20.500.12556/RUP-23014 Surface colonization of Aureobasidium pullulans a multimodal microscopy study toward living coating development Recent advances in engineered living materials (ELMs), which integrate living cells into functional structural and protective systems, have accelerated interest in understanding microbial surface interactions at fundamental scales. In this context, Aureobasidium pullulans, a polymorphic and polyextremotolerant black yeast fungus, has emerged as a promising candidate for diverse biotechnological applications, including engineered living coatings. However, its microscale surface colonization dynamics remain insufficiently characterized, limiting predictive control over colonization of various surfaces. Complementary optical approaches, including VHX digital imaging, fluorescence observation, and quantitative fluorescence measurements, were used to characterize temporal colonization of fungus on plastic coverslips and pine wood, representing two contrasting substrates used in the building sector. On plastic substrates, initial attachment involved dispersed cells that rapidly proliferated and merged into a continuous layer. Quantitative fluorescence revealed a significant increase in signal from Day 1 to Day 3 (from 25,328 to 42,510 RFU; P = 5.4 × 10⁻⁴) followed by a decrease by Day 6 (10,555 RFU). This decline may be associated with reduced dye penetration into the compact, melanized matrix rather than a reduction in biomass. On wood, colonization followed the native fiber orientation and progressed into cohesive multicellular structures. The results provide a new understanding of substrate-dependent colonization of A. pullulans and highlight methodological limitations in quantifying biomass on porous, heterogeneous materials. The multimodal microscopy framework established a robust comparative platform for analysing fungal-material interactions and enabled the rational development of fungal-based living coatings for protective and functional applications. Nedavni napredek na področju inženirsko izdelanih živih materialov (ELM), ki vključujejo žive celice v funkcionalne strukturne in zaščitne sisteme, je pospešil zanimanje za razumevanje interakcij mikrobov s površinami na temeljnih ravneh. V tem kontekstu se je Aureobasidium pullulans, polimorfna in poliekstremotolerantna črna kvasovka, izkazala kot obetaven kandidat za različne biotehnološke aplikacije, vključno z inženirsko izdelanimi živimi premazi. Vendar pa njena dinamika kolonizacije površin na mikroravni še vedno ni dovolj raziskana, kar omejuje napovedno nadzorovanje kolonizacije različnih površin. Za karakterizacijo časovne kolonizacije glive na plastičnih krovnih stekelcih in borovem lesu, ki predstavljata dva kontrastna substrata, ki se uporabljata v gradbenem sektorju, so bili uporabljeni dopolnilni optični pristopi, vključno z digitalnim slikanjem VHX, opazovanjem fluorescence in kvantitativnimi meritvami fluorescence. Aureobasidium pullulans surface colonization plastic pine wood living coating Aureobasidium pullulans kolonizacija površin plastika borov les živi premaz true false true Angleški jezik Slovenski jezik Članek v reviji 2026-05-06 08:55:31 2026-05-06 08:55:36 2026-05-07 03:08:26 0000-00-00 00:00:00 2026 0 0 str. 1-45 iss. , article no. Vol. 2026 0000-00-00 PreprintVObjavi NiDoloceno NiDoloceno 0000-00-00 0000-00-00 2026-04-08 69:60 2045-2322 10.1038/s41598-026-48101-5 275603459 https://www.nature.com/articles/s41598-026-48101-5 1 810666f9-4918-11f1-9e8d-005056ac49c0 https://repozitorij.upr.si/Dokument.php?lang=slv&id=33629 RAZ_Malat_Ihab_2026.pdf RAZ_Malat_Ihab_2026.pdf 1 BC43421060D3647E0B1C23DA79A8A169 8c7cb5acb5914f6f6fe6827701f93e3895927927381a418c1b236558edab8deb b69fa281-4918-11f1-9e8d-005056ac49c0 https://repozitorij.upr.si/Dokument.php?lang=slv&id=33630 Inštitut Andrej Marušič 0 0 0