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Liquid chromatography high-resolution mass spectrometry fingerprinting together with pattern recognition techniques was used to determine the metabolites involved in the susceptibility of apple cultivars to rosy apple aphid (RAA). Preprocessing of ultra-high-performance liquid chromatography coupled to lectrospray ionization and quadrupole time-of-flight mass spectrometry raw data of resistant and susceptible apple cultivars was carried out with XCMS and CAMERA packages. Univariate statistical tools and multivariate data analysis highlighted significant different profiles of the apple metabolomes according to their tolerance to RAA. Optimized and cross-validated Partial least squares discriminant analysis and orthogonal projections to latent structures discriminant analysis models confirmed trans-4-caffeoylquinic acid and 4-p-coumaroylquinic acid as biomarkers for the identification of resistant and susceptible apple cultivars to RAA and disclosed that only hydroxycinnamic acids are involved in the disease susceptibility of cultivars. In this sense, the final steps of the biosynthesis of caffeoylquinic acid (CQA) and p-coumaroylquinic acid (p-CoQA) become decisive because the isomerization of 5-CQA to 4-CQA is favored in resistant cultivars, whereas the isomerization of 5-p-CoQA to 4-p-CoQA is favored in susceptible cultivars.

Most apple trees (Malus domestica Borkh.) are self-incompatible and fruit yield depends on cross-pollination between genetically compatible cultivars with synchronous flowering. Flowering intensity can vary strongly among years due to the biennial bearing habit of the cultivars. The knowledge of the phenological stages and floral and pollen characteristics is essential to select suitable pollen donors. We evaluated the phenotypic variability of flowering-related traits (i.e., flowering phenology, flowering intensity, pollen production and pollen quality) in 45 apple cultivars over two successive flowering seasons. Large phenotypic variability was found among the studied cultivars indicating that the local germplasm collection provides a good source of genetic and phenotypic diversity. However, low correlations were observed between floral biology traits and, consequently, the improvement in one trait seems not to affect other traits. Some of the cultivars such as ‘Perurico’ and ‘Raxila Dulce’ regularly produced copious amounts of high-quality pollen which can improve the pollen load dispersion leading to a most effective pollination process. We did not identify statistically significant correlations between pollen attributes and the biennial bearing phenomenon. The large variation in bloom dates from year-to-year observed under a typical Oceanic climate makes it advisable to combine cultivars in new plantings.

Comprehensive chemical characterization of nine mono-varietal apple pomaces obtained from the production of ciders with PDO is described. They were rich in essential minerals, fibers (35–52.9 %), and polyphenols. High levels in GalA (11.8–21.6 %), revealed the suitability of these apple pomaces as efficient sources of pectins. Extracted pectins showed high variability in monomer composition, with degrees of methylesterification, strongly associated with pectins functional properties, ranging from 58 to 88 %. For a subset of apple pomace varieties, pectin extraction was accomplished by conventional acid heat treatment or ultrasound. Despite ultrasound-assisted extraction did not improve pectin yield, it minimized levels of “non-pectin” components as revealed by the low content of Glc/Man, leading to the obtainment of high-purity pectin. Our work highlights the key role played by the selection of the apple variety to streamline the potential food applications (gelling/thickening agents or prebiotics) of the extracted pectins that largely depend on their structural features.

The phenolic profiles of apple cultivars from the SERIDA Asturian cider apple breeding program, including parents and progenies, were determined by ultrahigh-performance liquid chromatography-diode array detector-electrospray ionization-quadrupole time of flight/mass spectrometer in order to study the relationship between phenols and the resistance of apple tree cultivars to rosy apple aphid (RAA). A pattern recognition technique named partial least square discriminant analysis (PLS-DA) was used to classify apple cultivars based on resistance to RAA, resistant and susceptible, reaching scores with accuracy higher than 97% and 91% respectively. Hydroxycinnamic acids, particularly 4-caffeoylquinic acid (4-CQA) and 4-p-coumaroylquinic acid (4-pCoQA), were identified as the major player in RAA resistance by the PLS-DA model. Indeed, the isomerisation 5-CQA → 4-CQA is favoured in resistant cultivars, whereas the isomerisation 5-pCoQA → 4-pCoQA is favoured in susceptible cultivars. As a result, resistant cultivars accumulate higher amounts of 4-CQA than susceptible ones, and the opposite occurs for 4-pCoQA. Also, minor isomerisations of 5-CQA to 1-CQA or 3-CQA show opposite behaviour for resistant and susceptible cultivars. Cultivar resistance to RAA is concluded to be related with the phenylpropanoid pathway, the isomerisation reactions being the key metabolic reaction for a cultivar to be resistant or susceptible to RAA.