Searches of sequenced genomes of diverse organisms revealed that the moss Physcomitrella patens is the most primitive organism possessing oleosin genes. Microscopy examination of Physcomitrella revealed that oil bodies (OBs) were abundant in the photosynthetic vegetative gametophyte and the reproductive spore. Chromatography illustrated the neutral lipids in OBs isolated from the gametophyte to be largely steryl esters and triacylglycerols, and SDS-PAGE showed the major proteins to be oleosins. Reverse transcription-PCR revealed the expression of all three oleosin genes to be tissue specific. This tissue specificity was greatly altered via alternative splicing, a control mechanism of oleosin gene expression unknown in higher plants. During the production of sex organs at the tips of gametophyte branches, the number of OBs in the top gametophyte tissue decreased concomitant with increases in the number of peroxisomes and level of transcripts encoding the glyoxylate cycle enzymes; thus, the OBs are food reserves for gluconeogenesis. In spores during germination, peroxisomes adjacent to OBs, along with transcripts encoding the glyoxylate cycle enzymes, appeared; thus, the spore OBs are food reserves for gluconeogenesis and equivalent to seed OBs. The one-cell-layer gametophyte could be observed easily with confocal microscopy for the subcellular OBs and other structures. Transient expression of various gene constructs transformed into gametophyte cells revealed that all OBs were linked to the endoplasmic reticulum (ER), that oleosins were synthesized in extended regions of the ER, and that two different oleosins were colocated in all OBs.Eukaryotes and prokaryotes contain neutral lipids in subcellular droplets as food reserves and/or for other purposes (Hsieh and Huang, 2004;Martin and Parton, 2006;Goodman, 2008;Rajakumari et al., 2008). These lipid droplets are present in seeds, pollens, fruits, and flowers of higher plants; the vegetative and reproductive organs of lower plants, algae, fungi, and nematodes; mammalian organs/tissues, such as mammalian glands and adipose tissues; and bacteria. Among all these lipid droplets, oil bodies (OBs) in seeds are the most prominent and have been extensively studied.Seeds of diverse plant species store oils (triacylglycerols [TAGs]) as food reserves for germination and postgermination growth (Napier et al., 1996;Frandsen et al., 2001;Murphy, 2001;Hsieh and Huang, 2004).The TAGs are present in small subcellular, spherical OBs of approximately 0.5 to 2 mm in diameter. Each OB has a matrix of TAGs surrounded by a layer of phospholipids (PLs) and the structural protein oleosins. The massive oleosins completely cover the surface of the OBs and prevent them from coalescence; so, a large surface area per unit TAG is available for lipase binding and catalysis during germination. Each oleosin molecule has a characteristic long central hydrophobic stretch, which forms a hairpin penetrating into the matrix TAGs for stable anchorage.Other than being present in the seeds of plants, oleos...