Cell permeability and nuclear DNA staining by propidium iodide in basidiomycetous yeasts
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Non-model yeasts within basidiomycetes have considerable importance in agriculture, industry, and environment, but they are not as well studied as ascomycetous yeasts. Serving as a basic technique, nuclear DNA staining is widely used in physiology, ecology, cell biology, and genetics. However, it is unclear whether the classical nuclear DNA staining method for ascomycetous yeasts is applicable to basidiomycetous yeasts. In this study, 5 yeasts ineffectively stained by the classical propidium iodide (PI) staining method were identified from 23 representative basidiomycetous yeasts. Pretreatment of cells using dimethyl sulfoxide (DMSO) or snailase markedly improved cell penetration to PI and thus enabled DNA content determination by flow cytometry on the recalcitrant yeasts. The pretreatments are efficient, simple, and fast, avoiding tedious mutagenesis or genetic engineering used in previous reports. The heterogeneity of cell penetration to PI among basidiomycetous yeasts was attributed to the discrepancy in cell wall polysaccharides instead of capsule or plasma membrane. This study also indicated that care must be taken in attributing PI-negative staining as viable cells when studying non-model microorganisms.
KeywordsBasidiomycetous yeast Cell permeability Cell wall Dimethyl sulfoxide DNA staining
We thank the Natural Science Foundation of China and the Chinese Academy of Sciences for financial support.
This study was funded by the Natural Science Foundation of China (No. 31670054, No. 51561145015, and No. 21676159) and Youth Innovation Promotion Association (2013137), Chinese Academy of Sciences.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Borgel D, van den Berg M, Huller T, Andrea H, Liebisch G, Boles E, Schorsch C, van der Pol R, Arink A, Boogers I, van der Hoeven R, Korevaar K, Farwick M, Köhler T, Schaffer S (2012) Metabolic engineering of the non-conventional yeast Pichia ciferrii for production of rare sphingoid bases. Metab Eng 14(4):412–426CrossRefPubMedGoogle Scholar
- Burke D, Dawson D, Stearns T (2000) Flow cytometry of yeast DNA. In: Methods in yeast genetics. A Cold Spring Harbor laboratory course manual. Laboratory Press, Cold Spring Harbor, NY, pp 155–156Google Scholar
- Flores ML, Stortz CA, Cerezo AS (2000) Studies on the skeletal cell wall of the cystocarpic stage of the red seaweed Iridaea undulosa B. Part II Fractionation of the cell wall and methylation analysis of the inner core-fibrillar polysaccharides. Inter J Bio Macromol 27(1):21–27CrossRefGoogle Scholar
- Gh MS, Wilhelm MJ, Sheffield JB, Dai HL (2015) Living E. coli is permeable to propidium iodide: a study by time-resolved second-harmonic scattering and fluorescence microscopy. Biophys J 108(2):148a–149aGoogle Scholar
- Maekawa E (1974) Effect of dimethyl-sulfoxide as solvent of cell-wall polysaccharides. J Agr Chem Soc Jpn 48(1):75–77Google Scholar
- Russell JJ, Theriot JA, Sood P, Marshall WF, Landweber LF, Fritz-Laylin L, Polka JK, Oliferenko S, Gerbich T, Gladfelter A, Umen J, Bezanilla M, Lancaster MA, He S, Gibson MC, Goldstein B, Tanaka EM, Hu CK, Brunet A (2017) Non-model model organisms. BMC Biol 15(1):55CrossRefPubMedPubMedCentralGoogle Scholar
- Sharma R, Gassel S, Steiger S, Xia X, Bauer R, Sandmann G, Thines M (2015) The genome of the basal agaricomycete Xanthophyllomyces dendrorhous provides insights into the organization of its acetyl-CoA derived pathways and the evolution of Agaricomycotina. BMC Genomics 16(1):233CrossRefPubMedPubMedCentralGoogle Scholar
- Williams SC, Hong Y, Danavall DCA, Howard-Jones MH, Gibson D, Frischer ME, Verity PG (1998) Distinguishing between living and nonliving bacteria: evaluation of the vital stain propidium iodide and its combined use with molecular probes in aquatic samples. J Microbiol Meth 32(3):225–236CrossRefGoogle Scholar