Comparative evaluation of DNA extraction from rice’s root-associated bacterial consortium for population structure study

• Main Authors: Kittiborwornkul Nichapat, Anne Sahithi S.T, Phetsom Jirapa, Phusantisampan Theerawut, Pongprayoon Wasinee, Sriariyanun Malinee
• Format: Article
• Language: English
• Published: EDP Sciences 2021-01-01
• Series: E3S Web of Conferences
• Subjects: dna extraction method; bacterial consortium; rice; root-associated microbiome; population analysis
• Online Access: https://www.e3s-conferences.org/articles/e3sconf/pdf/2021/78/e3sconf_ri2c2021_02014.pdf

Understanding in population structure of a plant’s root-associated microbiome is applied in good practices in agricultural activities to improve production yields and enhance plant immune responses. The molecular analysis of bacterial populations inhabited in soil faces difficulties in obtaining high yield and high purity of DNA, and different commercial DNA extraction kits have been developed for this purpose. This study focuses on the comparison of DNA extraction of six different rice root-associated bacterial consortium using three commercial kits with two key technologies, spin-column adsorption and magnetic bead adsorption. The quality and quantity of genomic DNA obtained from these extraction methods were analyzed and compared based on DNA concentration, DNA purity and efficiency to be used as a template for 16sRNA amplification. The results showed that the extraction kit with magnetic bead adsorption technology showed the highest concentration (101.32 ng/μl) compared to other DNA extraction kits (32.67 and 1.89 ng/μl). The purity values (A260/A280) were assessed by using Nano-drop spectrophotometer and resulted in purities of nucleic acids in the range of 1.4-1.7. Thus, it was concluded that the extracted DNA obtained from the extraction kit with magnetic bead adsorption technology can be valuable for molecular analysis of microbial communities present in the soil.