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Cell permeability and nuclear DNA staining by propidium iodide in basidiomycetous yeasts

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Abstract

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.

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Acknowledgments

We thank the Natural Science Foundation of China and the Chinese Academy of Sciences for financial support.

Funding

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.

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Authors and Affiliations

  1. Shandong Provincial Key Laboratory of Synthetic Biology, Qingdao Engineering Laboratory of Single Cell Oil, Key Laboratory of Biofuels, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, No.189 Songling Road, Laoshan District, Qingdao, Shandong, 266101, China

    Ning Zhang, Yuxuan Fan, Fuli Li & Shi’an Wang

  2. University of Chinese Academy of Sciences, Beijing, 100039, China

    Ning Zhang & Yuxuan Fan

  3. Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity, Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, 266071, China

    Chen Li

  4. Function Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266071, China

    Chen Li

  5. State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China

    Qiming Wang

  6. School of Agro-Industry, Chiang Mai University, Chiang Mai, 50100, Thailand

    Noppol Leksawasdi

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  1. Ning Zhang
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Correspondence to Shi’an Wang.

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Zhang, N., Fan, Y., Li, C. et al. Cell permeability and nuclear DNA staining by propidium iodide in basidiomycetous yeasts. Appl Microbiol Biotechnol 102, 4183–4191 (2018). https://doi.org/10.1007/s00253-018-8906-8

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