Fungal Mating Research Articles

Tracing the Origin of the Fungal α1 Domain Places Its Ancestor in the HMG-Box Superfamily: Implication for Fungal Mating-Type Evolution

BackgroundFungal mating types in self-incompatible Pezizomycotina are specified by one of two alternate sequences occupying the same locus on corresponding chromosomes. One sequence is characterized by a gene encoding an HMG protein, while the hallmark of the other is a gene encoding a protein with an α1 domain showing similarity to the Matα1p protein of Saccharomyces cerevisiae. DNA-binding HMG proteins are ubiquitous and well characterized. In contrast, α1 domain proteins have limited distribution and their evolutionary origin is obscure, precluding a complete understanding of mating-type evolution in Ascomycota. Although much work has focused on the role of the S. cerevisiae Matα1p protein as a transcription factor, it has not yet been placed in any of the large families of sequence-specific DNA-binding proteins.Methodology/Principal FindingsWe present sequence comparisons, phylogenetic analyses, and in silico predictions of secondary and tertiary structures, which support our hypothesis that the α1 domain is related to the HMG domain. We have also characterized a new conserved motif in α1 proteins of Pezizomycotina. This motif is immediately adjacent to and downstream of the α1 domain and consists of a core sequence Y-[LMIF]-x(3)-G-[WL] embedded in a larger conserved motif.Conclusions/SignificanceOur data suggest that extant α1-box genes originated from an ancestral HMG gene, which confirms the current model of mating-type evolution within the fungal kingdom. We propose to incorporate α1 proteins in a new subclass of HMG proteins termed MATα_HMG.

Molecular epidemiology and virulence assessment of Aspergillus fumigatus isolates from white stork chicks and their environment

Aspergillus fumigatus is a common pathogen in poultry and captive wild birds and an emerging opportunistic fungal pathogen in immunocompromised humans. Although invasive aspergillosis is frequently reported in free-ranging wild birds, the incidence and epidemiology of the disease in a natural setting is unknown. We recently reported endemic outbreaks of invasive aspergillosis at white stork nesting sites close to human habitation in Germany with significant subsequent breeding losses. Therefore, we hypothesized that A. fumigatus strains with higher virulence in birds may have evolved in this environment and performed the first epidemiological analysis of invasive aspergillosis in free-ranging wild birds. Sixty-one clinical and environmental A. fumigatus isolates from six affected nesting sites were genotyped by microsatellite analysis using the STR Af-assay. The isolates showed a remarkable high genomic diversity and, contrary to the initial hypothesis, clinical and environmental isolates did not cluster significantly. Interestingly, storks were infected with two to four different genotypes and in most cases both mating types MAT-1.1 and MAT-1.2 were present within the same specimen. The majority of selected clinical and environmental strains exhibited similar virulence in an in vivo infection model using embryonated chicken eggs. Noteworthy, virulence was not associated with one distinct fungal mating type. These results further support the assumption that the majority of A. fumigatus strains have the potential to cause disease in susceptible hosts. In white storks, immaturity of the immune system during the first three weeks of age may enhance susceptibility to invasive aspergillosis.

The Pneumocystis life cycle

First recognised as 'schizonts' of Trypanosoma cruzi, Pneumocystis organisms are now considered as part of an early-diverging lineage of Ascomycetes. As no robust long-term culture model is available, most data on the Pneumocystis cell cycle have stemmed from ultrastructural images of infected mammalian lungs. Although most fungi developing in animals do not complete a sexual cycle in vivo, Pneumocystis species constitute one of a few exceptions. Recently, the molecular identification of several key players in the fungal mating pathway has provided further evidence for the existence of conjugation and meiosis in Pneumocystisorganisms. Dynamic follow-up of stage-to-stage transition as well as studies of stage-specific proteins and/or genes would provide a better understanding of the still hypothetical Pneumocystislife cycle. Although difficult to achieve, stage purification seems a reasonable way forward in the absence of efficient culture systems. This mini-review provides a comprehensive overview of the historical milestones leading to the current knowledge available on the Pneumocystis life cycle.

First Report of Anthracnose Disease of Ornamental Feather Reed Grass (Calamagrostis × acutifolia 'Karl Foerster') Caused by Colletotrichum cereale.

Calamagrostis × acutifolia 'Karl Foerster' (feather reed grass) is a cool-season grass grown extensively as an ornamental plant throughout the United States. In July 2005, severe foliar damage was observed in feather reed grasses in a residential garden in Barrington, NJ. Symptoms were observed as small, yellowish brown, oval to irregularly shaped spots on the blades, with spread and coalescence of spots leading to eventual necrosis and plant death. Numerous acervuli with black setae diagnostic of fungi in the genus Colletotrichum were present on necrotic lesions. Two distinct fungi were isolated from diseased tissue by plating small sections of infested leaf tissue on potato dextrose agar (PDA) supplemented with 40 μg/liter each of penicillin, ampicillin, gentomycin, and streptomycin. The first fungus was identified as Colletotrichum gloeosporioides based on morphological, cultural (2), and molecular characteristics. Variable colonies of gray-white mycelia with masses of pink-to-salmon hued conidia formed at 25°C on PDA under constant light. Conidia were hyaline, aseptate, straight, and cylindrical with rounded apices (11.0 to 18.5 × 2.5 to 5.0 μm). Sequencing of the internal transcribed spacer (ITS) rDNA region showed the fungus to be most similar to C. gloeosporioides (GenBank Accession No. EU979125). The second fungus was identified as C. cereale based on morphological, cultural, and molecular characteristics (1). Variable colonies of gray-tan-white mycelia formed at 25°C on PDA under constant light and salmon-colored conidial masses surrounded numerous setae. Conidia were hyaline, aseptate, falcate, fusiform, and guttulate (15.0 to 21.5 × 2.5 to 4.5 μm). Hyphal appressoria were ovoid, sometimes lobate or multilobate (10.5 to 13.5 × 7.5 to 10.0 μm). Maximum likelihood phylogenetic analysis of the ITS sequence (GenBank Accession No. DQ126193) and fungal mating type idiomorph Mat1-2 HMG box (GenBank Accession No. DQ131962) identified the fungus as C. cereale (1). Pathogenicity was determined by inoculating healthy feather reed grasses (11.4-liter pots) established 61 cm off-center in a mulched bed. Three replicate plants per treatment were sprayed with a 20-ml conidial solution (5 × 104 conidia/ml in 0.1 potato dextrose broth) of either C. cereale or C. gloeosporioides and an uninoculated control. Temperatures ranged from 19 to 36°C and humidity varied between 31 and 79%. No symptoms were observed in uninoculated controls or plants inoculated with C. gloeosporioides. Plants inoculated with C. cereale developed disease symptoms within 21 days; the fungus was subsequently reisolated from symptomatic leaves and confirmed as C. cereale. To our knowledge, this is the first report of anthracnose of feather reed grass caused by C. cereale (formerly known as C. graminicola). Although C. cereale is known to inhabit numerous cool-season grass hosts, this is the first description of this fungus as a pathogen of an ornamental grass. Given the recent emergence of anthracnose epidemics caused by C. cereale on golf course turfgrass, the identification of this fungus as a pathogen of Calamagrostis × acutifolia highlights the need for nurseries and regulatory personnel to screen ornamental grasses such as feather reed grass for the presence of C. cereale so that the disease does not become problematic.

Using Neurospora to demonstrate the unidirectional nature of fungal mating.

Different fungi have been used to demonstrate mating in fungi including Neurospora (Griffiths, 1979), Aspergillus (Denison, 2000), and Sordaria (Fields, 1970) and kits for demonstrating mating in Sordaria are available from Carolina Biological, Inc. The present demonstration allows students to see both that fungi have different mating types and that one strain acts as the female parent. It takes advantage of the rapidity of Neurospora growth and the ease of using fluffy strains (Table 1). Strains carrying the fluffy lesion are mated with fl per-1 strains and the pattern of perethecial color on the resulting plates shows which strain was the female parent.

Pneumocystis carinii STE11, an HMG-box protein, is phosphorylated by the mitogen activated protein kinase PCM

A pheromone-induced mitogen activated protein kinase (MAPK) pathway controls mating in fungi by regulating gene transcription. In the opportunistic fungus Pneumocystis carinii, we have identified a protein containing a high-mobility group (HMG) motif which is homologous to the transcriptional activators STE11 of Schizosaccharomyces pombe and STE12 of Saccharomyces cerevisiae. In fungi, this transcriptional activator functions in sexual development, filamentous growth, and pathogenicity. The fungal pheromone-activated MAPK phosphorylates the transcriptional activator to allow binding to pheromone-response elements in the promoter regions of certain genes. We have previously identified a P. carinii MAPK, PCM, which has significant homology to fungal MAPKs involved in mating. As an initial step in understanding the downstream molecules which interact with the PCM kinase, we have cloned a STE11 homologue in P. carinii. PCSTE11 has an open-reading frame of 1.5 kb which encodes a protein of 501 amino acids with a molecular weight of 56 kDa. Greatest homology was to S. pombe STE11 (52%). We have expressed a His-tag fusion of PCSTE11 and purified the protein with nickel affinity resin. PCM phosphorylates the purified protein indicating that PCSTE11 is associated with the MAPK cascade in P. carinii.

INTRASPECIFIC COMPETITION AND MATING BETWEEN FUNGAL STRAINS OF THE ANTHER SMUT MICROBOTRYUM VIOLACEUM FROM THE HOST PLANTS SILENE LATIFOLIA AND S. DIOICA

We studied intraspecific competition and assortative mating between strains of the anther smut fungus Microbotryum violaceum from two of its host species, Silene latifolia and S. dioica. Specifically, we investigated whether strains from allopatric host populations have higher competitive ability on their native host species and show positive assortative mating. In general, strains isolated from S. latifolia outcompeted strains isolated from S. dioica on both host species, but in female hosts, heterotypic dikaryons (i.e., dikaryons composed of a haploid strain originating from S. latifolia and a haploid strain originating from S. dioica) were most successful in competition. Furthermore, the latency period was significantly shorter for heterokaryons that contained at least one strain originating from S. latifolia, compared to heterokaryons that only contained strains originating from S. dioica. The frequencies of conjugations between strains originating from S. latifolia were much higher than conjugation frequencies between strains originating from S. dioica. A significant positive correlation was detected between the relative success of strains in competition and in conjugation, suggesting that success of a strain in competition might be partly determined by its swiftness of mating. In addition, reciprocal differences within heterotypic crosses revealed a significant effect of fungal mating type, with mating type a1 being the main determinant of mating pace. The observed differences in infection success, conjugation rate, and latency period in favor of strains from S. latifolia relative to strains from S. dioica on both host species are discussed in an evolutionary context of opportunities for the maintenance of differentiation between different formae speciales upon secondary contact.

INTRASPECIFIC COMPETITION AND MATING BETWEEN FUNGAL STRAINS OF THE ANTHER SMUT MICROBOTRYUM VIOLACEUM FROM THE HOST PLANTS SILENE LATIFOLIA AND S. DIOICA

We studied intraspecific competition and assortative mating between strains of the anther smut fungus Microbotryum violaceum from two of its host species, Silene latifolia and S. dioica. Specifically, we investigated whether strains from allopatric host populations have higher competitive ability on their native host species and show positive assortative mating. In general, strains isolated from S. latifolia outcompeted strains isolated from S. dioica on both host species, but in female hosts, heterotypic dikaryons (i.e., dikaryons composed of a haploid strain originating from S. latifolia and a haploid strain originating from S. dioica) were most successful in competition. Furthermore, the latency period was significantly shorter for heterokaryons that contained at least one strain originating from S. latifolia, compared to heterokaryons that only contained strains originating from S. dioica. The frequencies of conjugations between strains originating from S. latifolia were much higher than conjugation frequencies between strains originating from S. dioica. A significant positive correlation was detected between the relative success of strains in competition and in conjugation, suggesting that success of a strain in competition might be partly determined by its swiftness of mating. In addition, reciprocal differences within heterotypic crosses revealed a significant effect of fungal mating type, with mating type a1 being the main determinant of mating pace. The observed differences in infection success, conjugation rate, and latency period in favor of strains from S. latifolia relative to strains from S. dioica on both host species are discussed in an evolutionary context of opportunities for the maintenance of differentiation between different formae speciales upon secondary contact.

Inactivation of Kex2p Diminishes the Virulence of Candida albicans

Deletion of the kexin gene (KEX2) in Candida albicans has a pleiotropic effect on phenotype and virulence due partly to a defect in the expression of two major virulence factors: the secretion of active aspartyl proteinases and the formation of hyphae. kex2/kex2 mutants are highly attenuated in a mouse systemic infection model and persist within cultured macrophages for at least 24 h without causing damage. Pathology is modest, with little disruption of kidney matrix. The infecting mutant cells are largely confined to glomeruli, and are aberrant in morphology. The complex phenotype of the deletion mutants reflects a role for kexin in a wide range of cellular processes. Taking advantage of the specificity of Kex2p cleavage, an algorithm we developed to scan the 9168 open reading frames in Assembly 6 of the C. albicans genome identified 147 potential substrates of Kex2p. These include all previously identified substrates, including eight secreted aspartyl proteinases, the exoglucanase Xog1p, the immunodominant antigen Mp65, and the adhesin Hwp1p. Other putative Kex2p substrates identified include several adhesins, cell wall proteins, and hydrolases previously not implicated in pathogenesis. Kexins also process fungal mating pheromones; a modification of the algorithm identified a putative mating pheromone with structural similarities to Saccharomyces cerevisiae alpha-factor.

Molecular evolution of Sry and Sox gene

The mammalian Sry on the short arm of the Y chromosome encodes a nuclear factor-like protein harboring a DNA-binding domain known as the HMG box. The Sox genes encode similar factor like proteins, but the sequence similarity of the HMG box to that of Sry is variable as being at least 60%. The functional relationship of Sox to Sry genes with special reference to sex determination is unclear except for a few items such as human autosomal Sox9. Thus, it is significant to know more about the evolutionary in addition to the functional relationship between Sry and Sox genes for deepening and broadening our understanding concerning primary sex determination. Therefore, to clarify the ancestry and molecular evolution of the mammalian sex determining gene Sry with its evolutionary relationships to the Sox gene, a molecular phylogenetic tree for the HMG box superfamily was constructed and analyzed, and the following conclusions were reached: (1) The nuclear non histone HMG proteins are supposedly the oldest, appearing at least more than one billion years ago, before the divergence of animals and plants. They diverged into two subgroups: one contains HMG14 and HMG17, and the other one contains HMG1 and HMG2 with various other genes. Subsequent divergences include the nucleolar UBF, nuclear SSRP as well as fungal mating protein Mc, MAT and Ste11. (2) The Sox and Sry genes diverged following the diversification of lymphoid transcription factors TCF and LEF. The Sry gene might have definitely evolved from the Sox gene cluster a few hundred million years ago. Additionally, the marsupial Sry, e.g. from Wallabie's and Dunnart's, is distinguished by being distant from eutherian Sry, but being closely related to the Sox gene cluster. (3) Molecular evolutionary rates estimated in mammalian Sry as the divergent rate per 100 million years are much higher than in Sox genes or other genes from the HMG box superfamily. This rapid evolution of Sry might agree with the fact that the Srys are present not on the pseudoautosomal region but on the distal region with no recombination of the Y chromosomal short arm.

The signature of balancing selection: fungal mating compatibility gene evolution.

A key problem in evolutionary biology has been distinguishing the contributions of current and historical processes to the maintenance of genetic variation. Because alleles at self-recognition genes are under balancing selection, they exhibit extended residence times in populations and thus may provide unique insight into population demographic history. However, evidence for balancing selection and extended residence times has almost exclusively depended on identification of transspecific polymorphisms; polymorphisms retained in populations through speciation events. We present a broadly applicable approach for detecting balancing selection and apply it to the b1 mating type gene in the mushroom fungus Coprinus cinereus. The comparison of neutral molecular variation within and between allelic classes was used to directly estimate the strength of balancing selection. Different allelic classes are defined as encoding different mating compatibility types and are thus potentially subject to balancing selection. Variation within an allelic class, where all alleles have the same mating compatibility type, provided an internal standard of neutral evolution. Mating compatibility in this organism is determined by the complex A mating type locus, and b1 is one of several redundantly functioning genes. Consequently, we conducted numerical simulations of a model with two subloci and varying levels of recombination to show that balancing selection should operate at each sublocus. Empirical data show that strong balancing selection has indeed occurred at the b1 locus. The widespread geographic distribution of identical b1 alleles suggests that their association with differing A mating types is the result of recent recombination events.

Cloning of the mating type loci from Pyrenopeziza brassicae reveals the presence of a novel mating type gene within a discomycete MAT 1‐2 locus encoding a putative metallothionein‐like protein

This article was published in Mol Microbiol (1998) 30: 799–806. In order to move the fungal mating type field towards a standard nomenclature, as suggested by Arie et al. (1997), the mating type designations for Pyrenopeziza brassicae have been reversed. The MAT 1-1 locus described by Singh and Ashby (1998), which was based on the genetic nomenclature proposed by Courtice and Ingram (1987), becomes MAT-2 (EMBL AJ006072). As a consequence the PHB1 gene, encoding the HMG protein, becomes PHB2 and vice versa. The MAT 1-2 locus described by Singh and Ashby (1998) becomes MAT-1 (EMBL AJ006073) and encodes three genes designated PAD1, PMT1 and PHB1.

Ligand Recognition in Multiallelic Pheromone Receptors from the Basidiomycete Schizophyllum commune Studied in Yeast

The homobasidiomycete Schizophyllum commune encodes a multiallelic pheromone receptor system that distinguishes more than 20 nonself from at least 2 self pheromones. The well-investigated pheromone response system of the yeast Saccharomyces cerevisiae was used to link the FUS1::lacZ reporter system to the heterologous pheromone receptors from S. commune. To investigate yeast G-protein binding, the unchanged heterologous receptor was compared to constructs carrying an exchange of the 3rd cytoplasmatic loop for the Ste2 sequence. A better coupling could be achieved with the altered constructs. In order to examine activation by single pheromones, an artificial peptide based on the sequence of a new putative pheromone gene, bap2(1), in the Bα2 mating-type locus encoding the shortest pheromone found so far in fungal mating types was used. Thus, we have reassembled the pheromone recognition of the basidiomycete S. commune and constructed a system ideal for specificity analysis in the yeast S. cerevisiae.

Selfing versus outcrossing propensity of the fungal pathogen Microbotryum violaceum across Silene latifolia host plants: O. KALTZ AND J. A. SHYKOFF

Abstract In the fungal pathogen Microbotryumviolaceum mating (i.e. conjugation between cells of opposite mating type) is indispensable for infection of its host plant Silenelatifolia. Since outcrossing opportunities are potentially rare, selfing may be appropriate to ensure reproduction. On the other hand, outcrossing may create genetic variability necessary in the coevolutionary arms race with its host. We investigated the propensity of M. violaceum to outcross vs. self in different host environments. We used haploid sporidia from each of three strains from five fungal populations for pairwise mixtures of opposite mating type, representing either selfing or outcrossing combinations. Mixtures were exposed to leaf extract from seven S. latifolia plants. The proportion of conjugated sporidia quantified mating propensity. The identity of both fungal strains and host influenced conjugation. First, individual strains differed in conjugation frequency by up to 30%, and strains differed in their performance across the different hosts. Second, selfing combinations produced, on average, more conjugations than did outcrossing combinations. Selfing appears to be the predominant mode of reproduction in this fungus, and selfing preference may have evolved as a mechanism of reproductive assurance. Third, individual strains varied considerably in conjugation frequency in selfing and outcrossing combinations across different hosts. This indicates that conjugation between outcrossing partners could be favoured at least in some hosts. Since the dikaryon resulting from conjugation is the infectious unit, conjugation frequency may correspond with infection probability. This assumption was supported by an inoculation experiment, where high infectious sporidial dosage resulted in higher infections success than did low dosage. We therefore predict that sexual recombination can provide this pathogen with novel genotypes able to infect local resistant hosts.

The distribution of mating-type bias in natural populations of the anther-smut Ustilago violacea on Silene alba in Virginia

Complete individual-wide mating-type bias (retrieval of sporidia of only one mating type from germinated teliospores of one fungal individual) was observed to be a common and widespread feature of the anther-smut fungus, Ustilago violacea, collected from natural populations of its host, Silene alba. The bias was usually to mating type A1, but the frequency of bias and its spatial distribution varied from region to region. Populations with high frequencies of bias still showed high rates of disease transmission. Crosses between A1 mating type sporidial lines from completely biased individuals and A2 mating types from unbiased individuals showed no bias in the progeny. During teliospore germination, biased individuals often showed conjugation among adjacent cells of the promycelium, suggesting that both mating types are present in the germinating teliospore but one mating type is unable to grow as free-living sporidia. The complete bias was most readily interpreted as evidence of “haploid lethals” linked to mating type that cause poor survival or growth of the sporidial stage. The results show that such “haploid lethals” may be a common occurrence in natural populations, and that fungal mating systems may vary considerably over short distances.

The Distribution of Mating-Type Bias in Natural Populations of the Anther-Smut Ustilago violacea on Silene alba in Virginia

Complete individual-wide mating-type bias (retrieval of sporidia of only one mating type from germinated teliospores of one fungal individual) was observed to be a common and widespread feature of the anther-smut fungus, Ustilago violacea, collected from natural populations of its host, Silene alba. The bias was usually to mating type A1, but the frequency of bias and its spatial distribution varied from region to region. Populations with high frequencies of bias still showed high rates of disease transmission. Crosses between A1 mating type sporidial lines from completely biased individuals and A2 mating types from unbiased individuals showed no bias in the progeny. During teliospore germination, biased individuals often showed conjugation among adjacent cells of the promycelium, suggesting that both mating types are present in the germinating teliospore but one mating type is unable to grow as free-living sporidia. The complete bias was most readily interpreted as evidence of “haploid lethals” linked to mating type that cause poor survival or growth of the sporidial stage. The results show that such “haploid lethals” may be a common occurrence in natural populations, and that fungal mating systems may vary considerably over short distances.

Functional consequence of mutating conserved residues of the yeast farnesyl-protein transferase beta-subunit Ram1(Dpr1).

Ras proteins, fungal mating pheromones, and other proteins terminating in the sequence CaaX (where C is Cys, a is any aliphatic amino acid, and X is the C-terminal residue) are posttranslationally prenylated. Farnesyl-protein transferase (FPTase) transfers the farnesyl moiety of farnesyl pyrophosphate (FPP) to the thiol of the CaaX box cysteine in a reaction that requires Zn2+ and Mg2+. We have created mutations in conserved amino acids of the yeast Ram1 protein to identify residues important for Zn2+-dependent FPTase activity. Wild-type and mutant Ram1 proteins were expressed as operon fusions in bacteria, and FPTase activity was measured. Mutations in conserved residues Glu256, His258, Asp307, Cys309, Asp360, and His363 reduce FPTase activity. Asp307, Cys309, and His363 correspond to the residues that have been shown to coordinate Zn2+ in mammalian FPTase. The H258N mutant enzyme exhibited an increased sensitivity to the Zn2+ chelator 1,10-phenanthroline, required higher concentrations of Zn2+ to restore activity to the apoenzyme, and had a 10-fold reduction in catalytic efficiency. The decreases in FPTase activity observed do not appear to be caused by major structural perturbations because the mutants were stably expressed and retained the ability to interact with Ram2p during purification. The FPTase activity of the mutants measured in vitro correlated well with their ability to complement the mating and growth defects of a ram1Delta strain in vivo.

Molecular recognition in fungal mating.

Fungal cells need to recognize that they are genetically different in order to mate. Cells signal to each other by secreting small peptide pheromones which are detected by cell surface receptors. New transcriptional regulators are formed that alter the pattern of gene transcription. Basidiomycete fungi are unusual in having several thousands of different mating types. In these fungi, mating partner recognition demands a remarkable degree of specificity on the part of the proteins and peptides involved.

Protein prenylation: molecular mechanisms and functional consequences.

Prenylation is a class of lipid modification involving covalent addition of either farnesyl (15-carbon) or geranylgeranyl (20-carbon) isoprenoids to conserved cysteine residues at or near the C-terminus of proteins. Known prenylated proteins include fungal mating factors, nuclear lamins, Ras and Ras-related GTP-binding proteins (G proteins), the subunits of trimeric G proteins, protein kinases, and at least one viral protein. Prenylation promotes membrane interactions of most of these proteins, which is not surprising given the hydrophobicity of the lipids involved. In addition, however, prenylation appears to play a major role in several protein-protein interactions involving these species. The emphasis in this review is on the enzymology of prenyl protein processing and the functional significance of prenylation in cellular events. Several other recent reviews provide more detailed coverage of aspects of prenylation that receive limited attention here owing to length restrictions (1-4).

Tetrapolar fungal mating types: sexes by the thousands.

In order to achieve genetic rearrangement in a sexual cycle, eukaryotes go through the processes of meiosis and mating. Different mating types assure that mating is only possible between two genetically diverse individuals. Basidiomycetous fungi display thousands of different mating types that are determined by two genetically unlinked loci. One locus is multiallelic and contains genes for homeodomain transcription factors which are able to form heterodimers. The activation of target genes is dependent on heterodimers formed from the monomeric transcription factor proteins originating from different alleles of this genetic locus. The interactions between the two monomeric transcription factors and the activation of target genes by the heterodimeric proteins make this regulatory system both complex and interesting. The second locus contains a pheromone receptor system: the pheromone receptor is similar to the G protein-linked serpentine receptors in Saccharomyces cerevisiae that activate the pheromone response via a phosphorylation signal transduction cascade in S. cerevisiae. This pheromone perception is a trigger of sexual development and not, as with yeast, itself under control of mating type genes. Rather it directly senses diversity at the mating type loci. Whereas heterobasidiomycetes display a bi-allelic structure for this locus with recognition between one receptor and the opposite pheromone, homobasidiomycetes contain multiple specificities for pheromone receptors and pheromones.