Ethylene has long been regarded as the main regulator of ripening in climacteric fruits. The characterization of a few tomato mutants, unable to produce climacteric ethylene and to ripen their fruits even following treatments with exogenous ethylene, has shown that other factors also play an important role in the control of climacteric fruit ripening. In climacteric peach and tomato fruits it has been shown that, concomitant with ethylene production, increases in the amount of auxin can also be measured. In this work a genomic approach has been used in order to understand if such an auxin increase is functional to an independent role played by the hormone during ripening of the climacteric peach fruits. Besides the already known indirect activity on ripening due to its up-regulation of climacteric ethylene synthesis, it has been possible to show that auxin plays a role of its own during ripening of peaches. In fact, the hormone has shown the ability to regulate the expression of a number of different genes. Moreover, many genes involved in biosynthesis and transport and, in particular, the signalling (receptors, Auxin Response Factors and Aux/IAA) of auxin had increased expression in the mesocarp during ripening, thus strengthening the idea that this hormone is actively involved in the ripening of peaches. This study has also demonstrated the existence of an important cross-talk between auxin and ethylene, with genes in the auxin domain regulated by ethylene and genes in the ethylene domain regulated by auxin.

Suberin, a polymer composed of both aliphatic and aromatic domains, is deposited as a rough matrix upon plant surface damage and during normal growth in the root endodermis, bark, specialized organs (e.g., potato [Solanum tuberosum] tubers), and seed coats. To identify genes associated with the developmental control of suberin deposition, we investigated the chemical composition and transcriptomes of suberized tomato (Solanum lycopersicum) and russet apple (Malus x domestica) fruit surfaces. Consequently, a gene expression signature for suberin polymer assembly was revealed that is highly conserved in angiosperms. Seed permeability assays of knockout mutants corresponding to signature genes revealed regulatory proteins (i.e., AtMYB9 and AtMYB107) required for suberin assembly in the Arabidopsis thaliana seed coat. Seeds of myb107 and myb9 Arabidopsis mutants displayed a significant reduction in suberin monomers and altered levels of other seed coat-associated metabolites. They also exhibited increased permeability, and lower germination capacities under osmotic and salt stress. AtMYB9 and AtMYB107 appear to synchronize the transcriptional induction of aliphatic and aromatic monomer biosynthesis and transport and suberin polymerization in the seed outer integument layer. Collectively, our findings establish a regulatory system controlling developmentally deposited suberin, which likely differs from the one of stressinduced polymer assembly recognized to date.

MYB10.1 and MYB10.3, with bHLH3, are the likely regulators of anthocyanin biosynthesis in peach fruit. MYB10.1/2/3 forms a cluster on the same genomic fragment where the Anther color ( Ag ) trait is located. Anthocyanins are bioactive compounds responsible for the pigmentation of many plant parts such as leaves, flowers, fruits and roots, and have potential benefits to human health. In peach [Prunus persica (L.) Batsch], peel color is a key determinant for fruit quality and is regulated by flavonoids including anthocyanins. The R2R3 MYB transcription factors (TFs) control the expression of anthocyanin biosynthetic genes with the help of co-activators belonging to the basic-helix-loop-helix (bHLH) and WD40 repeat families. In the peach genome six MYB10-like and three bHLH-like TFs were identified as candidates to be the regulators of the anthocyanin accumulation, which, in yellow flesh fruits, is highest in the peel, abundant in the part of the mesocarp surrounding the stone and lowest in the mesocarp. The expression of MYB10.1 and MYB10.3 correlates with anthocyanin levels of different peach parts. They also have positive correlation with the expression of key structural genes of the anthocyanin pathway, such as CHS, F3H, and UFGT. Functions of peach MYB10s were tested in tobacco and shown to activate key genes in the anthocyanin pathway when bHLHs were co-expressed as partners. Overexpression of MYB10.1/bHLH3 and MYB10.3/bHLH3 activated anthocyanin production by up-regulating NtCHS, NtDFR and NtUFGT while other combinations were not, or much less, effective. As three MYB10 genes are localized in a genomic region where the Ag trait, responsible for anther pigmentation, is localized, it is proposed they are key determinant to introduce new peach cultivars with higher antioxidant level and pigmented fruit.

Notwithstanding the economic importance of non-climacteric fruits like grape and strawberry, little is known about the mechanisms that regulate their ripening. Up to now no growth regulator has emerged with a primary role similar to that played by ethylene in the ripening of the climacteric fruits. Strawberries can produce ethylene, although in limited amounts. Two cDNAs coding for enzymes of the ethylene biosynthetic pathway (i.e. FaACO1 and FaACO2), and three cDNAs encoding different ethylene receptors have been isolated. Two receptors (i.e. FaEtr1 and FaErs1) belong to the type-I while the third (i.e. FaEtr2) belongs to the type-II group. The expression of both the ACO and the receptor-encoding genes has been studied in fruits at different stages of development and in fruits treated with hormones (i.e. ethylene and the auxin analogue NAA). All the data thus obtained have been correlated to the known data about ethylene production by strawberry fruits. Interestingly, a good correlation has resulted between the expression of the genes described in this work and the data of ethylene production. In particular, similarly to what occurs during climacteric fruit ripening, there is an increased synthesis of receptors concomitant with the increased synthesis of ethylene in strawberries as well. Moreover, the receptors mostly expressed in ripening strawberries are the type-II ones, that is those with a degenerate histidine-kinase domain. Since the latter domain is thought to establish a weaker link to the CTR1 proteins, even the little ethylene produced by ripening strawberries might be sufficient to trigger ripening-related physiological responses.

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During ripening, fleshy fruits undergo textural changes that lead to loss of tissue firmness and consequent softening. It is a common idea that this process is the consequence of cell wall dismantling carried out by different and orderly expressed enzymes. For this purpose, by using a single enzyme family approach many enzymes and related genes have been characterized in different fruits. In this work, the softening of the climacteric peach fruits (Prunus persica (L.) Batsch.) has been studied by using a genomic approach, and the results obtained are novel and partly unexpected. The genes analysed encode proteins involved in the main metabolic aspects of a primary cell wall: degradation, synthesis, structure. In addition, some genes encoding cell-wall-related proteins with an unknown function have been studied. The gene expression profiles show that the softening actually begins well before the climacteric rise and continues thereafter. Genes whose expression starts before the climacteric rise are mostly down-regulated by ethylene, while genes with a ripening-specific expression are mostly up-regulated by the hormone. A few other genes are apparently insensitive to ethylene. Besides the expected parietal degradation, the softening that results from this study also comprises some repairing of the cell wall performed by enzymes involved in the synthesis of parietal polysaccharides and, especially, by proteins with structural functions. The newly synthesized polysaccharides and the structural proteins would thus help to hold together the fruit cell wall while not preventing the softening.

Ethylene promotes the abscission of leaves and the ripening of fruits in pepper plants, and in both events an increase in cellulase activity is observed. However, two enzyme isoforms (pI 7.2 and 8.5, respectively) are differentially involved in the two physiological phenomena. The pI 8.5 form has been purified from ripe fruits. It is a glycoprotein with an apparent molecular mass of 54 kDa. Two short peptides were sequenced and a very high homology to a tomato cellulase was observed. Polyclonal antibodies, raised against the purified enzyme, have allowed us to demonstrate that the observed ethylene-induced increase in cellulase activity is paralleled by de novo synthesis of protein. Three cDNAs (CX1, CX2 and CX3), encoding different cellulases, were obtained and characterized and their expression investigated. Accumulation of all three mRNAs is induced by ethylene treatment, though to different levels. CX1 is mainly expressed in ripe fruits while CX2 is especially found in abscission zones. CX3 accumulates at very low levels in activated abscission zones. Comparisons with other known cellulases demonstrate clear heterogeneity within the higher plant cellulases. Differences in ethylene inducibility and molecular structure suggest different physiological roles for cellulase in pepper plants.