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

Nichapat Kittiborwornkul,S.T Anne Sahithi,Malinee Sriariyanun,Theerawut Phusantisampan,Jirapa Phetsom,Wasinee Pongprayoon,M Sriariyanun,M.P Gundupalli,W Rodiahwati,Y.S Cheng

doi:10.1051/e3sconf/202130202014

Abstract

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.

Highlights

Cereals play an essential role in the human daily diet, which is rich in nutrients and calorie composition [1]
Several microorganisms are involved for the growth and can cause disease resulting in reduce yields [5], [6,7]
The DNA was extracted from 3 different soil samples (Root associate soil, Root proximity soil and Bulky soil)

Summary

Introduction

Cereals play an essential role in the human daily diet, which is rich in nutrients and calorie composition [1]. Rice is one of the most economically and nutritionally important cereals, with about 60% of the world's population consuming rice as a basic diet. Some rice cultivars were characterized to contain beneficial nutrition to improve human health [4]. Several microorganisms are involved for the growth and can cause disease resulting in reduce yields [5], [6,7]. The study of microorganisms present in the soil helps to improve their productivity. Some species of these microbes provide the benefit to rice by activating the defense mechanism to protect plants from pathogens [8]

Methods

Results

Conclusion