The outer wall of Ornithogalum umbellatum ovary and the fruit epidermis are covered with a thick cuticle and contain lipotubuloids incorporating 3H-palmitic acid. This was earlier evidenced by selective autoradiographic labelling of lipotubuloids. After post-incubation in a non-radioactive medium, some marked particles insoluble in organic solvents (similar to cutin matrix) moved to the cuticular layer. Hence, it was hypothesised that lipotubuloids participated in cuticle synthesis. It was previously suggested that cutinsomes, nanoparticles containing polyhydroxy fatty acids, formed the cuticle. Thus, identification of the cutinsomes in O. umbellatum ovary epidermal cells, including lipotubuloids, was undertaken in order to verify the idea of lipotubuloid participation in cuticle synthesis in this species. Electron microscopy and immunogold method with the antibodies recognizing cutinsomes were used to identify these structures. They were mostly found in the outer cell wall, the cuticular layer and the cuticle proper. A lower but still significant degree of labelling was also observed in lipotubuloids, cytoplasm and near plasmalemma of epidermal cells. It seems that cutinsomes are formed in lipotubuloids and then they leave them and move towards the cuticle in epidermal cells of O. umbellatum ovary. Thus, we suggest that (1) cutinsomes could take part in the synthesis of cuticle components also in plant species other than tomato, (2) the lipotubuloids are the cytoplasmic domains connected with cuticle formation and (3) this process proceeds via cutinsomes.

Lipid bodies present in lipotubuloids of Ornithogalum umbellatum ovary epidermis take the form of a lens between leaflets of ER (endoplasmic reticulum) membrane filled with a highly osmiophilic substance. The two enzymes, DGAT1 [DAG (diacylglycerol) acyltransferase 1] and DGAT2 (DAG acyltransferase 2), involved in this process are synthesized on rough ER and localized in the ER near a monolayer surrounding entities like lipid bodies. After reaching the appropriate size, newly formed lipid bodies transform into mature spherical lipid bodies filled with less osmiophilic content. They appear to be surrounded by a half-unit membrane, with numerous microtubules running adjacently in different directions. The ER, no longer continuous with lipid bodies, makes contact with them through microtubules. At this stage, lipid synthesis takes place at the periphery of lipid bodies. This presumption, and a hypothesis that microtubules are involved in lipid synthesis delivering necessary components to lipid bodies, is based on strong arguments: (i) silver grains first appear over microtubules after a short [3H]palmitic acid incubation and before they are observed over lipid bodies; (ii) blockade of [3H]palmitic acid incorporation into lipotubuloids by propyzamide, an inhibitor of microtubule function; and (iii) the presence of gold grains above the microtubules after DGAT1 and DGAT2 reactions, as also near microtubules after an immunogold method that identifies phospholipase D1.

A metabolon is a temporary, structural-functional complex formed between sequential metabolic enzymes and cellular elements. Cytoplasmic domains called lipotubuloids are present in Ornithogalum umbellatum ovary epidermis. They consist of numerous lipid bodies entwined with microtubules, polysomes, rough endoplasmic reticulum (RER), and actin filaments connected to microtubules through myosin and kinesin. A few mitochondria, Golgi structures, and microbodies are also observed and also, at later development stages, autolytic vacuoles. Each lipotubuloid is surrounded by a tonoplast as it invaginates into a vacuole. These structures appear in young cells, which grow intensively reaching 30-fold enlargement but do not divide. They also become larger due to an increasing number of lipid bodies formed in the RER by the accumulation of lipids between leaflets of the phospholipid bilayer. When a cell ceases to grow, the lipotubuloids disintegrate into individual structures. Light and electron microscope studies using filming techniques, autoradiography with [(3)H]palmitic acid, immunogold labelling with antibodies against DGAT2, phospholipase D1 and lipase, and double immunogold labelling with antibodies against myosin and kinesin, as well as experiments with propyzamide, a microtubule activity inhibitor, have shown that lipotubuloids are functionally and structurally integrated metabolons [here termed lipotubuloid metabolons (LMs)] occurring temporarily in growing cells. They synthesize lipids in lipid bodies in cooperation with microtubules. Some of these lipids are metabolized and used by the cell as nutrients, and others are transformed into cuticle whose formation is mediated by cutinsomes. The latter were discovered in planta using specific anti-cutinsome antibodies visualized by gold labelling. Moreover, LMs are able to rotate autonomously due to the interaction of microtubules, actin filaments, and motor proteins, which influence microtubules by changing their diameter.

In the cells of Haemanthus albiflos leaf epidermis there are structures containing lipids analogous to ''elaioplasts'' (Wakker in Jahrb F Wiss Bot 19:423-496, 1888). Ultrastructural analysis has shown that they are cytoplasmic domains-lipotubuloids, since they exhibit all the features of Ornithogalum umbellatum lipotubuloids. They are composed of numerous lipid bodies surrounded by microtubules, ER cisternae and vesicles, some mitochondria, Golgi structures, and microbodies. In the center of some lipotubuloids there are also autolytic vacuoles. Microtubules adjacent to H. albiflos lipid bodies were revealed only when taxol preincubation was used before fixing the epidermis in the mixture of glutaraldehyde and OsO 4 . The presence of tubulin in H. albiflos and O. umbellatum lipotubuloids was confirmed with use of the immunogold method involving antibodies against tubulin a. It is possible that the association of microtubules with lipid bodies may be more common than originally thought, but it is difficult to reveal due to the methodological problems.

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The cuticle commonly appears as a continuous lipophilic layer located at the outer epidermal cell walls of land plants. Cutin and waxes are its main components. Two methods for cutin synthesis are considered in plants. One that is based on enzymatic biosynthesis, in which cutin synthase (CUS) is involved, is well-known and commonly accepted. The other assumes the participation of specific nanostructures, cutinsomes, which are formed in physicochemical self-assembly processes from cutin precursors without enzyme involvement. Cutinsomes are formed in ground cytoplasm or, in some species, in specific cytoplasmic domains, lipotubuloid metabolons (LMs), and are most probably translocated via microtubules toward the cuticle-covered cell wall. Cutinsomes may additionally serve as platforms transporting cuticular enzymes. Presumably, cutinsomes enrich the cuticle in branched and cross-linked esterified polyhydroxy fatty acid oligomers, while CUS1 can provide both linear chains and branching cutin oligomers. These two systems of cuticle formation seem to co-operate on the surface of aboveground organs, as well as in the embryo and seed coat epidermis. This review focuses on the role that cutinsomes play in cuticle biosynthesis in S. lycopersicum, O. umbellatum and A. thaliana, which have been studied so far; however, these nanoparticles may be commonly involved in this process in different plants.

Spermiogenesis in Chara vulgaris and in animals share many common features, including exchange of nucleohistones into nucleoprotamines, remodeling and extreme condensation of chromatin, formation of flagellae and of microtubule manchette, and decrease in cytoplasm volume. In C. vulgaris, spermiogenesis is not preceded by meiosis since this alga is a haplobiont. In the present work we showed that in early spermiogenesis characterized by a significant metabolic activity of spermatids, the inhibitors of proteasomes did not visibly change their ultrastructure but significantly prolonged this process. At late stages of spermiogenesis, MG-132 and epoxomicin dramatically changed the structure of nuclei: regular fibrillar and lamellar structure of chromatin was disturbed and clusters of grains corresponding to aggresomes appeared, but the nucleus shape and cytoplasm structure were the same as in the controls. Immunocytochemical studies revealed that these inhibitors blocked disappearance of histones from nuclei while the structures corresponding to aggresomes were clusters of undegraded ubiquitinated histones, since they gave positive immunosignals indicating the presence of ubiquitin and histones.

Abstract"Elaioplasts" observed in Vanilla planifolia, Funkia Sieboldiana and Althaea rosea exhibit all the features characteristic of lipotubuloids earlier described in Ornithogalum umbellatum. They are cytoplasmic domains containing aggregates of lipid bodies connected with microtubules. The immunogold technique confirmed the presence of tubulin in this domain. These structures do not have their own membranes but they are surrounded by a tonoplast at the side of a vacuole since they invaginate into it. In cytoplasm of this domain among lipid bodies there are numerous ribosomes, ER cisternae and vesicles as well as few mitochondria, Golgi structures and microbodies while at older developmental stages there are also autolytic vacuoles. The fact that they are so similar to O. umbellatum lipotubuloids suggest that "elaioplasts" of V. planifolia, F. Sieboldiana and A. rosea can also be named lipotubuloids.