Cutinsomes and CUTIN SYNTHASE1 Function Sequentially in Tomato Fruit Cutin Deposition.

The aerial parts of plants, including the leaves, fruits and non-lignified stems, are covered with a protective cuticle, largely composed of the polyester cutin. Two mechanisms of cutin deposition have been identified in tomato (Solanum lycopersicum) fruit. The contribution of each mechanism to cutin synthesis and deposition has shown a temporal and coordinated sequence that correlates with the two periods of organ growth, cell division and cell expansion. Cutinsomes, self-assembled particles composed of esterified cutin monomers, are involved in the synthesis of the procuticle during cell division and provide a template for further cutin deposition. CUTIN SYNTHASE1 (CUS1), an acyl transferase enzyme that links cutin monomers, contributes to massive cuticle deposition during the early stages of the cell expansion period by incorporating additional cutin to the procuticle template. However, cutin deposition and polymerization appear to be part of a more complex biological scenario, which is yet not fully understood. CUS1 is also associated with the coordinated growth of the cutinized and non-cutinized domains of the outer epidermal wall, and affects cell size. A dynamic and complex interplay linking cutin synthesis with cell wall development and epidermal cell size has been identified.

Ultrastructure of the Epidermal Cell Wall and Cuticle of Tomato Fruit (Solanum lycopersicum L.) during Development.

The epidermis plays a pivotal role in plant development and interaction with the environment. However, it is still poorly understood, especially its outer epidermal wall: a singular wall covered by a cuticle. Changes in the cuticle and cell wall structures are important to fully understand their functions. In this work, an ultrastructure and immunocytochemical approach was taken to identify changes in the cuticle and the main components of the epidermal cell wall during tomato fruit development. A thin and uniform procuticle was already present before fruit set. During cell division, the inner side of the procuticle showed a globular structure with vesicle-like particles in the cell wall close to the cuticle. Transition between cell division and elongation was accompanied by a dramatic increase in cuticle thickness, which represented more than half of the outer epidermal wall, and the lamellate arrangement of the non-cutinized cell wall. Changes in this non-cutinized outer wall during development showed specific features not shared with other cell walls. The coordinated nature of the changes observed in the cuticle and the epidermal cell wall indicate a deep interaction between these two supramolecular structures. Hence, the cuticle should be interpreted within the context of the outer epidermal wall.

Biomechanical properties of the tomato (Solanum lycopersicum) fruit cuticle during development are modulated by changes in the relative amounts of its components.

In this study, growth-dependent changes in the mechanical properties of the tomato (Solanum lycopersicum) cuticle during fruit development were investigated in two cultivars with different patterns of cuticle growth and accumulation. The mechanical properties were determined in uniaxial tensile tests using strips of isolated cuticles. Changes in the functional groups of the cuticle chemical components were analysed by attenuated total reflectance-Fourier transform infrared (ATR-FTIR). The early stages of fruit growth are characterized by an elastic cuticle, and viscoelastic behaviour only appeared at the beginning of cell enlargement. Changes in the cutin:polysaccharide ratio during development affected the strength required to achieve viscoelastic deformation. The increase in stiffness and decrease in extensibility during ripening, related to flavonoid accumulation, were accompanied by an increase in cutin depolymerization as a result of a reduction in the overall number of ester bonds. Quantitative changes in cuticle components influence the elastic/viscoelastic behaviour of the cuticle. The cutin:polysaccharide ratio modulates the stress required to permanently deform the cuticle and allow cell enlargement. Flavonoids stiffen the elastic phase and reduce permanent viscoelastic deformation. Ripening is accompanied by a chemical cleavage of cutin ester bonds. An infrared (IR) band related to phenolic accumulation can be used to monitor changes in the cutin esterification index.

Genome-wide QTL analysis of tomato fruit cuticle deposition and composition.

Genetics of traits related to fruit cuticle deposition and composition was studied in two red-fruited tomato species. Two mapping populations derived from the cross between the cultivated tomato (Solanum lycopersicum L.) and its closest relative wild species Solanum pimpinellifolium L. were employed to conduct a QTL analysis. A combination of fruit cuticle deposition, components and anatomical traits were investigated and the individual effect of each QTL evaluated. A total of 70 QTLs were identified, indicating that all the cuticle traits analyzed have a complex polygenic nature. A combination of additive and epistatic interactions was observed for all the traits, with positive contribution of both parental lines to most of them. Colocalization of QTLs for various traits uncovered novel genomic regions producing extensive changes in the cuticle. Cuticle density emerges as an important trait since it can modulate cuticle thickness and invagination thus providing a strategy for sustaining mechanical strength without compromising palatability. Two genomic regions, located in chromosomes 1 and 12, are responsible for the negative interaction between cuticle waxes and phenolics identified in tomato fruit. Several candidate genes, including transcription factors and structural genes, are postulated and their expression analyzed throughout development.

Pectin-cellulose nanocrystal biocomposites: Tuning of physical properties and biodegradability.

The fabrication of pectin-cellulose nanocrystal (CNC) biocomposites has been systematically investigated by blending both polysaccharides at different relative concentrations. Circular free-standing films with a diameter of 9 cm were prepared by simple solution of these carbohydrates in water followed by drop-casting and solvent evaporation. The addition of pectin allows to finely tune the properties of the biocomposites. Textural characterization by AFM showed fibrous morphology and an increase in fiber diameter with pectin content. XRD analysis demonstrated that pectin incorporation also reduced the degree of crystallinity though no specific interaction between both polysaccharides was detected, by ATR-FTIR spectroscopy. The optical properties of these biocomposites were characterized for the first time and it was found that pectin in the blend reduced the reflectance of visible light and increased UV absorbance. Thermal stability, analyzed by TGA, was improved with the incorporation of pectin. Finally, pectin-cellulose nanocrystal biocomposites showed a good biodegradability in seawater, comparable to other common bioplastics such as cellulose and low-molecular weight polylactide, among others.

Cutinisation of tomato fruit epidermis: Structural and morphological details.

In tomato, the ovary is covered with a thin, electron-dense and uniform cuticle. The first 10 d after anthesis are critical in the cutinisation of the outer epidermal wall. During this period, singular cytoplasmic domains have been identified in the epidermal cells which seem to be involved in lipid biosynthesis. Moreover, the inner side of the procuticle showed a globular structure with vesicle-like particles of different size that seemed to migrate from the cytoplasm to the procuticle. These electron-dense particles are postulated to play an important role in early cutin synthesis.