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Cochliobolus lunatus (anamorph: Curvularia lunata) is a major pathogenic fungus that causes the Curvularia leaf spot of maize. ClMAT1-1-1 and ClMAT1-2-1, the C. lunatus orthologs of C. heterostrophus ChMAT1-1-1 and ChMAT1-2-1, were investigated in the present study to uncover their functions in C. lunatus. Southern blot analysis showed that these mating-type MAT genes exist in the C. lunatus genome as a single copy. ClMAT1-1-1 and ClMAT1-2-1 were knocked out and complemented to generate ΔClmat1-1-1 and ΔClmat1-2-1 and ΔClmat1-1-1-C and ΔClmat1-2-1-C, respectively. The mutant strains had defective sexual development and failed to produce pseudothecia. There were no significant differences in growth rate or conidia production between the mutant and wild-type strains. However, the aerial mycelia and mycelial dry weight of ΔClmat1-1-1 and ΔClmat1-2-1 were lower than those of wild type, suggesting that MAT genes affect asexual development. ClMAT genes were involved in the responses to cell wall integrity and osmotic adaptation. ΔClmat1-2-1 had a lower conidial germination rate than the wild-type strain CX-3. The virulence of ΔClmat1-2-1 and ΔClmat1-1-1 was also reduced compared with the wild-type. Complementary strains could restore all the phenotypes.

Rice false smut (RFS), caused by Villosiclava virens, is an important fungal disease in panicles of rice. V. virens is a heterothallic ascomycete controlled by two opposite idiomorphs, MAT1-1 and MAT1-2. Previous study showed that sexual reproduction of V. virens plays an important role in the epidemic of RFS. In this study, we developed a loop-mediated isothermal amplification (LAMP) assay to detect the mating type of V. virens easily and rapidly by using specific primers based on the mating type genes MAT1-1-2 and MAT1-2-1, respectively. The LAMP assay used only a water/dry bath and could recognize the mating type of V. virens in just 45 min. The LAMP assay was so sensitive that it could detect small amounts of V. virens genomic DNA (as low as 2.0 pg of MAT1-1 and 200.0 pg of MAT1-2) and was 10 times more sensitive than PCR. In addition, we demonstrated the application of mating type via LAMP assay by assessing the genomic DNA of V. virens isolated from rice fields. The high efficiency and specificity of this LAMP assay suggest that it can be used as a rapid testing tool in mating type recognition of V. virens isolates in the field.

Venturia effusa (syn. Fusicladium effusum), causal agent of pecan scab, is the most prevalent pathogen of pecan (Carya illinoinensis), causing severe yield losses in the southeastern United States. V. effusa is currently known only by its asexual (conidial) stage. However, the degree and distribution of genetic diversity observed within and among populations of V. effusa are typical of a sexually reproducing fungal pathogen, and comparable with other dothideomycetes with a known sexual stage, including the closely related apple scab pathogen, V. inaequalis. Using the mating type (MAT) idiomorphs from V. inaequalis, we identified a single MAT gene, MAT1-1-1, in a draft genome of V. effusa. The MAT1-1-1 locus is flanked by two conserved genes encoding a DNA lyase (APN2) and a hypothetical protein. The MAT locus spanning the flanking genes was amplified and sequenced from a subset of 14 isolates, of which 7 contained MAT1-1-1 and the remaining samples contained MAT1-2-1. A multiplex polymerase chain reaction screen was developed to amplify MAT1-1-1, MAT1-2-1, and a conserved reference gene encoding β-tubulin, and used to screen 784 monoconidial isolates of V. effusa collected from 11 populations of pecan across the southeastern United States. A hierarchical sampling protocol representing region, orchard, and tree allowed for analysis of MAT structure at different spatial scales. Analysis of this collection revealed the frequency of the MAT idiomorphs is in a 1:1 equilibrium of MAT1-1:MAT1-2. The apparent equilibrium of the MAT idiomorphs provides impetus for a renewed effort to search for the sexual stage of V. effusa. [Formula: see text] Copyright © 2018 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Citrus black spot (CBS) is a disease caused by the ascomycetous fungus Phyllosticta (formerly Guignardia) citricarpa (Botryosphaeriales, Pezizomycotina) currently present in citrus groves in five counties in southwest Florida. Within Florida, P. citricarpa shows limited reproduction via asexual sporulation due to the presence of only one (MAT1-2-1) of the two required mating types for sexual reproduction. Here, we present two novel PCR assays standardized in quantitative PCR (qPCR) and digital PCR (dPCR) platforms to distinguish both mating types (MAT1-1-1 and MAT1-2-1) of P. citricarpa to monitor for the potential introduction of the MAT1-1-1 mating type into Florida and a novel protocol for DNA extraction from asymptomatic leaves. During citrus harvesting season 2018 − 19, fruit lesions as well as asymptomatic leaves adjacent to symptomatic fruit and asymptomatic trees in CBS-infected groves were surveyed for P. citricarpa presence and mating types. Results support the presence of only the MAT1-2-1 mating type in Florida, after surveying more than 1,145 citrus fruit lesions. We also confirmed the limited dispersal ability of the asexual state of P. citricarpa in Florida in 10 groves using the enhanced capabilities of the dPCR platform in the detection of P. citricarpa directly from asymptomatic leaves with low pathogen inoculum. [Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Sporisorium scitamineum is the causative agent of smut disease in sugarcane. The tricky life cycle of S. scitamineum consists of three distinct growth stages: diploid teliospores, haploid sporidia and dikaryotic mycelia. Compatible haploid sporidia representing opposite mating types (MAT-1 and MAT-2) of the fungus fuse to form infective dikaryotic mycelia in the host tissues, leading to the development of a characteristic whip shaped sorus. In this study, the transition of distinct stages of in vitro life cycle and in planta developmental stages of S. scitamineum are presented by generating stable GFP transformants of S. scitamineum. Haploid sporidia were isolated from the teliospores of Ss97009, and the opposite mating types (MAT-1 and MAT-2) were identified by random mating assay and mating type-specific PCR. Both haploid sporidia were individually transformed with pNIIST plasmid, harboring an enhanced green fluorescent protein (eGFP) gene and hygromycin gene by a modified protoplast-based PEG-mediated transformation method. Thereafter, the distinct in vitro developmental stages including fusion of haploid sporidia and formation of dikaryotic mycelia expressing GFP were demonstrated. To visualize in planta colonization, transformed haploids (MAT-1gfp and MAT-2gfp) were fused and inoculated onto the smut susceptible sugarcane cultivar, Co 97009 and examined microscopically at different stages of colonization. GFP fluorescence-based analysis presented an extensive fungal colonization of the bud surface as well as inter- and intracellular colonization of the transformed S. scitamineum in sugarcane tissues during initial stages of disease development. Noticeably, the GFP-tagged S. scitamineum led to the emergence of smut whips, which established their pathogenicity, and demonstrated initial colonization, active sporogenesis and teliospore maturation stages. Overall, for the first time, an efficient protoplast-based transformation method was employed to depict clear-cut developmental stages in vitro and in planta using GFP-tagged strains for better understanding of S. scitamineum life cycle development.

Sporothrix schenckii and allied species are thermodimorphic fungi widely distributed in nature which causes human and animal sporotrichosis, the most common subcutaneous mycosis globally. Sporotrichosis is acquired after a traumatic inoculation of soil or plant material contaminated with Sporothrix propagules or through bites and scratches from diseased cats. In Ascomycota, the master regulators of sex are MAT genes that lie in a single mating-type locus, in Sporothrix these are determined by two nonhomologous alleles, MAT1-1 and MAT1-2. We assessed the whole-genome sequences of medically relevant Sporothrix to develop a single-tube duplex PCR assay to screen S. brasiliensis, S. schenckii, S. globosa, and S. luriei idiomorphs (MAT1-1 or MAT1-2) and understand the distribution and incidence of mating-type strains from natural populations. Using our duplex PCR assay, a 673 bp amplicon (α-box protein) was consistently amplified from all MAT1-1 isolates, while a 291 bp fragment was only amplified from the isolates harboring MAT1-2 (HMG box). Molecular evidence suggests heterothallism (self-sterility) as the unique mating strategy among the species evaluated. The mating-type identity of 93 isolates revealed a nearly equal distribution (1:1 ratio) of mating type alleles within species but deviating between different outbreak areas. Remarkably, for S. brasiliensis in Rio de Janeiro, we report an overwhelming occurrence of MAT1-2 (1:13 ratio; χ2 = 10.286, P = 0.0013) opposing the high prevalence MAT1-1 in the Rio Grande do Sul (10:1 ratio; χ2 = 7.364, P = 0.0067). Therefore, the population structure of Sporothrix species refers from paucity to regular cycles of sexual recombination in most of the studied regions. Our PCR-based mating-type diagnostic assay is proposed here as an important marker to track the geographical expansion during the long-lasting outbreak of cat-transmitted sporotrichosis driven by S. brasiliensis.

Paramecium spp. is types of free-living protists that live in freshwater environments. They are ciliates with high motility and phagocytosis and have been used to analyze cell motility and as a host model for pathogens. Besides such biological characteristics, apart from the usual morphological and genetic classification of species, the existence of taxonomies (such as syngens) and mating types related to Paramecium’s unique reproduction is known. In this study, we attempted to develop a simple method to identify Paramecium strains, which are difficult to distinguish morphologically, using random amplified polymorphic DNA (RAPD) analysis. Consequently, we can observe strain-specific band patterns. We also confirm that the presence of endosymbiotic Chlorella cells affects the band pattern of P. bursaria. Furthermore, the results of the RAPD analysis using several P. caudatum strains with different syngens show that it is possible to detect a band specific to a certain syngen. By improving the reaction conditions and random primers, based on the results of this study, RAPD analysis can be applied to the identification of Paramecium strains and their syngen confirmation tests.

Citrus black spot (CBS), caused by Phyllosticta citricarpa, is a disease that affects citrus worldwide. In different regions of the world where both mating types occur, reports differ as to whether asexually produced pycnidiospores play an important role in the epidemiology of CBS and fruit infections. Therefore, we investigated the potential role of pycnidiospores in two lemon orchards in South Africa by using microsatellite-based analysis of fruit populations over time (two seasons) and space (distance). The two orchards were situated in the semiarid North West province (NW) and subtropical Mpumalanga province (MP). Each population contained both mating types in 1:1 ratios, and linkage disequilibrium analysis indicated a random mating population. A total of 109 and 94 multilocus genotypes (MLGs) were detected across the two seasons in the NW and MP orchards, respectively. Psex analyses indicated that most MLGs probably resulted from sexual reproduction, but there were six predominant MLGs in each orchard that were probably replicated via asexual reproduction. Each of the predominant MLGs was monomorphic for mating type. In the NW, five predominant and widespread MLGs caused 46 and 44% of the fruit infections in the two seasons, whereas in MP, three MLGs caused 34 and 48% of the infections. Asexual reproduction in both orchards was supported by low MLG evenness values in all populations. In both orchards, distance was not a reliable predictor of population genetic substructuring or season. Populations of P. citricarpa in the MP and NW orchards were significantly genetically differentiated from each other.

A total of 707 isolates of Botrytis were collected from plastic tunnel-grown strawberry and tomato in the Hubei province of China. They were identified based on the specific molecular markers. Diversity of the B. cinerea (Bc) isolates was evaluated by typing the transposable elements (Boty, Flipper) and the mating types (MAT1-1, MAT1-2), as well as by determining virulence on tobacco (Nicotiana benthamiana) and fenhexamid sensitivity in agar medium. The results showed that 706 isolates (99.9%) were Bc and 1 isolate (0.1%) was B. pseudocinerea. The Bc isolates (n = 706) were classified into four transposable element types, Vacuma (3.1%), Boty (9.6%), Flipper (18.4%), and Transposa (68.8%). The strawberry and tomato subpopulations of Bc had significantly different (P < 0.05) compositions of the four transposable element types. The overall ratio of MAT1-1 to MAT1-2 deviated from 1:1 (n = 706; P = 0.0002), and MAT1-2 (56.9%) predominated over MAT1-1 (43.1%). In 7 of 12 geographic subpopulations, the ratio of MAT1-1 to MAT1-2 matched 1:1; however, in the remaining five geographic subpopulations, the ratio of MAT1-1 to MAT1-2 did not match 1:1. Results of the biological characterizations showed that most Bc isolates were highly sensitive or sensitive to fenhexamid, and the majority of Bc isolates were highly virulent or virulent on tobacco. Moreover, the relationship between genetic diversity and biological characteristics was analyzed. The results achieved during this study are helpful for understanding of the populations of B. cinerea.

Mating type diversity has been recently studied in Diaporthe/Phomopsis isolates from different hosts and geographical origin. Two idiomorphic mating genes, MAT1-1-1 and MAT1-2-1, each related to different mating types, have been used for developing PCR markers for mating type determination. The limited number of Phomopsis viticola isolates from grapevine has been identified as heterothallic, carrying one of these mating genes. Mating types of P. viticola have not been studied in a broad sense at the population level, in any countries, including Turkey. The objective of this study is to determine mating types of P. viticola isolates collected from the major grape-producing areas of the Aegean Region. A total of 150 P. viticola isolates obtained from grapevine shoots were subjected to PCR assay with the two pairs of specific primers for mating type determination. All of the isolates produced a single amplicon approximately 229 bp in size, which is known to belong to the MAT1-2-1 gene. No amplicon product of mating type 1 was found. With this background, sexual reproduction of P. viticola is unlikely in our area of study. It can be assumed that P. viticola reproduces asexually and its population is mainly clonal in western Turkey.

In recent years, there has been an apparent Europe-wide emerging presence of brown spot needle blight, a disease of pine species caused by the fungus Lecanosticta acicola. In this study, we report ...

Powdery mildew caused by Erysiphe necator, is one of the most economically damaging diseases of grapevine throughout the world. This study aimed to identify the mating types of this pathogen in Syria using molecular markers. Three DNA extraction methods: Promega Kit, phenol chloroform isoamyl alcohol (PCI) and SDS were compared. The amount and quality of DNAs obtained by the SDS method were suitable for PCR amplification and other molecular assays. PCR amplification using specific primers (Uncin 144 and Uncin 511) was performed and the expected amplicon of 300-400 bp was obtained from 29 isolates of E. necator collected from different geographical locations and from different grapevine cultivars. Results obtained showed that E. necator may cause atypical symptoms similar to those of downy mildew, in some grapevine cultivars according to the environmental conditions and training system used. Isolates of E. necator were classified into four groups according to geographical locations and grapevine cultivar, based on the observed variation in banding pattern with E07 primer and the constructed phenogram by using UPGMA. RAPD analysis of 39 E. necator isolates using the primer E07 showed the presence of two mating types, and the frequency of each varied depending on the geographical location, with a predominance of the - mating type which did not produce a 1000 bp band using the PCR primer E07. Keywords: Powdery mildew, Erysiphe necator, RAPD, mating type, DNA extraction.

The fungal pathogen, Calonectria pauciramosa, has caused serious diseases of many important plants worldwide. Understanding the genetic diversity and mating type distribution of this pathogen provides an essential step toward the development of disease control measures. In this study, we designed 15 polymorphic microsatellite markers by using genome sequences of two Ca. pauciramosa isolates having opposite mating type and from different countries. These markers were used to determine the genetic diversity of 145 isolates representing 13 different hosts (12 plant hosts residing in 12 genera, and soil) from 10 countries. In addition, mating type genes were amplified to investigate the reproduction mode of the pathogens in these populations by using mating type primers designed for Calonectria spp. Results revealed that a single dominant genotype, isolated from 11 plant genera residing in eight families, was present in seven countries across five continents. Only mating type MAT1-1 or MAT1-2 was amplified in each of the isolates, confirming that Ca. pauciramosa is heterothallic. Both mating types were detected in isolates from Eucalyptus in South Africa and Uruguay. The MAT1-2 phenotype was widely distributed in isolates from 12 different hosts (11 plant hosts and soil) collected in 10 countries. Overall, the results suggest that there has been substantial global movement of Ca. pauciramosa and that this has shaped its current population structure.

While the process of meiosis is highly conserved across eukaryotes, the sexual systems that govern life cycle phase transitions are surprisingly labile. Switches between sexual systems have profound evolutionary and ecological consequences, in particular for plants, but our understanding of the fundamental mechanisms and ultimate causes underlying these transitions is still surprisingly incomplete. We explore here the idea that brown and green algae may be interesting comparative models that can increase our understanding of relevant processes in plant reproductive biology, from evolution of gamete dimorphism, gametogenesis, sex determination and transitions in sex-determining systems.

Many biological processes require cell fusion, therefore defects in cell fusion result in many diseases. The unicellular green alga Chlamydomonas reinhardtii is an excellent model organism to study cell-cell fusion. The objective of this study was to identify genes that are involved in C. reinhardtii mating type minus (MT_) gamete fusion. A forward genetics approach was taken in our work. We created several MT_ fusion defective mutants using DNA insertional mutagenesis. These mutants were normal in the early stages of mating; they agglutinated with mating type plus (MT+), removed their walls, adhered to their mating partner through their mating structures, but the cells did not fuse, indicating that the DNA insertional mutants were defective in the latest stages of fusion. The number of insertions was confirmed by Southern blots. Mutant J1 had one insertion and the flanking genomic DNA was cloned by TAIL-PCR and RESDA-PCR. The insertion is in a gene predicted to be involved in 5-deoxystrigol biosynthesis.

Phyllosticta citriasiana is the causal agent of citrus tan spot, an important pomelo disease in Asia. At present, there is little or no information on the epidemiology or population structure of P. citriasiana. By using simple sequence repeat markers, we analyzed 94 isolates from three pomelo production regions in southern and southeastern China. The analyses showed high genetic diversity in each of the three geographic populations. A STRUCTURE analysis revealed two genetic clusters among the 94 isolates; one geographic population was dominated by genotypes in one cluster, and the other two geographic populations were dominated by genotypes of the second cluster. P. citriasiana has a heterothallic mating system with two idiomorphs, MAT1-1 and MAT1-2. Analyses using mating type–specific primers revealed that both mating types were present in all three geographic populations, and in all three populations the mating type ratios were in equilibrium. Although the sexual stage of the fungus has not been discovered yet, analyses of allelic associations indicated evidence for sexual and asexual reproduction within and between populations. Despite the observed genetic differentiation between the three geographic populations, evidence for long-distance gene flow was found.

Haploid cells of the budding yeast Saccharomyces cerevisiae communicate using secreted pheromones and mate to form diploid zygotes. Mating is monogamous, resulting in the fusion of precisely one cell of each mating type. Monogamous mating in crowded conditions, where cells have access to more than one potential partner, raises the question of how multiple-mating outcomes are prevented. Here we identify mutants capable of mating with multiple partners, revealing the mechanisms that ensure monogamous mating. Before fusion, cells develop polarity foci oriented toward potential partners. Competition between these polarity foci within each cell leads to disassembly of all but one focus, thus favoring a single fusion event. Fusion promotes the formation of heterodimeric complexes between subunits that are uniquely expressed in each mating type. One complex shuts off haploid-specific gene expression, and the other shuts off the ability to respond to pheromone. Zygotes able to form either complex remain monogamous, but zygotes lacking both can re-mate.

AIMS To characterize the geo-distinct isolates of Ustilaginoidea virens for morpho-molecular and mating-type locus diversity. METHODS AND RESULTS Sixty-one isolates of U. virens collected from Southern India exhibited significant diversity in mycelial width (3·45-5·50 µm), colony colour (yellow, pale yellow, and white), and growth pattern (thick leather mat, raised-fluffy, flat-fluffy, and raised). Field-borne chlamydospores of each isolate were significantly smaller in size (3·34-5·26 µm2 ) compared to those formed on culture media (18·6-100·89 µm2 ). The phylogenetic study based on internal transcribed sequences revealed two clusters; however, most isolates (n = 54) were grouped in cluster-I, indicating common ancestral origin. We also identified 42 haplotypes; among them, Hap_3 has the highest number of isolates (n = 19). Mating-type locus (MAT1) analysis revealed all sixty-one isolates as heterothallic, wherein 37 and 24 isolates belonging to MAT1-1-1 and MAT1-2-1 heterothallic mating types, respectively. The microsynteny analysis of MAT1 loci of one of the Indian strain (Uv-Gvt) along with Uv-8b (China) strain revealed synteny conservation at MAT1 locus, which is flanked by conserved genes SLA2 and a hypothetical protein in the upstream and APN2, COX12 and APC5 in the downstream of the locus. CONCLUSIONS Morpho-molecular study revealed the significant diversity among geo-distinct isolates, and MAT1 loci analysis indicated the distribution of heterothallic mating types in south Indian paddy fields. And also, complete synteny conservation between Indian and Chinese strain was observed at the MAT1 locus. SIGNIFICANCE AND IMPACT OF THE STUDY This is the first report describing the sexuality of Indian strains of the U. virens, which would help better understand the genetic diversity of the U. virens prevailing in Southern India and aid in developing resistant rice cultivars against this pathogen population.

Zygosaccharomyces sp. is an industrially important yeast for the production traditional fermented foods in Japan. At present, however, there is no easy method for mating Zygosaccharomyces sp. strains in the laboratory; furthermore, little is known about the expression of mating-type-specific genes in this yeast. Here, mating was observed when Zygosaccharomyces sp. was subjected to nitrogen-starvation conditions. The expression of mating-type-specific genes, Zygo STE6 and Zygo MFα1, was induced under nitrogen-starvation conditions, as confirmed by lacZ reporter assay. This expression was mating-type-specific: Zygo STE6 was expressed specifically for mating-type a, whereas and Zygo MFα1 was expressed specifically for mating-type α. Yeast strains Zygosaccharomyces rouxii DL2 and DA2, derived from type strain Z. rouxii CBS732, did not show mating even under nitrogen-starvation conditions. Gene sequencing revealed that the Zygo STE12 in Z. rouxii CBS732 has a frameshift mutation. Under nitrogen starvation, mating was observed in both DL2 and DA2 transformed with the wild-type Zygo STE12. The expression of Zygo STE6 in Z. rouxii DL2 transformed with wild-type Zygo STE12 under nitrogen-starvation conditions was confirmed by lacZ reporter assay. Collectively, these results revealed that, under nitrogen-starvation conditions, Zygosaccharomyces sp. can mate and mating-type-specific genes are expressed. Furthermore, Zygo Ste12 is essential for both mating and the expression of mating-type-specific genes in Zygosaccharomyces sp.

Clones of two morphotypes corresponding to two species of pennate araphid diatoms, Ulnaria ulna and U. acus, were isolated from samples taken in Europe. Clonal cultures entered heterothallic sexual reproduction in combinations appropriate to their mating types. Following the concept of biological species, we tested reproductive compatibility between all representatives of geographically distant populations. Isolated clones proved to be reproductively compatible within the limits of each of the species; the progeny (generation F1) were viable and fertile and freely entered into backcrossing. No cases of interspecies hybridization were found. The absence of intraspecific reproductive isolation between distant populations in these two species suggests that the populations of both U. ulna and U. acus are conspecific.

Black Sigatoka, caused by Pseudocercospora fijiensis, is a major foliar disease of banana and plantain worldwide. There are few available data regarding the genetic diversity and population structure of the pathogen in East Africa, which are needed to design effective and durable disease management strategies. We genotyped 319 single-spore isolates of P. fijiensis collected from seven regions in Uganda and Tanzania and five isolates from Nigeria using 16 simple sequence repeat markers and mating type-specific primers. Isolates from each country and region within the country were treated as populations and subpopulations, respectively. A total of 296 multilocus genotypes (MLGs) were recovered, representing a clonal fraction of 7%. Subpopulations had a moderate level of genetic diversity (Hexp = 0.12 to 0.31; mean, 0.29). Mating type distribution did not deviate from equilibrium (MAT1-1: MAT1-2, 1:1 ratio) in Uganda; however, in Tanzania the mating types were not in equilibrium (4:1 ratio). The index of association tests (IA and r̄d) showed that all populations were at linkage equilibrium (P > 0.05), thus supporting the hypothesis of random association of alleles. These findings are consistent with a pathogen that reproduces both clonally and sexually. Low and insignificant levels of population differentiation were detected, with 90% of the variation occurring among isolates within subpopulations. The high intrapopulation variation has implications in breeding for resistance to P. fijiensis because isolates differing in aggressiveness and virulence are likely to exist over small spatial scales. Diverse isolates will be required for resistance screening to ensure selection of banana cultivars with durable resistance to Sigatoka in East Africa. [Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY 4.0 International license .

Sorghum growing fields surveyed for the infection by Fusarium stalk and head rots throughout five locations in Egypt were screened depending on section Liseola characteristics. A number of 137 isolates from total recovered in average 87.8% isolates belonged to this section. These isolates selected from the whole number recovered from this group of fungi were subjected to the identified biologically and screened for fumonisin FB1 production. A number of 111 isolates produced fumonisin FB1 in average 81% from total isolates belonged to section Liseola. Four mating populations were identified, a number of 70 isolates mating population (MATP-A) F. verticillioides, 29 (MATP-D) F. proliferatum, 21 (MATP-F) F. thapsinum and 17 (MATP-G) F. nygamai. Mating population, A F. verticillioides consisted of 46 mating type 1 (MAT-1) and 24 mating type 2 (MAT-2), (MATP-D) F. proliferatum consisted of only 18 strains of (MAT-1) and 11 of (MAT-2). Whereas (MATP-F) F. thapsinum consisted of 14 strains of (MAT-1) and 7 of (MAT-2) and (MATP-G) F. nygamai consisted of only 12 strains of (MAT-1) and 5 of (MAT-2). Isolates belong to MATP-A produced considerable amounts of fumonisin compared to those belong to MATP-D,F and G. Female fertile strains could produce higher amounts of the toxin than female sterile. Measuring the parameters of the effective population number in four biological species recovered from sorghum, it was found that the female sterility was lesser than 50% in mating population A (21.4%) and were high in MATP- D, F and G (58.6%, 61.9% and 64.7%, respectively). This caused decrease in effective population number based on female fertility (Nef) overall locations in MATP- D, F and G, but it was high in MATP-A. Ration between Mating types (Mat-1, Mat-2) were differed from location to other also differed in population, it was high in all Mating populations than (Nef). These results in MATP-A led to an advantage during sexual reproduction occurs. Whereas MAT-D, F and G population vegetative propagation were significant component of the fungus especially in Assiut MATP-D, based on (Nemt) and population of Minia MATP-G, Sohag MATP-F based on (Nef).

Unisexuality in fungi is the result of sexual reproduction in a single isolate that harbors genes associated with only a single mating type. To date, unisexual reproduction has been described in only three genera of filamentous fungi. Consequently, our understanding of this unusual pathway is limited. In this critical review, we compare genetic, genomic and transcriptomic data from a variety of unisexual species to similar data from their primary homothallic and heterothallic relatives. These analyses show that unisexual reproduction is likely derived from heterothallism via the mutation of genes involved in the initiation of sexual reproduction. We show that significant changes in mating-type genes, pheromone precursor genes and pheromone receptor genes are common in unisexual species, but that similar changes are not evident in their primary homothallic or heterothallic relatives. These findings are particularly notable because the unisexual species are accommodated in unrelated genera, illustrating that a similar transition to unisexuality has likely occurred independently in their lineages.

Eyespot, caused by the related fungal pathogens Oculimacula acuformis and O. yallundae, is an important cereal stem-base disease in temperate parts of the world. Both species are dispersed mainly by splash-dispersed conidia but are also known to undergo sexual reproduction, yielding apothecia containing ascospores. Field diagnosis of eyespot can be challenging, with other pathogens causing similar symptoms, which complicates eyespot management strategies. Differences between O. acuformis and O. yallundae (e.g., host pathogenicity and fungicide sensitivity) require that both be targeted for effective disease management. Here, we develop and apply two molecular methods for species-specific and mating-type (MAT1-1 or MAT1-2) discrimination of O. acuformis and O. yallundae isolates. First, a multiplex PCR-based diagnostic assay targeting the MAT idiomorph region was developed, allowing simultaneous determination of both species and mating type. This multiplex PCR assay was successfully applied to type a global collection of isolates. Second, the development of loop-mediated isothermal amplification (LAMP) assays targeting β-tubulin sequences, which allow fast (

Candida albicans is a harmless commensal resident in the human gut and a prevalent opportunistic pathogen. A key part of its commensalism and pathogenesis is its ability to switch between different morphological forms, including white‐to‐opaque switching. The Wor1 protein was previously identified as a master regulator of white‐to‐opaque switching in mating type locus (MTL) homozygous cells. The mechanisms by which the dark color of the opaque colonies is controlled and the pimpled surface of opaque cells is formed remain unknown. Candida albicans produces melanin pigment in vitro and during infection. However, the molecular mechanism underlying the regulation of melanin production is unclear. In this study, we demonstrated that ferroxidases (Fets) function as pigment multicopper oxidases and regulate the production of dark‐pigmented melanin in opaque cells. The FET genes presented distinct regulation patterns in response to different extracellular stimuli. In YPD (1% yeast extract, 2% peptone and 2% dextrose)‐rich medium, four of the five FET genes were up‐regulated by Wor1, especially at the human body temperature of 37 °C. In minimal medium with low ammonium concentrations, all five FET genes were up‐regulated by Wor1. However, at high ammonium concentrations, some FET genes were down‐regulated by Wor1. Wor1‐up‐regulated Fets contributed to dark pigment formation in opaque colonies, but not to the elongated shape of these opaque cells. Increased melanin externalization was associated with the pimpled surface of the opaque cells. Melanized C. albicans cells were more resistant to fungal clearance. Deletion of the five FET genes completely blocked melanin production in opaque cells and resulted in the generation of white elongated ‘opaque’ cells. In addition, the up‐regulated Fets are important for defense against oxidant attacks. The functional diversity of Fets may reflect the multiple strategies of C. albicans to rapidly adapt to diverse host niches., In Candida albicans opaque cells, the master regulator Wor1 up‐regulates Fets, which function as pigment multicopper oxidases and promote the production of dark‐pigmented melanin. Increased melanin externalization is associated with the pimpled surface and dark color of opaque cells. The melanized C. albicans cells were more resistant to fungal clearance, including bone marrow‐derived macrophage phagocytosis and oxidative killing.

Sex chromosomes often carry large nonrecombining regions that can extend progressively over time, generating evolutionary strata of sequence divergence. However, some sex chromosomes display an incomplete suppression of recombination. Large genomic regions without recombination and evolutionary strata have also been documented around fungal mating-type loci, but have been studied in only a few fungal systems. In the model fungus Podospora anserina (Ascomycota, Sordariomycetes), the reference S strain lacks recombination across a 0.8-Mb region around the mating-type locus. The lack of recombination in this region ensures that nuclei of opposite mating types are packaged into a single ascospore (pseudohomothallic lifecycle). We found evidence for a lack of recombination around the mating-type locus in the genomes of ten P. anserina strains and six closely related pseudohomothallic Podospora species. Importantly, the size of the nonrecombining region differed between strains and species, as indicated by the heterozygosity levels around the mating-type locus and experimental selfing. The nonrecombining region is probably labile and polymorphic, differing in size and precise location within and between species, resulting in occasional, but infrequent, recombination at a given base pair. This view is also supported by the low divergence between mating types, and the lack of strong linkage disequilibrium, chromosomal rearrangements, transspecific polymorphism and genomic degeneration. We found a pattern suggestive of evolutionary strata in P. pseudocomata. The observed heterozygosity levels indicate low but nonnull outcrossing rates in nature in these pseudohomothallic fungi. This study adds to our understanding of mating-type chromosome evolution and its relationship to mating systems.

Phytophthora capsici is a soilborne oomycete plant pathogen that causes severe vegetable crop losses in New York (NY) state and worldwide. This pathogen is difficult to manage, in part due to its production of long-lasting sexual spores and its tendency to quickly evolve fungicide resistance. We single nucleotide polymorphism (SNP) genotyped 252 P. capsici isolates, predominantly from NY, in order to conduct a genome-wide association study for mating type and mefenoxam sensitivity. The population structure and extent of chromosomal copy number variation in this collection of isolates were also characterized. Population structure analyses showed isolates largely clustered by the field site where they were collected, with values of FST between pairs of fields ranging from 0.10 to 0.31. Thirty-three isolates were putative aneuploids, demonstrating evidence for having up to four linkage groups present in more than two copies, and an additional two isolates appeared to be genome-wide triploids. Mating type was mapped to a region on scaffold 4, consistent with previous findings, and mefenoxam sensitivity was associated with several SNP markers at a novel locus on scaffold 62. We identified several candidate genes for mefenoxam sensitivity, including a homolog of yeast ribosome synthesis factor Rrp5, but failed to locate near the scaffold 62 locus any subunits of RNA polymerase I, the hypothesized target site of phenylamide fungicides in oomycetes. This work expands our knowledge of the population biology of P. capsici and provides a foundation for functional validation of candidate genes associated with epidemiologically important phenotypes.

International audience; Mating type is a critical trait in heterothallic organisms. In plant pathogenic oomycetes, like the late blight pathogen Phytophthora infestans, it is usually identified through pairing between tester and candidate isolates, a method that is both laborious and applicable to live isolates only. Therefore, developing simple and fast PCR tests to reliably identify P. infestans mating types is of great interest for population genetic studies. A multiplex PCR assay combining the amplification of a locus diagnostic for P. infestans and of one diagnostic for the A1 mating type was developed and validated on a collection of 1,441 samples, covering the current and past diversity of European P. infestans populations. These samples were obtained from either freeze-dried mycelium or FTA cards on which diseased leaflets had been pressed. The multiplex assay correctly identified mating types in 97.4% of these samples. The main source of incorrect assignment was the lack of amplification of the A1 diagnostic allele, due to insufficient DNA quality and/or quantity in the reaction mix. This multiplex PCR, applicable to both live and stored material, thus constitutes a useful addition to the set of molecular tools available for population typing in P. infestans.

Sexual agglutinins of the budding yeast Saccharomyces cerevisiae are proteins mediating cell aggregation during mating. Complementary agglutinins expressed by cells of opposite mating types “a” and “α” bind together to promote agglutination and facilitate fusion of haploid cells. By means of an innovative single-cell manipulation assay combining fluidic force microscopy with force spectroscopy, we unravel the strength of single specific bonds between a- and α-agglutinins (~100 pN) which require pheromone induction. Prolonged cell–cell contact strongly increases adhesion between mating cells, likely resulting from an increased expression of agglutinins. In addition, we highlight the critical role of disulfide bonds of the a-agglutinin and of histidine residue H273 of α-agglutinin. Most interestingly, we find that mechanical tension enhances the interaction strength, pointing to a model where physical stress induces conformational changes in the agglutinins, from a weak-binding folded state, to a strong-binding extended state. Our single-cell technology shows promises for understanding and controlling the complex mechanism of yeast sexuality., Mathelié-Guinlet et al. investigate the molecular binding mechanisms of sexual agglutinins in budding yeast Saccharomyces cerevisiae. They report that mechanical tension enhances the strength of agglutinin interactions, supporting a new model in which physical stress induces conformational changes in the binding sites of agglutinins.

Obligately biotrophic plant pathogens pose challenges in population genetic studies due to their genomic complexities and elaborate culturing requirements with limited biomass. Hop powdery mildew (Podosphaera macularis) is an obligately biotrophic ascomycete that threatens sustainable hop production. P. macularis populations of the Pacific Northwest (PNW) United States differ from those of the Midwest and Northeastern United States, lacking one of two mating types needed for sexual recombination and harboring two strains that are differentially aggressive on the cultivar Cascade and able to overcome the Humulus lupulus R-gene R6 (V6), respectively. To develop a high-throughput marker platform for tracking the flow of genotypes across the United States and internationally, we used an existing transcriptome of diverse P. macularis isolates to design a multiplex of 54 amplicon sequencing markers, validated across a panel of 391 U.S. samples and 123 international samples. The results suggest that P. macularis from U.S. commercial hop yards form one population closely related to P. macularis of the United Kingdom, while P. macularis from U.S. feral hop locations grouped with P. macularis of Eastern Europe. Included in this multiplex was a marker that successfully tracked V6-virulence in 65 of 66 samples with a confirmed V6-phenotype. A new qPCR assay for high-throughput genotyping of P. macularis mating type generated the highest resolution distribution map of P. macularis mating type to date. Together, these genotyping strategies enable the high-throughput and inexpensive tracking of pathogen spread among geographical regions from single-colony samples and provide a roadmap to develop markers for other obligate biotrophs.

The contemporary dominant clonal lineage of heterothallic Phytophthora infestans in Wisconsin, US-23, is classified as sensitive to the systemic fungicide mefenoxam and is of the A1 mating type. With the sporadic appearance of clonal lineage US-8, classified as resistant to mefenoxam and of the A2 mating type, there is a need for ongoing monitoring and characterization. Isolates of P. infestans collected from Wisconsin during the 2017 and 2018 growing seasons were tested for sensitivity to mefenoxam with discriminatory dose of 100 ppm. In 2017, both US-23 and US-8 were isolated. On average, isolates of US-23 were significantly more sensitive to mefenoxam than were US-8 isolates (P = 8e-04). There were significant differences in the sensitivity levels among the US-8 isolates (P = 2.02e-06), with a single isolate testing sensitive at 100 ppm of mefenoxam based on the one-way ANOVA. There were significant differences in the sensitivity levels among US-23 isolates (P = 3.75e-09), with two isolates showing resistance. In 2018 only US-23 was found, and isolates were tested for mefenoxam response at 0, 0.1, 1, 10, and 100 ppm. At 0.1 ppm, isolates showed significantly different levels of sensitivity (P = 2.1e-09), and a single isolate showed complete resistance. Isolates from both clonal lineages and years that exhibited moderate levels of resistance had greater variability among replicates. The phenotype of this multigenic trait comes through in the variability seen in isolates that are showing more resistance. Continued screening of P. infestans for mefenoxam sensitivity will help track the development and mechanism of resistance, as well as aid in development of best management approaches.

Rhynchosporium commune, the cause of barley scald, is an important disease of barley in Canada. Multiple studies have indicated that cryptic sexual reproduction may occur between opposite mating ty...

International audience; Genomic regions determining sexual compatibility often display recombination suppression, as occurs in sex chromosomes, plant self‐incompatibility loci and fungal mating‐type loci. Regions lacking recombination can extend beyond the genes determining sexes or mating types, by several successive steps of recombination suppression. Here we review the evidence for recombination suppression around mating‐type loci in fungi, sometimes encompassing vast regions of the mating‐type chromosomes. The suppression of recombination at mating‐type loci in fungi has long been recognized and maintains the multiallelic combinations required for correct compatibility determination. We review more recent evidence for expansions of recombination suppression beyond mating‐type genes in fungi (‘evolutionary strata’), which have been little studied and may be more pervasive than commonly thought. We discuss testable hypotheses for the ultimate (evolutionary) and proximate (mechanistic) causes for such expansions of recombination suppression, including (1) antagonistic selection, (2) association of additional functions to mating‐type, such as uniparental mitochondria inheritance, (3) accumulation in the margin of nonrecombining regions of various factors, including deleterious mutations or transposable elements resulting from relaxed selection, or neutral rearrangements resulting from genetic drift. The study of recombination suppression in fungi could thus contribute to our understanding of recombination suppression expansion across a broader range of organisms.

Random amplified polymorphic DNA (RAPD) markers were used to estimate genetic relationship and diversity of Fusarium species associated with pink ear rot of maize. Three Fusarium species – F. verticillioides, F. temperatum and F. subglutinans sensu stricto were collected from contaminated corn kernels. A collection of 41 randomly selected Fusarium isolates (16 F. verticillioides, 16 F. temperatum and nine F. subglutinans sensu stricto) was screened for polymorphism with thirty decamer primers. Seventeen primers that had generated a high number of polymorphic loci were used for the analysis of genetic diversity. The primers yielded a total of 146 polymorphic loci, an average of 8.59 polymorphic bands per primer. The percentage of polymorphic bands (%P) was 46.1%. The RAPD-based UPGMA analysis demonstrated that although F. temperatum and F. subglutinans s.s. form two distinct groups they are more similar to each other than to F. verticillioides. Fusarium verticillioides was the most heterogenous species (Nei’s genetic diversity 0.180), while the least heterogenous one was F. subglutinans s.s. whose genetic diversity was 0.098. The analysis of the translation elongation factor 1-α and β-tubulin gene sequences as well as the mating type ratio were employed to support the RAPD data.

Fusarium isolates obtained as endophytes from healthy leaves of Handroanthus chrysotrichus (Bignoniaceae) in the Atlantic Forest of north-eastern Brazil were characterised morphologically and phylogenetically on the basis of partial rDNA sequences encompassing the second largest subunit of RNA polymerase II, translation elongation factor EF-1 alpha, and β-tubulin genes. A novel species within the Fusarium lateritium species complex (FLSC) is described as Fusarium massalimae. Isolates of F. pseudocircinatum of the Fusarium fujikuroi species complex (FFSC) were also isolated as endophytes from leaves of H. chrysotrichus. Mating type gene analyses suggested that obtained isolates of F. massalimae and F. pseudocircinatum are heterothallic.

Zymoseptoria tritici is the pathogen responsible for Septoria tritici blotch (STB) disease in wheat, causing significant yield losses during some years. This disease is observed in wheat production areas in our country as in many regions of the world. In this study, STB diseased wheat leaves were collected from six geographical regions in wheat cultivation areas of Turkey during 2013 and 2014 production seasons. A total of 103 single spore isolates obtained from the collection of Z. tritici population were chosen as representative for testing of their mating type. The isolates were evaluated for mating type (MAT) idiomorph frequency by multiplex PCR using Z. tritici specific MAT primers. The prevalence of mating types was found to be 48.5% (50) and 51.5% (53) for Mat1-1 and Mat1-2, respectively. Chi-square test was used to assess that two mating type scores are at equal frequencies. The study affirms the hypothesis that Z. tritici is reproducing sexually and spread via airborne ascospores aside from rain splash dispersed asexual spores in Turkey. Genetic recombination during sexual mating has a potential to generate more virulent pathotypes of Z. tritici.

Neonectria ditissimaandN. faginataare canker pathogens involved in an insect-fungus disease complex of American beech (Fagus grandifolia) commonly known as beech bark disease (BBD). In Europe, bothN. ditissimaandN. coccineaare involved in BBD on European beech (Fagus sylvatica). Field observations across the range of BBD indicate that new infections occur primarily via ascospores. Both heterothallic (self-sterile) and homothallic (self-fertile) mating strategies have been reported forNeonectriafungi. As such, investigations into mating strategy are important for understanding both the disease cycle and population genetics ofNeonectria. This is particularly important in the U.S. given that over timeN. faginatadominates the BBD pathosystem despite high densities of non-beech hosts forN. ditissima. This study utilized whole-genome sequences of BBD-associatedNeonectriaspp. along with other publicly availableNeonectriaandCorinectriagenomes andin vitromating assays to characterize mating type (MAT) loci and confirm thallism for select members ofNeonectriaandCorinectria. MAT gene-specific primer pairs were developed to efficiently characterize the mating types of additional single ascospore strains ofN. ditissima,N. faginata, andN. coccineaand several other related species lacking genomic data.In vitromating assays were used in combination with molecular results to confirm thallism. These assays also comfirmed the sexual compatibility amongN. ditissimastrains from different plant hosts. Maximum likelihood phylogenetic analysis of both MAT1-1-1 and MAT1-2-1 sequences recovered trees with similar topology to previously published phylogenies ofNeonectriaandCorinectria. The results of this study indicate that allNeonectriaandCorinectriatested are heterothallic based on our limited sampling and, as such, thallism cannot help explain the inevitable dominance ofN. faginatain the BBD pathosystem.

Background: Late blight caused by Oomycete Phytophthora infestans remains a huge problem in potato production and one of the most severe crop diseases worldwide. In Cameroon, the populations characterization of this pathogen remain very little known. Aims: This study aims to characterize isolates of P. infestans collected in twelve localities of Highlands zone of Cameroon. Study Design: Twelve localities in three main potato production areas of the Highlands agro-ecological zone (HAZ) in Cameroon were selected for sample collection. The phenotypic parameters of sporangia were measured. The presence of mating types and pathogenicity tests on detached leaflets of two potato varieties (MANATE and CIPIRA) were assessed. Results: A total of 36 isolates were collected. Height (08) Sporangia shape of P. infestans were obtain and 02 new different shape were pip form and oval to ellipsoid. Sporangia length/width ratio ranged from 1.62 to 2.18. Mating types A1 and A2 were present in the studied areas. Pathogenicity test on detached potato leaflets was positive with all the 36 isolates. The isolate HPBT02 from Tsela locality was more aggressive (P

Sex offers advantages even in primarily asexual species. Some ciliates appear to utilize such reproductive strategy with many mating types. However, the factors determining the composition of mating types in the unicellular ciliate Tetrahymena thermophila are poorly understood, and this is further complicated by non-Mendelian determination of mating type in the offspring. We therefore developed a novel population genetics model to predict how various factors influence the dynamics of mating type composition, including natural selection. The model predicted either the coexistence of all seven mating types or fixation of a single mating type in a population, depending on parameter combinations, irrespective of natural selection. To understand what factor(s) may be more influential and to test the validity of theoretical prediction, five replicate populations were maintained in laboratory such that several factors could be controlled or measured. Whole-genome sequencing was used to identify newly arising mutations and determine mating type composition. Strikingly, all populations were found to be driven by strong selection on newly arising beneficial mutations to fixation of their carrying mating types, and the trajectories of speed to fixation agreed well with our theoretical predictions. This study illustrates the evolutionary strategies that T. thermophila can utilize to optimize population fitness.

Parkinson’s disease (PD) is one of the most prevailing aging diseases around the world. The present study was to investigate the potential effect of hydrogen sulfide (H(2)S) and silent mating type information regulation 2 homolog 1 (SIRT1) in MPP~+ induced SH-SY5Y cells and its underlying mechanisms in PD. SH-SY5Y cells were induced by MPP~+ and treated with the H(2)S donor NaHS to detect the effect of H(2)S on the molecular behaviors of MPP~+ induced SH-SY5Y cells. NaHS reduced the apoptosis rate and expressions of MDA, 4-HNE and p62, while increased cell viability, autophagy flux and expressions of LC3 II/I and Beclin1 in MPP~+ induced SH-SY5Y cells. Then, levels of autophagy-related proteins and inflammation-related proteins (TNF-α, IL-Iβ) were detected, indicating that Chloroquine and Sirtinol reversed the protective effect of H(2)S on SH-SY5Y cells induced by MPP~+. We further explored the particular function of H(2)S, SH-SY5Y cells treated with MPP~+, NaHS chloroquine, and SIRT1 inhibitor (Sirtinol). The results showed that H(2)S increased SIRT1 expression and sulfhydration. Finally, a PD mouse model verified the above results. In a word, H(2)S ameliorated SIRT1 activity through acceleration of SIRT1 sulfhydration to increase the autophagy flux and attenuate damage of SH-SY5Y cells induced by MPP~+. H(2)S and SIRT1 activator might be a target in the treatment of PD patients.

Molecular genetic study of Saccharomyces yeasts of different species was carried out using molecular karyotyping and comparative analysis of a number of nuclear and mitochondrial genes. Multigene phylogenetic analysis showed that the recently described species S. jurei was phylogenetically closest to S. mikatae, while S. bayanus and S. arboricola were the most divergent species within the genus Saccharomyces. Comparative analysis of amino acid sequences of the SUC β-fructosidase genes also indicated a close genetic relationship between the species S. jurei and S. mikatae. The biological species S. cerevisiae, S. kudriavzevii, and S. paradoxus possessed collinear karyotypes. Two reciprocal translocations were found in the S. jurei karyotype, one of which was unique (between chromosomes I and XIII), and the second was common with S. mikatae: VI/VII. Reciprocal translocations in the karyotypes of S. arboricola, S. bayanus, and S. cariocanus were species-specific. It was established that only chromosome III carrying the mating type locus MAT was of approximately the same size in all eight species of the genus Saccharomyces. The adaptive role of reciprocal translocations of chromosomes is discussed.

Brown rot disease caused by fungal species from theMonilinia genus has recently been observed as one of the most importantlimiting factors for yield and quality of peach fruits in Turkey. During Juneand July of 2018, field trips to different peach orchards were performed in sixprovinces located in four different geographical regions of Turkey. One hundredand twenty-nine isolates were obtained from the diseased fruits, and of those,109 were identified as Monilinia fructicola, whereas 20 were M. laxa accordingto the species-specific molecular markers. Mating types of the isolates weredetected by PCR assays using primers designed in this study. Each isolaterepresented only one of the mating type genes. Overall data sets presented 1:1ratio of mating types for both species, indicating a possible sexualreproduction. No polymorphisms were detected in partial sequences of matingtype genes. The isolates were in vitro tested for their pathogenicity using applefruits as a host and aggressiveness were evaluated at different time pointsusing lesion sizes. Monilinia laxa was found more aggressive than M.fructicola. Moreover, M. fructicola isolates displayed larger colony growthcompared to M. laxa isolates. However, no significant correlation was detectedbetween the colony growth rates and virulence. This study is the first to demonstratebrown rot pathogens of peach fruit in Turkey and identifies different aspectsof the pathogens, which would be useful in containment of pathogen spread andcomparison with other populations of these pathogens worldwide, and suggests anew set of mating type markers for these species.

The purpose of this study was to characterize Phytophthora infestans isolates obtained from diseased tomato tissue in Oman from 2013 to 2015. Late blight caused severe epidemics on tomatoes in Oman in 2013–2014 and 2014–2015. Prior to 2013, late blight had not previously been reported from Oman. Isolates collected from diseased tomato plants in commercial production fields in Oman were assayed for a series of traits. All fifty-five isolates tested for mating type were determined to be A1. A subset of nine isolates analysed for allozymes, sensitivity to mefenoxam, DNA fingerprint as detected with random genome probe fifty-seven (RG57), and microsatellite genotypes revealed a diverse population. A range of sensitivity to mefenoxam was observed and one multi-locus genotype (MLG) was detected in four of the nine isolates. The A2 strain Blue_13, which was recently identified in India and thought to be a possible source of the pathogen, was not detected.

Agrocybe cylindracea, an edible mushroom, is widely cultivated for its abundance of nutrients and flavor, and many of its metabolites are reported to have beneficial roles, such as medicinal effects on tumors and chronical illnesses. However, the lack of genomic information has hindered further molecular studies on this fungus. Here, we present a genome assembly of A. cylindracea together with comparative genomics and pathway analyses of Agaricales species. The draft, generated from both next-generation sequencing (NGS) and single-molecule real-time (SMRT) sequencing platforms to overcome high genetic heterozygosity, is composed of a 56.5 Mb sequence and 15,384 predicted genes. This mushroom possesses a complex reproductive system, including tetrapolar heterothallic and secondary homothallic mechanisms, and harbors several hydrolases and peptidases for gradual and effective degradation of various carbon sources. Our pathway analysis reveals complex processes involved in the biosynthesis of polysaccharides and other active substances, including B vitamins, unsaturated fatty acids, and N-acetylglucosamine. RNA-seq data show that A. cylindracea stipes tend to synthesize carbohydrate for carbon sequestration and energy storage, whereas pilei are more active in carbon utilization and unsaturated fatty acid biosynthesis. These results reflect diverse functions of the two anatomical structures of the fruiting body. Our comprehensive genomic and transcriptomic data, as well as preliminary comparative analyses, provide insights into the molecular details of the medicinal effects in terms of active compounds and nutrient components.

Pecan scab, caused by Venturia effusa, is the most prevalent disease of pecan in the southeastern United States. Recent characterization of the mating type (MAT) distribution of V. effusa revealed that the MAT idiomorphs are in equilibrium at various spatial scales, indicative of regular sexual recombination. However, the occurrence of the sexual stage of V. effusa has never been observed, and the pathogen was previously considered to rely entirely on asexual reproduction. We were able to generate the sexual stage by pairing isolates of opposite mating types on oatmeal culture media. Cultures were incubated at 24 C for 2 mo to allow hyphae from isolates of each mating type to interact. Culture plates were then incubated at 4 C for 4 mo, after which immature pseudothecia were observed. Following exposure to a 12-h photoperiod for 2 wk at 24 C, asci and ascospores readily developed. Pseudothecium and ascospore production was optimal when incubated for 4 mo at 4 C. We utilized progeny from a cross of an albino isolate and wild-type (melanized) isolates to determine that recombination had occurred. Multilocus genotyping using 32 microsatellite markers confirmed that progeny were the result of recombination, which was further supported by segregation of mating types and culture pigmentation. Albino progeny were all confirmed to contain the same mutation in the polyketide synthase (PKS1) melanin biosynthesis gene as the albino parent. The results of this study demonstrate the heterothallic nature of V. effusa. The impact of determining the source of the overwintering ascostroma will aid in management decisions to reduce the primary inoculum in the disease cycle.