Mating Compatibility Research Articles

Two non-target recessive genes confer resistance to the anti-oomycete microtubule inhibitor zoxamide in Phytophthora capsici.

This study characterized isolates of P. capsici that had developed a novel mechanism of resistance to zoxamide, which altered the minimum inhibition concentration (MIC) but not the EC50. Molecular analysis revealed that the β-tubulin gene of the resistant isolates contained no mutations and was expressed at the same level as in zoxamide-sensitive isolates. This suggested that P. capsici had developed a novel non-target-site-based resistance to zoxamide. Analysis of the segregation ratio of zoxamide-resistance in the sexual progeny of the sensitive isolates PCAS1 and PCAS2 indicated that the resistance to zoxamide was controlled by one or more recessive nuclear genes. Furthermore, the segregation of resistance in the F1, F2, and BC1 progeny was in accordance with the theoretical ratios of the χ2 test (P>0.05), which suggested that the resistance to zoxamide was controlled by two recessive genes, and that resistance to zoxamide occurred when at least one pair of these alleles was homozygous. This implies that the risk of zoxamide-resistance in P. capsici is low to moderate. Nevertheless this potential for resistance should be monitored closely, especially if two compatible mating types co-exist in the same field.

Clonality, recombination, and hybridization in the plumbing-inhabiting human pathogen Fusarium keratoplasticum inferred from multilocus sequence typing.

BackgroundRecent work has shown that Fusarium species and genotypes most commonly associated with human infections, particularly of the cornea (mycotic keratitis), are the same as those most commonly isolated from plumbing systems. The species most dominant in plumbing biofilms is Fusarium keratoplasticum, a cosmopolitan fungus known almost exclusively from animal infections and biofilms. To better understand its diversity and population dynamics, we developed and utilized a nine-locus sequence-based typing system to make inferences about clonality, recombination, population structure, species boundaries and hybridization.ResultsHigh levels of genetic diversity and evidence for recombination and clonality were detected among 75 clinical and 156 environmental isolates of Fusarium keratoplasticum. The multilocus sequence typing system (MLST) resolved 111 unique nine-locus sequence types (STs). The single locus bifactorial determinants of mating compatibility (mating types MAT1-1 and MAT1-2), were found in a ratio of 70:30. All but one of the 49 isolates of the most common ST (FSSC 2d-2) came from human infections, mostly of the cornea, and from biofilms associated with contact lenses and plumbing surfaces. Significant levels of phylogenetic incongruence were found among loci. Putative clonal relationships among genotypes were estimated, showing a mixture of large clonal complexes and unrelated singletons. Discordance between the nuclear ribosomal rRNA and other gene genealogies is consistent with introgression of ribosomal RNA alleles of phylogenetic species FSSC 9 into F. keratoplasticum. No significant population subdivision based on clinical versus non-clinical sources was found.ConclusionsIncongruent phylogenetic trees and the presence of both mating types within otherwise identical STs were observed, providing evidence for sexuality in F. keratoplasticum. Cryptic speciation suggested in a published three-locus MLST system was not supported with the addition of new loci, but evidence of introgression of ribosomal RNA genes from another strongly supported phylogenetic species (FSSC 9), also known from plumbing systems and human infections, was detected in two isolates. Overall, F. keratoplasticum is a diverse and geographically unstructured species with a mixed clonal and recombinant life history.

Evaluation of mating behaviour and mating compatibility methods for the Old World screwworm fly, Chrysomya bezziana

The effectiveness of the Sterile Insect Technique program (SIT) to eradicate pest insects relies on the success of mating competitiveness between irradiated male flies and wild type males for the wild type females. It has been successfully applied for the New World screwworm fly (NWSF), Cochliomyia hominivorax but remains unproven for the Old World screwworm fly (OWSF), Chrysomya bezziana. The aim of the study was to develop methods for investigating mating behaviour and mating compatibility of C. bezziana under laboratory conditions. Two methods were used for studying mating: individual mating (method 1) and group mating (method 2). The flies used in this study were 5-7 days old. Twenty four hours after emergence, adult flies were sexed and placed into different cages until studied. The female : male ratio in the group mating was 1 : 5 and the males were marked by painting a dot on the thorax using different oil colours. Observation of mating behaviour was investigated every 30 minutes through 10-20 replications for all methods depending on the availability of flies. Data were analysed using ANOVA and the Student’s t-test, with significance demonstrated at the 95% confidence level. The results demonstrated that the frequency of contacts between males and females at different ages was a significantly different (p 0.05) and method 2 (p > 0.05). Copulation was only initiated following longer periods of contact, mainly in the range of 270-449 seconds. The highest frequency of copulation occurred between 7-8 days, but the duration of mating was similar between 5-8 days old. The study demonstrated that the methods developed were suitable for a mating compatibility study of C. bezziana. Key Words: Chrysomya bezziana, Mating Behaviour, Copulation, Myiasis

Unseen sex in ancient virgin fungi

Unseen sex in ancient virgin fungi

A Brave New World for an Old World Pest: Helicoverpa armigera (Lepidoptera: Noctuidae) in Brazil

The highly polyphagous Old World cotton bollworm Helicoverpa armigera is a quarantine agricultural pest for the American continents. Historically H. armigera is thought to have colonised the American continents around 1.5 to 2 million years ago, leading to the current H. zea populations on the American continents. The relatively recent species divergence history is evident in mating compatibility between H. zea and H. armigera under laboratory conditions. Despite periodic interceptions of H. armigera into North America, this pest species is not believed to have successfully established significant populations on either continent. In this study, we provide molecular evidence via mitochondrial DNA (mtDNA) cytochrome oxidase I (COI) and cytochrome b (Cyt b) partial gene sequences for the successful recent incursion of H. armigera into the New World, with individuals being detected at two sites (Primavera do Leste, Pedra Preta) within the State of Mato Grosso in Brazil. The mtDNA COI and Cyt b haplotypes detected in the Brazilian H. armigera individuals are common throughout the Old World, thus precluding identification of the founder populations. Combining the two partial mtDNA gene sequences showed that at least two matrilines are present in Brazil, while the inclusion of three nuclear DNA Exon-Primed Intron-Crossing (EPIC) markers identified a further two possible matrilines in our samples. The economic, biosecurity, resistance management, ecological and evolutionary implications of this incursion are discussed in relation to the current agricultural practices in the Americas.

Experimental hybridization and backcrossing reveal forces of reproductive isolation in Microbotryum

BackgroundHybridization and reproductive isolation are central to the origin and maintenance of species, and especially for sympatric species, gene flow is often inhibited through barriers that depend upon mating compatibility factors. The anther-smut fungi (genus Microbotryum) serve as models for speciation in the face of sympatry, and previous studies have tested for but not detected assortative mating. In addition, post-mating barriers are indicated by reduced fitness of hybrids, but sources of those barriers (i.e. ecological maladaptation or genetic incompatibilities) have not yet been detected. Here, backcrossing experiments, specifically controlling for the fungal species origins of the mating compatibility factors, were used to investigate reproductive isolation in the recently-derived species Microbotryum lychnidis-dioicae and Microbotryum silenes-dioicae.ResultsAssortative mating was detected during backcrossing and was manifested by the preferential conjugation of the hybrid-produced gametes with non-hybrid gametes containing mating compatibility factors from the same parental species. Patterns of post-mating performance supported either a level of extrinsic isolation mechanism, where backcross progeny with a higher proportion of the pathogen genome adapted to the particular host environment were favored, or an infection advantage attributed to greater genetic contribution to the hybrid from the M. lychnidis-dioicae genome.ConclusionThe use of controlled backcrossing experiments reveals significant species-specific mating type effects on conjugations between recently-derived sister species, which are likely to play important roles in both maintaining species separation and the nature of hybrids lineages that emerge in sympatry between Microbotryum species.

The Implementation of the Golden Ratio to the Healthy Human Heart

Abstract Sexual development is a key evolutionary innovation of eukaryotes. In many species, mating involves interaction between compatible mating partners that can undergo cell and nuclear fusion and subsequent steps of development including meiosis. Mating compatibility in fungi is governed by mating type determinants, which are localized at mating type (MAT) loci. In basidiomycetes, the ancestral state is hypothesized to be tetrapolar (bifactorial), with two genetically unlinked MAT loci containing homeodomain transcription factor genes (HD locus) and pheromone and pheromone receptor genes (P/R locus), respectively. Alleles at both loci must differ between mating partners for completion of sexual development. However, there are also basidiomycete species with bipolar (unifactorial) mating systems, which can arise through genomic linkage of the HD and P/R loci. In the order Tremellales, which is comprised of mostly yeast-like species, bipolarity is found only in the human pathogenic Cryptococcus species. Here, we describe the analysis of MAT loci from the Trichosporonales, a sister order to the Tremellales. We analyzed genome sequences from 29 strains that belong to 24 species, including two new genome sequences generated in this study. Interestingly, in all of the species analyzed, the MAT loci are fused and a single HD gene is present in each mating type. This is similar to the organization in the pathogenic Cryptococci, which also have linked MAT loci and carry only one HD gene per MAT locus instead of the usual two HD genes found in the vast majority of basidiomycetes. However, the HD and P/R allele combinations in the Trichosporonales are different from those in the pathogenic Cryptococcus species. The differences in allele combinations compared to the bipolar Cryptococci as well as the existence of tetrapolar Tremellales sister species suggest that fusion of the HD and P/R loci and differential loss of one of the two HD genes per MAT allele occurred independently in the Trichosporonales and pathogenic Cryptococci. This finding supports the hypothesis of convergent evolution at the molecular level towards fused mating-type regions in fungi, similar to previous findings in other fungal groups. Unlike the fused MAT loci in several other basidiomycete lineages though, the gene content and gene order within the fused MAT loci are highly conserved in the Trichosporonales, and there is no apparent suppression of recombination extending from the MAT loci to adjacent chromosomal regions, suggesting different mechanisms for the evolution of physically linked MAT loci in these groups. Author summary Sexual development in fungi is governed by genes located within a single mating type (MAT) locus or at two unlinked MAT loci. While the latter is thought to be the ancestral state in basidiomycetes, physical linkage of the two MAT loci has occurred multiple times during basidiomycete evolution. Here, we show that physically linked MAT loci are present in all analyzed species of the basidiomycete order Trichosporonales. In contrast to previously studied basidiomycetes, the fused MAT loci in the Trichosporonales have highly conserved gene order, suggesting that this fusion might date back to the common ancestor of this lineage.

Pheromone Analyses of theAnastrepha fraterculus(Diptera: Tephritidae) Cryptic Species Complex

ABSTRACT The South American fruit fly Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae) cryptic species complex is presently one of the most studied pest models in terms of speciation and population mating compatibility. The improvement of pest-control techniques has strongly relied on successful implementation of laboratory strains into wild populations. Pheromone communication plays an important role in the mating process in the South American fruit fly. Therefore, the main goal of the present study was to investigate the pheromone composition of 7 different populations, originating from geographically distant locations in Brazil and Argentina. Fourteen volatile compounds were identified in calling male emanations by GC×GC/TOF-MS and the data obtained were subsequently analyzed by multivariate statistics. The pheromone composition varied both quantitatively and qualitatively among the studied populations. These results will serve as the basis for further electrophysiological analyses.

Rapid genotyping by low-coverage resequencing to construct genetic linkage maps of fungi: a case study in Lentinula edodes

BackgroundGenetic linkage maps are important tools in breeding programmes and quantitative trait analyses. Traditional molecular markers used for genotyping are limited in throughput and efficiency. The advent of next-generation sequencing technologies has facilitated progeny genotyping and genetic linkage map construction in the major grains. However, the applicability of the approach remains untested in the fungal system.FindingsShiitake mushroom, Lentinula edodes, is a basidiomycetous fungus that represents one of the most popular cultivated edible mushrooms. Here, we developed a rapid genotyping method based on low-coverage (~0.5 to 1.5-fold) whole-genome resequencing. We used the approach to genotype 20 single-spore isolates derived from L. edodes strain L54 and constructed the first high-density sequence-based genetic linkage map of L. edodes. The accuracy of the proposed genotyping method was verified experimentally with results from mating compatibility tests and PCR-single-strand conformation polymorphism on a few known genes. The linkage map spanned a total genetic distance of 637.1 cM and contained 13 linkage groups. Two hundred sequence-based markers were placed on the map, with an average marker spacing of 3.4 cM. The accuracy of the map was confirmed by comparing with previous maps the locations of known genes such as matA and matB.ConclusionsWe used the shiitake mushroom as an example to provide a proof-of-principle that low-coverage resequencing could allow rapid genotyping of basidiospore-derived progenies, which could in turn facilitate the construction of high-density genetic linkage maps of basidiomycetous fungi for quantitative trait analyses and improvement of genome assembly.

Colonized Aedes albopictus and its sexual performance in the wild: implications for SIT technology and containment

BackgroundMating is a physiological process of crucial importance underlying the size and maintenance of mosquito populations. In sterile and incompatible insect technologies (SIT and IIT), mating is essential for mass production, persistence, and success of released individuals, and is a central parameter for judging the effectiveness of SIT/IIT programs. Some mosquitoes have an enormous reproductive potential for both themselves and pathogens and mating may contribute to persistence of infection in nature. As Aedes albopictus can transmit flaviviruses both sexually and horizontally, and as infected insects are usually derived from laboratory colonies, we investigated the implications of mating between a long-term laboratory colony of Ae. albopictus and wild populations.MethodsThrough a series of mating experiments, we examined the reproductive outcomes of sexual cross-affinity between laboratory-raised and wild adults of Ae. albopictus.ResultsThe results indicated appreciable mating compatibility between laboratory-reared and wild adults, and equivalent levels of egg production among reciprocal crosses. We also observed comparable larval eclosion in lab females mated with wild males, and increased adult longevity in female offspring from wild females|×|laboratory males crosses.ConclusionsTaken together, these data suggest that Ae. albopictus can preserve its reproductive fitness over a long period of time in the laboratory environment and has valuable attributes for SIT application. These observations together with the ability to successfully inseminate heterospecific females indicate the potential of Ae. albopictus to act as an ecological barrier if non-sterilized males are massively released in areas occupied by Aedes aegypti. The observed substantial reproductive fitness combined with the capability to reproduce both, itself and viruses illustrates the potential of Ae. albopictus to pose a serious threat if infected and released accidentally.

Evolution of uni- and bifactorial sexual compatibility systems in fungi

Mating systems, that is, whether organisms give rise to progeny by selfing, inbreeding or outcrossing, strongly affect important ecological and evolutionary processes. Large variations in mating systems exist in fungi, allowing the study of their origin and consequences. In fungi, sexual incompatibility is determined by molecular recognition mechanisms, controlled by a single mating-type locus in most unifactorial fungi. In Basidiomycete fungi, however, which include rusts, smuts and mushrooms, a system has evolved in which incompatibility is controlled by two unlinked loci. This bifactorial system probably evolved from a unifactorial system. Multiple independent transitions back to a unifactorial system occurred. It is still unclear what force drove evolution and maintenance of these contrasting inheritance patterns that determine mating compatibility. Here, we give an overview of the evolutionary factors that might have driven the evolution of bifactoriality from a unifactorial system and the transitions back to unifactoriality. Bifactoriality most likely evolved for selfing avoidance. Subsequently, multiallelism at mating-type loci evolved through negative frequency-dependent selection by increasing the chance to find a compatible mate. Unifactoriality then evolved back in some species, possibly because either selfing was favoured or for increasing the chance to find a compatible mate in species with few alleles. Owing to the existence of closely related unifactorial and bifactorial species and the increasing knowledge of the genetic systems of the different mechanisms, Basidiomycetes provide an excellent model for studying the different forces that shape breeding systems.

Distribution of chestnut blight and diversity of Cryphonectria parasitica in chestnut forests in Bulgaria

SummaryWe surveyed chestnut stands at 18 sites in 11 locations in Bulgaria in 2005 and 2007 for the presence of chestnut blight. We found chestnut blight in seven locations (Belogradchik, Berkovitsa, Brezhani, Barziya, Govezhda, Petrich and Petrovo) but not in four others (Tsaparevo, Kresna, Dupnica and Botevgrad). We successfully isolated Cryphonectria parasitica from cankers on 606 trees with symptoms of chestnut blight and assayed them for vegetative compatibility (vc) types and mating type. Three vc types were identified among the 606 isolates; all three were among the European vc types with known vegetative incompatibility (vic) genotypes. Vc type EU‐12 was the most common, representing 80% of the isolates, and was found at all locations with blight, with the exception of Belogradchik in north‐west Bulgaria, where all isolates were vc type EU‐2. Only one population (Barziya) had more than one vc type, with a combination of EU‐12 and EU‐10 in almost equal frequencies. Similarly, the diversity of mating types was very low. All but three of 536 isolates assayed were in mating‐type MAT‐1; MAT‐2 was only found in one population in the north‐west (Berkovitsa). We inspected 671 bark samples from chestnut blight cankers with stromata of C. parasitica and found perithecia in only 33, of which 28 were from Berkovitsa where MAT‐2 was present. We did not detect hypoviruses in any of the 270 isolates screened using the standard double‐stranded RNA extraction protocol. Similar to results from previous studies in south‐eastern Europe, the diversity of vc types and mating type of C. parasitica in Bulgaria is low, and reproduction of the fungus is mainly asexual. Unfortunately, naturally occurring hypovirulence was not detected. Nevertheless, we observed a small number of superficial cankers typical of those caused by C. parasitica isolates infected with a hypovirus.

A central role for Ras1 in morphogenesis of the basidiomycete Schizophyllum commune.

Fungi have been used as model systems to define general processes in eukaryotes, for example, the one gene-one enzyme hypothesis, as well as to study polar growth or pathogenesis. Here, we show a central role for the regulator protein Ras in a mushroom-forming, filamentous basidiomycete linking growth, pheromone signaling, sexual development, and meiosis to different signal transduction pathways. ras1 and Ras-specific gap1 mutants were generated and used to modify the intracellular activation state of the Ras module. Transformants containing constitutive ras1 alleles (ras1(G12V) and ras1(Q61L)), as well as their compatible mating interactions, did show strong phenotypes for growth (associated with Cdc42 signaling) and mating (associated with mitogen-activated protein kinase signaling). Normal fruiting bodies with abnormal spores exhibiting a reduced germination rate were produced by outcrossing of these mutant strains. Homozygous Δgap1 primordia, expected to experience increased Ras signaling, showed overlapping phenotypes with a block in basidium development and meiosis. Investigation of cyclic AMP (cAMP)-dependent protein kinase A indicated that constitutively active ras1, as well as Δgap1 mutant strains, exhibit a strong increase in Tpk activity. Ras1-dependent, cAMP-mediated signal transduction is, in addition to the known signaling pathways, involved in fruiting body formation in Schizophyllum commune. To integrate these analyses of Ras signaling, microarray studies were performed. Mutant strains containing constitutively active Ras1, deletion of RasGap1, or constitutively active Cdc42 were characterized and compared. At the transcriptome level, specific regulation highlighting the phenotypic differences of the mutants is clearly visible.

Mating Compatibility Among Four Pest Members of the <I>Bactrocera dorsalis</I> Fruit Fly Species Complex (Diptera: Tephritidae)

Bactrocera dorsalis (Hendel), Bactrocera papayae Drew & Hancock, Bactrocera philippinensis Drew & Hancock, and Bactrocera carambolae Drew & Hancock are pest members within the B. dorsalis species complex of tropical fruit flies. The species status of these taxa is unclear and this confounds quarantine, pest management, and general research. Mating studies carried out under uniform experimental conditions are required as part of resolving their species limits. These four taxa were collected from the wild and established as laboratory cultures for which we subsequently determined levels of prezygotic compatibility, assessed by field cage mating trials for all pair-wise combinations. We demonstrate random mating among all pair-wise combinations involving B. dorsalis, B. papayae, and B. philippinensis. B. carambolae was relatively incompatible with each of these species as evidenced by nonrandom mating for all crosses. Reasons for incompatibility involving B. carambolae remain unclear; however, we observed differences in the location of couples in the field cage for some comparisons. Alongside other factors such as pheromone composition or other courtship signals, this may lead to reduced interspecific mating compatibility with B. carambolae. These data add to evidence that B. dorsalis, B. papayae, and B. philippinensis represent the same biological species, while B. carambolae remains sufficiently different to maintain its current taxonomic identity. This poses significant implications for this group's systematics, impacting on pest management, and international trade.

민자주방망이버섯(Lepista nuda )의 형태적 특성 및 교배에 관한 연구

In this study, twelve of Lepista nuda were collected from various localities in Korea. Also thirteen exotic L. nuda species were collected from Japan, France, Switzerland and Portugal. Spores were isolated under optical microscope. These spores were placed on the surface of YM medium for inducing to germination. Eleven mating-groups were selected by morpho- logical characters of fruit body such as size, color and stipe patterns. Intra-isolate crosses were made between two single-spore isolates derived from mating-groups. Also, dikaryotic crossing using the isolates from L. nuda were carried out to evaluated tetrakaryon formation. Cross-mating compatibility tests also verified its dikaryotic state by microscopic or molecular genetic observation of clamp connection and Random Amplified Polymorphic DNA (RAPD) band pattern. To analyze the growth rate of hybrids and parents mycelium in dikaryons obtained from compatible mating groups were placed on PDA medium. Intra-isolate crosses determined eleven mating-groups within L. nuda. The typical clamp connection were mostly observed in mating-groups of Korean L. nuda in K1xK2, K1xK3, K1xK4, K1xK6, K1xK5, K2xK4, K2xK3, K2xK6, K3xK4, K4xK5 and K4xK6. Korean L. nuda type of dikaryon, shown to cross-incompatibility with L. sordida, it seemed that mating induce more rapidly than wild types in a view of growth rate. In conclusion, it would be useful to improve mass production with better mor- phological characteristics through a special mating of L. nuda.

Mating compatibility of wild and sterile melon flies, Bactrocera cucurbitae (Diptera: Tephritidae) treated with entomopathogenic fungi

AbstractField cage tests were carried out to determine the mating compatibility of sterile and wild melon fly, Bactrocera cucurbitae (Coquillett), males that were treated or not treated with entomopathogenic fungi. The melon flies were irradiated at 70 Gy, and the adults were treated with dry conidia of either Beauveria bassiana (Balsamo) or Metarhizium anisopliae (Metchnikoff) Sorokin 1 day before the mating test. Our results demonstrate that sterile melon flies treated with either B. bassiana or M. anisopliae showed mating compatibility with the wild flies. Inoculation of the flies with the fungi did not significantly affect the mean time of copulation (323–567 min) as compared to mating involving only wild flies (407–518 min). Treatment of the flies with either B. bassiana or M. anisopliae did not significantly affect the propensity of mating. Wild females mated indiscriminately with sterile, sterile B. bassiana‐treated or sterile M. anisopliae‐treated males. Irrespective of the treatment combinations, mean percentage remating was significantly higher for sterile females with sterile males (7.72–27.12%) as compared to wild females with sterile males (0.76–14.90%). The possibility of using sterile melon flies as vectors of the entomopathogenic fungi to enhance an SIT programme is discussed.

Alternative Mating Tactics as Potential Prezygotic Barriers to Gene Flow Between Two Sister Species of Frugivorous Fruit Flies

Rhagoletis completa Cresson and Rhagoletis zoqui Bush, are two sister species in the suavis group that were thought to occupy non-overlapping geographical ranges. Recent discovery of a contact zone in North Eastern Mexico where natural interspecific hybrids with intermediate morphotypes can be found has led to laboratory studies in small enclosures that failed to identify the existence of reproductive barriers capable of maintaining the genetic integrity of both species. Because the experimental approach used in such studies could have interfered with natural behavior of flies, a series of additional mating compatibility tests under much less restrictive conditions were performed. Our observations, confirmed the fact that males and females of both species engage in interspecific matings. However, R. completa females were more reluctant to engage in heterospecific matings than R. zoqui females, revealing asymmetries in sexual isolation. Also, careful scrutiny of male guarding behavior and species specific partitioning of mating location unveiled subtle differences that could result in reproductive isolation if reinforced during secondary contact.

Phytopathogenic, morphological, genetic and molecular characterization of a Verticillium dahliae population from Crete, Greece

A population of 84 V. dahliae isolates mainly originating from Crete, Greece, was characterized in terms of pathogenicity and virulence on different hosts, in parallel with morphological/physiological characterization, vegetative compatibility grouping and mating type determination. Tomato race 2 was found to have supplanted race 1 and was more virulent on a tomato-susceptible cultivar than race 1. Using a differential host classification system which tests pathogenicity to tomato, eggplant, sweet pepper and turnip, 59 isolates were assigned to tomato, 19 to eggplant, one to sweet pepper and five to tomato-sweet pepper pathogenicity groups. All isolates from Crete fell into VCG subgroups 2A, 2B and 4B, while a remarkably high incidence of bridging isolates (compatible with two or more VCGs) was recorded. The tomato-sweet pepper pathogenicity group was morphologically quite distinct from the others, while conidial length and pigment intensity were discriminatory parameters among VCGs 2A, 2B and 4B. PCR-based molecular marker Tr1/Tr2 was reliable in race prediction among tomato-pathogenic isolates, except for members of VCG 4B, while the application of markers Tm5/Tm7 and 35-1/35-2 was highly successful for tomato-pathogenic isolates. E10 marker was related to VCG 2B, rather than to pathogenicity groups. A single nucleotide polymorphism in the ITS2 region, and two novel molecular markers, M1 and M2, proved useful for the fast and accurate determination of major VCGs 2A, 2B and 4B, and can be used for high-throughput population analyses in future studies. The mating type was unrelated to VCG classification and probably does not control heterokaryon incompatibility in V. dahliae.

Survival and Germination of Overwintering Oospores of <i>Phytophthora infestans</i> in the Field

Survival and germination of overwintering oospores of two strains from Gansu,China compared with two standard from Wageningen, the Netherlands of Phytophthora infestans were determined in 2010-2011. It was found that compatible mating strains of P. infestans A1 and A2 produce oospores abundantly in paired cultures on tomato-rye agar medium.The survival rate of overwintering was ranged from19.57% to 24.59%.The germination rate of overwintering was ranged from 3.58% to 8.49%, and there were no significant differences in germination rate of oospores between alternating light at day/night and whole darkness at day/night. Furthermore, the soil extract liquid was more suitable for oospore germination than distilled water.

Genetic variation and reproductive mode in two epiphytic lichens of conservation concern: a transatlantic study ofEvernia divaricataandUsnea longissima

North European epiphytic lichens are often genetically impoverished compared with their North American counterparts. This has been hypothesized to impede sexual reproduction due to reduced chances of finding compatible mating type partners. We compared genetic variation and reproductive mode in two threatened Scandinavian lichens, Evernia divaricata and Usnea longissima, with more viable populations in North America to see (i) if these species also show genetical depletion in northern Europe and (ii) if the occurrence of sexual propagules (ascospores in apothecia) is more prevalent in genetically diverse populations. Genetic variation of the fungal component was assessed by sequencing two nuclear rDNA gene regions (ITS and IGS) in 1005 and 1477 thalli, collected from 92 and 160 localities of E. divaricata and U. longissima, respectively. Scandinavian populations of both species were almost devoid of genetic variation compared with much higher genetic diversity in North America. We found no support for the proposed relationship between genetic diversity and fertility. Fertile thalli were found in several genetically invariable populations. Fertility increased with population size and regional abundance in E. divaricata, but not in U. longissima. In Scandinavia, E. divaricata was more fertile than previously recorded, whereas all sampled populations of U. longissima were sterile and possibly clonal.