{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T05:16:36Z","timestamp":1788498996724,"version":"build-2803163510"},"reference-count":32,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T00:00:00Z","timestamp":1788307200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NSF","award":["# MCB2125018"],"award-info":[{"award-number":["# MCB2125018"]}]},{"name":"U.S. Department of Energy, Office of Science, Basic Energy Sciences, Chemical Sciences, Geosciences and Biosciences Division","award":["#DE-SC0015662"],"award-info":[{"award-number":["#DE-SC0015662"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J. Fungi","JoF"],"abstract":"<jats:p>We carried out carbohydrate composition and carbohydrate linkage analyses on Neurospora crassa cell walls from vegetative hyphae, conidia (asexual spores), perithecia (female mating structures), and ascospores (sexual spores). We show that the composition and structure of the cell wall undergo dramatic changes during the N. crassa life cycle. All the cell walls contain \u03b2-1,3-glucan and mixed \u03b2-1,3-\/\u03b2-1,4-glucan (lichenin). The conidia cell walls and perithecia cell walls also contain \u03b1-1,3-glucan. \u03b2-1,3-glucan was the most abundant polysaccharide in the vegetative hyphae cell walls and mixed \u03b2-1,3-\/\u03b2-1,4-glucan was the most abundant polysaccharide in the perithecia and ascospore cell walls. Chitin was a major polysaccharide in the ascospore cell walls. Ascospore and perithecia cell walls also contained melanin as a structural element. We conclude that the composition and structure of the cell wall changes dramatically as cells proceed through the various stages of the N. crassa life cycle.<\/jats:p>","DOI":"10.3390\/jof12090657","type":"journal-article","created":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T07:53:11Z","timestamp":1788335591000},"page":"657","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Characterization of the Polysaccharide and Protein Composition of Neurospora crassa Conidium, Perithecium, and Ascospore Cell Walls"],"prefix":"10.3390","volume":"12","author":[{"given":"Protyusha","family":"Dey","sequence":"first","affiliation":[{"name":"Department of Biological Sciences, SUNY University at Buffalo, Buffalo, NY 14260, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ian","family":"Black","sequence":"additional","affiliation":[{"name":"Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christian","family":"Heiss","sequence":"additional","affiliation":[{"name":"Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liyanage Devthilini","family":"Fernando","sequence":"additional","affiliation":[{"name":"Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Parastoo","family":"Azadi","sequence":"additional","affiliation":[{"name":"Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Piotr","family":"Shtyk","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, SUNY University at Buffalo, Buffalo, NY 14260, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen J.","family":"Free","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, SUNY University at Buffalo, Buffalo, NY 14260, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"28513415","DOI":"10.1128\/microbiolspec.FUNK-0035-2016","article-title":"The Fungal Cell Wall: Structure, Biosynthesis, and Function","volume":"5","author":"Gow","year":"2017","journal-title":"Microbiol. Spectr."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Patel, P.K., and Free, S.J. (2019). The Genetics and Biochemistry of Cell Wall Structure and Synthesis in Neurospora crassa, a Model Filamentous Fungus. Front. Microbiol., 10.","DOI":"10.3389\/fmicb.2019.02294"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"103991","DOI":"10.1016\/j.fgb.2025.103991","article-title":"Fungal cell wall biogenesis: Structural complexity, regulation and inhibition","volume":"179","author":"Gow","year":"2025","journal-title":"Fungal Genet. Biol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/B978-0-12-407677-8.00002-6","article-title":"Fungal cell wall organization and biosynthesis","volume":"81","author":"Free","year":"2013","journal-title":"Adv. Genet."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Beauvais, A., and Latge, J.P. (2018). Special Issue: Fungal Cell Wall. J. Fungi., 4.","DOI":"10.3390\/jof4030091"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1259","DOI":"10.1016\/j.funbio.2023.05.001","article-title":"Cell wall of Aspergillus fumigatus: Variability and response to stress","volume":"127","author":"Latge","year":"2023","journal-title":"Fungal Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"11569","DOI":"10.1016\/S0021-9258(18)37997-3","article-title":"Chemical composition of the yeast ascospore wall. The second outer layer consists of chitosan","volume":"263","author":"Briza","year":"1988","journal-title":"J. Biol. Chem."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"4384","DOI":"10.1128\/jb.169.9.4384-4387.1987","article-title":"Tunicamycin inhibition of epispore formation in Saccharomyces cerevisiae","volume":"169","author":"Weinstock","year":"1987","journal-title":"J. Bacteriol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e0349523","DOI":"10.1128\/spectrum.03495-23","article-title":"A comparative cell wall analysis of Trichoderma spp. confirms a conserved polysaccharide scaffold and suggests an important role for chitosan in mycoparasitism","volume":"12","author":"Kappel","year":"2024","journal-title":"Microbiol. Spectr."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1618","DOI":"10.1099\/mic.0.080002-0","article-title":"Neurospora crassa 1,3-alpha-glucan synthase, AGS-1, is required for cell wall biosynthesis during macroconidia development","volume":"160","author":"Fu","year":"2014","journal-title":"Microbiology"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/0076-6879(71)17168-6","article-title":"Genetic and microbiological research techniques for Neurospora crassa","volume":"27","author":"Davis","year":"1970","journal-title":"Meth. Enzymol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"103826","DOI":"10.1016\/j.fgb.2023.103826","article-title":"Characterization of the need for galactofuranose during the Neurospora crassa life cycle","volume":"168","author":"Schaff","year":"2023","journal-title":"Fungal Genet. Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1016\/j.fgb.2017.03.002","article-title":"Genetic and biochemical characterization of the GH72 family of cell wall transglycosylases in Neurospora crassa","volume":"101","author":"Ao","year":"2017","journal-title":"Fungal Genet. Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.fgb.2016.07.003","article-title":"A proteomic and genetic analysis of the Neurospora crassa conidia cell wall proteins identifies two glycosyl hydrolases involved in cell wall remodeling","volume":"94","author":"Ao","year":"2016","journal-title":"Fungal Genet. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1766","DOI":"10.1128\/EC.00134-10","article-title":"alpha-1,6-Mannosylation of N-linked oligosaccharide present on cell wall proteins is required for their incorporation into the cell wall in the filamentous fungus Neurospora crassa","volume":"9","author":"Maddi","year":"2010","journal-title":"Eukaryot. Cell"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Maddi, A., Fu, C., and Free, S.J. (2012). The Neurospora crassa dfg5 and dcw1 genes encode alpha-1,6-mannanases that function in the incorporation of glycoproteins into the cell wall. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0038872"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/j.fgb.2014.05.009","article-title":"The Neurospora crassa CPS-1 polysaccharide synthase functions in cell wall biosynthesis","volume":"69","author":"Fu","year":"2014","journal-title":"Fungal Genet. Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"12851","DOI":"10.1021\/acs.analchem.3c02056","article-title":"Acetylation in Ionic Liquids Dramatically Increases Yield in the Glycosyl Composition and Linkage Analysis of Insoluble and Acidic Polysaccharides","volume":"95","author":"Black","year":"2023","journal-title":"Anal. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"100140","DOI":"10.1016\/j.tcsw.2024.100140","article-title":"Characterization of the Neurospora crassa GH72 family of Laminarin\/Lichenin transferases and their roles in cell wall biogenesis","volume":"13","author":"Chatrath","year":"2025","journal-title":"Cell Surf."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e00619-17","DOI":"10.1128\/mBio.00619-17","article-title":"The Dual Activity Responsible for the Elongation and Branching of beta-(1,3)-Glucan in the Fungal Cell Wall","volume":"8","author":"Aimanianda","year":"2017","journal-title":"mBio"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"872","DOI":"10.1080\/15572536.2006.11832778","article-title":"Mannosyltransferase is required for cell wall biosynthesis, morphology and control of asexual development in Neurospora crassa","volume":"97","author":"Bowman","year":"2005","journal-title":"Mycologia"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3621","DOI":"10.1073\/pnas.0505704103","article-title":"Structural basis for rodlet assembly in fungal hydrophobins","volume":"103","author":"Kwan","year":"2006","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1016\/j.fgb.2010.11.009","article-title":"Perithecium morphogenesis in Sordaria macrospora","volume":"48","author":"Lord","year":"2011","journal-title":"Fungal Genet. Biol."},{"key":"ref_24","first-page":"17","article-title":"Different cell types in Neurospora crassa","volume":"50","author":"Bistis","year":"2003","journal-title":"Fungal Genet. Newsl."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/0012-1606(76)90283-9","article-title":"Analysis of pattern formation in Neurospora perithecial development using genetic mosaics","volume":"54","author":"Johnson","year":"1976","journal-title":"Dev. Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.funbio.2018.10.005","article-title":"Characterization of the Neurospora crassa DHN melanin biosynthetic pathway in developing ascospores and peridium cells","volume":"123","author":"Ao","year":"2019","journal-title":"Fungal Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2386","DOI":"10.1074\/jbc.M109.077545","article-title":"Characterization of a new beta(1-3)-glucan branching activity of Aspergillus fumigatus","volume":"285","author":"Gastebois","year":"2010","journal-title":"J. Biol. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"768","DOI":"10.1016\/j.fgb.2009.06.005","article-title":"Trifluoromethanesulfonic acid-based proteomic analysis of cell wall and secreted proteins of the ascomycetous fungi Neurospora crassa and Candida albicans","volume":"46","author":"Maddi","year":"2009","journal-title":"Fungal Genet. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"859","DOI":"10.1038\/nature01554","article-title":"The genome sequence of the filamentous fungus Neurospora crassa","volume":"422","author":"Galagan","year":"2003","journal-title":"Nature"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1128\/EC.00248-13","article-title":"Global gene expression and focused knockout analysis reveals genes associated with fungal fruiting body development in Neurospora crassa","volume":"13","author":"Wang","year":"2014","journal-title":"Eukaryot. Cell"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"E7756","DOI":"10.1073\/pnas.1702591114","article-title":"A-to-I RNA editing is developmentally regulated and generally adaptive for sexual reproduction in Neurospora crassa","volume":"114","author":"Liu","year":"2017","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"e0226424","DOI":"10.1128\/mbio.02264-24","article-title":"VE-1 regulation of MAPK signaling controls sexual development in Neurospora crassa","volume":"15","author":"Gutierrez","year":"2024","journal-title":"mBio"}],"container-title":["Journal of Fungi"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2309-608X\/12\/9\/657\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T04:27:20Z","timestamp":1788496040000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2309-608X\/12\/9\/657"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,2]]},"references-count":32,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["jof12090657"],"URL":"https:\/\/doi.org\/10.3390\/jof12090657","relation":{},"ISSN":["2309-608X"],"issn-type":[{"value":"2309-608X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,2]]}}}