Open Access
Comparative enzymatic browning transcriptome analysis of three apple cultivars unravels a conserved regulatory network related to stress responses
(BMC, 2025-04-12) Bielsa González, Francisco Javier; Grimplet, Jérôme; Irisarri, Patricia; Miranda Jiménez, Carlos; Errea, Pilar; Pina, Ana; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura; Institute for Multidisciplinary Research in Applied Biology - IMAB
Enzymatic browning (EB) endangers the adaptation of apple fruit cultivars to new markets, affecting organoleptic properties and producing economic losses. Polyphenol oxidases and polyphenol compounds play a key role in EB development in apple. However, the regulation of apple response to EB remains to be uncovered. In this study, three apple cultivars with different EB phenotypes ranging from low to high browning in apple pulp were used to study transcriptomic changes over time after fresh cutting (0, 30 and 60 min). This study allowed the identification of 1448 differentially expressed genes (DEGs), revealing both shared and genotype-specific responses, particularly in the affected metabolic pathways associated with EB. At 60 min (T60 vsT0), 77 DEGs were shared by all genotypes, suggesting a conserved regulatory network. This network included genes encoding for protein families such as calcium-binding proteins, heat-shock proteins, redox-responsive transcription factors, WRKY family transcription factors, zinc finger family proteins and disease resistance proteins among others. A co-expressed gene cluster, identified through Weighed Gene Co-Expression Network Analysis (WGCNA), was found to correlate with EB and included 323 genes enriched in several biological terms according to Gene Ontology analysis. Moreover, a more detailed analysis of identified WGCNA gene cluster regulatory sequences allowed the detection of cis-regulatory elements belonging to CAMTA, WRKY and WUSCHEL transcription factor families. The identification of these sequences alongside with an abundant and diverse amount of overexpressed transcription factors from various families (WRKY, ERF, GRAS, GATA, etc.) point out to a highly regulated stress-response that is strictly connected to innate plant immunity. These findings provide valuable insights into the molecular mechanism involved in apple fresh-cut browning and offer new potential targets for EB regulation.
Open Access
Genetic diversity of Spanish Prunus domestica L. germplasm reveals a complex genetic structure underlying
(Public Library of Science, 2018) Urrestarazu Vidart, Jorge; Errea, Pilar; Miranda Jiménez, Carlos; Santesteban García, Gonzaga; Pina, Ana; Producción Agraria; Nekazaritza Ekoizpena
European plum (Prunus domestica L.) is an ancient domesticated species cultivated in temperate areas worldwide whose genetic structure has been scarcely analyzed to date. In this study, a broad representation of Spanish European plum germplasm collected in Northeastern Spain and a representative set of reference cultivars were compared using nuclear and chloroplast markers. The number of alleles per locus detected with the SSR markers ranged from 8 to 39, with an average of 23.4 alleles, and 8 haplotypes were identified. Bayesian model-based clustering, minimum spanning networks, and the analysis of molecular variance showed the existence of a hierarchical structure. At the first level, two genetic groups were found, one containing 'Reine Claude' type reference cultivars altogether with ca. 25% of local genotypes, and a second one much more diverse. This latter group split in two groups, one containing most (ca. 70%) local genotypes and some old Spanish and French reference cultivars, whereas the other included 24 reference cultivars and only six local genotypes. A third partition level allowed a significant finer delineation into five groups. As a whole, the genetic structure of European plum from Northeastern Spain was shown to be complex and conditioned by a geographical proximity factor. This study not only contributes to genetic conservation and breeding for this species at the national level, but also supports the relevance of undertaking similar tasks of collection and characterization in other unexplored areas. Moreover, this kind of research could lead to future coordinated actions for the examination of the whole European plum diversity, to define conservation strategies, and could be used to better understand the genetic control of traits of horticultural interest through association mapping.
Open Access
Unlocking Spanish pear genetic diversity: strategies for construction of a national core collection
(Springer Nature, 2024-11-04) Irisarri, Patricia; Urrestarazu Vidart, Jorge; Ramos-Cabrer, Ana; Pereira-Lorenzo, Santiago; Velázquez-Barrera, María Encarnación; Díaz-Hernández, María Belén; Dapena, Enrique; Urbina, Valero; Dalmases, Josep; Ríos-Mesa, Domingo; Crespo Martínez, Sara; Loidi Erviti, Maite; Santesteban García, Gonzaga; Ascasíbar-Errasti, Javier; Errea, Pilar; Miranda Jiménez, Carlos; Pina, Ana; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura; Institute for Multidisciplinary Research in Applied Biology - IMAB
Spanish pear germplasm collections are crucial for preservation, research, and breeding efforts. However, genetic diversity and structure is unknown at national level. A coordinated national project analyzed 1251 accessions from 7 Spanish pear collections using an internationally recognized set of 14 SSRs to enhance the utilization of these collections. Key findings included the identification of 760 unique genotypes (490 diploids and 270 triploids). Notably, genotypes represented by a single accession accounted for 49% of the total, indicating high vulnerability of this material. Using a Bayesian clustering method revealed two main genetic groups, G1 containing most foreign cultivars and G2 retaining local Spanish cultivars, which were further divided into two other subgroups using a nested approach, revealing moderate but significant differentiation among them. The populations were renamed according to the origin of the reference samples assigned to each group as 'South' (G1.1), 'Western Europe-1' (G1.2), 'Western Europe-2' (G2.1) and 'No-Pyrus communis' (G2.2). The results led to the creation of a 'generalist' collection, aiming to maximize genetic diversity representativeness, starting with 68 genotypes but expanding to 111 to achieve better allele recovery. This core collection is a valuable resource for genetic studies and conservation, enhancing efforts to preserve pear biodiversity.
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.

Miranda Jiménez, Carlos

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