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首页> 外文期刊>Proceedings of the Arkansas Academy of Science >Application of a Modified Ziehl-Neelsen's Carbol-Fuchsin Stain for Observing Nuclei in Vegetative Hyphae of the Puffball Species Lycoperdon pyriforme
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Application of a Modified Ziehl-Neelsen's Carbol-Fuchsin Stain for Observing Nuclei in Vegetative Hyphae of the Puffball Species Lycoperdon pyriforme

机译：改良的Ziehl-Neelsen's Carbol-Fuchsin染色剂在观察马勃菌Lycoperdon pyriforme菌丝营养菌丝中的核的应用

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摘要

Nuclei of vegetative hyphae of the puffball species Lycoperdon pyriforme are small and difficult to stain. A simple mitot-ic staining technique utilizing a modified version of Ziehl-Neelsen's carbol-fuchsin stain, originally developed for examining bacteria and small chromosomes of plants, has proven useful for observing fungal nuclei in this gasteromycete species. Cells of vegetative hyphae were pre-treated for 2 hours with an aqueous solution saturated with naphthalene, fixed in a 3:1 mixture of 100% ethanol and glacial acetic acid, and hydrolyzed for 10 minutes in a 1:1 mixture of 100% ethanol and 12 M hydrochloric acid. Cells were rinsed with distilled water before exposure to the carbol-fuchsin stain. Microscopic examination of hyphae revealed that the nuclei were darkly stained, whereas the rest of the tissue remained relatively clear. Examination of 800 apical and adjacent subapical cells from six isolates revealed that the hyphae are predominantly binucleate. This nuclear staining technique may prove useful in studies of the life cycle and genetics of this and other fungal species.
机译：马勃球菌Lycoperdon pyriforme的营养菌丝核小，难以染色。一种简单的线粒体染色技术，利用Ziehl-Neelsen的carbol-fuchsin染色剂的改良版，最初开发用于检查植物的细菌和小染色体，现已证明可用于观察该胃菌种的真菌核。营养性菌丝细胞用萘饱和水溶液预处理2小时，固定在100％乙醇和冰醋酸的3：1混合物中，并在100％乙醇的1：1混合物中水解10分钟和12 M盐酸。用蒸馏水冲洗细胞，然后再暴露于碳水化合物-品红染色剂中。菌丝的显微镜检查显示细胞核被染成深色，而其余组织保持相对清晰。从六个分离物中检查了800个顶端细胞和相邻的顶端细胞，发现菌丝主要为双核。这种核染色技术可能在研究这种真菌和其他真菌物种的生命周期和遗传学方面很有用。

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