Non-specific lipid transfer proteins (nsLTPs) play a crucial role in the transport and secretion of lipids in plants. In this study, two type II nsLTP genes were cloned, namely nsLTP2-1 and nsLTP2-2, from Lavandula angustifolia. The analysis of sequence characteristics revealed that the nsLTP2-1 and nsLTP2-2 genes encode 119 and 117 amino acids, respectively, which exhibit conserved lipid transfer proteins (LTP) domains and 8 highly conserved cysteine residues. The phylogenetic analysis revealed that these genes are located in separate branches and share the closest genetic relationship with Perilla frutescens, a member of the Labiatae family. The analysis of gene expression showed that both genes exhibit high expression levels in flower buds, while exhibiting comparatively reduced levels. However, notable disparities in expression were observed in the calyx, with nsLTP2-1 and nsLTP2-2 demonstrating higher expression levels in mature and young calyxes, respectively. The expression of two genes in flower buds and leaves was significantly inhibited by the treatment involving gibberellin and methyl jasmonate. Specifically,Dark treatment and gibberellin treatment significantly inhibited the expression of the two genes in bud and leaf. In the buds, abscisic acid and light treatment induced their expression, In the leaf, methyl jasmonate and ethylene induced the expression of nsLTP2-1 and nsLTP2-2, respectively. The subcellular localization of the yellow fluorescent protein (EYFP) fusion protein demonstrated that both nsLTPs were localized on the cell membrane and cell wall, indicating they may be related to the transport of secondary metabolites. These findings provide a fundamental basis for elucidating the role of lavender lipid transfer protein in lipid and terpenoid transportation.