Department of Biology, McMaster University, Hamilton, ON, L8S 4K1, Canada.
Institute of Plant and Environment Protection, Beijing Academy of Agriculture and Forestry Sciences, and Beijing Engineering Research Center for Edible Mushrooms, Beijing, 100097, China.
Sci Rep. 2017 Jun 7;7(1):3008. doi: 10.1038/s41598-017-03570-7.
Several methods have been reported for drying mushroom specimens for population genetic, taxonomic, and phylogenetic studies. However, most methods have not been directly compared for their effectiveness in preserving mushroom DNA. In this study, we compared silica gel drying at ambient temperature and oven drying at seven different temperatures. Two mushroom species representing two types of fruiting bodies were examined: the fleshy button mushroom Agaricus bisporus and the leathery shelf fungus Trametes versicolor. For each species dried with the eight methods, we assessed the mushroom water loss rate, the quality and quantity of extracted DNA, and the effectiveness of using the extracted DNA as a template for PCR amplification of two DNA fragments (ITS and a single copy gene). Dried specimens from all tested methods yielded sufficient DNA for PCR amplification of the two genes in both species. However, differences among the methods for the two species were found in: (i) the time required by different drying methods for the fresh mushroom tissue to reach a stable weight; and (ii) the relative quality and quantity of the extracted genomic DNA. Among these methods, oven drying at 70 °C for 3-4 h seemed the most efficient for preserving field mushroom samples for subsequent molecular work.
已有几种方法被报道用于干燥蘑菇标本,以进行种群遗传、分类和系统发育研究。然而,大多数方法并没有直接比较其在保存蘑菇 DNA 方面的有效性。在这项研究中,我们比较了在环境温度下用硅胶干燥和在七种不同温度下用烤箱干燥的效果。我们检查了两种代表两种子实体类型的蘑菇物种:肉质纽扣菇 Agaricus bisporus 和革质架子菌 Trametes versicolor。对于用八种方法干燥的每个物种,我们评估了蘑菇的失水率、提取 DNA 的质量和数量,以及使用提取的 DNA 作为模板进行 PCR 扩增两个 DNA 片段(ITS 和单拷贝基因)的有效性。所有测试方法的干燥标本都为两种物种的两个基因的 PCR 扩增提供了足够的 DNA。然而,在两种物种中,不同方法之间存在差异:(i)不同干燥方法使新鲜蘑菇组织达到稳定重量所需的时间;和(ii)提取基因组 DNA 的相对质量和数量。在这些方法中,70°C 烤箱干燥 3-4 小时似乎是最有效的方法,可用于保存后续分子工作的野外蘑菇样本。