Highly pathogenic avian influenza (HPAI) H5 viruses have been found to have a substantial geographic distribution since they were first reported in Guangdong Province, China. The emergence of new genotypes threatens the poultry industry and human health worldwide. Here, we report five HPAI H5N1 variants isolated from Anser indicus in Yunnan Province, China. A phylogenetic analysis of the hemagglutinin (HA) gene showed that all isolates belong to the highly pathogenic H5 clade 2.3.4.4b and formed two distinct genetic clusters. Bayesian phylogenetic analysis also revealed that the viruses were initially disseminated from wild birds to Anser indicus, implying that infected birds most likely contributed to viral transmission in the region. Genomic sequence analysis revealed several amino acid substitutions, also implying that the infected birds contributed to the spread of the virus throughout the region. Substitutions in the HA glycoprotein increased the virus´s binding affinity to human α-2,6 sialic acid residues. Substitutions in the PB1, PA, and PB2 motifs increased viral polymerase activity and replication in hosts, whereas substitutions in the NP, M1, and NS motifs increased viral pathogenicity in chickens and mice.