Abstract A full-length cDNA ( RlemLTP ) from rough lemon that encodes a nonspecific lipid transfer protein was isolated. Results of subcellular localization of RlemLTP fused to a green fluorescence protein indicated that RlemLTP is located in the chloroplast. Transcripts of RlemLTP were detected in leaves and seeds but not in stems and roots. The transcription of RlemLTP was higher in young leaves than in old leaves. Wounding and fungal inoculation induced RlemLTP expression, and transcription levels were increased 6–15 times by these treatments. RlemLTP obtained using an E. coli expression system showed anti-fungal activity against Alternaria alternata and Fusarium oxysporum .