Highly pathogenic avian influenza (HPAI) viruses of the H5N1 subtype have been identified as a potential pandemic threat by the World Health Organization (WHO). Since 1997, these viruses have been spreading from Asia to Europe and Africa with increasing genetic and antigenic diversities. Vaccination is the preferred strategy for the prevention and control of influenza infections and the availability of a system for the rapid engineering and production of vaccines is required in the event of an influenza pandemic. In this study, we engineered and produced recombinant hemagglutinin (HA) from A/Bar-headed Goose/Qinghai/1A/05 (clade 2.2) and A/Anhui/1/2005 (clade 2.3) in Nicotiana benthamiana plants. Immunization of mice with these plant-derived HA antigens elicited serum hemagglutination inhibition (HI) and virus neutralization (VN) antibodies. These results suggest the utility of our plant-expression system for recombinant influenza vaccine production.