title: Metagenomic sequencing of microbial community from healthy silage maize
design: Silage samples were randomly collected from each treatment group (HCXB, GCXB, HXB, GXB) (20 g  3 per group, i.e., three biological replicates). Each sample was extracted with 180 mL of sterile distilled water at 4C for over 4 h, filtered through sterile four-layer gauze, and subjected to high-speed centrifugation to isolate microbial cells. DNA Extraction and Library Preparation:Genomic DNA was extracted using the Mag-Bind Soil DNA Kit (Omega Bio-tek, USA). DNA fragmentation (~350 bp) was performed using Covaris M220 (Gene Company, China), and paired-end (PE) libraries were constructed using the NEXTFLEX Rapid DNA-Seq Kit (Bioo Scientific, USA). Sequencing was conducted on an Illumina NovaSeq platform (Illumina, USA) by Shanghai Majorbio Bio-Pharm Technology Co., Ltd. Bioinformatics and Statistical Analysis:Raw sequencing data were quality-filtered using fastp, and adapter trimming was performed with BWA. Sequence assembly and taxonomic annotation were conducted using MEGAHIT, Diamond, and multiple reference databases (NR, eggNOG, KEGG, and CAZy). Alpha () and beta () diversity analyses were performed using R software, while differential community analysis was conducted using LEfSe.All statistical analyses were performed using the Majorbio Cloud Platform (https://www.majorbio.com/web/www/index). Group differences were assessed using one-way ANOVA or the Kruskal-Wallis test, with P < 0.05 considered statistically significant.