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Multiscale imaging of Pleurotus ostreatus fruiting bodies – from 3D anatomy to the ultrastructure
ID Mrak, Polona (Author), ID Conti, Jacopo (Author), ID Bogataj, Urban (Author), ID Skočaj, Matej (Author), ID Kostanjšek, Rok (Author), ID Žnidaršič, Nada (Author)

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Abstract
Oyster mushroom, Pleurotus ostreatus, is economically important and widely used as a model organism, yet a comprehensive characterisation of its fruiting body structure across multiple levels of biological organisation is lacking. We applied a multiscale imaging, combining micro-computed tomography (micro-CT), light microscopy of paraffin, cryostat and semithin sections, and transmission electron microscopy to investigate the structure of P. ostreatus young fruiting bodies, and we provide a direct comparison of different sample preparation procedures. Micro-CT enabled 3D anatomy visualisation, revealing distinct tissue regions: a loose trama in the stipe and pileus medulla, a dense peripheral layer of pileus and gills, and a thin, loose layer covering pileus and gills. Histological analyses provided detailed information on tissue organisation, complementing the micro-CT images, including hyphal arrangement, gill architecture, hymenial layer, and basidia. Cryosections preserved overall tissue morphology and proved suitable for future molecular localisation studies. Transmission electron microscopy of Agar 100 and LR White embedded samples resolved the ultrastructure of hyphae and basidia, revealing nuclei, mitochondria, endoplasmic reticulum, vacuoles, cell walls, and dolipore septa with perforated pore cap. The study demonstrates the value of integrating imaging techniques to characterise the mushroom structure across scales and provides a toolbox for future investigations of fruiting bodies.

Language:English
Keywords:micro-CT, microscopy, fungi, hyphae, cell wall, septal pore
Work type:Article
Typology:1.01 - Original Scientific Article
Organization:BF - Biotechnical Faculty
Publication status:Published
Publication version:Version of Record
Year:2026
Number of pages:Str. 64-86
Numbering:Vol. 69, no. 3
PID:20.500.12556/RUL-185464 This link opens in a new window
UDC:543.456
ISSN on article:1854-3073
DOI:10.14720/abs.69.3.28176 This link opens in a new window
COBISS.SI-ID:286891523 This link opens in a new window
Publication date in RUL:06.08.2026
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Downloads:9
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Record is a part of a journal

Title:Acta biologica slovenica
Publisher:Društvo biologov Slovenije, Društvo biologov Slovenije, Založba Univerze v Ljubljani, Društvo biologov Slovenije
ISSN:1854-3073
COBISS.SI-ID:221921024 This link opens in a new window

Licences

License:CC BY-SA 4.0, Creative Commons Attribution-ShareAlike 4.0 International
Link:http://creativecommons.org/licenses/by-sa/4.0/
Description:This Creative Commons license is very similar to the regular Attribution license, but requires the release of all derivative works under this same license.

Secondary language

Language:Slovenian
Title:Slikovna aAnaliza trosnjakov glive Pleurotus ostreatus – od 3D anatomije do ultrastrukture
Abstract:
Bukov ostrigar (Pleurotus ostreatus) je gospodarsko pomembna goba in pogosto uporabljen kot modelni organizem. Kljub temu je morfologija trosnjaka ostrigarja slabo opredeljena in podrobnih raziskav strukture na več ravneh biološke organizacije ni. V raziskavi smo uporabili kombinacijo slikovnih tehnik, in sicer mikro-računalniško tomografijo (mikro-CT), svetlobno mikroskopijo parafinskih, kriostatskih in poltankih rezin ter presevno elektronsko mikroskopijo, za analizo strukture mladega trosnjaka vrste P. ostreatus. Poleg tega raziskava vključuje neposredno primerjavo različnih postopkov priprave vzorcev. Tehnika mikro-CT je omogočila tridimenzionalno vizualizacijo anatomije trosnjakov in prikazala različna tkivna območja: rahlo tkivo v betu in sredici klobuka, gosta robna plast klobuka in lističev ter tanko rahlo plast, ki prekriva klobuk in lističe. S histološko analizo smo podrobno pokazali organizacijo tkiv, kar dopolnjuje rezultate iz mikro-CT analiz, vključno z razporeditvijo hif, arhitekturo lističev, trosovnice in bazidijev. Kriostatske rezine so podale splošno morfologijo tkiva, primerno za nadaljnje študije molekularne lokalizacije. S presevno elektronsko mikroskopijo vzorcev vklopljenih v smolo Agar 100 in v smolo LR White smo pokazali ultrastrukturo hif in bazidijev, vidna so bila jedra, mitohondriji, endoplazemski retikulum, vakuole, celične stene in doliporne septe s perforirano porno kapico. Raziskava kaže pomen integracije slikovnih tehnik, s katerimi lahko opredelimo strukturo gobe na različnih prostorskih ravneh, in zagotavlja orodje za prihodnje raziskave trosnjakov.

Keywords:mikro-CT, mikroskopija, glive, hife, celična stena, septalna pora

Projects

Funder:ARIS - Slovenian Research and Innovation Agency
Project number:P1-0184-2020
Name:Integrativna zoologija in speleobiologija

Funder:ARIS - Slovenian Research and Innovation Agency
Project number:I0-0022-0481-0481-07
Name:IC Mikroskopija bioloških vzorcev
Acronym:ICMBV

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