Cell permeability and nuclear DNA staining by propidium iodide in basidiomycetous yeasts
Cell permeability and nuclear DNA staining by propidium iodide in basidiomycetous yeasts
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担子菌酵母中碘化丙啶的细胞通透性和核 DNA 染色
DOI：
10.1007/s00253-018-8906-8
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发表时间：
2018-03
影响因子：
5
通讯作者：
Wang Shi'an
中科院分区：
文献类型：
--
作者：
Zhang Ning;Fan Yuxuan;Li Chen;Wang Qiming;Leksawasdi Noppol;Li Fuli;Wang Shi'an
担子菌中的非模式酵母在农业、工业和环境中具有相当重要的意义，但它们的研究不如子囊酵母那么充分。核DNA染色作为一项基础技术，在生理学、生态学、细胞生物学和遗传学等领域有着广泛的应用。然而，经典的子囊酵母核DNA染色方法是否适用于担子酵母尚不清楚。本研究从23株代表性担子菌酵母中鉴定出5株经典碘化丙啶（PI）染色无效的酵母。使用二甲基亚砜（DMSO）或蜗牛酶对细胞进行预处理，可以显著提高细胞对PI的穿透能力，从而可以用流式细胞术测定顽固酵母的DNA含量。预处理高效、简单、快速，避免了以往报道中使用的繁琐的诱变或基因工程。担子酵母细胞对PI渗透的不均匀性主要归因于细胞壁多糖含量的差异，而不是荚膜或质膜的差异。本研究还表明，在研究非模式微生物时，必须注意将pi阴性染色归为活细胞。
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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影响因子：
5.2
作者：
Ciani M;Morales P;Comitini F;Tronchoni J;Canonico L;Curiel JA;Oro L;Rodrigues AJ;Gonzalez R
通讯作者：
Gonzalez R
影响因子：
16.5
作者：
Liu XZ;Wang QM;Göker M;Groenewald M;Kachalkin AV;Lumbsch HT;Millanes AM;Wedin M;Yurkov AM;Boekhout T;Bai FY
通讯作者：
Bai FY
影响因子：
7.7
作者：
Leqian Liu;H. Redden;H. Alper
通讯作者：
Leqian Liu;H. Redden;H. Alper
影响因子：
2.6
作者：
Carrie Baker Brachmann;A. Davies;G. Cost;E. Caputo;Joachim Li;P. Hieter;J. Boeke
通讯作者：
Carrie Baker Brachmann;A. Davies;G. Cost;E. Caputo;Joachim Li;P. Hieter;J. Boeke
影响因子：
2.6
作者：
M. Sentandreu;M. Elorza;R. Sentandreu
通讯作者：
M. Sentandreu;M. Elorza;R. Sentandreu