To explore how Asplenium nidus responds to drought stress and to find V-PPase (AVP1) homologous genes of this species, we conducted greenhouse experiments. We measured a series of physiological and biochemical indices after drought treatment and analyzed the expression of AVP1 homologous genes in A. nidus by RT-qPCR. The results indicated that A. nidus can adapt to drought by limiting stomatal conductance, thus inhibiting plant photosynthesis and reducing physiological activities. In addition, A. nidus can also change its osmotic potential by increasing proline content to maintain normal metabolic activities, and prevent the destruction of reactive oxygen species by increasing the activities of superoxide dismutase (SOD) and peroxidase (POD). According to analysis of the relative expression levels of genes, AVP1-2 and AVP1-4 may be the drought-resistant genes of A. nidus. This study lays a foundation for further exploration of the drought tolerance mechanism and drought-resistant genes of A. nidus.