Suberin is a natural biopolymer found in a variety of specialized tissues, including seed coat integuments, root endodermis, tree bark, potato tuber skin and the russeted and reticulated skin of fruits. The suberin polymer consists of polyaliphatic and polyphenolic domains. The former is made of very long chain fatty acids, primary alcohols and a glycerol backbone, while the latter consists of p-hydroxycinnamic acid derivatives, which originate from the core phenylpropanoid pathway. In the current review, we survey the current knowledge on genes/enzymes associated with the suberin biosynthetic pathway in plants, reflecting the outcomes of considerable research efforts in the last two decades. We discuss the function of these genes/enzymes with respect to suberin aromatic and aliphatic monomer biosynthesis, suberin monomer transport, and suberin pathway regulation. We also delineate the consequences of the altered expression/accumulation of these genes/enzymes in transgenic plants.

The hydrophobic cuticle encasing the fruit skin surface plays critical roles during fruit development and post-harvest. Skin failure often results in the fruit surface cracking and forming a wound-periderm tissue made of suberin and lignin. The factors that make the fruit skin susceptible to cracking have yet to be fully understood. Herein, we investigated two varieties of chili peppers (Capsicum annuum L.), Numex Garnet, whose fruit has intact skin, and Vezena Slatka, whose fruit has cracked skin. Microscopical observations, gas chromatography–mass spectrometry, biochemical and gene expression assays revealed that Vezena Slatka fruit form a thicker cuticle with greater levels of cutin monomers and hydroxycinnamic acids, and highly express key cutin-related genes. The skin of these fruit also had a lower epidermal cell density due to cells with very large perimeters, and highly express genes involved in epidermal cell differentiation. We demonstrate that skin cracking in the Vezena Slatka fruit is accompanied by a spatial accumulation of lignin-like polyphenolic compounds, without the formation of a typical wound-periderm tissues made of suberized cells. Lastly, we establish that skin cracking in chili-type pepper significantly affects fruit quality during post-harvest storage in a temperature-dependent manner. In conclusion, our data highlight cuticle thickness and epidermal cell density as two critical factors determining fruit skin susceptibility to cracking in chili-type pepper fruit.

The epidermal cells of petunia (Petunia × hybrida) flowers are the main site of volatile emission. However, the mechanisms underlying the release of volatiles into the environment are still being explored. Here, using cell-layer-specific transcriptomic analysis, reverse genetics by virus-induced gene silencing and clustered regularly interspaced short palindromic repeat (CRISPR), and metabolomics, we identified EPIDERMIS VOLATILE EMISSION REGULATOR (EVER)—a petal adaxial epidermis-specific MYB activator that affects the emission of volatiles. To generate ever knockout lines, we developed a viral-based CRISPR/Cas9 system for efficient gene editing in plants. These knockout lines, together with transient-suppression assays, revealed EVER's involvement in the repression of low-vapor-pressure volatiles. Internal pools and annotated scent-related genes involved in volatile production and emission were not affected by EVER. RNA-Seq analyses of petals of ever knockout lines and EVER-overexpressing flowers revealed enrichment in wax-related biosynthesis genes. Liquid chromatography/gas chromatography–MS analyses of petal epicuticular waxes revealed substantial reductions in wax loads in ever petals, particularly of monomers of fatty acids and wax esters. These results implicate EVER in the emission of volatiles by fine-tuning the composition of petal epicuticular waxes. We reveal a petunia MYB regulator that interlinks epicuticular wax composition and volatile emission, thus unraveling a regulatory layer in the scent-emission machinery in petunia flowers.

The necrotrophic fungus Botrytis cinerea, is considered a major cause of postharvest losses in a wide range of crops. The common fungal extracellular membrane protein (CFEM), containing a conserved eight-cysteine pattern, was found exclusively in fungi. Previous studies in phytopathogenic fungi have demonstrated the role of membrane-bound and secreted CFEM-containing proteins in different aspects of fungal virulence. However, non-G protein-coupled receptor (non-GPCR) membrane CFEM proteins have not been studied yet in phytopathogenic fungi. In the present study, we have identified a non-GPCR membrane-bound CFEM-containing protein, Bcin07g03260, in the B. cinerea genome, and generated deletion mutants, ΔCFEM-Bcin07g03260, to study its potential role in physiology and virulence. Three independent ΔCFEM-Bcin07g03260 mutants showed significantly reduced progression of a necrotic lesion on tomato (Solanum lycopersicum) leaves. Further analysis of the mutants revealed significant reduction (approximately 20–30%) in conidial germination and consequent germ tube elongation compared with the WT. Our data complements a previous study of secreted ΔCFEM1 mutants of B. cinerea that showed reduced progression of necrotic lesions on leaves, without effect on germination. Considering various functions identified for CFEM proteins in fungal virulence, our work illustrates a potential new role for a non-GPCR membrane CFEM in pathogenic fungi to control virulence in the fungus B. cinerea.

Plants synthesize ascorbate (ASC) via the D-mannose/L-galactose pathway whereas animals produce ASC and H₂O₂via the UDP-glucose pathway, with Gulono-1,4 γ-lactone oxidases (GULLO) as the last step. A. thaliana has seven isoforms, GULLO1-7; previous in silico analysis suggested that GULLO2, mostly expressed in developing seeds, might be involved in iron (Fe) nutrition. We isolated atgullo2-1 and atgullo2-2 mutants, quantified ASC and H₂O₂ in developing siliques, Fe(III) reduction in immature embryos and seed coats. Surfaces of mature seed coats were analysed via atomic force and electron microscopies; suberin monomer and elemental compositions of mature seeds, including Fe, were profiled via chromatography and inductively coupled plasma-mass spectrometry. Lower levels of ASC and H₂O₂ in atgullo2 immature siliques are accompanied by an impaired Fe(III) reduction in seed coats and lower Fe content in embryos and seeds; atgullo2 seeds displayed reduced permeability and higher levels of C18:2 and C18:3 ω-hydroxyacids, the two predominant suberin monomers in A. thaliana seeds. We propose that GULLO2 contributes to ASC synthesis, for Fe(III) reduction into Fe(II). This step is critical for Fe transport from endosperm into developing embryos. We also show that alterations in GULLO2 activity affect suberin biosynthesis and accumulation in the seed coat.

Fruit cuticles control water and gas diffusion and protect against biotic and environmental stresses. The cuticle is built from the cutin polymer—a composite of C16 and C18 ω-hydroxy fatty acids that are linked via ester bonds, embedded polysaccharides, and phenolics—as well as waxes made primarily from very-long-chain fatty acids that are deposited on the cuticle and incorporated within the cutin matrix. Considerable progress toward understanding fruit cuticle function has been achieved in recent years, but knowledge gaps remain regarding the biosynthesis and assembly of the cuticular constituents and how these processes are linked to the cuticle's macromolecular architecture and nanomechanical properties. Herein, we generated transgenic tomatoes (Solanum lycopersicum) that express, in an exocarp-specific manner, the Fusarium oxysporum f.sp. lycopersici fungal cutinase gene that degrades the host plant cuticle during infection via breakdown of ester bonds between cutin monomers. To track the consequences of this genetic manipulation on fruit cuticle ultrastructure, chemistry and surface behavior, we coordinated information from microscopy, chemical profiling, nanomechanics, biophysical measurements, and transcriptomics. The transgenic fruits deposited unexpectedly thicker cuticles with elevated cutin and wax content as well as distinctive surface cracks and suberized wound periderms; they had enhanced roughness, elastic modulus, stiffness, and adhesion of the cuticle surface; and they showed wide-ranging alterations in gene expression, phase state, and permeability. Taken together, our findings provide valuable knowledge on how the shifted balance among wax, cutin, and suberin that is triggered by exocarp-specific genetic modifications can remodel the fruit cuticle ultrastructure, chemistry and nanomechanics.