Agriculture and Agri-Food Canada, 960 Carling Ave, Ottawa K1A 0C6, Canada.
Mycorrhiza. 2013 May;23(4):333-40. doi: 10.1007/s00572-012-0472-9. Epub 2013 Jan 23.
The use of microwave irradiation as a source of energy to clear and stain intra-radical arbuscular mycorrhizal fungi propagules has been tested on a variety of indigenous and cultivated herbaceous plants. The aim of the study was to evaluate the efficiency of microwave irradiation on root softening, fungi tissue staining, and preservation of DNA integrity for subsequent molecular analyses. The proposed methodology has been adapted from the standard procedures used to detect and quantify mycorrhizal root colonization levels. Using a domestic microwave oven, tissue clearing and staining required together between 30 s and 1.5 min of microwave treatment to be completed, depending the diameter size of the roots. The well-performing chemical stains tested were acid fuchsin, trypan blue, and aniline blue. The acid fuchsin clearing and staining processes, as performed, were also demonstrated to preserve DNA integrity for further molecular analyses. Irradiation by microwaves has been used with success in our laboratory within the frame of several studies. It offers considerable time saving over traditional method, reducing processing times from several hours to a few minutes while decreasing considerably the amount of chemicals and energy required to perform analyses.
微波辐射作为一种能源，已被用于清除和染色内生丛枝菌根真菌孢子，在各种本地和栽培草本植物上进行了测试。该研究的目的是评估微波辐射对根软化、真菌组织染色和 DNA 完整性保存的效率，以便随后进行分子分析。所提出的方法学是从用于检测和量化菌根根定植水平的标准程序中改编而来的。使用家用微波炉，根据根的直径大小，组织透明化和染色总共需要 30 秒到 1.5 分钟的微波处理时间。测试的性能良好的化学染料是酸性品红、台盼蓝和苯胺蓝。如前所述，酸性品红的透明化和染色过程也被证明可以保持 DNA 完整性，以进行进一步的分子分析。在我们实验室的几项研究中，微波辐射已成功应用。与传统方法相比，它节省了大量时间，将处理时间从数小时缩短到几分钟，同时大大减少了进行分析所需的化学品和能源的数量。
