Biology subjects

Arrones, A.

Publications and source records attributed to Arrones, A..

12 recordsLinked to original sources

bioRxiv2026

CRISPR/Cas12a-mediated allele engineering of SmAPRR2 and SmGLK2 reveals complementary control of fruit peel and flesh chlorophyll pigmentation in eggplant

Eggplant (Solanum melongena L.) displays extensive fruit color diversity, in which chlorophyll-related pigmentation contributes to both external appearance and market value. Previous genetic studies identified SmAPRR2 and SmGLK2 as major candidate genes controlling uniform green pigmentation and green netting in fruit, respectively, but their individual and combined functional contributions had not been validated through targeted mutagenesis in a common genetic background. Here, we established a multiplex CRISPR/Cas12a system in eggplant accession MEL3, representing, to our knowledge, the first application of this nuclease for genome editing in eggplant. Transformation efficiency was 2.0%, but all 15 genotyped regenerants were edited, yielding four SmAPRR2 and six SmGLK2 alleles. Segregation and crossing enabled the recovery of six transgene-free lines carrying single or combined edited alleles. Disruption of SmGLK2 abolished the reticulated green netting pattern while preserving a uniformly green peel and the internal green ring. Conversely, disruption of SmAPRR2 reduced the background uniform peel pigmentation and eliminated the green ring while retaining green netting. Double mutants carrying disruptive alleles at both loci produced white fruits lacking internal green pigmentation, whereas putatively hypomorphic SmAPRR2 and SmGLK2 alleles generated intermediate phenotypes. Whole-genome resequencing identified only two predicted off-target sites under a canonical TTTV PAM search allowing up to four mismatches. Both were fully covered, and no edited-line-specific candidate variants were detected. These findings establish complementary and partially separable roles for SmAPRR2 and SmGLK2 in fruit peel and flesh chlorophyll pigmentation and demonstrate the potential of Cas12a for functional genomics, allele engineering, and precision breeding in eggplant.

plant biology↗

bioRxiv2026

Multi-trait evaluation of a tomato MAGIC population identifies promising lines with improved nitrogen use efficiency (NUE)

Nitrogen-use efficiency (NUE) is a pivotal breeding target in tomato (Solanum lycopersicum L.) to sustain production under reduced N inputs. Here, we leveraged a recently developed tomato multi-parent advanced generation inter-cross (ToMAGIC) population to identify lines with superior performance under reduced N availability. The eight founders and a core subset of 118 ToMAGIC lines were characterized with 10,684 SNP markers and evaluated under optimal (opN, 15 mM) and suboptimal (subN, 8 mM) N supply in an experiment totalling 1,576 plants, generating 48,068 data points across 61 phenotypic variables. Under both N treatments, ToMAGIC lines exhibited transgressive segregation for most traits, confirming the value of this population as a reservoir of untapped variation. Notably, under subN conditions, harvest index (Hi) increased by 29-44%, suggesting adaptive resource redistribution toward reproductive sinks. Variance partitioning revealed that agronomic and NUE-related traits were largely under genetic control, with heritability estimates frequently above 0.80 and broadly conserved across N treatments. Multivariate trait analysis identified fruit yield N concentration (NUE component, CN,y), shoot biomass N content (NAb), and shoot growth-related traits as the main drivers of treatment differentiation. Finally, proxy traits were prioritized by integrating response magnitude, heritability, trait correlations, and treatment-discriminatory power into multi-trait selection indices. This strategy generated favorable predicted genetic gains, reaching 158% for high-performance lines and 170% for subN-adapted lines, and consistently identified lines 402, 428, 518, 800, and 816 as promising pre-breeding materials. Overall, this study supports ToMAGIC as a powerful resource for developing N-efficient cultivars suited for sustainable agriculture.

plant biology↗

bioRxiv2026

A neofunctionalized flowering antagonist created an evolutionary contingency that channeled Solanaceae adaptation

Neofunctionalization is a rare fate of gene duplication, classically defined as the acquisition of novel functions that potentiate the emergence of new traits. Rather than evolving to function autonomously, neofunctionalized genes may also remain embedded within their ancestral regulatory networks, potentially reshaping the genetic trajectories through which phenotypic change occurs. Testing this hypothesis, we leveraged a pan-genetic platform comprising ten Solanaceae species and show that a paralog of the flowering hormone florigen neofunctionalized into a flowering antagonist and was repeatedly selected during crop domestication and adaptation of wild plants across 50 million years of evolution. Independent selection of cis-regulatory and coding mutations in SELF-PRUNING 5G (SP5G) enabled rapid flowering in the wild ancestor of domesticated tomato from Central America as well as major and indigenous eggplant crop lineages domesticated in Asia and Africa. We further found that cis-regulatory sequence changes reduced SP5G expression and flowering time in wild species native to distinct environments in the Americas and Australia, relationships that we validated by genome editing. Together with similar patterns observed across diverse species and developmental networks, we propose that antagonistic neofunctionalized paralogs create evolutionary contingencies that channel adaptive trajectories across plant lineages.

plant biology↗

bioRxiv2025

Characterization of a dominant SmNac-like gene as a candidate for photosensitivity in the fruit peel of eggplant

Anthocyanins in the fruit peel of photosensitive eggplants exhibit a different distribution pattern compared to the photo-insensitive ones. The latter exhibits a uniform anthocyanin content, whereas photosensitive eggplants lack anthocyanin accumulation in areas not exposed to light, such as under the calyx, or have lower concentrations in less-exposed areas. In the current research work, genetic analysis of F1 and F2 populations revealed that the photo-insensitive phenotype in eggplants follows an autosomal dominant inheritance with a 3:1 ratio, indicating that the photosensitive trait is regulated by a single dominant gene. To locate and narrow down the genomic region underlying photosensitivity, a segregating F2 population was used for bulked segregant analysis sequencing (BSA-seq) and compared with previous QTLs identified in previous developed eggplants populations (ILs and MAGIC population). The accumulation of QTLs at the end of chromosome 10 postulate that chromose region as a hot spot for anthocyanin related traits. In our population all the QTLs considered overlap between the genomic region 84,1-87,9 Mb. Moreover, no DNA mutations in the progenitors of the eggplant accessions used were found. A RNA-seq analysis of bagged photosensitive and photo- insensitive eggplants was performed, as a result we identified the SmNAC1-like protein gene as a promising gene to be involved in fruit photosensitivity trait. In the photo-insensitive accession (IVIA- 371) SmNAC1-like protein was depply repressed compare to the photosensitive accession (ASI-S-1). No consistent mutations in the coding sequences (CDS) of SmNac-like protein locus among all the different eggplants accessions used were found, suggesting that other layer of regulation maybe acting in our eggplant accessions. These findings provide new insight into the regulation of the molecular mechanisms of anthocyanin biosynthesis in eggplant as point out for the first time the possible role of NAC transcription factors in the anthocyanin biosynthesis in eggplant.

plant biology↗

bioRxiv2025

Water Stress Tolerance, Genomic Selection and Identification of Genomic Regions in a MAGIC Population of Eggplant

Horticultural crops are increasingly affected by water stress due to climate change, making the development of stress-tolerant varieties an urgent need. In this study, we evaluated a set of 184 Multi-parent Advanced Generation Inter-Cross (MAGIC) eggplant lines under water stress conditions, consisting of irrigation at 30% of field capacity. After 21 days of stress, we assessed traits related to growth, water content, plant pigments, and proline content. The MAGIC population displayed a high variability for water stress tolerance, as transgressive lines over parental values were found for most of the traits. Key traits associated with water stress tolerance were identified, including increased root growth and elevated proline, and flavonoid content. Genome-Wide Association Study (GWAS) analysis allowed identification of three genomic regions associated with total dry weight, water content and flavonoids, traits that contribute significantly to stress tolerance. Additionally, a linear genomic selection index was constructed based on total dry weight, dry weight increase during the stress period, root dry weight, water content, and proline content to identify water stress-tolerant and susceptible lines. The models predictive ability was calculated, and the value of the selection index was predicted for 141 unevaluated MAGIC lines, with prediction accuracy for the index traits ranging from 0.11 to 0.53. This study presents a comprehensive analysis, identifying critical traits, genomic regions and lines with genetic potential, providing valuable information for future breeding programmes to improve water stress tolerance in eggplant.

genetics↗

bioRxiv2025

Resequencing and phenotyping of the first highly inbred eggplant multiparent population reveal SmLBD13 as a key gene associated with root morphology

The MEGGIC (Magic EGGplant InCanum) population here presented is the first highly inbred eggplant (Solanum melongena) multiparent advanced generation intercross (MAGIC) population developed so far, derived from seven cultivated accessions and one wild S. incanum from arid regions. The final 325 S5 lines were high-throughput genotyped using low-coverage whole-genome sequencing (lcWGS) at 3X, yielding 293,783 high-quality SNPs after stringent filtering. Principal component analysis (PCA) and neighbour-joining clustering revealed extensive genetic diversity, lack of genetic structure, and the distinct genetic profile of the wild founder. The eight founders and a core subset of 212 lines were phenotyped for above- and below-ground traits, revealing wide phenotypic diversity. Root morphology traits displayed moderate heritability values, and strong correlation were found between root and aerial traits, suggesting that a well-developed root system supports greater above-ground growth. Genome-wide association studies (GWAS) identified a genomic region on chromosome 6 associated with root biomass (RB), total root length (RL), and root surface area (SA). Within this region, SmLBD13, a LOB-domain protein involved in lateral root development, was identified as a candidate gene. The S. incanum haplotype in this region was linked to reduced lateral root branching density, a trait that may enhance deeper soil exploration and resource uptake. These findings provide key insights into root genetics in eggplant, demonstrating MEGGIC potential for high-resolution trait mapping. Furthermore, they highlight the role of exotic wild germplasm in breeding more resilient cultivars and rootstocks with improved root architecture and enhanced nutrient uptake efficiency.

genomics↗

bioRxiv2024

Benchmarking of Low Coverage Sequencing Workflows for Precision Genotyping in Eggplant

Low-coverage whole-genome sequencing (lcWGS) presents a cost-effective solution for genotyping, particularly in applications requiring high marker density and reduced costs. In this study, we evaluated lcWGS for eggplant genotyping using eight founder accessions from the first eggplant MAGIC population (MEGGIC), testing various sequencing coverages and minimum depth of coverage (DP) thresholds with two SNP callers, Freebayes and GATK. Reference SNP panels were used to estimate the percentage of common biallelic SNPs (i.e, true positives, TP) relative to the low coverage datasets (accuracy) and the SNP panels themselves (sensitivity), along with the percentage of TP with the same genotype across the two datasets (genotypic concordance). Sequencing coverages as low as 1X and 2X achieved high accuracy but lacked sufficient sensitivity and genotypic concordance. However, 3X sequencing reached approximately 10% less sensitivity than 5X while maintaining genotypic concordance above 90% at any DP threshold. Freebayes outperformed GATK in terms of sensitivity and genotypic concordance. Therefore, we used this software to conduct a pilot test with some MEGGIC lines from the fifth generation of selfing (S5), comparing their datasets with a gold standard (GS). Sequencing coverages as low as 1X identified a substantial number of TP, with 3X significantly increasing the yield, particularly at moderate DP thresholds. Additionally, at least 30% of the TP were consistently genotyped in all lines when using coverages greater than 2X, regardless of the DP threshold applied. This study highlights the importance of using a GS to reduce false positives and demonstrates that lcWGS, with proper filtering, is a valuable alternative to high-coverage sequencing for eggplant genotyping, with potential applications to other crops.

genomics↗

bioRxiv2024

A novel tomato inter-specific (Solanum lycopersicum var. cerasiforme and S. pimpinellifolium) MAGIC population facilitates trait association and candidate gene discovery in untapped exotic germplasm

We developed a novel eight-way tomato multi-parental advanced generation inter-cross (MAGIC) population to improve the accessibility of the genetic resources of tomato relatives to geneticists and breeders. The inter-specific MAGIC population (ToMAGIC) was obtained by inter-crossing four accessions each of Solanum lycopersicum var. cerasiforme (SLC) and S. pimpinellifolium (SP), which respectively are the weedy relative and the ancestor of cultivated tomato. The eight exotic ToMAGIC founders were selected based on a representation of the genetic diversity and geographical distribution of the two taxa. The resulting MAGIC population comprises 354 lines which were genotyped using a new 12k tomato Single Primer Enrichment Technology (SPET) panel and yielded 6,488 high-quality SNPs. The genotyping data revealed a high degree of homozygosity (average 93.69%), an absence of genetic structure, and a balanced representation (11.62% to 14.16%) of the founder genomes. To evaluate the potential of the ToMAGIC population for tomato genetics and breeding, a proof-of-concept was conducted by phenotyping it for fruit size, plant pigmentation, leaf morphology, and earliness traits. Genome-wide association studies (GWAS) identified strong associations for the studied traits, pinpointing both previously identified and novel candidate genes near or within the linkage disequilibrium blocks. Domesticated alleles for fruit size were recessive and were found, at low frequencies, in wild/ancestral populations. Our findings demonstrate that the newly developed ToMAGIC population is a valuable resource for genetic research in tomato, offering significant potential for identifying new genes that govern key traits in tomato breeding. ToMAGIC lines displaying a pyramiding of traits of interest could have direct applicability for integration into breeding pipelines providing untapped variation for tomato breeding.

plant biology↗

bioRxiv2024

Convergent evolution of plant prickles is drivenby repeated gene co-option over deep time

An enduring question in evolutionary biology concerns the degree to which episodes of convergent trait evolution depend on the same genetic programs, particularly over long timescales. Here we genetically dissected repeated origins and losses of prickles, sharp epidermal projections, that convergently evolved in numerous plant lineages. Mutations in a cytokinin hormone biosynthetic gene caused at least 16 independent losses of prickles in eggplants and wild relatives in the genus Solanum. Strikingly, homologs promote prickle formation across angiosperms that collectively diverged over 150 million years ago. By developing new Solanum genetic systems, we leveraged this discovery to eliminate prickles in a wild species and an indigenously foraged berry. Our findings implicate a shared hormone-activation genetic program underlying evolutionarily widespread and recurrent instances of plant morphological innovation.

evolutionary biology↗

bioRxiv2023

The irregular fruit green netting: An eggplant domestication trait controlled by the SmGLK2 gene with implications in fruit colour diversification

The distribution of chlorophylls in the eggplant (Solanum melongena) fruit peel can be uniform or display an irregular green netting pattern. The fruit green netting phenotype, manifested as a gradient of dark green netting, more intense in the proximal part of the fruit on a pale green background, is commonly present in eggplant wild relatives as well as in some eggplant landraces. During domestication and modern breeding of eggplant, uniform fruit colour has been selected. However, the fruit green netting contributes to a greater diversity of fruit colours. Here, we have used over 2,300 individuals from several germplasm and experimental populations, including a multi-parental MAGIC population for candidate genomic region identification, an F2 population for BSA-Seq, and advanced backcrosses for edges-to-core fine mapping, to determine that SmGLK2 is the gene underlying the irregular netting in eggplant fruits. We have also analysed the gene sequence of 178 S. melongena accessions and 22 wild relative species for tracing the evolutionary changes that the gene has undergone over the course of domestication. Three different mutations were identified leading to the absence of netting. The main causative indel results in the appearance of a premature stop codon disrupting the protein conformation and function, which was confirmed by Western blotting analysis and confocal microscopy observations. SmGLK2 has a major role in the regulation of chlorophyll biosynthesis in eggplant fruit peel, and therefore in eggplant fruit photosynthesis.

genetics↗

bioRxiv2022

Mutations in the SmAPRR2 transcription factor suppressing chlorophyll pigmentation in the eggplant fruit peel are key drivers of a diversified colour palette

Understanding the mechanisms by which chlorophylls are synthesized in the eggplant (Solanum melongena) fruit peel is of great relevance for eggplant breeding. A multi-parent advanced generation inter-cross (MAGIC) population and a germplasm collection have been screened for green pigmentation in the fruit peel and used to identify candidate genes for this trait. A genome-wide association study (GWAS) performed with 420 MAGIC individuals revealed a major association on chromosome 8 close to a gene similar to APRR2. Two variants in SmAPRR2, predicted as having a high impact effect, were associated with the absence of fruit chlorophyll pigmentation in the MAGIC population, and a large deletion of 5.27 kb was found in two reference genomes of accessions without chlorophyll in the fruit peel. The validation of the candidate gene SmAPRR2 was performed by its sequencing in a set of MAGIC individuals and through its de novo assembly in 277 accessions from the G2P-SOL eggplant core collection. Two additional mutations in SmAPRR2 associated with the lack of chlorophyll were identified in the core collection set. The phylogenetic analysis of APRR2 reveals orthology within Solanaceae and suggests that specialization of APRR2-like genes occurred independently in Cucurbitaceae and Solanaceae. A strong geographical differentiation was observed in the frequency of predominant mutations in SmAPRR2, resulting in a lack of fruit chlorophyll pigmentation and suggesting that this phenotype may have arisen and been selected independently several times. This study represents the first identification of a major gene for fruit chlorophyll pigmentation in the eggplant fruit.

genetics↗

bioRxiv2021

Newly developed MAGIC population allows identification of strong associations and candidate genes for anthocyanin pigmentation in eggplant

MAGIC populations facilitate the genetic dissection of complex quantitative traits in plants and are valuable breeding materials. We report the development of the first eggplant MAGIC population (S3MEGGIC; 8-way), constituted by 420 S3 individuals developed from the intercrossing of seven cultivated eggplant (Solanum melongena) and one wild relative (S. incanum) parents. The S3MEGGIC recombinant population was genotyped with the eggplant 5k probes SPET platform and phenotyped for anthocyanins presence in vegetative plant tissues (PA) and fruit epidermis (FA), and for the light-sensitive anthocyanic pigmentation under the calyx (PUC). The 7,724 filtered high-confidence SNPs confirmed a low residual heterozygosity (6.87%) and a lack of genetic structure in the S3MEGGIC population, including no differentiation among subpopulations carrying cultivated or wild cytoplasm. Inference of haplotype blocks of the nuclear genome revealed an unbalanced representation of founder genomes, suggesting cryptic selection in favour or against specific parental genomes. GWAS analysis for PA, FA and PUC detected strong associations with two MYB genes similar to MYB113 involved in the anthocyanin biosynthesis pathway and with a COP1 gene, which encodes for a photo-regulatory protein and may be responsible for the PUC phenotype. Evidence was found of a duplication of an ancestral MYB113 gene with a translocation from chromosome 10 to chromosome 1. Parental genotypes for the three genes were in agreement with the candidate genes identification performed in the S3MEGGIC population. Our new eggplant MAGIC population is the largest recombinant population in eggplant and is a powerful tool for eggplant genetics and breeding studies.

genetics↗