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

文献类型： 外文期刊

作者： Zhang, Ning 1 ; Fan, Yuxuan 1 ; Li, Chen 3 ; Wang, Qiming 5 ; Leksawasdi, Noppol 6 ; Li, Fuli 1 ; Wang, Shi'an 1 ;

作者机构： 1.Chinese Acad Sci, Shandong Prov Key Lab Synthet Biol, Qingdao Engn Lab Single Cell Oil, Key Lab Biofuels,Qingdao Inst Bioenergy & Bioproc, 189 Songling Rd, Qingdao 266101, Shandong, Peoples R China

2.Univ Chinese Acad Sci, Beijing 100039, Peoples R China

3.Chinese Acad Fishery Sci, Qingdao Key Lab Mariculture Epidemiol & Biosecur, Key Lab Maricultural Organism Dis Control, Minist Agr,Yellow Sea Fisheries Res Inst, Qingdao 266071, Peoples R China

4.Qingdao Natl Lab Marine Sci & Technol, Funct Lab Marine Fisheries Sci & Food Prod Proc, Qingdao 266071, Peoples R China

5.Chinese Acad Sci, Inst Microbiol, State Key Lab Mycol, Beijing 100101, Peoples R China

6.Chiang Mai Univ, Sch Agroind, Chiang Mai 50100, Thailand

关键词： Basidiomycetous yeast; Cell permeability; Cell wall; Dimethyl sulfoxide; DNA staining

期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：4.813； 五年影响因子：4.697 ）

ISSN： 0175-7598

年卷期： 2018 年 102 卷 9 期

页码：

收录情况： SCI

摘要： 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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