Highlights d 35 fungal species were isolated from the ocean, sponge, coral, and coastal sediment d Time-lapse imaging reveals unusual cell cycles, cell-division patterns, and polarity

Fungi have been found in every marine habitat that has been explored, however, the diversity and functions of fungi in the ocean are poorly understood. In this study, fungi were cultured from the marine environment in the vicinity of Woods Hole, MA, USA including from plankton, sponge and coral. Our sampling resulted in 36 unique species across 20 genera. We observed many isolates by time-lapse differential interference contrast (DIC) microscopy and analyzed modes of growth and division. Several black yeasts displayed highly unconventional cell division cycles compared to those of traditional model yeast systems. Black yeasts have been found in habitats inhospitable to other life and are known for halotolerance, virulence, and stress-resistance. We find that this group of yeasts also shows remarkable plasticity in terms of cell size control, modes of cell division, and cell polarity. Unexpected behaviors include division through a combination of fission and budding, production of multiple simultaneous buds, and cell division by sequential orthogonal septations. These marine-derived yeasts reveal alternative mechanisms for cell division cycles that seem likely to expand the repertoire of rules established from classic model system yeasts.

Coral reefs are highly diverse ecosystems of immense ecological, economic, and aesthetic importance built on the calcium-carbonate-based skeletons of stony corals. The formation of these skeletons is threatened by increasing ocean temperatures and acidification, and a deeper understanding of the molecular mechanisms involved may assist efforts to mitigate the effects of such anthropogenic stressors. In this study, we focused on the role of the predicted bicarbonate transporter SLC4γ, which was suggested in previous studies to be a product of gene duplication and to have a role in coral-skeleton formation. Our comparative-genomics study using 30 coral species and 15 outgroups indicates that SLC4γ is present throughout the stony corals, but not in their non-skeleton-forming relatives, and apparently arose by gene duplication at the onset of stony-coral evolution. Our expression studies show that SLC4γ , but not the closely related and apparently ancestral SLC4β , is highly upregulated during coral development coincident with the onset of skeleton deposition. Moreover, we show that juvenile coral polyps carrying CRISPR/Cas9-induced mutations in SLC4γ are defective in skeleton formation, with the severity of the defect in individual animals correlated with their frequencies of SLC4γ mutations. Taken together, the results suggest that the evolution of the stony corals involved the neofunctionalization of the newly arisen SLC4γ for a unique role in the provision of concentrated bicarbonate for calcium-carbonate deposition. The results also demonstrate the feasibility of reverse-genetic studies of ecologically important traits in adult corals.

Fungi play a crucial role in biogeochemical cycling and shaping biological communities at macro‐ and microcosmic scales. However, fungi have been largely overlooked in studies of marine ecology and microbiology. Here we present protocols for preparing culture media, collecting and culturing fungi from several types of marine environments and animal hosts, and preparing microscopy slides for long‐term time lapse imaging of fungal isolates. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Solid media preparation Alternate Protocol 1: Liquid media preparation Basic Protocol 2: Sample collection and plating from ocean water Alternate Protocol 2: Sample collection and plating from coastal sediment Alternate Protocol 3: Sample collection and plating from animal holobionts Support Protocol 1: Making frozen stocks Support Protocol 2: Streaking cultures from frozen stocks Basic Protocol 3: Slide preparation and assembly for imaging © 2021 by John Wiley & Sons, Inc.

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