Open Access
Exploring phenotypic diversity and stability of key traits for apple breeding in northeastern Spanish germplasm
(Frontiers Media, 2025-09-16) Bielsa González, Francisco Javier; Errea, Pilar; Pina, Ana; Iturmendi Vizcay, Nerea; Irisarri, Patricia; Navarro Huidobro, Montserrat; Castel, Lourdes; Urrestarazu Vidart, Jorge; Santesteban García, Gonzaga; Miranda Jiménez, Carlos; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura; Institute for Multidisciplinary Research in Applied Biology - IMAB; Institute on Innovation and Sustainable Development in Food Chain - ISFOOD
Malus x domestica Borkh is a key temperate fruit crop globally, but climate change and market demands highlight the need to broaden its narrow genetic base. The collection and conservation of local germplasms support breeding efforts by enabling the development of resilient phenotypes with improved traits. Under this framework, a set of 130 apple accessions (recovered from Northeastern Spain and 14 commercial cultivars were analyzed in terms of 12 phenotypic traits (firmness, SSC, malic acid, enzymatic browning (EB) susceptibility, fructose and glucose content, and phenolic content in peel and pulp). A statistical framework was developed to assess trait stability under shifting climatic conditions and detect significant correlations between climatic variables and phenotypical performance of apple trees. The results revealed a wide phenotypical variation across the studied traits, with the distribution of physicochemical traits strongly influenced in a genotype-dependent manner. A correlation matrix was obtained for studied traits revealing positive correlations between harvest date and SSC, enzymatic browning and firmness. Furthermore, uni- and multifactorial ANOVA revealed that genetic group (GG) is a key factor influencing all measured traits, especially pH, SSC, EB, and browning speed. Several interaction effects were also statistically significant, especially those involving ploidy level, which strongly influenced malic acid content and EB responses. Additionally, significant differences in fructose content were observed depending on peel coloration, and year-to-year phenotypic variation appeared to be predominantly governed by genotypic response to environmental conditions. Moreover, obtention of a mean stability index (Di) for each trait allowed the identification of ‘Pinova’,’Fuji’ and ‘Gala’ as the most stable cultivars among 11 commercial cultivars. These findings will support future research on development of functional cultivars and selection of ideal genotypes under shifting conditions.
Open Access
Assessment of flesh browning diversity in apple germplasm collections phenotyped by image analysis
(ISHS, 2023) Miranda Jiménez, Carlos; Irisarri, Patricia; Arellano Zapatero, Julia; Bielsa González, Francisco Javier; Valencia Leoz, Ana; Urrestarazu Vidart, Jorge; Pina, Ana; Santesteban García, Gonzaga; Castel, L.; Errea, Pilar; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura; Institute for Multidisciplinary Research in Applied Biology - IMAB
Enzymatic flesh browning (EB) is one of the major problems affecting the quality and limiting the shelf life of minimally processed fruit. Traditionally, EB has been measured objectively using colourimeters. However, colourimeters are not suitable for phenotyping large quantities of fruit samples as they measure just one small area of a sample at a time, which hampers the acquisition of representative measurements and renders them time-consuming and costly. Previous research has shown that image analysis of digital photographs could be a viable alternative to obtain colour information of the entire surface of samples for large scale phenotyping, but to date there are no references for its practical application. The aim of this work was to assess the diversity in EB in a large set of cultivars phenotyped using digital photographs and a high-throughput analytical system based on image analysis developed by our team. A set of 143 cultivars from 104 genotypes, including modern references (16 cultivars) and traditional Spanish cultivars from UPNA (67 cultivars) and CITA (60 cultivars) germplasm collections was analysed in 2020 and 2021. The traditional cultivars were part of the core collection, which optimizes the representativeness of the genetic variation of apples preserved in Spanish collections. EB was evaluated in 10 fruits per cultivar and photographed at regular intervals from just after cutting to one hour later. A wide range of EB intensities was observed, with up to 20-fold differences between cultivars, which could be classified into five levels using two indices. The time at which EB was evaluated (30 or 60 min after slicing) had little influence on the classification. Traditional cultivars with low or very low EB were found to be comparable to those of references with less EB. The results show the potential of traditional germplasm to diversify the varietal offer and introduce new traits in apple breeding.
Open Access
Development and implementation of an affordable high-throughput imaging system for phenotyping enzymatic browning in apples
(Elsevier, 2025-11-18) Miranda Jiménez, Carlos; Irisarri, Patricia; Crespo Martínez, Sara; Bielsa González, Francisco Javier; Iturmendi Vizcay, Nerea; Romeo Los Arcos, Haizea; Urrestarazu Vidart, Jorge; Pina, Ana; Santesteban García, Gonzaga; Castel, Lourdes; Errea, Pilar; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura; Institute for Multidisciplinary Research in Applied Biology - IMAB; Institute on Innovation and Sustainable Development in Food Chain - ISFOOD; Universidad Pública de Navarra / Nafarroako Unibertistate Publikoa
Enzymatic browning (EB) substantially affects the visual quality and marketability of fresh-cut apples. This study aimed to develop an affordable high-throughput imaging system for phenotyping EB in apples. Browning was quantified using four CIELab-derived indices; a browning Index (BI), the difference in BI (ΔBI), a normalized CIE color difference (ΔE*); and a CIEDE2000 color difference (ΔE 00 ) at multiple time points post-cutting to evaluate browning speed (S EB ) and intensity (I EB ) in 142 apple cultivars, including commercial and traditional Spanish cultivars from germplasm collections. The image-based system has demonstrated high accuracy and practical relevance, overcoming limitations associated with traditional colorimeter-based approaches. A wide phenotypic range was observed, in which elite reference cultivars fell within a narrow band at the lower end of the range. Measurements taken at 30 min post-cutting were found to be nearly equivalent to those at 60 min, allowing to optimize the phenotyping protocol without compromising precision. EB has been shown to be an inherently stable trait, though different year effects were noted, particularly for BI and ΔBI. Among the indices evaluated, ΔE 00 proved less effective for cultivar differentiation, whereas ΔBI showed the highest discriminant capacity and strongest correlation with visual browning, making it the most suitable index for phenotyping purposes. These f indings provide a robust methodological basis for screening low-browning apple genotypes, establish a classif ication framework for EB expression levels, and highlight the potential of underutilized traditional cultivars in developing improved fresh-cut apple products.

Urrestarazu Vidart, Jorge

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