To characterize gut microbiome of the infant during the first year of life and assess the different contributions of delivery mode, feeding mode and infant gender to gut microbial development. We collected 314 faecal samples from 80 infants at 5 time points of 0, 1st, 3rd, 6th and 12th months prospectively, and finally 213 samples completed Miseq sequencing and analysis. We characterized gut microbiome of the infant at the different phases and evaluated the different contributions of delivery mode, feeding mode and gender to gut microbial development. Delivery mode, gender and feeding mode were the strongest factors determining gut microbiome colonization at 0 months, from 1 month to 6 months and 12 months, respectively. Four genera including Bifidobacterium, Bacteroides, Parabacteroides and Phascolarctobacterium were increased, whereas 10 genera e.g. Salmonella and Enterobacter were reduced, in vaginal delivery versus cesarean section. Two genera including Peptostreptococcaceae incertae sedis and Anaerococcus were increased, whereas 3 genera e.g. Coriobacteriaceae uncultured were reduced, in exclusive breastfeeding versus combined feeding. This study indicated the contribution degrees of delivery mode, feeding mode and gender to gut microbial initiation and evolvement, and reported microbial differences induced by the different delivery mode, feeding mode and gender.