Septoria Leaf Blotch In Wheat Research Articles

Bioactivity of Trichoderma harzianum A peptaibols against Zymoseptoria tritici causal agent of septoria leaf blotch of wheat

Bioactivity of Trichoderma harzianum A peptaibols against Zymoseptoria tritici causal agent of septoria leaf blotch of wheat

The Septoria leaf blotch of wheat in Central Kazakhstan: prognosis, evaluation and monitoring with remotely sensed data

Fungal diseases represent a widely spread natural phenomenon affecting many wild and domesticated plants. In nature, all plant species form plant communities of a mixed character, and the spatial pattern of dominant species is usually irregular and spotted. Some species are impregnable to a certain infection, which provides a kind of natural barrier to the infection spreading within the natural community. Under the agricultural environment, when a single plant species may occupy a huge area, the species-specific parasite takes a great advantage to develop focal outbreaks and fast spreading of the infection within the area. The concentration of vulnerable plants and the absence of natural barriers within the agricultural areas provoke outbreaks of fungal diseases that may have highly harmful consequences and result in significant yield losses. One of the purposes of the satellite optical data is an operative, cost-effective diagnostic tool and, in combination with climatic datasets and crop rotation information, a prognosis of fungal disease appearance and severity. This paper describes the system of prognostic and monitoring measures to control the fungal diseases of wheat in Central Kazakhstan, with particular attention to Septoria leaf blotch. The prognostic procedure provides a map of the probability of septoria leaf blotch appearance. The prognosis considers the combination of three main variables: the model of ecological niche for Septoria, the presence of wheat residue, and the vegetation condition index counted for the late spring (May) of the current year. The novel spectral-based approach introduced in this paper is the core component of monitoring activity. The SLBS-equation appears to have high sensitivity to Septoria leaf blotch severity in the middle to late (stages 8–11, accordingly, Feekes growth stages) periods of wheat development. Several other spectral indices (RETA, VSDI, and vegetation indices) may help provide information on the spatial unevenness of wheat crops that may indicate the presence of fungal infection.

The septoria leaf blotch of wheat in Central Kazakhstan: prognosis, evaluation and monitoring with remotely sensed data

Fungal diseases represent a widely spread natural phenomenon affecting many of wild and domesticated plants. In nature, all plant species forms plant communities of a mixed character, and the spatial pattern of dominant species is usually irregular and spotted. Some species are impregnable to a certain infection, which provides a kind of natural barrier to the infection spread within the natural community. Under the agricultural environment, when the single plant species may occupy a huge area, the species-specific parasite takes a great advantage to develop focal outbreaks and fast spreading of the infection within the area. The concentration of vulnerable plants and the absence of natural barriers within the agricultural areas provokes outbreaks of fungal diseases, that may have highly harmful consequences and result in significant yield losses. One of the purposes of the satellite optical data is an operative, cost-effective diagnostic and, in combination with climatic datasets and crop rotation information, a prognosis of fungal disease appearance and severity. In this paper, we describe the system of prognostic and monitoring measures to control the fungal diseases of wheat in Central Kazakhstan with special attention to septoria leaf blotch. The prognostic procedure provides a map of the probability of septoria leaf blotch appearance. The prognosis takes into consideration the combination of three main variables: the model of ecological niche for Septoria, the presence of wheat residue, and Vegetation Condition Index counted for the late spring (May) of the current year. The new spectral index, introduced in this paper, is the core component of monitoring activity. The index is sensitive to septoria leaf blotch severity at middle to late (stages 8-11, accordingly Feekes growth stages) periods of wheat development. Several other indices (RETA, VSDI, vegetation indices) may be of help in providing information on the spatial unevenness of wheat crops that may indicate the presence of fungal infection.

Influence of inorganic salts and chitosan as foliar spray on wheat septoria leaf blotch disease severity under field conditions

Tri sodium polyphosphate, Potassium silicate, sodium silicate, Glutathione and chitosan were evaluated as foliar spray for suppressing septoria leaf blotch (SLB) disease severity were performed under field conditions. Meanwhile, the fungicide Topsin M70 was used as comparison treatment. Field experiments conducted with cultivation of three wheat cultivars in two different fields at two successive dates. Spray treatments with tested chemicals were carried out twice, during the growing season. At all applied treatments, it was observed that the cultivar Giza 171 showed the least SLB disease severity followed by Misr 1 and Gemmiza 12, respectively. The applied silicate salts, potassium and sodium showed superior suppressing effect of SLB severity on all sown wheat cultivars followed by chitosan, tri-sodium polyphosphate, glutathione and the fungicide, in respective order. Furthermore, increase in wheat grains yield showed conversely trend with disease severity. Such promising treatments might be included in integrated management strategies in comparable crops.

A Review of the Interactions between Wheat and Wheat Pathogens: Zymoseptoria tritici, Fusarium spp. and Parastagonospora nodorum.

Zymoseptoria tritici is a hemibiotrophic pathogen which causes Septoria leaf blotch in wheat. The pathogenesis of the disease consists of a biotrophic phase and a necrotrophic phase. The pathogen infects the host plant by suppressing its immune response in the first stage of infection. Hemibiotrophic pathogens of the genus Fusarium cause Fusarium head blight, and the necrotrophic Parastagonospora nodorum is responsible for Septoria nodorum blotch in wheat. Cell wall-degrading enzymes in plants promote infections by necrotrophic and hemibiotrophic pathogens, and trichothecenes, secondary fungal metabolites, facilitate infections caused by fungi of the genus Fusarium. There are no sources of complete resistance to the above pathogens in wheat. Defense mechanisms in wheat are controlled by many genes encoding resistance traits. In the wheat genome, the characteristic features of loci responsible for resistance to pathogenic infections indicate that at least several dozen genes encode resistance to pathogens. The molecular interactions between wheat and Z. tritici, P. nodorum and Fusarium spp. pathogens have been insufficiently investigated. Most studies focus on the mechanisms by which the hemibiotrophic Z. tritici suppresses immune responses in plants and the role of mycotoxins and effector proteins in infections caused by P. nodorum and Fusarium spp. fungi. Trichothecene glycosylation and effector proteins, which are involved in defense responses in wheat, have been described at the molecular level. Recent advances in molecular biology have produced interesting findings which should be further elucidated in studies of molecular interactions between wheat and fungal pathogens. The Clustered Regularly-Interspaced Short Palindromic Repeats/ CRISPR associated (CRISPR/Cas) system can be used to introduce targeted mutations into the wheat genome and confer resistance to selected fungal diseases. Host-induced gene silencing and spray-induced gene silencing are also useful tools for analyzing wheat–pathogens interactions which can be used to develop new strategies for controlling fungal diseases.

Preceding crop and seasonal effects influence fungal, bacterial and nematode diversity in wheat and oilseed rape rhizosphere and soil

Crop rotation can have major influences on yield, which may be the result of changes in the composition of the rhizosphere microbiome. In particular there is evidence that yields of both oilseed rape and wheat can be influenced by the frequency in which they are grown in rotation with each other. In the current study we investigated the effect of preceding crops (either wheat or oilseed rape) on wheat and oilseed rape yield, with associated changes in the rhizosphere and bulk soil communities of fungi, bacteria and nematodes using terminal restriction fragment length polymorphism (TRFLP) of rRNA genes. Yield of wheat and oilseed rape were reduced by 11 and 10% respectively when grown two years consecutively. Rhizosphere populations were significantly different to bulk soil populations for all groups of organisms. Seasonal shifts in the communities were observed in the rhizosphere for all groups. Communities of fungi, bacteria and nematodes were all significantly influenced by the preceding crop in the wheat rhizosphere, while just the nematode population was affected by preceding crop in the oilseed rape rhizosphere. In particular when two consecutive crops of oilseed rape were grown, relative abundance of members of nematodes within the genus Eumonhystera increased markedly. The fungal foliar pathogen Mycosphaerella graminicola, the teleomorph of Zymoseptoria tritici which causes septoria leaf blotch in wheat, was identified in the rhizosphere of wheat and was significantly more abundant in wheat grown after oilseed rape. We conclude that overall, preceding crop had less impact on community composition than season or crop type, but that specific changes in communities at particular plant growth stages may have substantive impacts on crop growth.

Using Epidemiological Principles to Explain Fungicide Resistance Management Tactics: Why do Mixtures Outperform Alternations?

Whether fungicide resistance management is optimized by spraying chemicals with different modes of action as a mixture (i.e., simultaneously) or in alternation (i.e., sequentially) has been studied by experimenters and modelers for decades. However, results have been inconclusive. We use previously parameterized and validated mathematical models of wheat Septoria leaf blotch and grapevine powdery mildew to test which tactic provides better resistance management, using the total yield before resistance causes disease control to become economically ineffective ("lifetime yield") to measure effectiveness. We focus on tactics involving the combination of a low-risk and a high-risk fungicide, and the case in which resistance to the high-risk chemical is complete (i.e., in which there is no partial resistance). Lifetime yield is then optimized by spraying as much low-risk fungicide as is permitted, combined with slightly more high-risk fungicide than needed for acceptable initial disease control, applying these fungicides as a mixture. That mixture rather than alternation gives better performance is invariant to model parameterization and structure, as well as the pathosystem in question. However, if comparison focuses on other metrics, e.g., lifetime yield at full label dose, either mixture or alternation can be optimal. Our work shows how epidemiological principles can explain the evolution of fungicide resistance, and also highlights a theoretical framework to address the question of whether mixture or alternation provides better resistance management. It also demonstrates that precisely how spray tactics are compared must be given careful consideration. [Formula: see text] Copyright © 2018 The Author(s). This is an open access article distributed under the CC BY 4.0 International license .

Study of the Protective Activity of Chitosan Hydrolyzate Against Septoria Leaf Blotch of Wheat and Brown Spot of Tobacco

Chitosan hydrolyzate containing low-molecular-weight chitosan (≤24 kDa) and its oligomers (≤1.2 kDa) has been obtained via chemical depolimerization of high-molecular-weight chitosan by nitric acid. The fractions of the obtained chitosan hydrolyzate have been characterized by high performance gel permeation chromatography and proton magnetic resonance. The test performed on detached leaves of wheat has shown that the hydrolyzate completely inhibits the development of Stagonospora nodorum, a casual agent of the septoria leaf blotch, at a concentration of 200 μg/mL. A similar test with detached tobacco leaves has shown that the hydrolyzate at a concentration of 100 μg/mL also inhibits the development of Alternaria longipes, which causes brown spot of tobacco, by 75%.

A role for random, humidity-dependent epiphytic growth prior to invasion of wheat by Zymoseptoria tritici

Zymoseptoria tritici causes Septoria leaf blotch of wheat. The prevailing paradigm of the Z. tritici-wheat interaction assumes fungal ingress through stomata within 24–48h, followed by days of symptomless infection. This is extrapolated from studies testing the mode of fungal ingress under optimal infection conditions. Here, we explicitly assess the timing of entry, using GFP-tagged Z. tritici. We show that early entry is comparatively rare, and extended epiphytic growth possible. We test the hypotheses that our data diverge from earlier studies due to: i. random ingress of Z. tritici into the leaf, with some early entry events; ii. previous reliance upon fungal stains, combined with poor attachment of Z. tritici to the leaf, leading to increased likelihood of observing internal versus external growth, compared to using GFP; iii. use of exceptionally high humidity to promote entry in previous studies. We combine computer simulation of leaf-surface growth with thousands of in planta observations to demonstrate that while spores germinate rapidly on the leaf, over 95% of fungi remain epiphytic, growing randomly over the leaf for ten days or more. We show that epiphytic fungi are easily detached from leaves by rinsing and that humidity promotes epiphytic growth, increasing infection rates. Together, these results explain why epiphytic growth has been dismissed and early ingress assumed. The prolonged epiphytic phase should inform studies of pathogenicity and virulence mutants, disease control strategies, and interpretation of the observed low in planta growth, metabolic quiescence and evasion of plant defences by Zymoseptoria during symptomless infection.

First report of the Zymoseptoria tritici teleomorph stage causing septoria leaf blotch of wheat in Algeria

Durum wheat ( Triticum durum ) is one of the most important cereal crops in Algeria with an annual production of approximately 1.3 million tonnes in 2015-2016. Septoria leaf blotch caused by Zymoseptoria tritici (syn. Mycosphaerella …

Spatial prediction of wheat septoria leaf blotch (Septoria tritici) disease severity in Central Ethiopia

A number of studies have reported the presence of wheat septoria leaf blotch (Septoria tritici; SLB) disease in Ethiopia. However, the environmental factors associated with SLB disease, and areas under risk of SLB disease, have not been studied. Here, we tested the hypothesis that environmental variables can adequately explain observed SLB disease severity levels in West Shewa, Central Ethiopia. Specifically, we identified 50 environmental variables and assessed their relationships with SLB disease severity. Geographically referenced disease severity data were obtained from the field, and linear regression and Boosted Regression Trees (BRT) modeling approaches were used for developing spatial models. Moderate-resolution imaging spectroradiometer (MODIS) derived vegetation indices and land surface temperature (LST) variables highly influenced SLB model predictions. Soil and topographic variables did not sufficiently explain observed SLB disease severity variation in this study. Our results show that wheat growing areas in Central Ethiopia, including highly productive districts, are at risk of SLB disease. The study demonstrates the integration of field data with modeling approaches such as BRT for predicting the spatial patterns of severity of a pathogenic wheat disease in Central Ethiopia. Our results can aid Ethiopia's wheat disease monitoring efforts, while our methods can be replicated for testing related hypotheses elsewhere.

Selection and characterization of Argentine isolates of Trichoderma harzianum for effective biocontrol of Septoria leaf blotch of wheat.

Species of the genus Trichoderma are economically important as biocontrol agents, serving as a potential alternative to chemical control. The applicability of Trichoderma isolates to different ecozones will depend on the behavior of the strains selected from each zone. The present study was undertaken to isolate biocontrol populations of Trichoderma spp. from the Argentine wheat regions and to select and characterize the best strains of Trichoderma harzianum by means of molecular techniques. A total of 84 out of the 240 strains of Trichoderma were able to reduce the disease severity of the leaf blotch of wheat. Thirty-seven strains were selected for the reduction equal to or greater than 50% of the severity, compared with the control. The percentage values of reduction of the pycnidial coverage ranged between 45 and 80%. The same last strains were confirmed as T. harzianum by polymerase chain reaction amplification of internal transcribed spacers, followed by sequencing. Inter-simple sequence repeat was used to examine the genetic variability among isolates. This resulted in a total of 132 bands. Further numerical analysis revealed 19 haplotypes, grouped in three clusters (I, II, III). Shared strains, with different geographical origins and isolated in different years, were observed within each cluster. The origin of the isolates and the genetic group were partially related. All isolates from Paraná were in cluster I, all isolates from Lobería were in cluster II, and all isolates from Pergamino and Santa Fe were in cluster III. Our results suggest that the 37 native strains of T. harzianum are important in biocontrol programs and could be advantageous for the preparation of biopesticides adapted to the agroecological conditions of wheat culture.

A high-resolution melt (HRM) assay to characterize CYP51 haplotypes of the wheat pathogen Mycosphaerella graminicola

Mycosphaerella graminicola is a fungal pathogen that causes Septoria leaf blotch in wheat, resulting in substantial crop losses in years conducive to infection. The disease is mainly controlled by the use of sterol 14α-demethylation inhibitor (DMI) fungicides, which target the CYP51 protein involved in sterol biosynthesis in the fungus. The field efficacy of these chemicals has however been declining in the latest years, mainly due to single nucleotide polymorphisms (SNPs) in the target gene, leading to reduced binding of the resulting protein to these fungicides. Therefore, DMIs are nowadays used in mixes and alternate spraying schemes, together with new generation compounds belonging to the succinate dehydrogenase class of chemicals. To allow knowledgeable and sustainable control of the disease, monitoring of the field populations is paramount. Molecular characterization of field strains is currently mostly based on amplification of the CYP51 gene followed by classic Sanger sequencing. In this study, we optimized a simpler and more cost-effective genotyping method based on the high-resolution melting (HRM) technology. We demonstrate that this protocol can be used to detect and genotype the most common and resistance-determining SNPs in the M. graminicola CYP51 gene.

Transcriptional Reprogramming of Wheat and the Hemibiotrophic Pathogen Septoria tritici during Two Phases of the Compatible Interaction

The disease septoria leaf blotch of wheat, caused by fungal pathogen Septoria tritici, is of worldwide concern. The fungus exhibits a hemibiotrophic lifestyle, with a long symptomless, biotrophic phase followed by a sudden transition to necrotrophy associated with host necrosis. Little is known about the systematic interaction between fungal pathogenicity and host responses at specific growth stages and the factors triggering the transition. In order to gain some insights into global transcriptome alterations in both host and pathogen during the two phases of the compatible interaction, disease transition was monitored using pathogenesis-related gene markers and H2O2 signature prior to RNA-Seq. Transcriptome analysis revealed that the slow symptomless growth was accompanied by minor metabolic responses and slightly suppressed defences in the host, whereas necrotrophic growth was associated with enhanced host responses involving energy metabolism, transport, signalling, defence and oxidative stress as well as a decrease in photosynthesis. The fungus expresses distinct classes of stage-specific genes encoding potential effectors, probably first suppressing plant defence responses/facilitating the symptomless growth and later triggering life style transition and inducing host necrosis/facilitating the necrotrophic growth. Transport, signalling, anti-oxidative stress mechanisms and apoplastic nutrient acquisition play important roles in the entire infection process of S. tritici. Our findings uncover systematic S. tritici-induced expression profiles of wheat related to specific fungal infection strategies and provide a transcriptome resource for studying both hosts and pathogens in plant-Dothideomycete interactions.

Real-time PCR to study the effect of timing and persistence of fungicide application and wheat varietal resistance onMycosphaerella graminicolaand its sterol 14α-demethylation-inhibitor-resistant genotypes

Sterol 14α-demethylase inhibitors (DMIs) have been widely used for more than 20 years against wheat Septoria leaf blotch. However, resistance towards DMIs has increased in recent years. The objective of this study was to evaluate the effect of fungicide timing and persistence and wheat resistance varietal on Mycosphaerella graminicola and its DMI-resistant genotypes. Using qPCR, M. graminicola was detected 2 weeks later in the resistant cultivar than in the susceptible cultivar. A high proportion of DMI-moderate resistant genotypes (≥94%) was found in all samples, with an average of 74.2, 0.1 and 19.7% for R6, R7- and R7+ genotypes, respectively. Distribution of DMI-resistant genotypes was neither affected by different wheat cultivars nor by analysis dates. Electron microscopy coupled with qPCR analysis showed that the DMI fungicide prothioconazole had a significant inhibitive effect against spore germination and post-germination. However, the preventive treatment was the most effective, but it was affected strongly by fungicide persistence. Preventive fungicide applications are more effective against Septoria leaf blotch than the curative treatments, so persistence and wheat varietal resistance should be taken into account in the management of this disease. It would seem that none of the studied factors affect the frequency of DMI-resistant genotypes.

Risk assessment studies on succinate dehydrogenase inhibitors, the new weapons in the battle to control Septoria leaf blotch in wheat

Chemical control of Septoria leaf blotch, caused by Mycosphaerella graminicola, is essential to ensure wheat yield and food security in most European countries. Mycosphaerella graminicola has developed resistance to several classes of fungicide and, with the efficacy of azoles gradually declining over time, new modes of action and/or improvements in host varietal resistance are urgently needed to ensure future sustainable disease control. Several new-generation carboxamide fungicides with broad-spectrum activity have recently been introduced into the cereal market. Carboxamides inhibit succinate dehydrogenase (Sdh) of the mitochondrial respiratory chain (complex II) but, because of their single-site specificity, these fungicides may be prone to resistance development. The objective of this study was to assess the risk of resistance development to different Sdh inhibitor (SDHI) fungicides in M. graminicola. UV mutagenesis was conducted to obtain a library of carboxin-resistant mutants. A range of SDHI resistance-conferring mutations was found in Sdh subunits B, C and D. Pathogenicity studies with a range of Sdh variants did not detect any fitness costs associated with these mutations. Most of the amino acid residues identified (e.g. B-S221P/T, B-H267F/L/N/Y, B-I269V and D-D129E/G/T) are directly involved in forming the cavity in which SDHI fungicides bind. Docking studies of SDHI fungicides in structural models of wild-type and mutated Sdh complexes also indicated which residues were important for the binding of different SDHI fungicides and showed a different binding for fluopyram. The predictive power of the model was also shown. Further diagnostic development, enabling the detection of resistant alleles at low frequencies, and cross-resistance studies will aid the implementation of anti-resistance strategies to prolong the cost-effectiveness and lifetime of SDHI fungicides.

A novel substitution I381V in the sterol 14alpha-demethylase (CYP51) of Mycosphaerella graminicola is differentially selected by azole fungicides.

SUMMARY The recent reduction in the efficacy of azole fungicides in controlling Septoria leaf blotch of wheat, caused by Mycosphaerella graminicola, has prompted concerns over possible development of resistance, particularly in light of the recent emergence of widespread resistance to quinone outside inhibitors (QoIs). We have recently implicated alterations in the target-encoding sterol 14alpha-demethylase protein (CYP51), and over-expression of genes encoding efflux pumps, in reducing sensitivity to the azole class of sterol demethylation inhibitors (DMIs) in M. graminicola. Here we report on the prevalence and selection of two CYP51 alterations, substitution I381V and deletion of codons 459 and 460 (DeltaY459/G460), in populations of M. graminicola. Neither alteration has previously been identified in human or plant pathogenic fungi resistant to azoles. The presence of DeltaY459/G460 showed a continuous distribution of EC(50) values across isolates with either I381 or V381, and had no measurable effect on azole sensitivity. Data linking fungicide sensitivity with the presence of I381V in M. graminicola show for the first time that a particular CYP51 alteration is differentially selected by different azoles in field populations of a plant pathogen. Substitution I381V although not an absolute requirement for reduced azole sensitivity, is selected by tebuconazole and difenoconazole treatment, suggesting an adaptive advantage in the presence of these two compounds. Prochloraz treatments appeared to select negatively for I381V, whereas other azole treatments did not or only weakly impacted on the prevalence of this substitution. These findings suggest treatments with different members of the azole class of fungicides could offer a resistance management strategy.

Physiologic variation in Mycosphaerella graminicola from western Canada

Mycosphaerella graminicola (anamorph: Septoria tritici), the causal agent of septoria leaf blotch of wheat, has increased in prevalence in the prairie region of Canada in recent years. It is important to know if variation in virulence occurs in the local population as was reported in other countries. Seventy-four pathogen isolates were collected from hexaploid wheat fields in Manitoba and Saskatchewan and tested for virulence on a set of differential genotypes, including four hexaploid and two tetraploid wheat genotypes. Seedlings were inoculated with a conidial suspension and assessed 14 and 21 days later for the presence of necrosis, chlorosis, and pycnidia to identify virulence patterns. Reactions of the differential host genotypes to different isolates ranged from susceptible to near-immune. Adult-plant reactions were similar to seedling reactions for all genotypes used in this study. A difference in virulence patterns on one genotype suggested that physiological specialization occurs and that two virulence groups are present in the pathogen population from western Canada, based on the set of wheat genotypes used in this study

Position of inoculum in the canopy affects the risk of septoria tritici blotch epidemics in winter wheat

The spread of septoria leaf blotch in wheat caused by Mycosphaerella graminicola is widely reported to depend on the occurrence of splashy rainfall. Previous studies have also implicated an important effect of canopy architecture on the risk of disease spread. This is because architecture affects the proximity of the yield‐forming leaves to inoculum present on older diseased leaves within the crop. This study demonstrated that infection of the final three leaves of winter wheat could occur in the absence of splashy rainfall. For cvs Riband and Longbow, the final two leaf layers emerged at a position ≈ 8 cm below older leaves containing sporulating lesions. Under these conditions, infection of new leaves occurred in treatments that simulated dew and nonsplashy rainfall. These treatments resulted in disease incidences of 10–40% above the untreated control on the final two leaf layers. Within a season, the distance between diseased and healthy leaves during the period of stem extension varied substantially across a range of 30 cultivars. While the magnitude of these differences was not the same across seasons, the relative differences between cultivars were generally consistent, suggesting a strong genotype influence on lesion proximity. This study shows how knowledge of the distribution of lesion proximity within a crop can be used to estimate the risk of inoculum transfer for a given maximum splash height. A rapid crop‐monitoring method for estimating the distribution of lesion proximity was developed and tested. Lesion proximity was manipulated by plant growth regulator (PGR) treatments. Significant increases in disease, between 14 and 62%, were observed on the upper canopy leaves of plants treated with PGR. The largest differences were observed in treatments where lesion proximity was most affected.

Isolation and Characterization of the Mating-Type Idiomorphs from the Wheat Septoria Leaf Blotch Fungus Mycosphaerella graminicola

Both mating-type loci from the wheat septoria leaf blotch pathogen Mycosphaerella graminicola have been cloned and sequenced. The MAT1-2 gene was identified by screening a genomic library from the MAT1-2 isolate IPO94269 with a heterologous probe from Tapesia yallundae. The MAT1-2 idiomorph is 2772 bp and contains a single gene encoding a putative high-mobility-group protein of 394 amino acids. The opposite idiomorph was obtained from isolate IPO323, which has the complementary mating type, by long-range PCR using primers derived from sequences flanking the MAT1-2 idiomorph. The MAT1-1 locus is 2839 bp in size and contains a single open reading frame encoding a putative α1-domain protein of 297 amino acids. Within the nonidiomorphic sequences, homology was found with palI, encoding a membrane receptor from Aspergillus nidulans, and a gene encoding a putative component of the anaphase-promoting complex from Schizosaccharomyces pombe and a DNA-(apurinic or apyrimidinic) lyase from S. pombe. For each of the MAT genes specific primers were designed and tested on an F1 mapping population that was generated from a cross between IPO323 and IPO94269. An absolute correlation was found between the amplified allele-specific fragments and the mating type as determined by backcrosses of each F1 progeny isolate to the parental isolates. The primers were also used to screen a collection of field isolates in a multiplex PCR. An equal distribution of MAT1-1 and MAT1-2 alleles was found for most geographic origins examined.