A clearing technique for detecting the fungal endophyte Acremonium sp. in grasses.
A clearing technique for detecting the fungal endophyte Acremonium sp. in grasses.
复制标题
一种用于检测内生真菌顶孢霉属的清除技术。
DOI：
10.3109/10520299309104672
复制
发表时间：
1993
期刊：
影响因子：
--
通讯作者：
A. van Wijk
中科院分区：
文献类型：
--
作者：
K. Hignight;G. Muilenburg;A. van Wijk
高羊茅叶组织的研究。采用玫瑰红或苯胺蓝染色，对硬羊茅、红羊茅和多年生黑麦草进行内生真菌Acremonium sp.的检测。标本用水杨酸甲酯（一种光学清除剂）清除，并使用明场显微镜观察。在染色和清理之前，将组织作为干燥组织保存或保存在70%的乙醇水溶液中。在清除后2、4和12周观察组织以检查染色保留。用孟加拉玫瑰染色效果不如苯胺蓝，然后用水杨酸甲酯清净。与苯胺蓝染色菌丝体相比，玫瑰红染色菌丝体颜色较浅，检测难度较大。结果表明，结合染色和水杨酸甲酯清除对真菌内生菌的检测是有效的。
Leaf tissue of tall fescue Festuca arundinacea Schreb., hard fescue Festuca ovina L., red fescue Festuca rubra L. and perennial ryegrass Lolium perenne L. was stained with rose Bengal or aniline blue to detect the presence of the fungal endophyte Acremonium sp.. Specimens were cleared using methyl salicylate, an optical clearing agent, and viewed using bright field microscopy. Tissue was preserved as dried tissue or stored in 70% aqueous ethyl alcohol before staining and clearing. Tissue was observed at 2, 4 and 12 weeks following clearing to check for stain retention. Staining with rose Bengal was inferior to aniline blue when followed by the clearing agent methyl salicylate. Fungal mycelia stained lighter with rose Bengal and were more difficult to detect than mycelia stained with aniline blue. The results illustrate the usefulness of combining staining and methyl salicylate clearing for detecting fungal endophytes.