Soil bacterial community dynamics during different growth stages of strawberry Raw sequence reads

goals:To investigate the succession patterns of rhizosphere soil microbial communities during the growth and development of tomato (Solanum lycopersicum), this study used 16S rRNA gene high-throughput sequencing to analyze the soil bacterial community structure at different growth stages, including initial soil, seedling stage, vegetative growth stage, flowering stage, and fruiting stage.methoud:The bacterial 16S rRNA gene V4 region was amplified using primers 515F (GTGYCAGCMGCCGCGGTAA) and 806R (GGACTACNVGGGTWTCTAAT) (Yuan et al., 2020), yielding PCR products of approximately 292 bp. The total PCR reaction volume was 50 uL, containing 25 uL of 2x premixed Taq polymerase, 1 uL of forward and reverse primers (10 uM), 3 uL of DNA template (20 ng/uL), and 20 uL of sterile ultrapure water. The amplification conditions were set as follows: initial denaturation at 95C for 5 min, followed by 30 cycles of 94C for 30 s, 52C for 30 s, and 72C for 30 s, with a final extension at 72C for 10 min. PCR products were checked by 1% agarose gel electrophoresis to select clear bands with lengths between 290-310 bp for purification and sequencing. PCR products were quantified using GeneTools (version 4.03.05.0, Syngene) and purified with the Ezna Gel Extraction Kit (Omega, USA). Sequencing libraries were prepared using the NEBNext Ultra DNA Library Prep Kit for Illumina (New England Biolabs, USA), with index barcodes added. Library quality was assessed using a Qubit 2.0 Fluorometer (Thermo Scientific) and an Agilent 2100 Bioanalyzer. Finally, sequencing was performed on the Illumina HiSeq 2500 platform. Raw data were quality-filtered using Trimmomatic to obtain high-quality sequences and assigned to the corresponding samples based on sample-specific barcodes.