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.

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.

European canker, caused by Neonectria ditissima Bres., is an economically damaging fungal disease of apple. Breeding new cultivars with a high level of resistance to European canker is the main aim of apple breeding programs. Observations of symptoms in naturally infected trees were carried out in 400 apple accessions in Asturias (north-western Spain). Young and mature field planted trees were assessed under conditions highly conducive for N. ditissima development. The results demonstrated that juvenile trees (4-year-old) barely showed noticeable symptoms whereas a wide variability in the levels of resistance among accession was observed in mature trees (14-year-old). Around 28% of the locally maintained collection resulted to be highly resistant to this disease in the region. Field observations on mature trees were also compared to four rapid screening tests based on artificially induced lesions. Spearman correlation analysis using two resistance parameters revealed that none of the methods resulted in similar rankings of cultivar susceptibility as some accessions that were ranked as resistant for a given test turned out to be susceptible in the field. This study might suggest that whilst conventional resistance phenotyping techniques are time-consuming, the outcomes of this approach still seem the preferred option to assess the response to N. ditissima of apple accessions.

Fire blight, caused by Erwinia amylovora, is one of the most devastating apple diseases. The selection of cultivars of low susceptibility and the study of the genetic mechanisms of the disease play important roles in fire blight management. The susceptibility level to fire blight was evaluated in 102 accessions originating from Asturias, a cider-producing region located in the north of Spain with a wide apple germplasm. Evaluations took place under quarantine conditions using artificial inoculations of grafted plants. The results revealed wide variation in susceptibility responses and low-susceptible cultivars were identified. In addition, 91 cultivars were genotyped using the Affymetrix Axiom^® Apple 480 K SNP array to conduct genome-wide association studies (GWAS). A statistically significant signal was detected on chromosome 10 using the multi-locus mixed model (MLMM). Two genes were identified as major putative candidate genes: a TIR-NBS-LRR class disease protein and a protein containing a development and cell death (DCD) domain. The outcomes of this study provide a promising source of information, particularly in the context of cider apples, and set a starting point for future genetic and breeding approaches.