methane emission from rice paddy soils
This study investigated the bacterial communities in rice paddies subjected to different fertilizer types, namely swine manure, straw biochar, and straw compost, all mixed with inorganic fertilizer under 2 flooding conditions (continuous or alternate wetting and drying). The impacts of different fertilizer types on methane emission from rice stalks were analysed through the analysis of the species abundance and bacterial community structure. The microbial composition of these samples was evaluated by Illumina MiSeq shortgun sequencing of 16S rRNA library (V4 region, 253 bp) amplified using universal 16S primers (Forward primer: 5’- GTGTGCCAGCMGCCGCGGTAA-3’, Revert primer: 5’- CCGGACTACHVGGGTWTCTAAT-3’). Unique molecular identifier (MID) tags and linkers were incorporated at the 5’ end of both target-specific primers to allow identification of individual sequences from pooled amplicons.  Total DNA was extracted from 25 mg paddy soil using MOBIO Power Soil DNA isolation kit (MO BIO Laboratories Inc., CA USA). PCR was carried out in triplicate using Maxime PCR PreMix Kit (iNtRON Biotechnology Inc., Korea). Amplicons were extracted from agarose gel using the AccuPrep Gel Purification kit (Bioneer Inc., Korea). All obtained DNA was quantified using Qubit (Invitrogen, CA, USA), and equimolar purified amplicons were pooled, ligated adapters for Illumina MiSeq sequencing according to MiSeq protocols (Illumina Inc., USA).
Jeju National University
C2