorum exposed to serotonin revealed metabolites changes previously associated with sporulation, including trehalose and alternariol. Furthermore, we identified significantly lower concentrations of serotonin during infection compared with infiltration with Tox A, providing evidence that S. nodorum may suppress plant defence., This is the first study demonstrating induction of plant secondary metabolites in response to a necrotrophic effector that have significant antifungal potential against the pathogen. While it is generally accepted that necrotrophs exploit host cell responses, the current research strengthens the notion that necrotrophs require mechanisms to overcome plant defence to survive initial stages of infection. [ABSTRACT FROM AUTHOR]

This work was done to demonstrate the opportunities provided by application of meta-analysis in plant pathology. It was a case study used to determine the effectiveness of foliar fungicides in minimising yield loss from a complex of yellow spot ( Pyrenophora tritici- repentis) and septoria nodorum blotch ( Stagonospora nodorum, teleomorph: Phaeosphaeria nodorum) (YS-SNB disease complex) on wheat in the northern grain-belt of Western Australia. Forty-seven datasets of experimental results from 14 growing seasons, using 18 varieties sprayed one to three times, predominantly with tebuconazole or propiconazole fungicides, were analysed. Across the datasets, the wheat yield gain from fungicide application was 297 kg ha with a 95 % confidence interval of 11.6 kg ha. Significant yield gains resulted from single or multiple applications of fungicides. Both propiconazole and tebuconazole, increased yield of wheat affected by the YS-SNB disease complex, with yield gain from propiconazole being greater than that from tebuconazole. Yield response varied significantly among crop growing seasons. Meta-analysis was able to aggregate a large number of experimental results and answer important questions related to the variables that influenced those results; in this case the effectiveness of fungicides in minimising yield loss from the YS-SNB disease complex on wheat. It also identified areas where further research needs to be done. It is concluded that meta-analysis has the potential to contribute to similar analyses in other crop disease systems. [ABSTRACT FROM AUTHOR]

In two independent experiments set up in the greenhouse the seedling resistance to Stagonospora nodorum blotch was investigated in 92 varieties, breeding lines and genotypes with a known genetic background. The greatest area under the disease progress curve calculated from lesion type was 37.06, while in the case of the most resistant genotype this value was 0.38. Many of the lines and varieties bred in Martonvásár proved to have excellent resistance in terms of both percentage of infected leaf area and lesion type. Observations indicate that, depending on the aim of the experiment, the efficient selection of breeding lines is possible in the seedling stage either on the basis of the area under the disease progress curve calculated for lesion types, or on the basis of lesion types scored 7, 11 or 14 days after inoculation. [ABSTRACT FROM AUTHOR]

Stagonospora nodorum blotch (SNB) is a significant fungal disease of bread wheat (Triticum aestivum L.) caused by Stagonospora nodorum, in which reduced grain yield is caused when the pathogen infects flag leaves and glumes at the critical time of grain filling. Wheat breeding programs have made limited progress in improving resistance to SNB due to the underlying complexity of the pathogen, host resistance, and their interactions. There has been an increase in knowledge of components of host-pathogen interactions in the past five decades, including pathogen diversity, biological factors contributing toward pathogen infection, environmental conditions favoring disease progression, and the genetics of host resistance. This review captures major outcomes and assesses different approaches and methodologies for improving resistance to SNB. The review concludes by proposing strategies for deploying, selecting, and combining gene and trait combinations in genetic backgrounds and improving methods for evaluation and selection of SNB resistance in wheat breeding. [ABSTRACT FROM AUTHOR]

The Stagonospora nodorum-wheat interaction involves multiple pathogen-produced necrotrophic effectors that interact directly or indirectly with specific host gene products to induce the disease Stagonospora nodorum blotch ( SNB). Here, we used a tetraploid wheat mapping population to identify and characterize a sixth effector-host gene interaction in the wheat- S. nodorum system. Initial characterization of the effector SnTox5 indicated that it is a proteinaceous necrotrophic effector that induces necrosis on host lines harbouring the Snn5 sensitivity gene, which was mapped to the long arm of wheat chromosome 4B. On the basis of ultrafiltration, SnTox5 is probably in the size range 10-30 kDa. Analysis of SNB development in the mapping population indicated that the SnTox5- Snn5 interaction explains 37%-63% of the variation, demonstrating that this interaction plays a significant role in disease development. When the SnTox5- Snn5 and SnToxA- Tsn1 interactions occurred together, the level of SNB was increased significantly. Similar to several other interactions in this system, the SnTox5- Snn5 interaction is light dependent, suggesting that multiple interactions may exploit the same pathways to cause disease. [ABSTRACT FROM AUTHOR]

Abstract: The origin of the fungal wheat pathogen Phaeosphaeria nodorum remains unclear despite earlier intensive global population genetic and phylogeographical studies. We sequenced 1683bp distributed across three loci in 355 globally distributed Phaeosphaeria isolates, including 74 collected in Iran near the center of origin of wheat. We identified nine phylogenetically distinct clades, including two previously unknown species tentatively named P1 and P2 collected in Iran. Coalescent analysis indicates that P1 and P2 are sister species of P. nodorum and the other Phaeosphaeria species identified in our analysis. Two species, P. nodorum and P. avenaria f. sp. tritici 1 (Pat1), comprised ∼85% of the sampled isolates, making them the dominant wheat-infecting pathogens within the species complex. We designed a PCR-RFLP assay to distinguish P. nodorum from Pat1. Approximately 4% of P. nodorum and Pat1 isolates showed evidence of hybridization. Measures of private allelic richness at SSR and sequence loci suggest that the center of origin of P. nodorum coincides with its host in the Fertile Crescent. We hypothesize that the origin of this species complex is also in the Fertile Crescent, with four species out of nine found exclusively in the Iranian collections. [Copyright &y& Elsevier]

Stagonospora nodorum is a major necrotrophic pathogen of wheat that causes the diseases S. nodorum leaf and glume blotch. A series of tools and resources, including functional genomics, a genome sequence, proteomics and metabolomics, host-mapping populations, and a worldwide collection of isolates, have enabled the dissection of pathogenicity mechanisms. Metabolic and signaling genes required for pathogenicity have been defined. Interaction with the host is dominated by interplay of fungal effectors that induce necrosis on wheat lines carrying specific sensitivity loci. As such, the pathogen has emerged as a model for the Pleosporales group of pathogens. [ABSTRACT FROM AUTHOR]

SUMMARY The wheat pathogen Stagonospora nodorum, causal organism of the wheat disease Stagonospora nodorum blotch, has emerged as a model for the Dothideomycetes, a large fungal taxon that includes many important plant pathogens. The initial annotation of the genome assembly included 16 586 nuclear gene models. These gene models were used to design a microarray that has been interrogated with labelled transcripts from six cDNA samples: four from infected wheat plants at time points spanning early infection to sporulation, and two time points taken from growth in artificial media. Positive signals of expression were obtained for 12 281 genes. This represents strong corroborative evidence of the validity of these gene models. Significantly differential expression between the various time points was observed. When infected samples were compared with axenic cultures, 2882 genes were expressed at a higher level in planta and 3630 were expressed more highly in vitro. Similar numbers were differentially expressed between different developmental stages. The earliest time points in planta were particularly enriched in differentially expressed genes. A disproportionate number of the early expressed gene products were predicted to be secreted, but otherwise had no obvious sequence homology to functionally characterized genes. These genes are candidate necrotrophic effectors. We have focused attention on genes for carbohydrate metabolism and the specific biosynthetic pathways active during growth in planta. The analysis points to a very dynamic adjustment of metabolism during infection. Functional analysis of a gene in the coenzyme A biosynthetic pathway showed that the enzyme was dispensable for growth, indicating that a precursor is supplied by the plant. [ABSTRACT FROM AUTHOR]

SUMMARY In this study, proteomics and metabolomics were used to study the wheat response to exposure to the SnToxA effector protein secreted by the fungal pathogen Stagonospora nodorum during infection. Ninety-one different acidic and basic proteins and 101 metabolites were differentially abundant when comparing SnToxA- and control-treated wheat leaves during a 72-h time course. Proteins involved in photosynthesis were observed to increase marginally initially after exposure, before decreasing rapidly and significantly. Proteins and metabolites associated with the detoxification of reactive oxygen species in the chloroplast were also differentially abundant during SnToxA exposure, implying that the disruption of photosynthesis causes the rapid accumulation of chloroplastic reactive oxygen species. Metabolite profiling revealed major metabolic perturbations in central carbon metabolism, evidenced by significant increases in tricarboxylic acid (TCA) cycle intermediates, suggestive of an attempt by the plant to generate ATP and reducing equivalents in response to the collapse of photosynthesis caused by SnToxA. This was supported by the observation that the TCA cycle enzyme malate dehydrogenase was up-regulated in response to SnToxA. The infiltration of SnToxA also resulted in a significant increase in abundance of many pathogenicity-related proteins, even in the absence of the pathogen or other pathogen-associated molecular patterns. This approach highlights the complementary nature of proteomics and metabolomics in studying effector-host interactions, and provides further support for the hypothesis that necrotrophic pathogens, such as S. nodorum, appear to exploit existing host cell death mechanisms to promote pathogen growth and cause disease. [ABSTRACT FROM AUTHOR]

Stagonospora nodorum, the causal agent of wheat glume and leaf blotch can affect all aerial parts causing lens-shaped necrotic lesions surrounded by a yellow-greenish halo. The resistance of wheat to S. nodorum can be non-specific to race and durable, depending of several individual components of partial resistance that can be expressed in adult-plant or seedling. In this study, the infection degree in wheat seedlings of fi ve genotypes with different level of resistance to S. nodorum was estimated by a histopathological analysis considering the damage in tissues and amount of fungal colonization. The behavior of genotypes in seedlings and adult-plant was different. The BAU/TNMU and NG8675/CBRD genotypes showed the highest tissue damage and the largest colonization 24 h after inoculation and were considered the most susceptible. The THB/ MAYA, BATÁN F-96 and REBECA F2000 genotypes showed different resistance degrees, THB/MAYA was the less resistant. Based on different times in which fungal colonization was recorded, structural and chemical responses of resistance to the infection were not observed in any genotype, inferring that resistance in seedlings is carried out in the phase prior to penetration. [ABSTRACT FROM AUTHOR]

Stagonospora nodorum blotch (SNB) caused by Phaeosphaeria nodorum is an important foliar disease of wheat. Efficient breeding for SNB resistance requires detailed knowledge of the pathosystem so that targeted improvements can be made. The objective of this study was to examine the wheat–P. nodorum pathosystem for evidence of host specificity amongst Canadian isolates. To this end, 49 P. nodorum isolates were inoculated on 16 wheat lines in indoor seedling inoculation experiments. Biologically significant host × pathogen interactions were identified, indicating that host specificity exists in the Canadian isolates tested. This observation is consistent with an inverse gene-for-gene pathosystem based upon host-selective toxins. Unweighted Pair Group Method with Arithmetic Mean (UPGMA) analysis of disease reaction data suggested nine virulence clusters; however, additional sub-groupings could be argued based upon the reaction of specific wheat lines to isolates within virulence clusters. The common wheat lines 86ISMN 2137, RL5407 and W7984 had good resistance to the P. nodorum isolates in seedling tests, which will be useful for improvements in common wheat and possibly durum wheat. The development of a SNB differential set is discussed. P. nodorum isolates characterized in this study will be useful for future studies in the wheat–P. nodorum pathosystem. [ABSTRACT FROM AUTHOR]

Background: Monoculture, multi-cropping and wider use of highly resistant cultivars have been proposed as mechanisms to explain the elevated rate of evolution of plant pathogens in agricultural ecosystems. We used a mark-release-recapture experiment with the wheat pathogen Phaeosphaeria nodorum to evaluate the impact of two of these mechanisms on the evolution of a pathogen population. Nine P. nodorum isolates marked with ten microsatellite markers and one minisatellite were released onto five replicated host populations to initiate epidemics of Stagonospora nodorum leaf blotch. The experiment was carried out over two consecutive host growing seasons and two pathogen collections were made during each season. Results: A total of 637 pathogen isolates matching the marked inoculants were recovered from inoculated plots over two years. Genetic diversity in the host populations affected the evolution of the corresponding P. nodorum populations. In the cultivar mixture the relative frequencies of inoculants did not change over the course of the experiment and the pathogen exhibited a low variation in selection coefficients. Conclusions: Our results support the hypothesis that increasing genetic heterogeneity in host populations may retard the rate of evolution in associated pathogen populations. Our experiment also provides indirect evidence of fitness costs associated with host specialization in P. nodorum as indicated by differential selection during the pathogenic and saprophytic phases. [ABSTRACT FROM AUTHOR]

This is the first report of variability in sensitivity of Phaeosphaeria nodorum to the fungicide azoxystrobin, and also reports on sensitivity to propiconazole, prothioconazole and cyprodinil. An in vitro sensitivity test of 42 isolates from five Swedish winter wheat fields, collected in 2003–2005, was performed on malt extract agar amended with six concentrations of each fungicide. Four isolates collected during the early 1990s, before azoxystrobin had been commercially used in agriculture, were used as references. Fragments of DNA from 231 isolates, including the reference isolates, were sequenced for the genes of cytochrome b and CYP51 in search for amino acid substitutions known to cause loss of sensitivity to strobilurins and triazoles, respectively. The majority of the P. nodorum isolates possessed the amino acid substitution G143A, associated with loss of sensitivity in fungi to strobilurins, except in one field where only half of the isolates had the substitution. The EC50 values varied between 0·66 mg L−1 to estimations far above 1000 mg L−1, with an estimated median value of 366 mg L−1. The EC50 values of the reference isolates ranged from 0·02 to 80·72 mg L−1. The P. nodorum population is still sensitive to propiconazole, prothioconazole and cyprodinil, even though some isolates varied in sensitivity to triazoles. Part of the CYP51 gene, a possible target for triazole sensitivity, was sequenced but no nonsynonymous substitutions were found. [ABSTRACT FROM AUTHOR]

Abstract: Stagonospora nodorum is a necrotrophic fungal pathogen that is the causal agent of leaf and glume blotch on wheat. S. nodorum is a polycyclic pathogen, whereby rain-splashed pycnidiospores attach to and colonise wheat tissue and subsequently sporulate again within 2–3weeks. As several cycles of infection are needed for a damaging infection, asexual sporulation is a critical phase of its infection cycle. A non-targeted metabolomics screen for sporulation-associated metabolites identified that trehalose accumulated significantly in concert with asexual sporulation both in vitro and in planta. A reverse-genetics approach was used to investigate the role of trehalose in asexual sporulation. Trehalose biosynthesis was disrupted by deletion of the gene Tps1, encoding a trehalose 6-phosphate synthase, resulting in almost total loss of trehalose during in vitro growth and in planta. In addition, lesion development and pycnidia formation were also significantly reduced in tps1 mutants. Reintroduction of the Tps1 gene restored trehalose biosynthesis, pathogenicity and sporulation to wild-type levels. Microscopic examination of tps1 infected wheat leaves showed that pycnidial formation often halted at an early stage of development. Further examination of the tps1 phenotype revealed that tps1 pycnidiospores exhibited a reduced germination rate while under heat stress, and tps1 mutants had a reduced growth rate while under oxidative stress. This study confirms a link between trehalose biosynthesis and pathogen fitness in S. nodorum. [Copyright &y& Elsevier]

Abstract: The fungus Stagonospora nodorum is the causal agent of stagonospora nodorum blotch (syn. leaf and glume blotch) disease of wheat. The Gna1-encoded Gα protein is an important signal transduction component in the fungus, which is required for full pathogenicity, sporulation and extracellular depolymerase production. In this study, we sought to gain a better understanding of defects associated with the gna1 mutant by using two-dimensional gel electrophoresis to analyse the extracellular proteome for differences to the wildtype. Mass spectrometry analysis of altered abundant protein spots and peptide matching to the Stagonospora nodorum genome database have led to the identification of genes implicated in cell wall degradation, proteolysis, RNA hydrolysis and aromatic compound metabolism. In addition, quantitative RT-PCR has demonstrated that some of the encoding genes showed differential expression throughout host infection. Implications of these proteins and their corresponding genes in fungal virulence are discussed. [Copyright &y& Elsevier]

The article presents a study on the antifungal compound malayamycin. Malayamycin refers to the perhydrofuropyran C-nucleoside isolated from Streptomyces malayasiensis that demonstrates an antifungal activity. It is characterized by its broad-spectrum fungicide, which is a well-known combat crop disease. Examples of fungicides include mancozeb and copper, both prevent infection when apply correctly. Chemical structure of malayamycin is also presented within the study. Additional experimental discussion regarding fungus growth and sequencing is also determined.

Abstract: A non-targeted metabolomics approach was used to identify significant changes in metabolism upon exposure of the wheat pathogen Stagonospora nodorum to 0.5M NaCl. The polyol arabitol, and to a lesser extent glycerol, was found to accumulate in response to the osmotic stress treatment. Amino acid synthesis was strongly down-regulated whilst mannitol levels were unaffected. A reverse genetic approach was undertaken to dissect the role of arabitol metabolism during salt stress. Strains of S. nodorum lacking a gene encoding an l-arabitol dehydrogenase (abd1), a xylitol dehydrogenase (xdh1) and a double-mutant lacking both genes (abd1xdh1) were exposed to salt and the intracellular metabolites analysed. Arabitol levels were significantly up-regulated upon salt stress in the xdh1 strains but were significantly lower than the wild-type. Arabitol was not significantly different in either the abd1 or the abd1xdh1 strains during osmotic stress but the concentration of glycerol was significantly higher indicating a compensatory mechanism in operation. Genome sequence analysis identified a second possible enzyme capable of synthesizing arabitol explaining the basal level of arabitol present in the abd1xdh1 strains. This study identified that arabitol is the primary compatible solute in S. nodorum but in-built levels of redundancy are present allowing the fungus to tolerate osmotic stress. [Copyright &y& Elsevier]

Mating type ratios and SSR marker analysis were used to study the genetic structure of Phaeosphaeria nodorum, the causal agent of glume blotch in wheat. The study was based on leaf collections in five fields located in different regions in Sweden. In total 302 isolates of P. nodorum were obtained from 203 sampling sites (including eight ascospore isolates). Three strong indications of sexual recombination were found: (i) the two mating types were present at a 1:1 ratio; (ii) the genetic structure was diverse, with many unique genotypes, and 69 of the 93 genotypes were only found once; and (iii) random association of alleles indicated that genetic recombination was frequent. However, asexual reproduction could not be excluded since identical genotypes were found within the fields. The fungal population had experienced a demographic bottleneck, as indicated by a low ratio of number of alleles to microsatellite size range ( M-value) of 0·5. [ABSTRACT FROM AUTHOR]

Abstract: The production situation–injury profile paradigm can be used as a framework to assess the harmfulness of multiple-pest complexes in a changing agriculture. A mechanistic simulation model for wheat, WHEATPEST, was developed to incorporate drivers of (i) variable production situations and (ii) their related injury profiles. The model simulates the harmful effects of pathogens, pests, and weeds in a simple, open, generic manner. Simulation drivers were derived from published reports, in particular through a meta-analysis of highly detailed farmers’ field surveys in the United Kingdom and the Netherlands. Preliminary analysis of the model''s performances indicates that WHEATPEST conforms with available published reports in a range of production situations and injury profiles. While improvement on some components of the model are discussed, this work points at the need for the collection of cross-disciplinary, reasonably accurate, and standardised data at a system''s level, and at the usefulness of modelling tools for basic research and policy. [Copyright &y& Elsevier]

Abstract: We compared patterns of mitochondrial restriction fragment length polymorphism (RFLP) diversity with patterns of nuclear RFLP diversity to investigate the effects of selection, gene flow, and sexual reproduction on the population genetic structure and evolutionary history of the wheat pathogen Phaeosphaeria nodorum. A total of 315 fungal isolates from Texas, Oregon, and Switzerland were analyzed using seven nuclear RFLP probes that hybridized to discrete loci and purified mitochondrial DNA that hybridized to the entire mtDNA genome. Forty-two different mitochondrial haplotypes and 298 different nuclear haplotypes were detected. The two most frequent mtDNA haplotypes were present in every population and represented 32% of all isolates. High levels of gene flow, low levels of population subdivision, no evidence for either host specificity or cyto-nuclear disequilibrium were inferred from the analysis of both genomes. The concordance in estimates of these population genetic parameters from both genomes suggests that the two genomes experienced similar degrees of migration, genetic drift and selection. [Copyright &y& Elsevier]

The wheat tan spot fungus (Pyrenophora tritici-repentis) produces a well-characterized host-selective toxin (HST) known as Ptr ToxA, which induces necrosis in genotypes that harbor the Tsn1 gene on chromosome 5B. In previous work, we showed that the Stagonospora nodorum isolate Sn2000 produces at least 2 HSTs (SnTox1 and SnToxA). Sensitivity to SnTox1 is governed by the Snn1 gene on chromosome 1B in wheat. SnToxA is encoded by a gene with a high degree of similarity to the Ptr ToxA gene. Here, we evaluate toxin sensitivity and resistance to S. nodorum blotch (SNB) caused by Sn2000 in a recombinant inbred population that does not segregate for Snn1. Sensitivity to the Sn2000 toxin preparation cosegregated with sensitivity to Ptr ToxA at the Tsn1 locus. Tsn1-disrupted mutants were insensitive to both Ptr ToxA and SnToxA, suggesting that the 2 toxins are functionally similar, because they recognize the same locus in the host to induce necrosis. The locus harboring the tsn1 allele underlies a major quantitative trait locus (QTL) for resistance to SNB caused by Sn2000, and explains 62% of the phenotypic variation, indicating that the toxin is an important virulence factor for this fungus. The Tsn1 locus and several minor QTLs together explained 77% of the phenotypic variation. Therefore, the Tsn1–ToxA interaction in the wheat–S. nodorum pathosystem parallels that of the wheat–tan spot system, and the wheat Tsn1 gene serves as a major determinant for susceptibility to both SNB and tan spot. [ABSTRACT FROM AUTHOR]