Lee Yu-Ja, Phang Guan Jie, Chen Che-Chih, Ou Jie-Hao, Fan Yu-Hsuan, Huang Yin-Tse
Department of Biomedical Science and Environment Biology, Kaohsiung Medical University, Kaohsiung, 80708, Taiwan.
Department of Biology, National Museum of Natural Science, Taichung, 404605, Taiwan.
Heliyon. 2024 Aug 24;10(17):e36829. doi: 10.1016/j.heliyon.2024.e36829. eCollection 2024 Sep 15.
Preserving fungal tissue DNA in the field is essential for molecular ecological research, enabling the study of fungal biodiversity and community dynamics. This study systematically compares two liquid-based preservation solutions, RNAlater and DESS, for their effectiveness in maintaining macrofungi DNA integrity during field collection and storage. The research encompasses both controlled experiments and real-world field collections. In the controlled experiments, two fungal species were preserved in RNAlater and DESS at different temperatures and durations. DNA extraction success rates were high, but DNA quality and quantity metrics exhibited variations across samples. However, both preservation solutions demonstrated their viability for preserving fungal DNA, with no significant differences between them. In the field-collected macrofungi experiment, 160 paired fungal specimens were preserved in RNAlater and DESS, respectively. Including a drying process to facilitate tissue lysis for DNA extraction significantly impacted the outcomes. RNAlater showed a higher success rate and better DNA quality and quantity compared to DESS. Statistical analysis, including paired and independent t-tests, confirmed significant differences in DNA quality and quantity between the two preservation methods for field-collected samples. This study evaluates RNAlater and DESS for preserving macrofungi DNA in field conditions. Both methods are effective, but RNAlater is superior when a drying step is included in DNA extraction. Researchers can choose based on their specific needs without compromising DNA integrity. These findings advance fungal molecular ecology and DNA preservation strategies in ecological and environmental studies.
在野外保存真菌组织DNA对于分子生态学研究至关重要，有助于开展真菌生物多样性和群落动态研究。本研究系统比较了两种基于液体的保存溶液RNAlater和DESS在野外采集和储存过程中维持大型真菌DNA完整性的有效性。研究涵盖了对照实验和实际野外采集。在对照实验中，将两种真菌物种在不同温度和时长下保存在RNAlater和DESS中。DNA提取成功率较高，但样本间的DNA质量和数量指标存在差异。然而，两种保存溶液都证明了其保存真菌DNA的可行性，它们之间没有显著差异。在野外采集的大型真菌实验中，分别将160对真菌标本保存在RNAlater和DESS中。包括一个干燥过程以促进组织裂解用于DNA提取，这对结果有显著影响。与DESS相比，RNAlater显示出更高的成功率以及更好的DNA质量和数量。包括配对和独立t检验在内的统计分析证实，对于野外采集的样本，两种保存方法在DNA质量和数量上存在显著差异。本研究评估了RNAlater和DESS在野外条件下保存大型真菌DNA的效果。两种方法都有效，但在DNA提取中包含干燥步骤时，RNAlater更具优势。研究人员可以根据自身特定需求进行选择，而不会损害DNA完整性。这些发现推动了真菌分子生态学以及生态和环境研究中的DNA保存策略的发展。
