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In order to investigate the barotolerance of marine fungi and to elucidate their ecological role in the deep sea, high-pressure equipment was built and tested. Cultures of the marine yeasts Debaryomyces hansenii, Rhodotorula rubra and Rhodosporidium sphaerocarpum were inoculated into gas-permeable plastic foil bags and incubated in pressure vessels filled with hydraulic fluid that serves also as an oxygen reservoir. The equipment was used at temperatures from 7° to 34°C and pressures from 0·1 to 80·0 MPa. Comparison of five types of plastic foil showed effects on total cell number in batch culture. An exchange of the hydraulic fluid increased yield by replenishing oxygen, the growth-limiting factor. There were no detectable growth differences between buffered (TRIS, HEPES, imidazole, MES) and unbuffered media and thus unbuffered medium was used. Yeasts were best cultivated in an unbuffered sea-water medium (glucose–peptone–yeast extract) sealed in bags made from polyethylene foil. The fluorocarbon liquid FC-77 provided the best oxygen reservoir owing to its high oxygen and carbon dioxide solubility. An exchange of the hydraulic fluid is not necessary if sensitive growth determination methods are used. All marine yeasts cultivated under simulated deep sea conditions were able to grow at least up to a pressure of 20 MPa, with Rhodotorula rubra and Rhodosporidium sphaerocarpum growing at 40 MPa, corresponding to 4000 m depth. Thus, marine yeasts are able to grow under simulated deep sea conditions and may participate in the degradation of organic matter in the deep sea.