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Sterol composition of nystatin and amphotericin B resistant tobacco calluses
Author(s) -
Chiu PeiLu,
Bottino P. J.,
Patterson G. W.
Publication year - 1980
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/bf02534118
Subject(s) - nystatin , stigmasterol , callus , ergosterol , microbiology and biotechnology , subculture (biology) , amphotericin b , sterol , natamycin , polyene , filipin , nicotiana tabacum , biology , griseofulvin , agar , chemistry , food science , botany , antibiotics , chromatography , biochemistry , medicine , bacteria , cholesterol , antifungal , genetics , pathology , gene
The objective of this study was to select and characterize tobacco cell lines resistant to the polyene antibiotics Amphotericin B and Nystatin. Suspension culture cells of tobacco ( Nicotiana tabacum L. ‘Wisconsin 38’) were treated with 0.025% ethyl methane sulfonate (EMS) and suspended in melted agar medium containing 0.005 mg/ml Amphotericin B. Cells were also irradiated with UV from a germicidal lamp and plated in the melted agar plating medium. After 6–10 days growth, the cultures were overplated with medium containing 750 u/ml Nystatin. All cultures were grown at 26 C in the dark. After three weeks, a single callus was isolated from the Amphotericin B treatment. This callus was subcultured, and would grow at concentrations of the antibiotic as high as 0.1 mg/ml (a 20‐fold increase over the selection concentration). Six callus lines were isolated from the Nystatin treatment. After subculture and transfer to media containing higher concentrations of Nystatin, only two calli survived and would grow at 1250 u/uml. Gas liquid chromatographic analysis of the resistant and control callus lines revealed similar sterol content among all the lines. However, when comparing total sterols produced, all three polyene resistant calluses contained significantly more sterols than the control. The Amphotericin B resistant calli produced about three times as much sterol as the controls, and the Nystatin resistant calli produced about 1.5 times as much sterol as the control. Both Nystatin resistant lines showed significant increases in stigmasterol. It appears that resistance of the three tobacco cell lines to these polyene antibiotics is due principally to varied amounts of overproduction of sterols rather than qualitative differences in sterols.