A simple protocol for producing axenic seeds of Sorghum bicolor

Microbes within seeds can confound research on microbial colonization, symbiosis, and pathogenesis. Sterilization of both external and internal seed tissues is therefore essential in certain experiments, but the method must also preserve seed viability. Here, we present a reliable and simple protocol for sterilizing Sorghum bicolor seeds by submerging them in 95% ethanol for 2 minutes followed by 3.75% sodium hypochlorite for 20 minutes. This approach yielded a low contamination rate (2 out of 95 seeds) and a robust median germination rate (63%). Its simplicity, cost-effectiveness, and accessibility make it a practical option for experiments requiring axenic seeds.