The non-specific lipid transfer proteins (nsLTPs) are small,basic proteins characterized by a tunnel-like hydrophobic cavity,capable of transferring various lipid molecules between lipid bilayers.Most nsLTPs are synthesized with an N-terminal signal peptide that localizes the protein to the apoplastic space.The nsLTPs have only been identified in seed plants,where they are encoded by large gene families.We have initiated an analysis of the evolutionary history of the nsLTP family using genomic and EST information from non-seed land plants and green algae to determine:(1) when the nsLTP family arose,(2) how often new nsLTP subfamilies have been created,and (3) how subfamilies differ in their pattes of expansion and loss in different plant lineages.In this study,we searched sequence databases and found that genes and transcripts encoding nsLTPs are abundant in liverworts,mosses,and all other investigated land plants,but not present in any algae.The tertiary structures of representative liverwort and moss nsLTPs were further studied with homology modeling.The results indicate that the nsLTP family has evolved after plants conquered land.Only two of the four major subfamilies of nsLTPs found in flowering plants are present in mosses and liverworts.The additional subfamilies have arisen later,during land plant evolution.In this report,we also introduce a modified nsLTP classification system.