In order to study the functional basis of Phage shock protein A (PspA), the full-length coding sequence (CDS) of the PspA gene was isolated based on sequence homolog and its expression pattern of PspA from N. flagelliforme under drought stress was studied by RT-qPCR. We further generated a plasmid pCAM35s-GFP-PspA which were subjected for a subcellular localization analysis and a transformation in A. thaliana. PspA contained a 777-bp full-length coding sequence, and this gene was significantly up-regulated in N. flagelliforme upon drought stress treatment. A transient expression of PspA in N. benthamiana suggested a sublocalization on the plasma membrane. Moreover, by transforming this gene in A. thaliana, the transgenic plants showed low copies of the PspA gene using Southern hybridization and expressed the recombination protein by Western blot analysis. The transgenic plants expressing the PspA gene, if compared to the wild type plants, were found to be highly resistant against drought stress. Collectively, the results provided a foundation for further exploring the biological function of the N. flagelliforme PspA gene that interplays with drought stress.