Mating Type Ratio Research Articles

Effects of mating-type ratio imbalance on the degeneration of Cordyceps militaris subculture and preventative measures.

The rapid degeneration of Cordyceps militaris strains during subculture represents a bottleneck problem that affects production stability. This study explored the mechanism underlying this degeneration in three production and three wild-type strains of Cordyceps militaris, isolating single-conidium strains from each. The effects of subculturing on fructification in both original and single mating-type strains were compared. Changes in the ratio of the two mating types were analyzed in both original and degenerated strains. Based on these findings, the two mating strains were paired in different ratios to determine their effects on fruiting. The resulting five strains were heterokaryotic strains with both MAT1-1 and MAT1-2 mating-type genes. Strain jb-2 was a single mating type (MAT1-1) mutant strain that produced stable fruiting bodies but failed to produce ascospores. It was found that the loss of or imbalance in mating types was the main reason for the rapid degeneration of fruiting traits during subculture and that this occurred randomly in the MAT1-1 and MAT1-2 types. The strains differed significantly in their stability during subculture. Fruiting was stable in the single mating-type Jb-2 strain, and the eleventh-generation fruited normally. There were differences in yield between the production and wild strains after inoculation with spawn containing different proportions of mating types. The production strain was more stable when inoculated with strains with mating-type ratios of 1:9 to 9:1 without affecting the yield. However, the yield of the wild-type strain xf-1 was positively correlated with the proportion of the MAT1-2 type, while the other two strains showed no correlations. Subculturing single mating-type mycelia separately and mixing them before production effectively mitigated degeneration during subculture. For Cordyceps militaris breeding, selecting strains containing both mating types, which are insensitive to the proportion of mating-type genes, enhanced stability in subculture and reduced the risk of mating-type loss. Direct breeding of specific single-mating type strains to induce fruiting is thus an effective breeding strategy.

Multiannual patterns of genetic structure and mating type ratios highlight the complex bloom dynamics of a marine planktonic diatom

Understanding the genetic structure of populations and the processes responsible for its spatial and temporal dynamics is vital for assessing species’ adaptability and survival in changing environments. We investigate the genetic fingerprinting of blooming populations of the marine diatom Pseudo-nitzschia multistriata in the Gulf of Naples (Mediterranean Sea) from 2008 to 2020. Strains were genotyped using microsatellite fingerprinting and natural samples were also analysed with Microsatellite Pool-seq Barcoding based on Illumina sequencing of microsatellite loci. Both approaches revealed a clonal expansion event in 2013 and a more stable genetic structure during 2017–2020 compared to previous years. The identification of a mating type (MT) determination gene allowed to assign MT to strains isolated over the years. MTs were generally at equilibrium with two notable exceptions, including the clonal bloom of 2013. The populations exhibited linkage equilibrium in most blooms, indicating that sexual reproduction leads to genetic homogenization. Our findings show that P. multistriata blooms exhibit a dynamic genetic and demographic composition over time, most probably determined by deeper-layer cell inocula. Occasional clonal expansions and MT imbalances can potentially affect the persistence and ecological success of planktonic diatoms.

Study of mating type ratio and colony sectorization of asexual spores on the progress of Cordyceps militaris degeneration

Study of mating type ratio and colony sectorization of asexual spores on the progress of Cordyceps militaris degeneration

Population structure and mixed reproductive strategies in Bipolaris maydis from single and multiple corn cultivars in Fujian Province, China.

Bipolaris maydis is the pathogenic microorganism of southern corn leaf blight, a persistent biotic constraint responsible for substantial yield losses of corn worldwide. In the present study, 96 isolates from six representative fields growing single and multiple sweet corn cultivars in Pingnan, Fuqing, and Jian'ou in Fujian Province, which are characterized by different geographical characteristics and cropping patterns, were genetically analyzed using inter-simple sequence repeat (ISSR) markers to assess the impact of geographical origins and corn cultivars on B. maydis population differentiation. B. maydis isolates originated from diverse regions possessed higher genetic variety than those from single and multiple sweet corn cultivars. Phylogenetic analysis showed that the isolates from single and multiple sweet corn cultivars were randomly grouped into different clusters, with those from the same location tending to form clusters. A greater genetic differentiation among different geographical populations than between those from single and multiple sweet corn cultivars was observed by pairwise comparison. Hierarchical analysis indicated that among-population variation was higher when comparatively analyzed B. maydis populations from different locations than in those from single and multiple sweet corn cultivars. In conclusion, these results suggest that geographical origin acts a more considerable role in genetic differentiation of B. maydis than corn cultivar. Two divided genetic clusters were detected in the B. maydis populations from single and multiple sweet corn cultivars at the three locations in Fujian Province, with major genetic variation being derived within populations. The high haplotypic diversity and expected mating type ratio of 1:1 in combination with significant linkage disequilibrium suggested that a mixed reproductive strategy occurs in the B. maydis population in Fujian Province. This study will enrich the information on the role that geographical origins and corn cultivars play in the population structure of the pathogen as well as the reproductive strategies in B. maydis population in Fujian Province.

Indirect Evidence Based on Mating-Type Ratios for the Role of Sexual Reproduction in European and Chinese Populations of Plenodomus biglobosus (Blackleg of Oilseed Rape)

Blackleg (Phoma) disease, caused by the ascomycete fungi Plenodomus biglobosus and P. lingam, threatens oilseed rape (OSR; Brassica napus) crops internationally. In many parts of the world, both species co-occur, but in China only P. biglobosus has so far been reported. Plenodomus biglobosus reproduces asexually (pycnidiospores), but also sexually (pseudothecia-yielding ascospores), via a heterothallic mating system requiring MAT1-1 and MAT1-2 genotypes. However, the roles of airborne ascospore inoculum in driving blackleg disease outbreaks in China are less well understood compared to elsewhere in the world. This is despite the very different agronomic cropping practices in parts of China, in which paddy rice and OSR are often grown in rotation; OSR stubble is often submerged under water for long periods potentially affecting pseudothecial development. Here, we indirectly investigate the potential role of sexual reproduction by developing new polymerase chain reaction (PCR) -based mating-type diagnostics for P. biglobosus and subsequently screening an international collection of 59 European and 157 Chinese isolates. Overall, in both Europe and China, P. biglobosus mating types did not deviate from a 1:1 ratio, such as is generally thought to occur under frequency-dependent selection in sexually reproducing pathogen populations. Both mating types were balanced in all the individual European countries tested (Austria, France, Poland, UK). Conversely, in China, mating types were only balanced in the eastern region; in the northern and southwestern regions there were skewed ratios, more typical of predominantly asexual reproduction, towards MAT1-1 and MAT1-2, respectively. The implications of these findings and future research directions for improved understanding of P. biglobosus epidemiology on OSR, particularly in China, are considered.

Genetic diversity and structure of Bipolaris oryzae and Exserohilum rostratum populations causing brown spot of rice in Burkina Faso based on genotyping-by-sequencing.

In recent years, Brown spot disease of rice (BSR) has been observed on leaves and seeds of rice in all rice-growing areas of Burkina Faso. Bipolaris oryzae and Exserohilum rostratum are the main fungal species isolated from BSR infected tissues and they are frequently observed in the same field. However, we are lacking information on the genetic diversity and population structure of these fungi in Burkina Faso. The mode of reproduction is also unknown. The genetic diversity of isolates of B. oryzae (n=61) and E. rostratum (n=151), collected from major rice-growing areas of Burkina Faso, was estimated using genotyping-by-sequencing (GBS). The mean values for nucleotide diversity (π) were 1.9 x10-4 for B. oryzae and 4.8 x10-4 for E. rostratum. There is no genetic differentiation between the geographical populations of each species. The analysis of molecular variance revealed that 89% and 94% of the genetic variances were within the populations of B. oryzae and E. rostratum, respectively. For each species, four genetic clusters were identified by two clustering methods (DAPC and sNMF). The distribution of these genetic groups was independent of the geographical origin of the isolates. Evidence of recombination was detected in the populations of B. oryzae and E. rostratum. For B. oryzae balanced mating type ratios were supporting sexual reproduction. For E. rostratum overrepresentation of MAT1-2 isolates (79%) suggested a predominant asexual reproduction. This study provides important information on the biology and genetics of the two major fungi causing brown spot disease of rice in Burkina Faso.

Phylogenetic Analysis of Phaeosphaeria Species Using Mating Type Genes and Distribution of Mating Types in Iran.

Phaeosphaeria species are pathogenic on wheat, barley and a wide range of wild grasses. To analyze mating type loci of the Phaeosphaeria species and investigate mating type distribution in Iran, we sequenced mating type loci of 273 Phaeosphaeria isolates including 67 isolates obtained from symptomatic leaves and ears of wheat, barley, and wild grasses from two wheat-growing region in Iran as well as 206 isolates from our collection from other regions in Iran which were isolated in our previous studies. Mating type genes phylogeny was successfully used to determine the species identity and relationships among isolates within the Phaeosphaeria spp. complex. In this study, we reported seven new host records for Phaeosphaeria species and the Phaeosphaeria avenaria f. sp. tritici 3 group was first reported from Iran in this study. Mating type distribution among Phaeosphaeria species was determined. Both mating types were present in all sampling regions from Iran. We observed skewed distribution of mating types in one region (Kohgiluyeh va Boyer-Ahmad) and equal distribution in the other region (Bushehr). However, when considering our entire dataset of 273 Iranian Phaeosphaeria isolates, the ratio of mating types was not deviated significantly from 1:1 suggesting possibilities for isolates of opposite mating type to interact and reproduce sexually, although the sexual cycle may infrequently occur in some regions especially when the climatic conditions are unfavorable for teleomorph development.

Genetic Diversity of the Symbiotic Fungus Epichloë festucae in Naturally Occurring Host Grass Populations

Epichloë festucae is a common symbiont of the perennial and widely distributed cool season grass, Festuca rubra. The symbiosis is highly integrated involving systemic growth of the fungus throughout above-ground host parts and vertical transmission from plant to its offspring via host seeds. However, the nature of symbiosis is labile ranging from antagonistic to mutualistic depending on prevailing selection pressures. Both the loss of fungus in the maternal host lineage and horizontal transmission through sexual spores within the host population may partly explain the detected variation in symbiosis in wild grass populations. Epichloë species are commonly considered as pathogens when they produce sexual spores and partly castrate their host plant. This is the pathogenic end of the continuum from antagonistic to mutualistic interactions. Here we examined the population genetic structure of E. festucae to reveal the gene flow, importance of reproduction modes, and alkaloid potential of the symbiotic fungus in Europe. Epichloë-species are highly dependent on the host in survival and reproduction whilst benefits to the host are largely linked to defensive mutualism attributable to fungal-origin bioactive alkaloids that negatively affect vertebrate and/or invertebrate herbivores. We detected decreased genetic diversity in previously glaciated areas compared to non-glaciated regions during the last glacial maximum period and found three major genetic clusters in E. festucae populations: southern, northeastern and northwestern Europe. Sexual reproduction may have a higher role than expected in Spanish E. festucae populations due to the predominance of unique genotypes and presence of both mating types in the region. In contrast, asexual reproduction via host seeds predominates in the Faroe Island and Finland in northern Europe due to the presence of biased mating-type ratios and large dominant genotypes in the E. festucae populations within the region. A substantially larger variation of alkaloid genotypes was observed in the fungal populations than expected, although the variability of the alkaloid genotypes within populations is considerably lower in northern than Spanish populations in southern Europe. E. festucae populations consist of different combinations of alkaloid classes from the gene clusters of ergot alkaloid and indole-terpenes, and from pyrrolopyrazine alkaloid gene. We suggest that the postglacial distribution history of the host grass, prevailing reproduction strategies of E. festucae, and local selection pressures likely explain a large part of the genetic variation observed in fungal populations among geographic regions. The identified alkaloid genotypes can be used by turfgrass breeders to improve resistance against herbivores in red fescue varieties and to develop new sustainable cultivars in Europe.

On computing relative effective population size estimates and parameters from an equilibrium cycle of hermaphrodite frequency fluctuation due to mixed reproductive modes in filamentous fungi

Many filamentous ascomycete fungi reproduce primarily asexually, with only occasional sexual generations. This can lead to a departure from the 1:1 mating type ratio that is expected in obligate sexual populations. The relaxed selection on sexual traits also can lead to a decrease in the frequency of female fertile strains in field populations, while male fertility does not similarly decrease since male gametes also can serve as asexual spores. Both changes ultimately impact the strength of genetic drift in populations. The frequency of female sterility likely increases with the time since the last generation of sexual reproduction, such that it can be used to estimate the relative frequency of sexual reproduction. Here I provide additional details relevant to Leslie and Klein’s (1996) model of mixed sexual reproduction and vegetative propagation as related to the frequency of female sterility. This includes new or modified equations that allow for simpler calculations of i) two estimates of relative Ne, ii) the expected range of hermaphrodite frequencies during the cycles of mixed reproduction, and iii) the relative frequency of sex. These equations also are included in spreadsheet templates into which researchers can directly enter frequencies computed from their population data to estimate these parameters for their own populations. These resources will make the results of Leslie and Klein (1996) more accessible and should increase the use of this model in evaluating the frequency of sexual reproduction of filamentous fungi.

Phylogeography of the wide-host range panglobal plant pathogen Phytophthora cinnamomi.

Various hypotheses have been proposed regarding the origin of the plant pathogen Phytophthora cinnamomi. P.cinnamomi is a devastating, highly invasive soilborne pathogen associated with epidemics of agricultural, horticultural and forest plantations and native ecosystems worldwide. We conducted a phylogeographic analysis of populations of this pathogen sampled in Asia, Australia, Europe, southern and northern Africa, South America, and North America. Based on genotyping-by-sequencing, we observed the highest genotypic diversity in Taiwan and Vietnam, followed by Australia and South Africa. Mating type ratios were in equal proportions in Asia as expected for a sexual population. Simulations based on the index of association suggest a partially sexual, semi-clonal mode of reproduction for the Taiwanese and Vietnamese populations while populations outside of Asia are clonal. Ancestral area reconstruction provides new evidence supporting Taiwan as the ancestral area, given our sample, indicating that this region might be near or at the centre of origin for this pathogen as speculated previously. The Australian and South African populations appear to be a secondary centre of diversity following migration from Taiwan or Vietnam. Our work also identified two panglobal, clonal lineages PcG1-A2 and PcG2-A2 of A2mating type found on all continents. Further surveys of natural forests across Southeast Asia are needed to definitively locate the actual centre of origin of this important plant pathogen.

The Plot Thickens: Haploid and Triploid-Like Thalli, Hybridization, and Biased Mating Type Ratios in Letharia.

The study of the reproductive biology of lichen fungal symbionts has been traditionally challenging due to their complex lifestyles. Against the common belief of haploidy, a recent genomic study found a triploid-like signal in Letharia. Here, we infer the genome organization and reproduction in Letharia by analyzing genomic data from a pure culture and from thalli, and performing a PCR survey of the MAT locus in natural populations. We found that the read count variation in the four Letharia specimens, including the pure culture derived from a single sexual spore of L. lupina, is consistent with haploidy. By contrast, the L. lupina read counts from a thallus' metagenome are triploid-like. Characterization of the mating-type locus revealed a conserved heterothallic configuration across the genus, along with auxiliary genes that we identified. We found that the mating-type distributions are balanced in North America for L. vulpina and L. lupina, suggesting widespread sexual reproduction, but highly skewed in Europe for L. vulpina, consistent with predominant asexuality. Taken together, we propose that Letharia fungi are heterothallic and typically haploid, and provide evidence that triploid-like individuals are hybrids between L. lupina and an unknown Letharia lineage, reconciling classic systematic and genetic studies with recent genomic observations.

Genetic Differentiation and Mixed Reproductive Strategies in the Northern Corn Leaf Blight Pathogen Setosphaeria turcica From Sweet Corn in Fujian Province, China.

The northern corn leaf blight (NCLB) pathogen Setosphaeria turcica (Luttrell) Leonard and Suggs is one of the main biotic constraints on sweet corn (Zea mays L.) yield and quality in Fujian Province, China. Currently, however, there is comparatively little information available regarding the distribution of mating types, population genetics, and reproductive strategies of this pathogen in Fujian. In this study, we investigated the distribution of mating types and population genetics of 117 isolates of S. turcica collected from seven of the main sweet corn-growing regions in Fujian Province, based on multiple polymerase chain reaction analyses using two mating type-specific primer pairs and 11 inter-simple sequence repeat markers. Furthermore, we examined the mode of reproduction of Fujian S. turcica populations. Both MAT1-1 and MAT1-2 mating types were detected throughout all seven sampling locations. The majority of MAT1-2 isolates were detected from Dongyou, Jian’ou, Pingnan, Songxi, and Longyan, whereas a large proportion of the detected MAT1-1 isolates were among those collected from Dongfeng and Nanjing. Furthermore, we detected five shared multi-locus haplotypes among S. turcica isolates from Dongyou, Jian’ou, Pingnan, Nanjing, and Songxi, whereas no shared haplotypes were observed between the Dongfeng (or Longyan) population and these five populations. Pairwise comparisons of the indices ΦPT and Nm, and population structure and principal coordinate analyses indicated genetic differentiation between both the regional and the mating type populations of S. turcica in Fujian. The skewed mating type ratio associated with low a haplotypic diversity and evident linkage disequilibrium reveals a mixed reproductive strategy for S. turcica populations in Fujian Province. The findings of this study advance our current understanding of the genetic diversity, population structure, and reproductive strategies of S. turcica populations infecting sweet corn in Fujian Province, and will potentially contribute to further resistance breeding efforts.

Genetic Diversity and Population Structure of Phyllosticta citriasiana in China.

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.

Genetic variation and mating type distribution of Rhynchosporium commune in Turkey

Rhynchosporium commune is the causal agent of barley scald disease and is an important foliar disease of barley worldwide. During 2012, 2013, and 2014, Rhynchosporium commune single-spore isolates were collected from different parts of Turkey, and from those, 60 isolates were selected. The mating type distribution and prevalence of these selected isolates were determined. Twenty-one of the 60 single-spore isolates were found to be MAT1-1 type and 39 isolates were MAT1-2 type χ2 (1:1) types = 5.40 (P = 0.02). The mating type ratio showed that R. commune populations may have predominantly asexual reproduction in Turkey. To determine the genetic diversity of R. commune isolates, 5 single-spore isolates from different regions of Turkey were tested with 72 ISSR primers. Eleven primers were found to be polymorphic. UBC 826, UBC 861, UBC 886, UBC 887, UBC 890, UBC 848, UBC 884 and UBC 850 primers showed optimal band numbers and good band intensities. Sixty isolates were evaluated with these 8 ISSR primers. UBC826 and UBC886 primers amplified the largest numbers of bands. The genetic diversity of the isolates in the R. commune population in Turkey appeared to be low. There were no definite groupings of the mating type or geographical region. Turkey is one of the gene centers of barley and this study is the first one addressing the genetic variation and mating type distribution of R. commune in Turkey. The low genetic diversity and possible asexual reproduction observed in our study may contribute to the hypothesis that R. commune did not evolve in the origin of the barley plant.

Genetic diversity of Australian Fusarium pseudograminearum populations causing crown rot in wheat

Fusarium crown rot (FCR) caused by the fungus Fusarium pseudograminearum (Fp) is an important disease of wheat that reduces yield and grain quality in many countries, including Australia. In this study, we investigated mating type idiomorph composition, putative chemotype and population genotypic structure of 98 Fp isolates from Western Australia (WA) and the eastern Australian states of New South Wales, Victoria and South Australia. Our results revealed the expected 1:1 mating type ratio for isolates from the eastern states while there was significant variation to a 1:1 mating type composition with isolates from WA. A chemotype-specific PCR assay indicated that all Fp isolates from eastern states and WA segregated for the 3-ADON trichothecene chemotype. Genetic diversity assessed using 21 cleaved amplified polymorphic sequence markers revealed a high level of genotypic variation within and between Fp populations from eastern Australian states and WA. Analysis of molecular variance (AMOVA) showed significant difference between populations from eastern states and WA. The genetic diversity measured by Shannon index ranged from 0.95 to 2.30 with the lowest and highest values detected in Hart and Rowena populations, respectively in the eastern states. Index of association analysis showed no significant linkage of markers among isolates within 60% of the populations, suggesting that sexual reproduction may be occurring in the pathogen from those locations. These results improve understanding of Fp population dynamics across Australia and highlight the importance of monitoring for shifts in the population which could have implications for disease management.

Distribution and frequency of mating types of Rhynchosporium commune in central Alberta

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 types (MAT1-1 and MAT1-2). The occurrence of R. commune mating types in Canada has not been determined previously. Infected leaves from barley cultivars in seven commercial fields and three breeding nurseries in central Alberta were collected and subjected to multiplex PCR to assess the presence of scald mating types within the lesions. Mating types were determined in 410 scald lesions from 288 leaves. Over 10% of lesions contained both mating types, and 44% of leaves with more than one lesion analyzed were colonized by both mating types. The mating types significantly deviated from the 1:1 ratio that would have been expected with frequent sexual reproduction. MAT1-2 was more common in six of the ten locations and for the region as a whole. The presence of both mating types, often in close proximity to each other, suggests that the cryptic sexual stage could be present in Alberta if it does occur. The skewed mating type ratio does suggest, however, that asexual reproduction is predominant within the pathogen population.

Global Sexual Fertility in the Opportunistic Pathogen Aspergillus fumigatus and Identification of New Supermater Strains.

A sexual cycle in Aspergillus fumigatus was first described in 2009 with isolates from Dublin, Ireland. However, the extent to which worldwide isolates can undergo sexual reproduction has remained unclear. In this study a global collection of 131 isolates was established with a near 1:1 ratio of mating types. All isolates were crossed to MAT1-1 or MAT1-2 Irish strains, and a subset of isolates from different continents were crossed together. Ninety seven percent of isolates were found to produce cleistothecia with at least one mating partner, showing that sexual fertility is not limited to the Irish population but is a characteristic of global A. fumigatus. However, large variation was seen in numbers of cleistothecia produced per cross, suggesting differences in the possibility for genetic exchange between strains in nature. The majority of crosses produced ascospores with >50% germination rates, but with wide variation evident. A high temperature heat shock was required to induce ascospore germination. Finally, a new set of highly fertile MAT1-1 and MAT1-2 supermater strains were identified and pyrimidine auxotrophs generated for community use. Results provide insights into the potential for the A. fumigatus sexual cycle to generate genetic variation and allow gene flow of medically important traits.

Molecular diversity of the Fusarium fujikuroi species complex from maize

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.

Distribution and genetic diversity of Dothistroma septosporum in Pinus brutia forests of south‐western Turkey

AbstractDothistroma needle blight (DNB) is a serious disease of the Pinaceae, mainly Pinus species, caused by the fungi Dothistroma septosporum and D. pini. Both species are regarded as invasive forest pathogens worldwide, with rising incidence in central and northern Europe over the last three decades. In this work, 29 sites were investigated between 2013 and 2015 in south‐western Turkey. Morphological examination of needles confirmed DNB infection (i.e., Dothistroma conidiospores observed) at 18 sites, and a total of 108 Dothistroma sp. isolates were obtained from 11 of the sites. Host age seemed to be an important factor in both occurrence and severity of DNB in Pinus brutia forests. Continuous rainy days, especially in December, may increase severity of disease; however, extreme rain events may reduce available conidiospores on plant tissues or in the air. Species‐specific mating type primers showed that all isolates were D. septosporum; D. pini was not detected. The mating type ratio was close to 1:1, indicating sexual recombination was occurring. Eleven microsatellite markers revealed 59 unique multilocus haplotypes (MLHs) among the 73 isolates originating from different conidiomata. The majority of MLHs were represented by a single isolate (n = 52) and only one MLH was shared between two localities. Analyses showed high genetic diversity, isolation‐by‐distance, and clear population clusters. These findings suggest that D. septosporum is well established in south‐western Turkey and is probably not a recent introduction.

Species diversity, mating type assays and aggressiveness patterns of Monilinia pathogens causing brown rot of peach fruit in Turkey

Brown rot disease caused by fungal species from the Monilinia genus has recently been observed as one of the most important limiting factors for yield and quality of peach fruits in Turkey. During June and July of 2018, field trips to different peach orchards were performed in six provinces located in four different geographical regions of Turkey. One hundred and twenty-nine isolates were obtained from the diseased fruits, and of those, 109 were identified as Monilinia fructicola, whereas 20 were M. laxa according to the species-specific molecular markers. Mating types of the isolates were detected by PCR assays using primers designed in this study. Each isolate represented only one of the mating type genes. Overall data sets presented 1:1 ratio of mating types for both species, indicating a possible sexual reproduction. No polymorphisms were detected in partial sequences of mating type genes. The isolates were in vitro tested for their pathogenicity using apple fruits as a host and aggressiveness were evaluated at different time points using lesion sizes. Monilinia laxa was found more aggressive than M. fructicola. Moreover, M. fructicola isolates displayed larger colony growth compared to M. laxa isolates. However, no significant correlation was detected between the colony growth rates and virulence. This study is the first to demonstrate brown rot pathogens of peach fruit in Turkey and identifies different aspects of the pathogens, which would be useful in containment of pathogen spread and comparison with other populations of these pathogens worldwide, and suggests a new set of mating type markers for these species.