scholarly journals Characterization of Serine Proteinase Expression in Agaricus bisporus and Coprinopsis cinerea by Using Green Fluorescent Protein and the A. bisporus SPR1 Promoter

2008 ◽  
Vol 75 (3) ◽  
pp. 792-801 ◽  
Author(s):  
Mary N. Heneghan ◽  
Claudine Porta ◽  
Cunjin Zhang ◽  
Kerry S. Burton ◽  
Michael P. Challen ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Fruiting Body ◽  
Agaricus Bisporus ◽  
Fluorescent Protein ◽  
Serine Proteinase ◽  
Developmental Expression ◽  
Coprinopsis Cinerea ◽  
Body Development ◽  
Fruiting Body Development ◽  
Green Fluorescent

ABSTRACT The Agaricus bisporus serine proteinase 1 (SPR1) appears to be significant in both mycelial nutrition and senescence of the fruiting body. We report on the construction of an SPR promoter::green fluorescent protein (GFP) fusion cassette, pGreen_hph1_SPR_GFP, for the investigation of temporal and developmental expression of SPR1 in homobasidiomycetes and to determine how expression is linked to physiological and environmental stimuli. Monitoring of A. bisporus pGreen_hph1_SPR_GFP transformants on media rich in ammonia or containing different nitrogen sources demonstrated that SPR1 is produced in response to available nitrogen. In A. bisporus fruiting bodies, GFP activity was localized to the stipe of postharvest senescing sporophores. pGreen_hph1_SPR_GFP was also transformed into the model basidiomycete Coprinopsis cinerea. Endogenous C. cinerea proteinase activity was profiled during liquid culture and fruiting body development. Maximum activity was observed in the mature cap, while activity dropped during autolysis. Analysis of the C. cinerea genome revealed seven genes showing significant homology to the A. bisporus SPR1 and SPR2 genes. These genes contain the aspartic acid, histidine, and serine residues common to serine proteinases. Analysis of the promoter regions revealed at least one CreA and several AreA regulatory motifs in all sequences. Fruiting was induced in C. cinerea dikaryons, and fluorescence was determined in different developmental stages. GFP expression was observed throughout the life cycle, demonstrating that serine proteinase can be active in all stages of C. cinerea fruiting body development. Serine proteinase expression (GFP fluorescence) was most concentrated during development of young tissue, which may be indicative of high protein turnover during cell differentiation.

scholarly journals Strand-Specific RNA-Seq Analyses of Fruiting Body Development in Coprinopsis cinerea

PLoS ONE ◽  
2015 ◽  
Vol 10 (10) ◽  
pp. e0141586 ◽  
Author(s):  
Hajime Muraguchi ◽  
Kiwamu Umezawa ◽  
Mai Niikura ◽  
Makoto Yoshida ◽  
Toshinori Kozaki ◽  
...  
Keyword(s):  
Fruiting Body ◽  
Rna Seq ◽  
Coprinopsis Cinerea ◽  
Body Development ◽  
Fruiting Body Development

scholarly journals Discovery of microRNA-like RNAs during early fruiting body development in the model mushroom Coprinopsis cinerea

PLoS ONE ◽  
2018 ◽  
Vol 13 (9) ◽  
pp. e0198234 ◽  
Author(s):  
Amy Yuet Ting Lau ◽  
Xuanjin Cheng ◽  
Chi Keung Cheng ◽  
Wenyan Nong ◽  
Man Kit Cheung ◽  
...  
Keyword(s):  
Fruiting Body ◽  
Coprinopsis Cinerea ◽  
Body Development ◽  
Fruiting Body Development

Integrative, multifunctional plasmids for hypha-specific or constitutive expression of green fluorescent protein in Candida albicans

Microbiology ◽  
2003 ◽  
Vol 149 (10) ◽  
pp. 2977-2986 ◽  
Author(s):  
Janet F. Staab ◽  
Yong-Sun Bahn ◽  
Paula Sundstrom
Keyword(s):  
Candida Albicans ◽  
Green Fluorescent Protein ◽  
Germ Tube ◽  
Fluorescent Protein ◽  
Constitutive Expression ◽  
Tube Formation ◽  
Developmental Expression ◽  
Specific Gene ◽  
Expression Of Genes ◽  
Green Fluorescent

The authors have engineered plasmid constructs for developmental and constitutive expression of yeast-enhanced green fluorescent protein (yEGFP3) in Candida albicans. The promoter for the hyphae-specific gene Hyphal Wall Protein 1 (HWP1) conferred developmental expression of yEGFP3 in germ tubes and hyphae but not in yeasts or pseudohyphae when targeted to the ENO1 (enolase) locus in single copy. The pHWP1GFP3 construct allows for the easy visualization of HWP1 promoter activity in individual cells expressing true hyphae without having to prepare RNA for analysis. Constitutive expression of yEGFP was seen in all cell morphologies when the HWP1 promoter was replaced with the ENO1 promoter region. The use of the plasmids for expression of genes other than yEGFP3 was examined by substituting the putative C. albicans BCY1 (SRA1) gene, a component of the cAMP signalling pathway involved in yeast to hyphae transitions, for yEGFP3. Strains overexpressing BCY1 from the ENO1 promoter were inhibited in germ tube formation and filamentation in both liquid and solid media, a phenotype consistent with keeping protein kinase A in its inactive form by association with Bcy1p. The plasmids are suitable for studies of germ tube induction or assessing germ tube formation by measuring yEGFP3 expression, for inducible expression of genes concomitant with germ tube formation by the HWP1 promoter, for constitutive expression of genes by the ENO1 promoter, and for expressing yEGFP3 using a promoter of choice.

scholarly journals 5'-Serial Analysis of Gene Expression studies reveal a transcriptomic switch during fruiting body development in Coprinopsis cinerea

BMC Genomics ◽  
2013 ◽  
Vol 14 (1) ◽  
pp. 195 ◽  
Author(s):  
Chi Keung Cheng ◽  
Chun Hang Au ◽  
Sarah K Wilke ◽  
Jason E Stajich ◽  
Miriam E Zolan ◽  
...  
Keyword(s):  
Gene Expression ◽  
Fruiting Body ◽  
Coprinopsis Cinerea ◽  
Body Development ◽  
Expression Studies ◽  
Fruiting Body Development ◽  
Gene Expression Studies

scholarly journals Discovery of microRNA-like RNAs during early fruiting body development in the model mushroom Coprinopsis cinerea

2018 ◽  
Author(s):  
Amy Yuet Ting Lau ◽  
Xuanjin Cheng ◽  
Chi Keung Cheng ◽  
Wenyan Nong ◽  
Man Kit Cheung ◽  
...  
Keyword(s):  
Fruiting Body ◽  
Developmental Stages ◽  
Expression Patterns ◽  
Regulation Of Gene Expression ◽  
Evolutionary Relationship ◽  
Mirna Biogenesis ◽  
Coprinopsis Cinerea ◽  
Stem Loop ◽  
Body Development ◽  
Fruiting Body Development

AbstractCoprinopsis cinerea is a model mushroom particularly suited to study fungal fruiting body development and the evolution of multicellularity in fungi. While microRNAs (miRNAs) are extensively studied in animals and plants for their essential roles in post-transcriptional regulation of gene expression, miRNAs in fungi are less well characterized and their potential roles in controlling mushroom development remain unknown. To identify miRNA-like RNAs (milRNAs) in C. cinerea and explore their expression patterns during the early developmental transition of mushroom development, small RNA libraries of vegetative mycelium and primordium were generated and putative milRNA candidates were identified following the standards of miRNA prediction in animals and plants. Two out of 22 novel predicted milRNAs, cci-milR-12c and cci-milR-13e-5p, were validated by northern blot and stem-loop reverse transcription real-time PCR. Cci-milR-12c was differentially expressed whereas the expression levels of cci-milR-13e-5p were similar in the two developmental stages. Target prediction of the validated milRNAs resulted in genes associated with fruiting body development, including pheromone, hydrophobin, cytochrome P450, and protein kinase. Besides, essential genes for miRNA biogenesis, including three coding for Dicer-like (DCL), two for Argonaute-like (AGO-like) and one for quelling deficient-2 (QDE-2) proteins, were identified in the C. cinerea genome. Phylogenetic analysis showed that the DCL and AGO-like proteins of C. cinerea were more closely related to those in other basidiomycetes and ascomycetes than to animals and plants. Taken together, our findings provided the first evidence of milRNAs in the model mushroom and their potential roles in regulating fruiting body development. Information on the evolutionary relationship of milRNA biogenesis proteins across kingdoms has also provided new insights into further functional and evolutionary studies of miRNAs.

scholarly journals Transcriptional profiling elucidates the essential role of glycogen synthase kinase 3 to fruiting body formation in Coprinopsis cinerea

2018 ◽  
Author(s):  
Kathy PoLam Chan ◽  
Jinhui Chang ◽  
Yichun Xie ◽  
Man Kit Cheung ◽  
Ka Lee Ma ◽  
...  
Keyword(s):  
Fruiting Body ◽  
Glycogen Synthase ◽  
Glycogen Synthase Kinase 3 ◽  
Coprinopsis Cinerea ◽  
Fruiting Body Formation ◽  
Body Formation ◽  
Body Development ◽  
Fruiting Body Development ◽  
Control Samples ◽  
Time Point

The functions of glycogen synthase kinase 3 (GSK3) have been well-studied in animal, plant and yeast. However, information on its roles in basidiomycetous fungi is still limited. In this study, we used the model mushroom Coprinopsis cinerea to study the characteristics of GSK3 in fruiting body development. Application of a GSK3 inhibitor Lithium chloride (LiCl) induced enhanced mycelial growth and inhibited fruiting body formation in C. cinerea. RNA-Seq of LiCl-treated C. cinerea resulted in a total of 14128 unigenes. There were 1210 differentially expressed genes (DEGs) between the LiCl-treated samples and control samples in the mycelium stage (first time point), whereas 1402 DEGs were detected at the stage when the control samples formed hyphal knots and the treatment samples were still in mycelium (second time point). Kyoto Encyclopedia of Genes and Genome (KEGG) pathway enrichment analysis of the DEGs revealed significant associations between the enhanced mycelium growth in LiCl treated C. cinerea and metabolism pathways such as “biosynthesis of secondary metabolite” and “biosynthesis of antibiotics”. In addition, DEGs involved in cellular process pathways, including “cell cycle-yeast” and “meiosis-yeast”, were identified in C. cinerea fruiting body formation suppressed by LiCl under favorable environmental conditions. Our findings suggest that GSK3 activity is essential for fruiting body formation as it affects the expression of fruiting body induction genes and genes in cellular processes. Further functional studies of GSK3 in basidiomycetous fungi may help understand the relationships between environmental signals and fruiting body development.

scholarly journals The genome of Auriculariopsis ampla sheds light on fruiting body development and wood-decay of bark-inhabiting fungi

2019 ◽  
Author(s):  
Éva Almási ◽  
Neha Sahu ◽  
Krisztina Krizsán ◽  
Balázs Bálint ◽  
Gábor M. Kovács ◽  
...  
Keyword(s):  
Fruiting Body ◽  
Schizophyllum Commune ◽  
Wood Decay ◽  
High Rate ◽  
White Rot ◽  
Developmental Expression ◽  
Close Relative ◽  
Plant Materials ◽  
Body Development ◽  
Fruiting Body Development

AbstractThe Agaricomycetes are fruiting body forming fungi that produce some of the most efficient enzyme systems to degrade woody plant materials. Despite decades-long interest in the ecological and functional diversity of wood-decay types and in fruiting body development, the evolution of the genetic repertoires of both traits are incompletely known. Here, we sequenced and analyzed the genome of Auriculariopsis ampla, a close relative of the model species Schizophyllum commune. Comparative analyses of wood-decay genes in these and other 29 Agaricomycetes species revealed that the gene family composition of A. ampla and S. commune are transitional between that of white rot species and less efficient wood-degraders (brown rot, ectomycorrhizal). Rich repertoires of suberinase and tannase genes were found in both species, with tannases generally restricted to species that preferentially colonize bark-covered wood. Analyses of fruiting body transcriptomes in both A. ampla and S. commune highlighted a high rate of divergence of developmental gene expression. Several genes with conserved developmental expression were found, nevertheless, including 9 new transcription factors as well as small secreted proteins, some of which may serve as fruiting body-specific effector molecules. Taken together, the genome sequence and developmental transcriptome of Auriculariopsis ampla has highlighted novel aspects of wood-decay diversity and of fruiting body development in mushroom-forming fungi.

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