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Kato, Naoki, Wilhelmina Brooks, and Ana M. Calvo. "The Expression of Sterigmatocystin and Penicillin Genes in Aspergillus nidulans Is Controlled by veA, a Gene Required for Sexual Development." Eukaryotic Cell 2, no. 6 (December 2003): 1178–86. http://dx.doi.org/10.1128/ec.2.6.1178-1186.2003.
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ABSTRACT Secondary metabolism is commonly associated with morphological development in microorganisms, including fungi. We found that veA, a gene previously shown to control the Aspergillus nidulans sexual/asexual developmental ratio in response to light, also controls secondary metabolism. Specifically, veA regulates the expression of genes implicated in the synthesis of the mycotoxin sterigmatocystin and the antibiotic penicillin. veA is necessary for the expression of the transcription factor aflR, which activates the gene cluster that leads to the production of sterigmatocystin. veA is also necessary for penicillin production. Our results indicated that although veA represses the transcription of the isopenicillin synthetase gene ipnA, it is necessary for the expression of acvA, the key gene in the first step of penicillin biosynthesis, encoding the delta-(l-alpha-aminoadipyl)-l-cysteinyl-d-valine synthetase. With respect to the mechanism of veA in directing morphological development, veA has little effect on the expression of the known sexual transcription factors nsdD and steA. However, we found that veA regulates the expression of the asexual transcription factor brlA by modulating the α/β transcript ratio that controls conidiation.
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Zhang, Chenghua, Hong Huang, Wangqiu Deng, and Taihui Li. "Genome-Wide Analysis of the Zn(II)2Cys6 Zinc Cluster-Encoding Gene Family in Tolypocladium guangdongense and Its Light-Induced Expression." Genes 10, no. 3 (February 26, 2019): 179. http://dx.doi.org/10.3390/genes10030179.
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The Zn(II)2Cys6 zinc cluster gene family is a subclass of zinc-finger proteins, which are transcriptional regulators involved in a wide variety of biological processes in fungi. We performed genome-wide identification and characterization of Zn(II)2Cys6 zinc-cluster gene (C6 zinc gene) family in Tolypocladium guangdongense, Cordyceps militaris and Ophiocordyceps sinensis. Based on the structures of the C6 zinc domains, these proteins were observed to be evolutionarily conserved in ascomycete fungi. We focused on T. guangdongense, a medicinal fungus, and identified 139 C6 zinc genes which could be divided into three groups. Among them, 49.6% belonged to the fungal specific transcriptional factors, and 16% had a DUF3468 domain. Homologous and phylogenetic analysis indicated that 29 C6 zinc genes were possibly involved in the metabolic process, while five C6 zinc genes were supposed to be involved in asexual or sexual development. Gene expression analysis revealed that 54 C6 zinc genes were differentially expressed under light, including two genes that possibly influenced the development, and seven genes that possibly influenced the metabolic processes. This indicated that light may affect the development and metabolic processes, at least partially, through the regulation of C6 zinc genes in T. guangdongense. Our results provide comprehensive data for further analyzing the functions of the C6 zinc genes.
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Scherer, Mario, Huijun Wei, Ralf Liese, and Reinhard Fischer. "Aspergillus nidulans Catalase-Peroxidase Gene (cpeA) Is Transcriptionally Induced during Sexual Development through the Transcription Factor StuA." Eukaryotic Cell 1, no. 5 (October 2002): 725–35. http://dx.doi.org/10.1128/ec.1.5.725-735.2002.
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ABSTRACT Catalases, peroxidases, and catalase-peroxidases are important enzymes to cope with reactive oxygen species in pro- and eukaryotic cells. In the filamentous fungus Aspergillus nidulans three monofunctional catalases have been described, and a fourth catalase activity was observed in native polyacrylamide gels. The latter activity is probably due to the bifunctional enzyme catalase-peroxidase, which we characterized here. The gene, named cpeA, encodes an 81-kDa polypeptide with a conserved motif for heme coordination. The enzyme comprises of two similar domains, suggesting gene duplication and fusion during evolution. The first 439 amino acids share 22% identical residues with the C terminus. Homologous proteins are found in several prokaryotes, such as Escherichia coli and Mycobacterium tuberculosis (both with 61% identity). In fungi the enzyme has been noted in Penicillium simplicissimum, Septoria tritici, and Neurospora crassa (69% identical amino acids) but is absent from Saccharomyces cerevisiae. Expression analysis in A. nidulans revealed that the gene is transcriptionally induced upon carbon starvation and during sexual development, but starvation is not sufficient to reach high levels of the transcript during development. Besides transcriptional activation, we present evidence for posttranscriptional regulation. A green fluorescent protein fusion protein localized to the cytoplasm of Hülle cells. The Hülle cell-specific expression was dependent on the developmental regulator StuA, suggesting an activating function of this helix-loop-helix transcription factor.
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Klix, V., M. Nowrousian, C. Ringelberg, J. J. Loros, J. C. Dunlap, and S. Pöggeler. "Functional Characterization of MAT1-1-Specific Mating-Type Genes in the Homothallic Ascomycete Sordaria macrospora Provides New Insights into Essential and Nonessential Sexual Regulators." Eukaryotic Cell 9, no. 6 (April 30, 2010): 894–905. http://dx.doi.org/10.1128/ec.00019-10.
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ABSTRACT Mating-type genes in fungi encode regulators of mating and sexual development. Heterothallic ascomycete species require different sets of mating-type genes to control nonself-recognition and mating of compatible partners of different mating types. Homothallic (self-fertile) species also carry mating-type genes in their genome that are essential for sexual development. To analyze the molecular basis of homothallism and the role of mating-type genes during fruiting-body development, we deleted each of the three genes, SmtA-1 (MAT1-1-1), SmtA-2 (MAT1-1-2), and SmtA-3 (MAT1-1-3), contained in the MAT1-1 part of the mating-type locus of the homothallic ascomycete species Sordaria macrospora. Phenotypic analysis of deletion mutants revealed that the PPF domain protein-encoding gene SmtA-2 is essential for sexual reproduction, whereas the α domain protein-encoding genes SmtA-1 and SmtA-3 play no role in fruiting-body development. By means of cross-species microarray analysis using Neurospora crassa oligonucleotide microarrays hybridized with S. macrospora targets and quantitative real-time PCR, we identified genes expressed under the control of SmtA-1 and SmtA-2. Both genes are involved in the regulation of gene expression, including that of pheromone genes.
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Wellham, Peter A. D., Abdul Hafeez, Andrej Gregori, Matthias Brock, Dong-Hyun Kim, David Chandler, and Cornelia H. de Moor. "Culture Degeneration Reduces Sex-Related Gene Expression, Alters Metabolite Production and Reduces Insect Pathogenic Response in Cordyceps militaris." Microorganisms 9, no. 8 (July 22, 2021): 1559. http://dx.doi.org/10.3390/microorganisms9081559.
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Cordyceps militaris is an entomopathogenic ascomycete, known primarily for infecting lepidopteran larval (caterpillars) and pupal hosts. Cordycepin, a secondary metabolite produced by this fungus has anti-inflammatory properties and other pharmacological activities. However, little is known about the biological role of this adenosine derivate and its stabilising compound pentostatin in the context of insect infection the life cycle of C. militaris. During repeated subcultivation under laboratory conditions a degeneration of C. militaris marked by decreasing levels of cordycepin production can occur. Here, using degenerated and parental control strains of an isolate of C. militaris, we found that lower cordycepin production coincides with the decline in the production of various other metabolites as well as the reduced expression of genes related to sexual development. Additionally, infection of Galleria mellonella (greater wax moth) caterpillars indicated that cordycepin inhibits the immune response in host haemocytes. Accordingly, the pathogenic response to the degenerated strain was reduced. These data indicate that there are simultaneous changes in sexual reproduction, secondary metabolite production, insect immunity and infection by C. militaris. This study may have implications for biological control of insect crop pests by fungi.
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Murry, Reyna, Lea Traxler, Jessica Pötschner, Thomas Krüger, Olaf Kniemeyer, Katrin Krause, and Erika Kothe. "Inositol Signaling in the Basidiomycete Fungus Schizophyllum commune." Journal of Fungi 7, no. 6 (June 10, 2021): 470. http://dx.doi.org/10.3390/jof7060470.
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Intracellular signaling is conserved in eukaryotes to allow for response to extracellular signals and to regulate development and cellular functions. In fungi, inositol phosphate signaling has been shown to be involved in growth, sexual reproduction, and metabolic adaptation. However, reports on mushroom-forming fungi are lacking so far. In Schizophyllum commune, an inositol monophosphatase has been found up-regulated during sexual development. The enzyme is crucial for inositol cycling, where it catalyzes the last step of inositol phosphate metabolism, restoring the inositol pool from the monophosphorylated inositol monophosphate. We overexpressed the gene in this model basidiomycete and verified its involvement in cell wall integrity and intracellular trafficking. Strong phenotypes in mushroom formation and cell metabolism were evidenced by proteome analyses. In addition, altered inositol signaling was shown to be involved in tolerance towards cesium and zinc, and increased metal tolerance towards cadmium, associated with induced expression of kinases and repression of phosphatases within the inositol cycle. The presence of the heavy metals Sr, Cs, Cd, and Zn lowered intracellular calcium levels. We could develop a model integrating inositol signaling in the known signal transduction pathways governed by Ras, G-protein coupled receptors, and cAMP, and elucidate their different roles in development.
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Hallen, Heather E., and Frances Trail. "The L-Type Calcium Ion Channel Cch1 Affects Ascospore Discharge and Mycelial Growth in the Filamentous Fungus Gibberella zeae (Anamorph Fusarium graminearum)." Eukaryotic Cell 7, no. 2 (December 14, 2007): 415–24. http://dx.doi.org/10.1128/ec.00248-07.
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ABSTRACT Cch1, a putative voltage-gated calcium ion channel, was investigated for its role in ascus development in Gibberella zeae. Gene replacement mutants of CCH1 were generated and found to have asci which did not forcibly discharge spores, although morphologically ascus and ascospore development in the majority of asci appeared normal. Additionally, mycelial growth was significantly slower, and sexual development was slightly delayed in the mutant; mutant mycelia showed a distinctive fluffy morphology, and no cirrhi were produced. Wheat infected with Δcch1 mutants developed symptoms comparable to wheat infected with the wild type; however, the mutants showed a reduced ability to protect the infected stalk from colonization by saprobic fungi. Transcriptional analysis of gene expression in mutants using the Affymetrix Fusarium microarray showed 2,449 genes with significant, twofold or greater, changes in transcript abundance across a developmental series. This work extends the role of CCH1 to forcible spore discharge in G. zeae and suggests that this channel has subtle effects on growth and development.
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Schubert, Daniela, Marjatta Raudaskoski, Nicole Knabe, and Erika Kothe. "Ras GTPase-Activating Protein Gap1 of the Homobasidiomycete Schizophyllum commune Regulates Hyphal Growth Orientation and Sexual Development." Eukaryotic Cell 5, no. 4 (April 2006): 683–95. http://dx.doi.org/10.1128/ec.5.4.683-695.2006.
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ABSTRACT The white rot fungus Schizophyllum commune is used for the analysis of mating and sexual development in homobasidiomycete fungi. In this study, we isolated the gene gap1 encoding a GTPase-activating protein for Ras. Disruption of gap1 should therefore lead to strains accumulating Ras in its activated, GTP-bound state and to constitutive Ras signaling. Haploid Δgap1 monokaryons of different mating types did not show alterations in mating behavior in the four different mating interactions possible in fungi expressing a tetrapolar mating type system. Instead, the growth rate in Δgap1 monokaryons was reduced by ca. 25% and ca. 50% in homozygous Δgap1/Δgap1 dikaryons. Monokaryons, as well as homozygous dikaryons, carrying the disrupted gap1 alleles exhibited a disorientated growth pattern. Dikaryons showed a strong phenotype during clamp formation since hook cells failed to fuse with the peg beside them. Instead, the dikaryotic character of the hyphae was rescued by fusion of the hooks with nearby developing branches. Δgap1/Δgap1 dikaryons formed increased numbers of fruitbody primordia, whereas the amount of fruitbodies was not raised. Mature fruitbodies formed no or abnormal gills. No production of spores could be observed. The results suggest Ras involvement in growth, clamp formation, and fruitbody development.
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Pandit, Sandesh, Jessica Lohmar, Shawana Ahmed, Oier Etxebeste, Eduardo Espeso, and Ana Calvo. "UrdA Controls Secondary Metabolite Production and the Balance between Asexual and Sexual Development in Aspergillus nidulans." Genes 9, no. 12 (November 23, 2018): 570. http://dx.doi.org/10.3390/genes9120570.
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The genus Aspergillus includes important plant pathogens, opportunistic human pathogens and mycotoxigenic fungi. In these organisms, secondary metabolism and morphogenesis are subject to a complex genetic regulation. Here we functionally characterized urdA, a gene encoding a putative helix-loop-helix (HLH)-type regulator in the model fungus Aspergillus nidulans. urdA governs asexual and sexual development in strains with a wild-type veA background; absence of urdA resulted in severe morphological alterations, with a significant reduction of conidial production and an increase in cleistothecial formation, even in the presence of light, a repressor of sex. The positive effect of urdA on conidiation is mediated by the central developmental pathway (CDP). However, brlA overexpression was not sufficient to restore wild-type conidiation in the ΔurdA strain. Heterologous complementation of ΔurdA with the putative Aspergillus flavus urdA homolog also failed to rescue conidiation wild-type levels, indicating that both genes perform different functions, probably reflected by key sequence divergence. UrdA also represses sterigmatocystin (ST) toxin production in the presence of light by affecting the expression of aflR, the activator of the ST gene cluster. Furthermore, UrdA regulates the production of several unknown secondary metabolites, revealing a broader regulatory scope. Interestingly, UrdA affects the abundance and distribution of the VeA protein in hyphae, and our genetics studies indicated that veA appears epistatic to urdA regarding ST production. However, the distinct fluffy phenotype of the ΔurdAΔveA double mutant suggests that both regulators conduct independent developmental roles. Overall, these results suggest that UrdA plays a pivotal role in the coordination of development and secondary metabolism in A. nidulans.
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Turgeon, B. Gillian, Amir Sharon, Stefan Wirsel, Kenichi Yamaguchi, Solveig K. Christiansen, and Olen C. Yoder. "Structure and function of mating type genes in Cochliobolus spp. and asexual fungi." Canadian Journal of Botany 73, S1 (December 31, 1995): 778–83. http://dx.doi.org/10.1139/b95-322.
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Mating type (MAT) genes of Cochliobolus heterostrophus have homologs in other heterothallic Cochliobolus spp., in homothallic Cochliobolus spp., and in asexual fungi thought to be taxonomically related to Cochliobolus (e.g., Bipolaris spp.). To examine the cause of asexuality in B. sacchari, its homolog of C. heterostrophus MAT-2 was cloned. The B. sacchari sequence was 98% identical to that of C. heterostrophus MAT-2, the gene conferred homothallism when expressed in a C. heterostrophus MAT-1 strain, and transgenic strains mated with C. heterostrophus MAT-1. Thus the cause of asexuality in B. sacchari is not absence or lack of a functional MAT gene. When the C. heterostrophus MAT genes were expressed in B. sacchari, however, no sexual development occurred, suggesting that this asexual fungus lacks an attribute, other than the mating type gene, which is required for mating. Although cloned MAT genes function upon transformation into recipient strains, they do not confer full fertility. When an homologous or heterologous (e.g., from C. carbonum, C. victoriae, or B. sacchari) MAT gene is transferred into a C. heterostrophus strain of opposite mating type, the strain can self and cross to tester strains of either mating type. However, any transgenic strain carrying both a resident MAT gene and an homologous or heterologous MAT transgene develops normal perithecia but few ascospores in a cross that requires function of the transgene. To determine if the resident MAT gene interferes with function of the transgene, the MAT locus was deleted from the genome of C. heterostrophus and then replaced with the MAT gene of C. heterostrophus, C. carbonum, C. victoriae, or B. sacchari. Interference was eliminated and abundant ascospores were formed when the four transgenic strains were crossed to C. heterostrophus strains of opposite mating type. Key words: asexual fungi, DNA-binding proteins, heterologous expression, transformation.
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Nowrousian, Minou, Sandra Masloff, Stefanie Pöggeler, and Ulrich Kück. "Cell Differentiation during Sexual Development of the Fungus Sordaria macrospora Requires ATP Citrate Lyase Activity." Molecular and Cellular Biology 19, no. 1 (January 1, 1999): 450–60. http://dx.doi.org/10.1128/mcb.19.1.450.
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ABSTRACT During sexual development, mycelial cells from most filamentous fungi differentiate into typical fruiting bodies. Here, we describe the isolation and characterization of the Sordaria macrosporadevelopmental mutant per5, which exhibits a sterile phenotype with defects in fruiting body maturation. Cytological investigations revealed that the mutant strain forms only ascus precursors without any mature spores. Using an indexed cosmid library, we were able to complement the mutant to fertility by DNA-mediated transformation. A single cosmid clone, carrying a 3.5-kb region able to complement the mutant phenotype, has been identified. Sequencing of the 3.5-kb region revealed an open reading frame of 2.1 kb interrupted by a 66-bp intron. The predicted polypeptide (674 amino acids) shows significant homology to eukaryotic ATP citrate lyases (ACLs), with 62 to 65% amino acid identity, and the gene was named acl1. The molecular mass of the S. macrospora ACL1 polypeptide is 73 kDa, as was verified by Western blot analysis with a hemagglutinin (HA) epitope-tagged ACL1 polypeptide. Immunological in situ detection of the HA-tagged polypeptide demonstrated that ACL is located within the cytosol. Sequencing of the mutant acl1 gene revealed a 1-nucleotide transition within the coding region, resulting in an amino acid substitution within the predicted polypeptide. Further evidence that ACL1 is essential for fruiting body maturation comes from experiments in which truncated and mutated versions of theacl1 gene were used for transformation. None of these copies was able to reconstitute the fertile phenotype in transformed per5 recipient strains. ACLs are usually involved in the formation of cytosolic acetyl coenzyme A (acetyl-CoA), which is used for the biosynthesis of fatty acids and sterols. Protein extracts from the mutant strain showed a drastic reduction in enzymatic activity compared to values obtained from the wild-type strain. Investigation of the time course of ACL expression suggests that ACL is specifically induced at the beginning of the sexual cycle and produces acetyl-CoA, which most probably is a prerequisite for fruiting body formation during later stages of sexual development. We discuss the contribution of ACL activity to the life cycle of S. macrospora.
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Namekawa, Satoshi H., Kazuki Iwabata, Hiroko Sugawara, Fumika N. Hamada, Akiyo Koshiyama, Hiroyuki Chiku, Takashi Kamada, and Kengo Sakaguchi. "Knockdown of LIM15/DMC1 in the mushroom Coprinus cinereus by double-stranded RNA-mediated gene silencing." Microbiology 151, no. 11 (November 1, 2005): 3669–78. http://dx.doi.org/10.1099/mic.0.28209-0.
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The basidiomycete Coprinus cinereus has many advantages as a model organism for studying sexual development and meiosis, but it has been difficult to investigate using reverse-genetics methods, such as gene disruption by homologous recombination. Here, gene repression by dsRNA-mediated gene silencing was tried as an alternative method for reverse-genetics studies. It was shown that transformation of the LIM15/DMC1 dsRNA expression construct (LIM15dsRNA) resulted in genomic insertion of LIM15dsRNA and paucity of the LIM15/DMC1 transcript. First, LIM15dsRNA was transformed into the homothallic strain AmutBmut to generate a homozygote in which both nuclei had a copy of LIM15dsRNA. The LIM15/DMC1-repressed strain showed abnormal homologous chromosome synapsis during meiosis. Basidiospore production was reduced to 16 % by the induction of dsRNA. However, approximately 60 % of basidiospores were viable. Next, a heterozygote was generated in which one nucleus had a copy of LIM15dsRNA. The phenotype was similar to that of the homozygote. These results are not only the first demonstration of dsRNA-mediated gene silencing in a member of the homobasidiomycete fungi, to which 90 % of mushroom species belong, but also the first successful use of a reverse-genetics approach in C. cinereus research.
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Meister, Thieme, Thieme, Köhler, Schmitt, Valerius, and Braus. "COP9 Signalosome Interaction with UspA/Usp15 Deubiquitinase Controls VeA-Mediated Fungal Multicellular Development." Biomolecules 9, no. 6 (June 18, 2019): 238. http://dx.doi.org/10.3390/biom9060238.
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COP9 signalosome (CSN) and Den1/A deneddylases physically interact and promote multicellular development in fungi. CSN recognizes Skp1/cullin-1/Fbx E3 cullin-RING ligases (CRLs) without substrate and removes their posttranslational Nedd8 modification from the cullin scaffold. This results in CRL complex disassembly and allows Skp1 adaptor/Fbx receptor exchange for altered substrate specificity. We characterized the novel ubiquitin-specific protease UspA of the mold Aspergillus nidulans, which corresponds to CSN-associated human Usp15 and interacts with six CSN subunits. UspA reduces amounts of ubiquitinated proteins during fungal development, and the uspA gene expression is repressed by an intact CSN. UspA is localized in proximity to nuclei and recruits proteins related to nuclear transport and transcriptional processing, suggesting functions in nuclear entry control. UspA accelerates the formation of asexual conidiospores, sexual development, and supports the repression of secondary metabolite clusters as the derivative of benzaldehyde (dba) genes. UspA reduces protein levels of the fungal NF-kappa B-like velvet domain protein VeA, which coordinates differentiation and secondary metabolism. VeA stability depends on the Fbx23 receptor, which is required for light controlled development. Our data suggest that the interplay between CSN deneddylase, UspA deubiquitinase, and SCF-Fbx23 ensures accurate levels of VeA to support fungal development and an appropriate secondary metabolism.
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Yang, Qi, Sheven I. Poole, and Katherine A. Borkovich. "A G-Protein β Subunit Required for Sexual and Vegetative Development and Maintenance of Normal Gα Protein Levels in Neurospora crassa." Eukaryotic Cell 1, no. 3 (June 2002): 378–90. http://dx.doi.org/10.1128/ec.1.3.378-390.2002.
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ABSTRACT The genome of the filamentous fungus Neurospora crassa contains a single gene encoding a heterotrimeric G-protein β subunit, gnb-1. The predicted GNB-1 protein sequence is most identical to Gβ proteins from the filamentous fungi Cryphonectria parasitica and Aspergillus nidulans. N. crassa GNB-1 is also 65% identical to the human GNB-1 protein but only 38 and 45% identical to Gβ proteins from budding and fission yeasts. Previous studies in animal and fungal systems have elucidated phenotypes of Gβ null mutants, but little is known about the effects of Gβ loss on Gα levels. In this study, we analyzed a gnb-1 deletion mutant for cellular phenotypes and levels of the three Gα proteins. Δgnb-1 strains are female-sterile, with production of aberrant fertilized reproductive structures. Δgnb-1 strains conidiate more profusely and have altered mass on solid medium. Loss of gnb-1 leads to inappropriate conidiation and expression of a conidiation-specific gene during growth in submerged culture. Intracellular cyclic AMP levels are reduced by 60% in vegetative plate cultures of Δgnb-1 mutants. Loss of gnb-1 leads to lower levels of the three Gα proteins under a variety of conditions. Analysis of transcript levels for the gna-1 and gna-2 Gα genes in submerged cultures indicates that regulation of Gα protein levels by gnb-1 is posttranscriptional. The results suggest that GNB-1 directly regulates apical extension rate and mass accumulation. In contrast, many other Δgnb-1 phenotypes, including female sterility and defective conidiation, can be explained by altered levels of the three N. crassa Gα proteins.
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Min, Kyunghun, Hokyoung Son, Jae Yun Lim, Gyung Ja Choi, Jin-Cheol Kim, Steven D. Harris, and Yin-Won Lee. "Transcription Factor RFX1 Is Crucial for Maintenance of Genome Integrity in Fusarium graminearum." Eukaryotic Cell 13, no. 3 (January 24, 2014): 427–36. http://dx.doi.org/10.1128/ec.00293-13.
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ABSTRACT The survival of cellular organisms depends on the faithful replication and transmission of DNA. Regulatory factor X (RFX) transcription factors are well conserved in animals and fungi, but their functions are diverse, ranging from the DNA damage response to ciliary gene regulation. We investigated the role of the sole RFX transcription factor, RFX1, in the plant-pathogenic fungus Fusarium graminearum . Deletion of rfx1 resulted in multiple defects in hyphal growth, conidiation, virulence, and sexual development. Deletion mutants of rfx1 were more sensitive to various types of DNA damage than the wild-type strain. Septum formation was inhibited and micronuclei were produced in the rfx1 deletion mutants. The results of the neutral comet assay demonstrated that disruption of rfx1 function caused spontaneous DNA double-strand breaks (DSBs). The transcript levels of genes involved in DNA DSB repair were upregulated in the rfx1 deletion mutants. DNA DSBs produced micronuclei and delayed septum formation in F. graminearum . Green fluorescent protein (GFP)-tagged RFX1 localized in nuclei and exhibited high expression levels in growing hyphae and conidiophores, where nuclear division was actively occurring. RNA-sequencing-based transcriptomic analysis revealed that RFX1 suppressed the expression of many genes, including those required for the repair of DNA damage. Taken together, these findings indicate that the transcriptional repressor rfx1 performs crucial roles during normal cell growth by maintaining genome integrity.
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Berka, Randy M., and Christopher C. Barnett. "The development of gene expression systems for filamentous fungi." Biotechnology Advances 7, no. 2 (January 1989): 127–54. http://dx.doi.org/10.1016/0734-9750(89)90356-x.
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Wang, Lu, Yanzhi Liu, Jinliang Liu, Yanhua Zhang, Xianghui Zhang, and Hongyu Pan. "The Sclerotinia sclerotiorum FoxE2 Gene Is Required for Apothecial Development." Phytopathology® 106, no. 5 (May 2016): 484–90. http://dx.doi.org/10.1094/phyto-08-15-0181-r.
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Sclerotinia sclerotiorum is a widely dispersed plant pathogenic fungus causing many diseases such as white mold, Sclerotinia stem rot, stalk rot, and Sclerotinia head rot on many varieties of broadleaf crops worldwide. Previous studies have shown that the Forkhead-box transcription factors (FOX TFs) play key regulatory roles in the sexual reproduction of some fungi. Ss-FoxE2 is one of four FOX TF family member genes in S. sclerotiorum. Based on ortholog function in other fungi it is hypothesized to function in S. sclerotiorum sexual reproduction. In this study, the role of Ss-FoxE2 in S. sclerotiorum was identified with a gene knock-out strategy. Following transformation and screening, strains having undergone homologous recombination in which the hygromycin resistance gene replaced the gene Ss-FoxE2 from the genomic DNA were identified. No difference in hyphae growth, number, and weight of sclerotia and no obvious change in virulence was observed among the wild type Ss-FoxE2 knock-out mutant and genetically complemented mutant; however, following induction of sclerotia for sexual development, apothecia were not formed in Ss-FoxE2 knock-out mutant. The Ss-FoxE2 gene expressed significantly higher in the apothecial stages than in other developmental stages. These results indicate that Ss-FoxE2 appears to be necessary for the regulation of sexual reproduction, but may not affect the pathogenicity and vegetative development of S. sclerotiorum significantly.
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Rensing, Ludger, Christian Monnerjahn, and Ulf Meyer. "Differential stress gene expression during the development ofNeurospora crassaand other fungi." FEMS Microbiology Letters 168, no. 2 (November 1998): 159–66. http://dx.doi.org/10.1111/j.1574-6968.1998.tb13268.x.
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Fabritius, Anna-Liisa, Cristina Cvitanich, and Howard S. Judelson. "Stage-specific gene expression during sexual development in Phytophthora infestans." Molecular Microbiology 45, no. 4 (August 2002): 1057–66. http://dx.doi.org/10.1046/j.1365-2958.2002.03073.x.
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Wilson, Wilken, van der Nest, Wingfield, and Wingfield. "It’s All in the Genes: The Regulatory Pathways of Sexual Reproduction in Filamentous Ascomycetes." Genes 10, no. 5 (April 30, 2019): 330. http://dx.doi.org/10.3390/genes10050330.
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Sexual reproduction in filamentous ascomycete fungi results in the production of highly specialized sexual tissues, which arise from relatively simple, vegetative mycelia. This conversion takes place after the recognition of and response to a variety of exogenous and endogenous cues, and relies on very strictly regulated gene, protein, and metabolite pathways. This makes studying sexual development in fungi an interesting tool in which to study gene–gene, gene–protein, and protein–metabolite interactions. This review provides an overview of some of the most important genes involved in this process; from those involved in the conversion of mycelia into sexually-competent tissue, to those involved in the development of the ascomata, the asci, and ultimately, the ascospores.
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Nowrousian, Minou, and Ulrich Kück. "Comparative gene expression analysis of fruiting body development in two filamentous fungi." FEMS Microbiology Letters 257, no. 2 (April 2006): 328–35. http://dx.doi.org/10.1111/j.1574-6968.2006.00192.x.
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Ingleby, Fiona C., Ilona Flis, and Edward H. Morrow. "Sex-Biased Gene Expression and Sexual Conflict throughout Development: Table 1." Cold Spring Harbor Perspectives in Biology 7, no. 1 (November 6, 2014): a017632. http://dx.doi.org/10.1101/cshperspect.a017632.
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Burcea, Alexandru, Gina‐Oana Popa, Iulia E. Florescu (Gune), Sami Gharbia, Gisela Gaina, Andreea Dudu, Marilena Maereanu, Anca Hermenean, Sergiu E. Georgescu, and Marieta Costache. "Gene expression involved in the sexual development of Best Beluga hybrid sturgeons." Journal of Fish Biology 93, no. 5 (October 31, 2018): 1021–26. http://dx.doi.org/10.1111/jfb.13815.
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Casselton, Lorna A., and Natalie S. Olesnicky. "Molecular Genetics of Mating Recognition in Basidiomycete Fungi." Microbiology and Molecular Biology Reviews 62, no. 1 (March 1, 1998): 55–70. http://dx.doi.org/10.1128/mmbr.62.1.55-70.1998.
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SUMMARY The recognition of compatible mating partners in the basidiomycete fungi requires the coordinated activities of two gene complexes defined as the mating-type genes. One complex encodes members of the homeobox family of transcription factors, which heterodimerize on mating to generate an active transcription regulator. The other complex encodes peptide pheromones and 7-transmembrane receptors that permit intercellular signalling. Remarkably, a single species may have many thousands of cross-compatible mating types because the mating-type genes are multiallelic. Different alleles of both sets of genes are necessary for mating compatibility, and they trigger the initial stages of sexual development—the formation of a specialized filamentous mycelium termed the dikaryon, in which the haploid nuclei remain closely associated in each cell but do not fuse. Three species have been taken as models to describe the molecular structure and organization of the mating-type loci and the genes sequestered within them: the pathogenic smut fungus Ustilago maydis and the mushrooms Coprinus cinereus and Schizophyllum commune. Topics addressed in this review are the roles of the mating-type gene products in regulating sexual development, the molecular basis for multiple mating types, and the molecular interactions that permit different allelic products of the mating type genes to be discriminated. Attention is drawn to the remarkable conservation in the mechanisms that regulate sexual development in basidiomycetes and unicellular ascomycete yeasts, Saccharomyces cerevisiae and Schizosaccharomyces pombe, a theme which is developed in the general conclusion to include the filamentous ascomycetes Neurospora crassa and Podospora anserina.
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He, Wenxia, Xiangyan Dai, Xiaowen Chen, Jiangyan He, and Zhan Yin. "Zebrafish pituitary gene expression before and after sexual maturation." Journal of Endocrinology 221, no. 3 (April 7, 2014): 429–40. http://dx.doi.org/10.1530/joe-13-0488.
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Sexual maturation and somatic growth cessation are associated with adolescent development, which is precisely controlled by interconnected neuroendocrine regulatory pathways in the endogenous endocrine system. The pituitary gland is one of the key regulators of the endocrine system. By analyzing the RNA sequencing (RNA-seq) transcriptome before and after sexual maturation, in this study, we characterized the global gene expression patterns in zebrafish pituitaries at 45 and 90 days post-fertilization (dpf). A total of 15 043 annotated genes were expressed in the pituitary tissue, 3072 of which were differentially expressed with a greater than or equal to twofold change between pituitaries at 45 and 90 dpf. In the pituitary transcriptome, the most abundant transcript was gh. The expression levels of gh remained high even after sexual maturation at 90 dpf. Among the eight major pituitary hormone genes, lhb was the only gene that exhibited a significant change in its expression levels between 45 and 90 dpf. Significant changes in the pituitary transcripts included genes involved in the regulation of immune responses, bone metabolism, and hormone secretion processes during the juvenile–sexual maturity transition. Real-time quantitative PCR analysis was carried out to verify the RNA-seq transcriptome results and demonstrated that the expression patterns of the eight major pituitary hormone genes did not exhibit a significant gender difference at 90 dpf. For the first time, we report the quantitative global gene expression patterns at the juvenile and sexual maturity stages. These expression patterns may account for the dynamic neuroendocrine regulation observed in body metabolism.
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Nelson, M. A., and R. L. Metzenberg. "Sexual development genes of Neurospora crassa." Genetics 132, no. 1 (September 1, 1992): 149–62. http://dx.doi.org/10.1093/genetics/132.1.149.
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Abstract The filamentous fungus Neurospora crassa undergoes a complex program of sexual development to form a fruiting body composed of several kinds of specialized tissue. Subtractive hybridization was used to isolate genes that are expressed preferentially during this sexual phase. Many such sexual development (sdv) genes were identified in a cosmid library of Neurospora genomic DNA. Fourteen of the sdv genes were subcloned, and their expression in mutant strains and under crossing and vegetative growth conditions was examined. All of the regulated transcripts were less abundant (and in many cases not detectable) in strains grown under vegetative (high nitrogen) conditions, suggesting that nitrogen starvation is required for their synthesis. The expression of most of the sdv genes also required a functional A mating type product, even under crossing growth conditions, suggesting that this product functions as a master control in sexual development. To determine if the products of the sdv genes play essential roles in the sexual cycle, a reverse-genetic approach (based on RIP (repeat-induced point mutation)-mediated gene disruptions) was used to create mutations in the genes. A mutant strain (asd-1) with a recessive crossing defect (apparently caused by the RIP process) was isolated; in this strain, early development is normal and may asci are formed, but ascospores are never delineated. A second recessive mutant strain (asd-2) was apparently created by ectopic integration of the transforming DNA into a gene required for the sexual process; in this strain the sexual process was blocked at an early stage, and the ascogeneous tissue underwent little development.
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Töhönen, Virpi, Jessica Frygelius, Majid Mohammadieh, Ulrik Kvist, Lauri J. Pelliniemi, Kevin O'Brien, Katarina Nordqvist, and Anna Wedell. "Normal Sexual Development and Fertility in testatin Knockout Mice." Molecular and Cellular Biology 25, no. 12 (June 15, 2005): 4892–902. http://dx.doi.org/10.1128/mcb.25.12.4892-4902.2005.
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ABSTRACT The testatin gene was previously isolated in a screen focused on finding novel signaling molecules involved in sex determination and differentiation. testatin is specifically upregulated in pre-Sertoli cells in early fetal development, immediately after the onset of Sry expression, and was therefore considered a strong candidate for involvement in early testis development. testatin expression is maintained in the adult Sertoli cell, and it can also be found in a small population of germ cells. Testatin shows homology to family 2 cystatins, a group of broadly expressed small secretory proteins that are inhibitors of cysteine proteases in vitro but whose in vivo functions are unclear. testatin belongs to a novel subfamily among the cystatins, comprising genes that all show expression patterns that are strikingly restricted to reproductive tissue. To investigate a possible role of testatin in testis development and male reproduction, we have generated a mouse with targeted disruption of the testatin gene. We found no abnormalities in the testatin knockout mice with regard to fetal and adult testis morphology, cellular ultrastructure, body and testis weight, number of offspring, spermatogenesis, or hormonal parameters (testosterone, luteinizing hormone, and follicle-stimulating hormone).
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Tsitsigiannis, Dimitrios I., Terri M. Kowieski, Robert Zarnowski, and Nancy P. Keller. "Three putative oxylipin biosynthetic genes integrate sexual and asexual development in Aspergillus nidulans." Microbiology 151, no. 6 (June 1, 2005): 1809–21. http://dx.doi.org/10.1099/mic.0.27880-0.
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Oxylipins called psi factors have been shown to alter the ratio of asexual to sexual sporulation in the filamentous fungus Aspergillus nidulans. Analysis of the A. nidulans genome has led to the identification of three fatty acid oxygenases (PpoA, PpoB and PpoC) predicted to produce psi factors. Here, it is reported that deletion of ppoB (ΔppoB) reduced production of the oleic-acid-derived oxylipin psiBβ and increased the ratio of asexual to sexual spore development. Generation of the triple mutant ΔppoAΔppoBΔppoC resulted in a strain deficient in producing oleic- and linoleic-acid-derived 8′-hydroxy psi factor and caused increased and mis-scheduled activation of sexual development. Changes in asexual to sexual spore development were positively correlated to alterations in the expression of brlA and veA, respectively. PpoB and/or its products antagonistically mediate the expression levels of ppoA and ppoC, thus revealing regulatory feedback loops among these three genes. Phylogenetic analyses showed that ppo genes are present in both saprophytic and pathogenic Ascomycetes and Basidiomycetes, suggesting a conserved role for Ppo enzymes in the life cycle of fungi.
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Hull, Christina M., Marie-Josee Boily, and Joseph Heitman. "Sex-Specific Homeodomain Proteins Sxi1α and Sxi2a Coordinately Regulate Sexual Development in Cryptococcus neoformans." Eukaryotic Cell 4, no. 3 (March 2005): 526–35. http://dx.doi.org/10.1128/ec.4.3.526-535.2005.
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ABSTRACT Homeodomain proteins are central regulators of development in eukaryotes. In fungi, homeodomain proteins have been shown to control cell identity and sexual development. Cryptococcus neoformans is a human fungal pathogen with a defined sexual cycle that produces spores, the suspected infectious particles. Previously, only a single homeodomain regulatory protein involved in sexual development, Sxi1α, had been identified. Here we present the discovery of Sxi2a, a predicted but heretofore elusive cell-type-specific homeodomain protein essential for the regulation of sexual development. Our studies reveal that Sxi2a is necessary for proper sexual development and sufficient to drive this development in otherwise haploid α cells. We further show that Sxi1α and Sxi2a interact with one another and impart similar expression patterns for two key mating genes. The discovery of Sxi2a and its relationship with Sxi1α leads to a new model for how the sexual cycle is controlled in C. neoformans, with implications for virulence.
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Renfree, Marilyn B., Andrew J. Pask, and Geoff Shaw. "Sexual development of a model marsupial male." Australian Journal of Zoology 54, no. 3 (2006): 151. http://dx.doi.org/10.1071/zo05057.
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In eutherian mammals sexual differentiation occurs during fetal development, making experimental manipulation difficult, unlike in marsupials. We are investigating the roles of several key genes and hormones whose exact role in gonadal differentiation is still unclear using the tammar wallaby (Macropus eugenii) as a model. As in humans, unlike in mice, the testis-determining gene SRY is expressed in male tammar fetuses in many tissues over an extended period. Not all sexual differentiation depends on testicular hormones. Scrotum and mammary glands are under the control of X-linked gene(s). Our demonstration of DMRT1 expression in tammar and mouse ovaries suggests it has a wider role than previously thought. The Y-borne copy of ATRX (ATRY) is coexpressed with DMRT1 in developing testis. Gonadal sex reversal can be induced in males by neonatal oestrogen treatment and in females by grafting developing ovaries to males or culturing them in minimal medium. Treatments of developing young with various androgens, and studies of steroid metabolism have shown that the steroid androstenediol may have a previously unrecognised role in virilisation. Our studies using a marsupial model have given some surprising insights into the evolution and control of sexual development in all mammals.
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Greene, Andrew V., Nancy Keller, Hubertus Haas, and Deborah Bell-Pedersen. "A Circadian Oscillator in Aspergillus spp. Regulates Daily Development and Gene Expression." Eukaryotic Cell 2, no. 2 (April 2003): 231–37. http://dx.doi.org/10.1128/ec.2.2.231-237.2003.
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ABSTRACT We have established the presence of a circadian clock in Aspergillus flavus and Aspergillus nidulans by morphological and molecular assays, respectively. In A. flavus, the clock regulates an easily assayable rhythm in the development of sclerotia, which are large survival structures produced by many fungi. This developmental rhythm exhibits all of the principal clock properties. The rhythm is maintained in constant environmental conditions with a period of 33 h at 30°C, it can be entrained by environmental signals, and it is temperature compensated. This endogenous 33-h period is one of the longest natural circadian rhythms reported for any organism, and this likely contributes to some unique responses of the clock to environmental signals. In A. nidulans, no obvious rhythms in development are apparent. However, a free running and entrainable rhythm in the accumulation of gpdA mRNA (encoding glyceraldehyde-3-phosphate dehydrogenase) is observed, suggesting the presence of a circadian clock in this species. We are unable to identify an Aspergillus ortholog of frequency, a gene required for normal circadian rhythmicity in Neurospora crassa. Together, our data indicate the existence of an Aspergillus circadian clock, which has properties that differ from that of the well-described clock of N. crassa.
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Casas-Tintó, Sergio, Mercedes Arnés, and Alberto Ferrús. "Drosophila enhancer-Gal4 lines show ectopic expression during development." Royal Society Open Science 4, no. 3 (March 2017): 170039. http://dx.doi.org/10.1098/rsos.170039.
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In Drosophila melanogaster the most widely used technique to drive gene expression is the binary UAS/Gal4 system. We show here that a set of nervous system specific enhancers ( elav , D42/ Toll-6 , OK6/ RapGAP1 ) display ectopic activity in epithelial tissues during development, which is seldom considered in experimental studies. This ectopic activity is variable, unstable and influenced by the primary sequence of the enhancer and the insertion site in the chromosome. In addition, the ectopic activity is independent of the protein expressed, Gal4, as it is reproduced also with the expression of Gal80. Another enhancer, LN2 from the sex lethal ( Sxl ) gene, shows sex-dependent features in its ectopic expression. Feminization of LN2 expressing males does not alter the male specific pattern indicating that the sexual dimorphism of LN2 expression is an intrinsic feature of this enhancer. Other X chromosome enhancers corresponding to genes not related to sex determination do not show sexual dimorphism in their ectopic expressions. Although variable and unstable, the ectopic activation of enhancer-Gal4 lines seems to be regulated in terms of tissue and intensity. To characterize the full domain of expression of enhancer-Gal4 constructs is relevant for the design of transgenic animal models and biotechnology tools, as well as for the correct interpretation of developmental and behavioural studies in which Gal4 lines are used.
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Wang, Zheng, Francesc Lopez-Giraldez, Nina Lehr, Marta Farré, Ralph Common, Frances Trail, and Jeffrey P. Townsend. "Global Gene Expression and Focused Knockout Analysis Reveals Genes Associated with Fungal Fruiting Body Development in Neurospora crassa." Eukaryotic Cell 13, no. 1 (November 15, 2013): 154–69. http://dx.doi.org/10.1128/ec.00248-13.
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ABSTRACTFungi can serve as highly tractable models for understanding genetic basis of sexual development in multicellular organisms. Applying a reverse-genetic approach to advance such a model, we used random and multitargeted primers to assay gene expression across perithecial development inNeurospora crassa. We found that functionally unclassified proteins accounted for most upregulated genes, whereas downregulated genes were enriched for diverse functions. Moreover, genes associated with developmental traits exhibited stage-specific peaks of expression. Expression increased significantly across sexual development for mating type genemat a-1and format A-1specific pheromone precursorccg-4. In addition, expression of a gene encoding a protein similar to zinc finger,stc1, was highly upregulated early in perithecial development, and a strain with a knockout of this gene exhibited arrest at the same developmental stage. A similar expression pattern was observed for genes in RNA silencing and signaling pathways, and strains with knockouts of these genes were also arrested at stages of perithecial development that paralleled their peak in expression. The observed stage specificity allowed us to correlate expression upregulation and developmental progression and to identify regulators of sexual development. Bayesian networks inferred from our expression data revealed previously known and new putative interactions between RNA silencing genes and pathways. Overall, our analysis provides a fine-scale transcriptomic landscape and novel inferences regarding the control of the multistage development process of sexual crossing and fruiting body development inN. crassa.
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Alzan, Heba F., Audrey O. T. Lau, Donald P. Knowles, David R. Herndon, Massaro W. Ueti, Glen A. Scoles, Lowell S. Kappmeyer, and Carlos E. Suarez. "Expression of 6-Cys Gene Superfamily Defines Babesia bovis Sexual Stage Development within Rhipicephalus microplus." PLOS ONE 11, no. 9 (September 26, 2016): e0163791. http://dx.doi.org/10.1371/journal.pone.0163791.
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Watanabe, Y., and M. Yamamoto. "Schizosaccharomyces pombe pcr1+ encodes a CREB/ATF protein involved in regulation of gene expression for sexual development." Molecular and Cellular Biology 16, no. 2 (February 1996): 704–11. http://dx.doi.org/10.1128/mcb.16.2.704.
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The Schizosaccharomyces pombe pcr1 gene encodes a bZIP protein that apparently belongs to the cyclic AMP response element (CRE)-binding protein/activating transcription factor family. The deduced pcr1 gene product consists of 171 amino acid residues and is most similar to the mammalian CRE-BP1. A glutathione S-transferase-Pcr1 fusion protein produced in Escherichia coli was able to bind specifically to the CRE motif in vitro. Analysis with anti-Pcr1 serum suggested that Pcr1 is included in the major CRE-binding factors present in the S. pombe cell extract. Disruption of the pcr1 gene was not lethal, but the disruptant showed cold-sensitive growth on rich medium. The disruptant was also inefficient in mating and sporulation, though it was not completely sterile. Expression of the ste11 gene, which encodes a key transcription factor for sexual development, was greatly reduced in the disruptant, and overexpression of ste11+ suppressed the deficiency of the pcr1 disruptant in sexual development. It has been shown that expression of ste11 is negatively regulated by cyclic AMP-dependent protein kinase (PKA) and that the loss of PKA activity results in ectopic sexual development. Disruption of pcr1 blocked ectopic sexual development. Furthermore, disruption of pcr1 reduced expression of fbp1, a glucose-repressible gene negatively regulated by PKA. These results suggest that Pcr1 is a putative transcriptional regulator whose activity may be controlled by PKA. Alternatively, its activity may be independent of PKA, and full induction of ste11 and fbp1 expression requires the function of Pcr1 in addition to elimination of the repression by PKA.
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Yin, Chuntao, James E. Jurgenson, and Scot H. Hulbert. "Development of a Host-Induced RNAi System in the Wheat Stripe Rust Fungus Puccinia striiformis f. sp. tritici." Molecular Plant-Microbe Interactions® 24, no. 5 (May 2011): 554–61. http://dx.doi.org/10.1094/mpmi-10-10-0229.
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Rust fungi cause devastating diseases of wheat and other cereal species globally. Genetic resistance is the preferred method to control rusts but the effectiveness of race-specific resistance is typically transient due to the genetic plasticity of rust populations. The advent of RNA interference (RNAi) technology has shown promise for the engineering of resistance to some biotrophic pathogens in plants by altering the expression of essential pathogens' genes. Gene fragments from the rust fungi Puccinia striiformis f. sp. tritici or P. graminis f. sp. tritici were delivered to plant cells through the Barley stripe mosaic virus system, and some reduced the expression of the corresponding genes in the rust fungus. The ability to detect suppression was associated with the expression patterns of the fungal genes because reduction was only detected in transcripts with relatively high levels of expression in fungal haustoria. The results indicate that an in planta RNAi approach can be used in functional genomics research for rust fungi and that it could potentially be used to engineer durable resistance.
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Mukherjee, Arijit, and Jean-Michel Ané. "Germinating Spore Exudates from Arbuscular Mycorrhizal Fungi: Molecular and Developmental Responses in Plants and Their Regulation by Ethylene." Molecular Plant-Microbe Interactions® 24, no. 2 (February 2011): 260–70. http://dx.doi.org/10.1094/mpmi-06-10-0146.
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Arbuscular mycorrhizal (AM) fungi stimulate root development and induce expression of mycorrhization-specific genes in both eudicots and monocots. Diffusible factors released by AM fungi have been shown to elicit similar responses in Medicago truncatula. Colonization of roots by AM fungi is inhibited by ethylene. We compared the effects of germinating spore exudates (GSE) from Glomus intraradices in monocots and in eudicots, their genetic control, and their regulation by ethylene. GSE modify root architecture and induce symbiotic gene expression in both monocots and eudicots. The genetic regulation of root architecture and gene expression was analyzed using M. truncatula and rice symbiotic mutants. These responses are dependent on the common symbiotic pathway as well as another uncharacterized pathway. Significant differences between monocots and eudicots were observed in the genetic control of plant responses to GSE. However, ethylene inhibits GSE-induced symbiotic gene expression and root development in both groups. Our results indicate that GSE signaling shares similarities and differences in monocots versus eudicots, that only a subset of AM signaling pathways has been co-opted in legumes for the establishment of root nodulation with rhizobia, and that regulation of these pathways by ethylene is a feature conserved across higher land plants.
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Faccio, P., C. Vazquez-Rovere, E. Hopp, G. González, C. Décima-Oneto, E. Favret, A. D. Paleo, and P. Franzone. "Increased tolerance to wheat powdery mildew by heterologous constitutive expression of the Solanum chacoense Snakin-1 gene." Czech Journal of Genetics and Plant Breeding 47, Special Issue (October 20, 2011): S135—S141. http://dx.doi.org/10.17221/3268-cjgpb.
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Great efforts are currently being devoted to studying the use of transgenes to confer resistance to phytopathogenic fungi. Snakin-1 is a broad-spectrum antimicrobial peptide isolated from Solanum that is active in vitro against bacteria and fungi. Recently, it was reported that overexpression of the snakin-1 (SN1) gene in transgenic potato plants enhanced resistance to Rhizoctonia solani and Erwinia carotovora. In this work wheat transgenic plants that constitutively expressed the S. chacoense SN1 gene were challenged with Blumeria graminis f.sp. tritici. Enhanced resistance to the pathogen was observed in two transgenic lines in which the development of the disease was delayed and reduced compared with the wild type variety ProINTA Federal. An association between high resistance to the pathogen and a high level of snakin-1 transcripts in the plant was observed. This is the first report on SN1 gene expression in Gramineae and its effects on wheat powdery mildew development.
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Gao, Ling-Ling, Wolfgang Knogge, Gabriele Delp, F. Andrew Smith, and Sally E. Smith. "Expression Patterns of Defense-Related Genes in Different Types of Arbuscular Mycorrhizal Development in Wild-Type and Mycorrhiza-Defective Mutant Tomato." Molecular Plant-Microbe Interactions® 17, no. 10 (October 2004): 1103–13. http://dx.doi.org/10.1094/mpmi.2004.17.10.1103.
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The expression of defense-related genes was analyzed in the interactions of six arbuscular mycorrhizal (AM) fungi with the roots of wild-type tomato (Lycopersicon esculentum Mill.) cv. 76R and of the near-isogenic mycorrhiza-defective mutant rmc. Depending on the fungal species, wild-type tomato forms both major morphological AM types, Arum and Paris. The mutant rmc blocks the penetration of the root surface or invasion of the root cortex by most species of AM fungi, but one fungus has been shown to develop normal mycorrhizas. In the wild-type tomato, accumulation of mRNA representing a number of defense-related genes was low in Arum-type interactions, consistent with findings for this AM morphotype in other plant species. In contrast, Paris-type colonization, particularly by members of the family Gigasporaceae, was accompanied by a substantial transient increase in expression of some defense-related genes. However, the extent of root colonization did not differ significantly in the two wild-type AM morpho-types, suggesting that accumulation of defense gene products per se does not limit mycorrhiza development. In the mutant, interactions in which the fungus failed to penetrate the root lacked significant accumulation of defense gene mRNAs. However, phenotypes in which the fungus penetrated epidermal or hypodermal cells were associated with an enhanced and more prolonged gene expression. These results are discussed in relation to the mechanisms that may underlie the specificity of the interactions between AM fungi and the rmc mutant.
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Xiang, Quanju, Keyu Shen, Xiumei Yu, Ke Zhao, Yunfu Gu, Xiaoping Zhang, Xiaoqiong Chen, and Qiang Chen. "Analysis of the oligopeptide transporter gene family in Ganoderma lucidum: structure, phylogeny, and expression patterns." Genome 60, no. 4 (April 2017): 293–302. http://dx.doi.org/10.1139/gen-2016-0120.
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Oligopeptide transporters (OPTs) are believed to transport broad ranges of substrates across the plasma membrane from the extracellular environment into the cell and are thought to contribute to various biological processes. In the present study, 13 putative OPTs (Gl-OPT1 to Gl-OPT13) were identified through extensive search of Ganoderma lucidum genome database. Phylogenetic analysis with OPTs from other fungi and plants indicates that these genes can be further divided into five groups. Motif compositions of OPT members are highly conserved in each group, indicative of functional conservation. Expression profile analysis of the 13 Gl-OPT genes indicated that, with the exception of Gl-OPT7–Gl-OPT9, for which no transcripts were detected, all paralogues were differentially expressed, suggesting their potential involvement in stress response and functional development of fungi. Overall, the analyses in this study provide a starting point for elucidating the functions of OPT in G. lucidum, and for understanding the complexities of metabolic regulation.
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Chen, Biao, Guitao Liang, Xuenong Zhu, Yuwen Tan, Jiguo Xu, Hongxiang Wu, Huirong Mao, et al. "Gene Expression Profiling in Ovaries and Association Analyses Reveal HEP21 as a Candidate Gene for Sexual Maturity in Chickens." Animals 10, no. 2 (January 21, 2020): 181. http://dx.doi.org/10.3390/ani10020181.
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The age of onset of sexual maturity is an important reproductive trait in chickens. In this study, we explored candidate genes associated with sexual maturity and ovary development in chickens. We performed DGE RNA-sequencing analyses of ovaries of pre-laying (P-F-O1, L-F-O1) and laying (P-F-O2, L-F-O2) hens of two sub-breeds of Ningdu Yellow chicken. A total of 3197 genes were identified in the two comparisons, and 966 and 1860 genes were detected exclusively in comparisons of P-F-O1 vs. P-F-O2 and L-F-O1 vs. L-F-O2, respectively. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses showed that genes involved in transmembrane signaling receptor activity, cell adhesion, developmental processes, the neuroactive ligand–receptor interaction pathway, and the calcium signaling pathway were enriched in both comparisons. Genes on these pathways, including growth hormone (GH), integrin subunit beta 3 (ITGB3), thyroid stimulating hormone subunit beta (TSHB), prolactin (PRL), and transforming growth factor beta 3 (TGFB3), play indispensable roles in sexual maturity. As a gene unique to poultry, hen egg protein 21 kDa (HEP21) was chosen as the candidate gene. Differential expression and association analyses were performed. RNA-seq data and qPCR showed that HEP21 was significantly differentially expressed in pre-pubertal and pubertal ovaries. A total of 23 variations were detected in HEP21. Association analyses of single nucleotide polymorphisms (SNPs) in HEP21 and reproductive traits showed that rs315156783 was significantly related to comb height at 84 and 91 days. These results indicate that HEP21 is a candidate gene for sexual maturity in chickens. Our results contribute to a more comprehensive understanding of sexual maturity and reproduction in chickens.
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Brambl, Robert. "Fungal physiology and the origins of molecular biology." Microbiology 155, no. 12 (December 1, 2009): 3799–809. http://dx.doi.org/10.1099/mic.0.035238-0.
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Molecular biology has several distinct origins, but especially important are those contributed by fungal and yeast physiology, biochemistry and genetics. From the first gene action studies that became the basis of our understanding of the relationship between genes and proteins, through chromosome structure, mitochondrial genetics and membrane biogenesis, gene silencing and circadian clocks, studies with these organisms have yielded basic insight into these processes applicable to all eukaryotes. Examples are cited of pioneering studies with fungi that have stimulated new research in clinical medicine and agriculture; these studies include sexual interactions, cell stress responses, the cytoskeleton and pathogenesis. Studies with the yeasts and fungi have been effective in applying the techniques and insights gained from other types of experimental systems to research in fungal cell signalling, cell development and hyphal morphogenesis.
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PRADEL, G. "Proteins of the malaria parasite sexual stages: expression, function and potential for transmission blocking strategies." Parasitology 134, no. 14 (August 23, 2007): 1911–29. http://dx.doi.org/10.1017/s0031182007003381.
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SUMMARYThe sexual phase of the malaria pathogen,Plasmodium falciparum, culminates in fertilization within the midgut of the mosquito and represents a crucial step in the completion of the parasite's life-cycle and transmission of the disease. Two decades ago, the first sexual stage-specific surface proteins were identified, among themPfs230,Pfs48/45, andPfs25, which were of scientific interest as candidates for the development of transmission blocking vaccines. A decade later, gene information gained from the sequencing of theP. falciparumgenome led to the identification of numerous additional sexual-stage proteins with antigenic properties and novel enzymes that putatively possess regulatory functions during sexual-stage development. This review aims to summarize the sexual-stage proteins identified to date, to compare their stage specificities and expression patterns and to highlight novel regulative mechanisms of sexual differentiation. The prospective candidacy of select sexual-stage proteins as targets for transmission blocking strategies will be discussed.
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Santos, P. R. S., F. D. Oliveira, M. A. M. Arroyo, M. F. Oliveira, P. Castelucci, A. J. Conley, and A. C. Assis Neto. "Steroidogenesis during postnatal testicular development of Galea spixii." Reproduction 154, no. 5 (November 2017): 645–52. http://dx.doi.org/10.1530/rep-17-0075.
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The androgen/estrogen balance is essential for normal sexual development and reproduction in mammals. Studies performed herein investigated the potential for estrogen synthesis in cells of the testes of a hystricomorph rodent, Galea spixii. The study characterized the expression of the key enzymes responsible for estrogen and androgen synthesis, cytochromes P450 aromatase (P450arom), 17α-hydroxylase/17,20-lyase (P450c17) respectively, as well as the redox partner NADPH cytochrome P450 oxido-reductase (CPR) required to support electron transfer and catalysis of these P450s, by immunohistochemistry (IHC) and quantitative polymerase chain reaction (qPCR) analysis, throughout postnatal sexual development. Testes (immature, pre-pubertal, pubertal and post-pubertal) were collected, fixed for IHC (CYP19, CYP17 and CPR) and stored frozen for qPCR for the relevant gene transcripts (Cyp19a1 and Cyp17a1). Expression of P450c17 was significantly elevated at the pre-pubertal and pubertal stages. Based on IHC, P450c17 was expressed only in Leydig cell clusters. The expression of P450arom was detectable at all stages of sexual development of Galea spixii. IHC data suggest that estrogen synthesis was not restricted to somatic cells (Leydig cells/Sertoli cells), but that germ cells may also be capable of converting androgens into estrogens, important for testicular function and spermatogenesis.
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Barba-Ostria, Carlos, Fernando Lledías, and Dimitris Georgellis. "The Neurospora crassa DCC-1 Protein, a Putative Histidine Kinase, Is Required for Normal Sexual and Asexual Development and Carotenogenesis." Eukaryotic Cell 10, no. 12 (November 4, 2011): 1733–39. http://dx.doi.org/10.1128/ec.05223-11.
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ABSTRACT Two-component signaling pathways based on phosphoryl group transfer between histidine kinase and response regulator proteins regulate environmental responses in bacteria, archaea, plants, slime molds, and fungi. Here we characterize a mutant form of DCC-1, a putative histidine kinase encoded by the NCU00939 gene of the filamentous fungus Neurospora crassa . We show that this protein participates in the regulation of processes such as conidiation, perithecial development, and, to a certain degree, carotenogenesis. Furthermore, DCC-1 is suggested to exert its effect by promoting cyclic AMP production, thereby placing this protein within the context of a signaling pathway.
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Delemarre-van de Waal, H. A., K. A. Burton, E. B. Kabigting, D. K. Clifton, and R. A. Steiner. "SEXUAL DIMORPHISM OF GALANIN GENE EXPRESSION IN GROWTH HORMONE-RELEASING HORMONE NEURONS OF THE RAT DURING DEVELOPMENT." Pediatric Research 33 (May 1993): S27. http://dx.doi.org/10.1203/00006450-199305001-00145.
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Guenther, John C., Heather E. Hallen-Adams, Heike Bücking, Yair Shachar-Hill, and Frances Trail. "Triacylglyceride Metabolism by Fusarium graminearum During Colonization and Sexual Development on Wheat." Molecular Plant-Microbe Interactions® 22, no. 12 (December 2009): 1492–503. http://dx.doi.org/10.1094/mpmi-22-12-1492.
Full textAbstract:
Fusarium graminearum, a devastating pathogen of small grains, overwinters on crop residues and produces ephemeral perithecia. Accumulation of lipids in overwintering hyphae would provide reserves for overwinter survival and perithecium development. Fatty acid composition of cultures during perithecium development indicated a drop in neutral lipid levels during development but little change in fatty acid composition across stages. Microscopic examination of cultures early in sexual development revealed hyphal cells engorged with lipid bodies. In comparison, vegetative hyphae contained few lipid bodies. Microarray analysis was performed on wheat stems at stages of colonization through perithecium development. Gene expression analysis during stages of perithecium development both in planta and in vitro (previously published) supports the view that lipid biosynthesis occurs during early stages of wheat colonization leading to sexual development and that lipid oxidation occurs as perithecia are developing. Analysis of gene expression during the stages of wheat stem colonization also revealed sets of genes unique to these stages. These results support the view that lipids accumulate in hyphae colonizing wheat stalks and are subsequently used in perithecium formation on stalk tissue. These results indicate that extensive colonization of plant tissue prior to harvest is essential for subsequent sporulation on crop residues and, thus, has important implications for inoculum reduction.
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Wilson, Richard A., Harold W. Gardner, and Nancy P. Keller. "Cultivar-Dependent Expression of a Maize Lipoxygenase Responsive to Seed Infesting Fungi." Molecular Plant-Microbe Interactions® 14, no. 8 (August 2001): 980–87. http://dx.doi.org/10.1094/mpmi.2001.14.8.980.
Full textAbstract:
Maize kernels are highly susceptible to Aspergillus spp. infection and aflatoxin (AF) contamination. Fatty acid signaling molecules appear to mediate the plant-fungal interaction by affecting the growth, development, and AF production of the fungus. In particular, fatty acid derivatives of the plant lipoxygenase (LOX) pathway are implicated in the Aspergillus spp.-seed interaction. The 9(S)-hydroperoxide derivative of linoleic acid promotes transcription of AF genes, whereas the 13(S)-hydroperoxide derivative decreases AF gene expression and production; both are sporulation factors. Our goal was to identify LOX genes responsive to Aspergillus spp. colonization and determine their specificities, 9(S)- or 13(S)-. Screening maize LOX expressed sequence tags (ESTs) identified one clone, cssap 92, which is highly expressed in Aspergillus spp.-infected seed susceptible to AF contamination and repressed in lines with resistance to AF contamination. The accumulation of cssap 92 transcript was similar during Fusarium spp. infection. The cDNA clone has 94% identity to the previously described L2 LOX gene from maize. Product-specificity analysis of the CSSAP 92 protein shows that it preferentially adds oxygen to carbon 9 of linoleic acid. Because 9(S)-hydroperoxy linoleic acid has been implicated as an aflatoxin-signaling molecule, it is possible that cssap 92 could be used as a biomarker that is indicative of AF resistance in maize lines.
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Loubradou, Gabriel, Joel Bégueret, and Béatrice Turcq. "A Mutation in an HSP90 Gene Affects the Sexual Cycle and Suppresses Vegetative Incompatibility in the Fungus Podospora anserina." Genetics 147, no. 2 (October 1, 1997): 581–88. http://dx.doi.org/10.1093/genetics/147.2.581.
Full textAbstract:
Vegetative incompatibility is widespread in fungi but its molecular mechanism and biological function are still poorly understood. A way to study vegetative incompatibility is to investigate the function of genes whose mutations suppress this phenomenon. In Podospora anserina, these genes are known as mod genes. In addition to suppressing vegetative incompatibility, mod mutations cause some developmental defects. This suggests that the molecular mechanisms of vegetative incompatibility and development pathways are interconnected. The mod-E1 mutation was isolated as a suppressor of the developmental defects of the mod-D2 strain. We show here that mod-E1 also partially suppresses vegetative incompatibility, strengthening the link between development and vegetative incompatibility. mod-E1 is the first suppressor of vegetative incompatibility characterized at the molecular level. It encodes a member of the Hsp90 family, suggesting that development and vegetative incompatibility use common steps of a signal transduction pathway. The involvement of mod-E in the sexual cycle has also been further investigated.
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Ahrén, Dag, Margareta Tholander, Csaba Fekete, Balaji Rajashekar, Eva Friman, Tomas Johansson, and Anders Tunlid. "Comparison of gene expression in trap cells and vegetative hyphae of the nematophagous fungus Monacrosporium haptotylum." Microbiology 151, no. 3 (March 1, 2005): 789–803. http://dx.doi.org/10.1099/mic.0.27485-0.
Full textAbstract:
Nematode-trapping fungi enter the parasitic stage by developing specific morphological structures called traps. The global patterns of gene expression in traps and mycelium of the fungus Monacrosporium haptotylum were compared. The trap of this fungus is a unicellular spherical structure called the knob, which develops on the apex of a hyphal branch. RNA was isolated from knobs and mycelium and hybridized to a cDNA array containing probes of 2822 EST clones of M. haptotylum. Despite the fact that the knobs and mycelium were grown in the same medium, there were substantial differences in the patterns of genes expressed in the two cell types. In total, 23·3 % (657 of 2822) of the putative genes were differentially expressed in knobs versus mycelium. Several of these genes displayed sequence similarities to genes known to be involved in regulating morphogenesis and cell polarity in fungi. Among them were several putative homologues for small GTPases, such as rho1, rac1 and ras1, and a rho GDP dissociation inhibitor (rdi1). Several homologues to genes involved in stress response, protein synthesis and protein degradation, transcription, and carbon metabolism were also differentially expressed. In the last category, a glycogen phosphorylase (gph1) gene homologue, one of the most upregulated genes in the knobs as compared to mycelium, was characterized. A number of the genes that were differentially expressed in trap cells are also known to be regulated during the development of infection structures in plant-pathogenic fungi. Among them, a gas1 (mas3) gene homologue (designated gks1), which is specifically expressed in appressoria of the rice blast fungus, was characterized.