| Congresso Brasileiro de Microbiologia 2023 | Resumo: 1159-2 | ||||
Resumo:Advancements in understanding the intricate relationships between soil and plant-associated microbial communities have fueled the need for robust DNA extraction methods. This abstract presents a comprehensive overview of innovative, and techniques tailored for extracting microbial DNA from diverse soil and plant matrices. The extractions start with a good sample. Samples well-made and frozen right before the sampling will present higher fidelity with the regular microbiota from the studied environment. After that, we highlight the significance of choosing appropriate extraction methods based on the soil type, microbial community structure, and downstream applications. Classical methods involving bead-beating, phenol-chloroform, and CTAB extraction have laid the foundation for microbial DNA extraction. However, modern approaches, such as commercial kits, have gained popularity due to their efficiency and reproducibility. Insights about decrease temperature in some steps of the extraction and changes (or modifications) of the elution reagent could increase the final amount of DNA. The same is expected by reducing the shaking force in soils with high organic matter content. In plant-associated microorganisms, the microbes must be separated from the leaves (or litter) for a better output. One of the most promising methods relies on washing the plant material with a phosphate-buffered saline solution (PBS, pH 7.4) for twelve hours. After that, the filtered solution should be centrifuged at a high force for at least 10 minutes to take the whole pellet from the microbiota to subsequent procedures. We also point out that sample preparation, including soil homogenization, storage, and freeze-thaw cycles may affect DNA extraction efficiency. In conclusion, by reviewing and updating the most recommended practices for soil microbial DNA extraction, new insights can be brought up from the selection of appropriate methods when considering protocols for high-quality DNA extraction, with increased DNA concentration of more than 100%. By adhering to these practices, researchers can enhance the accuracy and reproducibility of soil microbial studies, ultimately contributing to a deeper understanding of soil ecosystems and their ecological functions. Palavras-chave: Bacterial DNA, Culture-Independent Methods, Endosphere, Microbial Ecology, Rhizosphere Agência de fomento:FAPESP; CAPES | |||||