Expression Of Mating Type Genes Research Articles

Differential transcription of mating-type genes during sexual reproduction of natural Cordyceps sinensis

Natural Cordyceps sinensis as an insect-fungal complex, which is developed after Ophiocordyceps sinensis infects a larva of Hepialidae family. Seventeen genotypes of O. sinensis have been identified in natural C. sinensis. This paper summarized the literature reports and GenBank database regarding occurrence and transcription of the mating-type genes of MAT1-1 and MAT1-2 idiomorphs in natural C. sinensis, in Hirsutella sinensis(GC-biased Genotype #1 of O. sinensis), to infer the mating pattern of O. sinensis in the lifecycle of natural C. sinensis. The mating-type genes and transcripts of MAT1-1 and MAT1-2 idiomorphs were identified in the metagenomes and metatranscriptomes of natural C. sinensis. However, their fungal sources are unclear because of co-colonization of several genotypes of O. sinensis and multiple fungal species in natural C. sinensis. The mating-type genes of MAT1-1 and MAT1-2 idiomorphs were differentially present in 237 H. sinensis strains, constituting the genetic control of the O. sinensis reproduction. Transcriptional control of the O. sinensis reproduction includes: differential transcription or silencing of the mating-type genes of MAT1-1 and MAT1-2 idiomorphs, and the MAT1-2-1 transcript with unspliced intron I that contains 3 stop codons. Research on the H. sinensis transcriptome demonstrated differential and complementary transcriptions of the mating-type genes of MAT1-1 and MAT1-2 idiomorphs in Strains L0106 and 1229, which may become mating partners to accomplish physiological heterothallism. The differential occurrence and transcription of the mating-type genes in H. sinensis are inconsistent with the self-fertilization hypothesis under homothallism or pseudohomothallism, but instead indicate the need of mating partners of the same H. sinensis species, either monoecious or dioecious, for physiological heterothallism, or heterospecific species for hybridization. Multiple GC-and AT-biased genotypes of O. sinensis were identified in the stroma, stromal fertile portion(densely covered with numerous ascocarps) and ascospores of natural C. sinensis. It needs to be further explored if the genome-independent O. sinensis genotypes could become mating partners to accomplish sexual reproduction. S. hepiali Strain FENG experienced differential transcription of the mating-type genes with a pattern complementary to that of H. sinensis Strain L0106. Additional evidence is needed to explore a hybridization possibility between S. hepiali and H. sinensis, whether they are able to break the interspecific reproductive isolation. Genotypes #13～14 of O. sinensis feature large DNA segment reciprocal substitutions and genetic material recombination between 2 heterospecific parental fungi, H. sinensis and an AB067719-type fungus, indicating a possibility of hybridization or parasexuality. Our analysis provides important information at the genetic and transcriptional levels regarding the mating-type gene expression and reproduction physiology of O. sinensis in the sexual life of natural C. sinensis and offers crucial reproductive physiology evidence, to assist in the design of the artificial cultivation of C. sinensis to supplement the increasing scarcity of natural resource.

Mating-type switching and mating-type gene array expression in the methylotrophic yeast Ogataea thermomethanolica TBRC656

The methylotrophic yeast, Ogataea thermomethanolica TBRC656, is an attractive host organism for heterologous protein production owing to the availability of protein expression vectors and a genome-editing tool. In this study, we focused on mating-type switching and gene expression in order to elucidate its sexual life cycle and establish genetic approaches applicable for the strain. A putative mating-type gene cluster was identified in TBRC656 that is syntenic to the cluster in Ogataea parapolymorpha DL-1 (previously named Hansenula polymorpha). Like DL-1, TBRC656 possesses two mating loci, namely MATa and MATα, and also shows flip-flop mating-type switching. Interestingly, unlike any other methylotrophic yeast, TBRC656 robustly switched mating type during late growth in rich medium (YPD). Under nutrient depletion, mating-type switching was observed within one hour. Transcription from both MATa and MATα mating loci was detected during growth in YPD, and possibly induced upon nitrogen depletion. Gene expression from MATα was detected as a single co-transcript from a three-gene array (α2-α1-a1S). Deletion of a putative a1S ORF at the MATα locus had no observed effect on mating-type switching but demonstrated significant effect on mating-type gene expression at both MATa and MATα loci.

Determining if Mating Type Proteins Play a Role in Immaturity of the Ciliate Tetrahymena thermophila

Tetrahymena thermophila is free‐living ciliate, commonly found in freshwater ponds, and is used as a model research organism. Mating type gene expression is induced in mature cells after they have been starved and put in the presence of cells of a different mating type (Tetrahymena Genome Database, November 10, 2018). T. thermophila are known to have developmental stages of maturity, and immature cells are unable to mate, although the scientific processes that prevent mating are unknown. It is known that mating type gene expression increases one‐hundred fold from logarithmic phase growth to three hours of starvation (Tetrahymena Genome Database, November 10, 2018). It is currently unknown if the mating type genes are expressed in immature progeny since immature cells do not mate.The goal of this study was to determine if immature progeny cells expressed mating type proteins under a variety of conditions. We hypothesized that mating type genes would not be expressed in immature progeny because T. thermophila will not mate until they reach an level of maturity, and the expression of the mating type genes is essential for mating (Rogers and Karrer, 1985). We plan to quantify the mating type gene expression using q‐RT PCR and RNA Seq transcriptase analysis.This study mated two strains, one mating type VI and II, of T. thermophila and isolated the progeny based on drug sensitivity differences from parental cells. For the q‐RT PCR experiment, mating type gene expression for the parental strain was tested at log phase, thirty minutes of starvation, and two hours of starvation. The mating type gene expression for the progeny strain was tested at the same three conditions as the parental strain. RNA samples from each of the six conditions were isolated and used to create cDNA. The cDNA was confirmed through a PCR containing a genomic control, mating type primers, starvation versus log primers, and negative control primers. Then qPCR was utilized to quantify the expression of mating type genes in the six previously described conditions. RNA Seq was utilized to quantify the expression of all genes for triplicate samples of parental cells and progeny, each starved for 24 hours.Mating type gene expression will be validated through the cross referencing of quantified results provided by RNA Seq and qPCR for T. thermophila and are hypothesized to be lower or absent in progeny samples. There may also be differences in expression levels of transcription factors between the progeny and mature cells. However, if there is no difference in either mating type gene or transcription factors, then prevention of mating in immature cells occurs after transcriptional processes and needs further investigation.Support or Funding InformationAlbion College Foundation for Undergraduate Research, Scholarship, and Creative ActivityThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Some evidence for skewed mating type distribution in Iranian populations of Rhynchosporium commune, the cause of barley scald disease

AbstractRhynchosporium commune(formerly known asRhynchosporium secalis), the causal agent of scald disease on barley, is known to spread asexually by splash dispersed conidia. However, there are multiple lines of evidence for the possibility of a clandestine sexual cycle occurrence in this species including extensive genotypic diversity, equal distribution of mating type alleles across the world and expression of mating type genes. In the current study, the potential for the occurrence of a sexual cycle amongst the Iranian population ofR. communewas assessed by analyzing distribution and frequency of the mating type alleles at both micro and macro-spatial scales. A total of 95 single-conidialR. communeisolates were obtained from different barley fields in Kurdistan province. Previously designed primers were applied in a multiplex PCR assay to study distribution and frequency of the mating type alleles within and between populations. Totally, 67 isolates were determined asMAT1-1and the remaining 28 isolates asMAT1-2throughout the sampling counties. The results obtained at a macro-spatial scale revealed that unlike Kamyaran county (bothMAT1-1andMAT1-2at an equal ratio), an unequal distribution of mating type genes was dominant amongR. communeisolates in both Mariwan and Dehgolan counties. Our findings support a predominantly asexual reproduction for Mariwan and Dehgolan counties and the possibility of sexual stage occurrence in Kamyarna county.

The Hos2 Histone Deacetylase Controls Ustilago maydis Virulence through Direct Regulation of Mating-Type Genes.

Morphological changes are critical for host colonisation in plant pathogenic fungi. These changes occur at specific stages of their pathogenic cycle in response to environmental signals and are mediated by transcription factors, which act as master regulators. Histone deacetylases (HDACs) play crucial roles in regulating gene expression, for example by locally modulating the accessibility of chromatin to transcriptional regulators. It has been reported that HDACs play important roles in the virulence of plant fungi. However, the specific environment-sensing pathways that control fungal virulence via HDACs remain poorly characterised. Here we address this question using the maize pathogen Ustilago maydis. We find that the HDAC Hos2 is required for the dimorphic switch and pathogenic development in U. maydis. The deletion of hos2 abolishes the cAMP-dependent expression of mating type genes. Moreover, ChIP experiments detect Hos2 binding to the gene bodies of mating-type genes, which increases in proportion to their expression level following cAMP addition. These observations suggest that Hos2 acts as a downstream component of the cAMP-PKA pathway to control the expression of mating-type genes. Interestingly, we found that Clr3, another HDAC present in U. maydis, also contributes to the cAMP-dependent regulation of mating-type gene expression, demonstrating that Hos2 is not the only HDAC involved in this control system. Overall, our results provide new insights into the role of HDACs in fungal phytopathogenesis.

The white collar complex is involved in sexual development of Fusarium graminearum.

Sexual spores (ascospores) of Fusarium graminearum, a homothallic ascomycetous fungus, are believed to be the primary inocula for epidemics of the diseases caused by this species in cereal crops. Based on the light requirement for the formation of fruiting bodies (perithecia) of F. graminearum under laboratory conditions, we explored whether photoreceptors play an important role in sexual development. Here, we evaluated the roles of three genes encoding putative photoreceptors [a phytochrome gene (FgFph) and two white collar genes (FgWc-1 and FgWc-2)] during sexual development in F. graminearum. For functional analyses, we generated transgenic strains lacking one or two genes from the self-fertile Z3643 strain. Unlike the wild-type (WT) and add-back strains, the single deletion strains (ΔFgWc-1 and ΔFgWc-2) produced fertile perithecia under constant light on complete medium (CM, an unfavorable medium for sexual development) as well as on carrot agar (a perithecial induction condition). The expression of mating-type (MAT) genes increased significantly in the gene deletion strains compared to the WT under both conditions. Deletion of FgFph had no significant effect on sexual development or MAT gene expression. In contrast, all of the deletion strains examined did not show significant changes in other traits such as hyphal growth, mycotoxin production, and virulence. A split luciferase assay confirmed the in vivo protein-protein interactions among three photoreceptors along with FgLaeA, a global regulator of secondary metabolism and fungal development. Introduction of an intact copy of the A. nidulans LreA and LreB genes, which are homologs of FgWc-1 and FgWc-2, into the ΔFgWc-1 and ΔFgWc-2 strains, respectively, failed to repress perithecia formation on CM in the gene deletion strains. Taken together, these results demonstrate that FgWc-1 and FgWc-2, two central components of the blue-light sensing system, negatively regulate sexual development in F. graminearum, which differs from the regulation pattern in A. nidulans.

Transcriptome analyses during fruiting body formation in Fusarium graminearum and Fusarium verticillioides reflect species life history and ecology

Fusarium graminearum and F. verticillioides are devastating cereal pathogens with very different life history and ecological characteristics. F. graminearum is homothallic, and sexual spores are an important component of its life cycle, responsible for disease initiation. F. verticilloides is heterothallic, and produces only modest numbers of fruiting bodies, which are not a significant source of inoculum. To identify corresponding differences in the transcriptional program underlying fruiting body development in the two species, comparative expression was performed, analyzing six developmental stages. To accompany the transcriptional analysis, detailed morphological characterization of F. verticillioides development was performed and compared to a previous morphological analysis of F. graminearum. Morphological development was similar between the two species, except for the observation of possible trichogynes in F. verticillioides ascogonia, which have not been previously reported for any Fusarium species. Expression of over 9000 orthologous genes were measured for the two species. Functional assignments of highly expressed orthologous genes at each time-point revealed the majority of highly expressed genes fell into the “unclassified proteins” category, reflecting the lack of characterization of genes for sexual development in both species. Simultaneous examination of morphological development and stage-specific gene expression suggests that degeneration of the paraphyses during sexual development is an apoptotic process. Expression of mating type genes in the two species differed, possibly reflecting the divergent roles they play in sexual development. Overall, the differences in gene expression reflect the greater role of fruiting bodies in the life cycle and ecology of F. graminearum versus F. verticillioides.

Complex Mechanisms Regulate Developmental Expression of thematA(HMG) Mating Type Gene in HomothallicAspergillus nidulans

Sexual reproduction is a fundamental developmental process that allows for genetic diversity through the control of zygote formation, recombination, and gametogenesis. The correct regulation of these events is paramount. Sexual reproduction in filamentous fungi, including mating strategy (self-fertilization/homothallism or outcrossing/heterothallism), is determined by the expression of mating type genes at mat loci. Aspergillus nidulans matA encodes a critical regulator that is a fungal ortholog of the hSRY/SOX9 HMG box proteins. In contrast to well-studied outcrossing systems, the molecular basis of homothallism and role of mating type genes during a self-fertile sexual cycle remain largely unknown. In this study the genetic model organism, A. nidulans, has been used to investigate the regulation and molecular functions of the matA mating type gene in a homothallic system. Our data demonstrate that complex regulatory mechanisms underlie functional matA expression during self-fertilization and sexual reproduction in A. nidulans. matA expression is suppressed in vegetative hyphae and is progressively derepressed during the sexual cycle. Elevated levels of matA transcript are required for differentiation of fruiting bodies, karyogamy, meiosis, and efficient formation of meiotic progeny. matA expression is driven from both initiator (Inr) and novel promoter elements that are tightly developmentally regulated by position-dependent and position-independent mechanisms. Deletion of an upstream silencing element, matA SE, results in derepressed expression from wild-type (wt) promoter elements and activation of an additional promoter. These studies provide novel insights into the molecular basis of homothallism in fungi and genetic regulation of sexual reproduction in eukaryotes.

Mating type influences chromosome loss and replicative senescence in telomerase‐deficient budding yeast by Dnl4‐dependent telomere fusion

As we age, the majority of our cells gradually lose the capacity to divide because of replicative senescence that results from the inability to replicate the ends of chromosomes. The timing of senescence is dependent on the length of telomeric DNA, which elicits a checkpoint signal when critically short. Critically short telomeres also become vulnerable to deleterious rearrangements, end-degradation and telomere-telomere fusions. Here we report a novel role of non-homologous end-joining (NHEJ), a pathway of double-strand break repair in influencing both the kinetics of replicative senescence and the rate of chromosome loss in telomerase-deficient Saccharomyces cerevisiae. In telomerase-deficient cells, the absence of NHEJ delays replicative senescence, decreases loss of viability during senescence, and suppresses senescence-associated chromosome loss and telomere-telomere fusion. Differences in mating-type gene expression in haploid and diploid cells affect NHEJ function, resulting in distinct kinetics of replicative senescence. These results suggest that the differences in the kinetics of replicative senescence in haploid and diploid telomerase-deficient yeast are determined by changes in NHEJ-dependent telomere fusion, perhaps through the initiation of the breakage-fusion-bridge cycle.

Mating Type and the Genetic Basis of Self-Fertility in the Model Fungus Aspergillus nidulans

Sexual reproduction occurs in two fundamentally different ways: by outcrossing, in which two distinct partners contribute nuclei, or by self-fertilization (selfing), in which both nuclei are derived from the same individual. Selfing is common in flowering plants, fungi, and some animal taxa. We investigated the genetic basis of selfing in the homothallic fungus Aspergillus nidulans. We demonstrate that alpha and high-mobility group domain mating-type (MAT) genes, found in outcrossing species, are both present in the genome of A. nidulans and that their expression is required for normal sexual development and ascospore production. Balanced overexpression of MAT genes suppressed vegetative growth and stimulated sexual differentiation under conditions unfavorable for sex. Sexual reproduction was correlated with significantly increased expression of MAT genes and key genes of a pheromone-response MAP-kinase signaling pathway involved in heterothallic outcrossing. Mutation of a component MAP-kinase mpkB gene resulted in sterility. These results indicate that selfing in A. nidulans involves activation of the same mating pathways characteristic of sex in outcrossing species, i.e., self-fertilization does not bypass requirements for outcrossing sex but instead requires activation of these pathways within a single individual. However, unlike heterothallic species, aspects of pheromone signaling appeared to be independent of MAT control.

Fungal Genomics: Forensic Evidence of Sexual Activity

The genome sequence of the 'asexual' human pathogenic fungus Aspergillus fumigatus suggests it has the capability to undergo mating and meiosis. That this organism engages in clandestine sexual activity is also suggested by observations of two equally distributed complementary mating types in nature, the expression of mating type genes and evidence of recent genome recombination events.

PKA and MAPK phosphorylation of Prf1 allows promoter discrimination in Ustilago maydis

Mating in Ustilago maydis requires cross-talk between cAMP and mitogen-activated protein kinase (MAPK) signalling. During this process, pheromone response factor 1 (Prf1) activates transcription of a and b mating type genes by binding to pheromone response elements (PREs) located in regulatory regions of these genes. Here, we show that PREs are also necessary and sufficient to mediate cAMP-induced gene expression. Prf1 interacts with cAMP-dependent protein kinase A (PKA) Adr1 as well as MAPK Kpp2 in vivo, and its central phosphorylation sites that are functionally important are modified by the respective kinases in vitro. PKA sites in Prf1 are essential for induced expression of a and b mating type genes. In contrast, MAPK sites are not required for pheromone-induced expression of a genes but are crucial for pheromone-responsive b gene expression. This illustrates how a single transcription factor can integrate signals from two pathways and how its phosphorylation status can determine different transcriptional responses.

THE CONTROL OF SEXUAL MORPHOGENESIS IN THE ASCOMYCOTINA

Summary(1) A series of factors controls sexual morphogenesis in the Ascomycotina, a process involving the formation of novel structures such as ascocarps (fruit bodies) and asci (sacs containing spores) during sexual reproduction.(2) Environmental and genetic factors must be correct before Ascomycetes may sexually reproduce. Compatibility in many heterothallic species is under polygenic control, with the mating type loci and also other genetic factors determining the productivity of sexual crosses.(3) Classical genetic studies have shown that sexual morphogenesis involves the expression of a series of developmentally regulated genes, and this has been confirmed by recent molecular studies which have demonstrated changes in patterns of mRNA and protein synthesis during ascocarp formation.(4) Hyphal differentiation leading to the formation of mature fruit bodies occurs in response to a series of signals, which include various physical and chemical factors.(5) Chemical sex factors have been identified which are believed to have important regulatory or nutritional roles in sexual morphogenesis. These include the following.(a) Diffusible sex hormones which may regulate developmental switching between asexual and sexual modes of reproduction, including (i) pheromones involved with the induction of gametangia and gamete attraction, and (ii) sex morphogens involved with triggering particular stages of fruit body formation. (b) Sexual growth substances which are required as nutrients, and may be precursors for the production of sex hormones, or metabolites used in the synthesis of novel sexual structures. Most of these sex factors are lipids.(6) Certain sex morphogens and sexual growth substances have been shown to exhibit activity in a variety of fungal species, suggesting that fungi of related phylogenetic descent may utilize similar metabolites or signalling factors during sexual reproduction.(7) Phenoloxidase enzymes may catalyse hyphal aggregation in developing fruit bodies.(8) Initial stages of ascocarp development may occur independently of the events of the sexual cycle. However, a link(s) with the functional ascogenous hyphae is needed for the formation of morphologically mature ascocarps.(9) Suitable environmental conditions are sufficient to trigger sexual morphogenesis in homothallic Ascomycetes. However, an extra level of control is present in heterothallic species, with a compatible partner required to complete sexual reproduction. This may be partly because novel regulatory products, formed by the combined action of the mating type loci of different partners, are required for further ascocarp development.(10) Further research is required to identify more fungal chemical sex factors and to determine the role of environmental stress in controlling sexual morphogenesis, and how this may be related to temporal patterns in the expression of mating type genes.