Horticulture Research

Climate change poses an increasing threat to the cultivation of deciduous fruit trees, placing greater demands on modern pear breeding. Using pear germplasm adapted to diverse environments, we assembled 11 chromosome-level genomes. In combination with 13 publicly accessible pear genomes, we analyzed presence-absence variations (PAVs) and constructed a graph-based pangenome for pear. By performing a PAV-eQTL analysis of the fruit of 123 pear accessions, we identified PAVs significantly associated with expression levels of genes that may be involved in regulating agronomic traits. Population analysis of 268 pear accessions revealed two stop-gained variants in DAM1 of independent origin, which may function in advancing the blooming date and reducing the chilling requirement. We detected complex PAVs at the NOR1 locus, including two copy-number variations and one deletion. These PAVs contributed to the rapid diversification of the NOR1 locus and the fruit development period through regulating ARF5 and other ripening-related genes. We revealed the selection history of the NOR1 locus and developed novel pear individuals that accumulated alleles for low chilling requirement, early blooming date, and short fruit development period. The results provide valuable resources for pear genomics research and offer a guideline for breeding modern pears with climate resilience.

Tomato wild relatives are valuable genetic resources for trait discovery and understanding the genetic basis of fruit metabolism and quality. Yet, only a fraction of naturally occurring variation has been exploited. Here, we performed metabolite profiling of two large Backcross Inbred Line populations derived from crosses between the wild species S. pennellii accession LA5240 (Lost) and cultivated genotypes LEA (determinate) and TOP (indeterminate), including [~]1400 and [~]500 lines, respectively. High-resolution mapping identified enormous metabolic quantitative trait loci (mQTL), including a new locus on chromosome 12 associated with fruit sucrose accumulation that harbours INVERTASE INHIBITOR 3 (SlINVINH3) protein. Comparative analysis indicated that SlINVINH3 is highly expressed in wild S. pennellii 0716 fruit, whereas a six-amino acid deletion is present in its coding sequence compared with S.pennellii LA5240 and S. lycopersicum. We further demonstrated that in SlINVINH3-overexpressing tomato plants, only the S. pennellii LA5240 allele led to increased sucrose, accompanied by reduced fructose and glucose levels. Furthermore, the large population size enabled us to assess the epistatic interactions, with approximately 40% of interactions being more-than-additive and 60% less-than-additive. Our results demonstrate the power of permanent exotic populations to reveal hidden metabolic diversity and provide an approach for improving fruit quality through targeted breeding and metabolic engineering.

Starch is the primary carbohydrate reserve in potato (Solanum tuberosum L.) tubers and a critical determinant of their industrial value. The rate of starch degradation during postharvest storage directly influences key traits such as endodormancy (ED) duration and cold-induced sweetening (CIS), which affect sprouting behavior. In this study, we used CRISPR/Cas9 genome editing to knockout StAMY23, a gene encoding -amylase involved in starch breakdown. stamy23 plants exhibited higher yield and extended tuber ED postharvest, without significantly altering CIS or starch granule content. To further reduce CIS, we knockout StAMY23 in VACUOLAR INVERTASE knockout (stvinv) backgrounds, generating stamy23/stvinv double-knockouts plants. These lines showed significantly reduced CIS, prolonged ED, and elevated starch content, along with altered starch granule content. Collectively, our findings demonstrate that simultaneous downregulation of StAMY23 and StVINV can additively enhance desirable postharvest traits, providing a promising strategy for improving potato storage quality through precision genome editing.

Apples (Malus x domestica) are popular fruits grown in temperate regions of the world. The various genotypes must meet a specific threshold amount of cold exposure before they are competent to break dormancy, a quantity approximated as "chill hours". Several varieties have been identified that exhibit an ultra-low-chill requirement, or more precisely shallow dormancy, breaking vegetative and floral buds early in spring in response to minimal cold exposure. These ultra-low-chill genotypes originated from the Bahamas ( Dorsett Golden,1960s), Israel ( Anna, 1950s) and Alabama, USA ( Shell of Alabama, 1880s). The separation in time and space implies that each would feature distinct genetic lesions that govern dormancy control, providing discrete mechanisms to incorporate a low-chill trait in variety improvement. However, analysis of microsatellites and ultimately genome sequence indicates that Dorsett Golden and Anna share strong concordance with the Shell of Alabama genotype, as well as other ultra-low-chill varieties. Kinship analysis confirms that all are closely related, despite differences in year and place of origin. All three low-chill genotypes share common mutations in the DORMANCY ASSOCIATED MADS-BOX1(DAM1) gene, a known repressor of vegetative growth during dormancy. Genomic sequence diversity is observed among Shell of Alabama individuals, including differences in DAM1 that match differences in flowering time. The results of this study call into question the pedigrees of the ultra-low-chill apple germplasm and indicate variation in an otherwise narrow genetic base for use in future breeding efforts.

This study integrated morphometric characterization and machine-learning modelling to identify key predictors of yield in Annona reticulata under semi-arid conditions. Thirty-one canopy, fruit, seed, and biochemical traits were evaluated across 62 genotypes, revealing substantial phenotypic diversity, particularly in structural attributes such as tree growth nature and branch angle. Principal Component Analysis and hierarchical clustering differentiated genotypes into three ideotypes representing high-yielding, structurally stable, and quality-oriented groups. Random Forest modelling and SHapley Additive exPlanations (SHAP) interpretation consistently highlighted leaf breadth, leaf length, fruit shape, and pulp-associated traits as dominant yield predictors, underscoring the coordinated influence of source-sink balance. Integration of SHAP importances with trait stability (CV%) further revealed that moderately variable traits provide reliable selection indices. These findings demonstrate that yield performance is governed by multivariate trait networks rather than isolated descriptors. The proposed framework provides a robust basis for precision phenotyping and strategic parent selection to develop high-yielding, nutritionally enriched, and climate-resilient custard apple cultivars.

Asparagus racemosus Wild., commonly known as Shatavari, is an important medicinal plant in Ayurveda and is valued for its steroidal saponins, particularly shatavarin compounds, which contribute to its adaptogenic, galactagogue, immunomodulatory, and therapeutic properties. Despite its medicinal and economic importance, genomic resources for this species have remained limited, restricting molecular breeding, pathway discovery, and comparative evolutionary studies within Asparagaceae. Here, we report a high quality chromosome scale reference genome assembly of A. racemosus var. CIM Shakti generated using PacBio HiFi long read sequencing and Omni C chromatin conformation scaffolding. The pseudo haploid assembly spans 817 Mb across 53 scaffolds, with a scaffold N50 of 98.50 Mb, L50 of 5, and a largest scaffold of 113.80 Mb. Ten major chromosome scale pseudomolecules were resolved, corresponding to the haploid chromosome complement of A. racemosus. The assembly showed high gene space completeness, with BUSCO completeness of 99.8% against the Eukaryota dataset and 98.0% against the Embryophyta dataset. BlobToolKit profiling further supported assembly quality, with GC content of approximately 39 to 40% and no major evidence of contamination. EDTA based repeat annotation identified 580.93 Mb of interspersed repetitive elements, accounting for 71.06% of the 817.57 Mb genome assembly. The repeat landscape was dominated by LTR retrotransposons, particularly Gypsy elements, which accounted for 25.01% of the assembly, followed by unclassified LTR elements at 26.58% and Copia elements at 4.84%. Structural and functional annotation identified 29,199 protein coding genes represented by 29,199 transcript models, 138,433 exons, and 125,201 CDS features. The annotation was structurally robust, with an average gene length of 4,605.1 bp, 4.74 exons per transcript, and 97.80% of transcripts containing multiple exons. The CIM Shakti reference genome provides a foundational genomic resource for investigating steroidal saponin biosynthesis, sex chromosome evolution, repeat driven genome expansion, and comparative genomics in Asparagaceae. This assembly will support future studies on medicinal trait improvement, conservation genomics, and genomics assisted breeding of climate resilient Shatavari cultivars.

The genetic basis of wine monoterpene variation and its sensory consequences remain poorly understood. We investigated how allelic variation at a chromosome 10 linalool QTL harboring a terpene synthase cluster, including VviTPS54, influences wine aroma in a Riesling x Cabernet Sauvignon mapping population. Twenty-six wines from progeny selected for contrasting genotypes were subjected to HS-SPME-GC-MS profiling and descriptive sensory analysis. Genotypes carrying the high-linalool Riesling allele produced substantially elevated monoterpene levels dominated by (3S)-linalool. Berry and wine monoterpene profiles were significantly correlated, and berry monoterpene glycosides showed broad correlations with wine monoterpenes. High-monoterpene genotypes were more closely associated with Floral, Tropical Fruit, and Apricot/Peach attributes, though perception was modulated by matrix effects from berry pigmentation. These results demonstrate that allelic variation at a terpene synthase locus drives coordinated changes in wine monoterpene composition and linked sensory attributes, while underscoring the complexity of translating genetic variation into sensory outcomes.

HighlightThe sweet fragrance of lettuce was attributed, for the first time, to the synthesis of 2-acetyl-1-pyrroline caused by a deficiency in the betaine aldehyde dehydrogenase gene. Fragrance is among the most valuable traits of high-quality crops and influences consumer preferences. Although 2-acetyl-1-pyrroline (2AP) is a key component of fragrant cultivars in several crops, its genetic mechanism in lettuce (Lactuca sativa L.) remains poorly understood. The betaine aldehyde dehydrogenase (BADH) gene has been identified as causative for 2AP-derived fragrance in rice and soybean cultivars. Hence, we conducted a linkage analysis using an F2 population derived from a cross between Kukichisya (fragrant) and Rennet (non-fragrant) for three candidate genes of BADH orthologs in the lettuce genome. Analysis linked LOC111877932 located in LG4 to the fragrance trait, and it was designated LsBADH1. Comparison among Kukichisya, Salinas, and candidate BADH of sunflower (Helianthus annuus L.) revealed three non-synonymous single-nucleotide polymorphisms (nsSNPs) in exons 1, 2, and 9, and suggested that nsSNP in exon 9 was strongly correlated with fragrance in Kukichisya. A premature stop codon introduced in exon 5 of LsBADH1 using Target-AID base-editing technology resulted in truncated BADH1 and higher 2AP levels. Our results indicated that LsBADH1 is responsible for the 2AP-derived fragrance. Our findings can be applied to select cultivars based on a novel concept for the cooking process, providing a transformative platform to breed fragrant lettuce as a high-value-added product.

Grapevine cultivars vary widely in monoterpenoid content, yet the genetic and regulatory mechanisms underlying this variation remain poorly characterized beyond highly aromatic Muscat types. We profiled free volatiles and monoterpenoid glycosides in a Riesling x Cabernet Sauvignon F1 mapping population, revealing extensive variation and transgressive segregation consistent with multigenic control. QTL mapping identified 59 significant loci associated with 47 volatile compounds and monoterpene glycosides, including two major QTLs explaining 33.8% and 32.4% of phenotypic variance in (3S)-linalool accumulation. Integration of haplotype-resolved transcriptomics with metabolite data, enabled by a chromosome-scale diploid Riesling genome assembly, resolved a (3S)-linalool/nerolidol synthase cluster on chromosome 10 and identified VviTPS54 as the strongest candidate underlying linalool variation. VviTPS54 exhibited haplotype-specific expression strongly correlated with (3S)-linalool accumulation across genotypes, while no QTL was detected at the VviDXS1 locus and VviDXS1 expression was not correlated with terpene levels, indicating that regulatory variation within terpene synthase clusters, rather than methylerythritol phosphate (MEP) pathway flux, drives monoterpenoid composition in this population. These results establish regulatory variation of terpene synthases as a key mechanism underlying monoterpenoid diversity in grapevine and demonstrate that resolving such variation requires haplotype-phased genome assemblies coupled with haplotype-resolved transcriptomics to detect allele-specific expression differences at complex, heterozygous loci.

Buckwheat (Fagopyrum spp.) is a climate-resilient pseudocereal, yet its global adoption is constrained by the distylous self-incompatibility of common buckwheat (F. esculentum) and grain bitterness of Tartary buckwheat (F. tataricum). While the S-locus early flowering 3 (FeS-ELF3) gene has been identified as a key regulator of self-compatibility, a stable genetic transformation of F. esculentum has not yet been developed. In this study, we developed an Agrobacterium-mediated transformation protocol of F. esculentum (27% transformation efficiency) and applied it to the agronomically relevant Panda cultivar. Inactivation of the FeS-ELF3 gene using the CRISPR/Cas9 system yielded self-compatible lines with long-homostylous flowers. fes-elf3 mutants showed a distinct architectural shift: mutant plants were shorter and had shorter inflorescences than wild-type plants. Notably, these traits did not compromise yield, as the mutants produced a similar number of seeds per plant in the greenhouse. In F. tataricum, we targeted the rutin-degrading enzyme (FtRDE2), which was suspected to be a driver of grain bitterness by hydrolysing rutin into the bitter quercetin. Metabolic profiling of seeds of two ftrde2 mutant lines revealed significantly lower quercetin levels in both. Analysis of enzyme extracts confirmed the loss of rutinosidase activity; the mutant samples maintained stable rutin concentrations without the characteristic increase in quercetin observed in the control. Furthermore, organoleptic sensory evaluation of flour demonstrated that respondents identified the control as significantly more bitter than the flour from the ftrde2 mutants. These precise edits show proof-of-concept of overcoming domestication barriers: self-incompatibility and palatability, establishing a framework for rapid improvement of buckwheat.

Targeted epigenetic modifications of specific gene regulatory regions have the potential to confer beneficial traits for crop improvement. Two recently developed CRISPR/Cas9-based epigenome editing tools were tested in transgenic cassava to target cytosine methylation to the promoter region of MeSWEET10a, a necessary gene for infection by the Cassava bacterial blight pathogen, Xanthomonas phaseoli pv. manihotis. The two systems leverage unique methyltransferases, and each induced distinct DNA methylation profiles at the targeted site, decreased effector-triggered MeSWEET10a expression, and attenuated water-soaking symptoms in inoculated leaves. Further, DNA methylation was simultaneously targeted, in addition to MeSWEET10a, to two susceptibility genes for Cassava brown streak virus. Relative levels of de novo DNA methylation at the three loci were inversely correlated with DNA methylation-antagonizing H3K4me3 marks. Finally, an initial assessment of DNA methylation after one generation indicated specific inheritance of CpG methylation that was unstable in the absence of the methyltransferase systems.

Plants produce specialized metabolites that function in plant defense and as attractants to pollinators and symbionts. One class of specialized metabolites are terpenoids that are synthesized from universal C5 building blocks via activities including terpene synthases, cytochromes P450, and glycosyl transferases. Some terpenes are highly valued for their use as insect repellants, fragrances, antimicrobial compounds, low calorie sweeteners, flavors, and medicines. Low abundance in target tissues, present in complex mixtures, as well as challenging extraction logistics are barriers to economic sustainable production of these compounds from their native species. While heterologous expression of terpenoid biosynthetic genes is feasible, the potential derivation of the products into conjugates via endogenous cytochromes P450 and glycosyl transferases limits this approach. In this project, we used multiplex gene editing technologies to overcome these challenges by creating novel tomato chassis with altered terpenoid biosynthetic capacity in fruit. Excluding central metabolic genes to minimalize impacts on growth and development, we selected 23 known and potential terpene-related genes expressed specifically in the fruit for gene editing. Fruit production and metabolic profiles of three chassis lines with alterations in the major classes of fruit specialized metabolites indicate loss of these genes is tolerated for fruit production. These combinatorial knockouts also showed modulation of native carbon reallocation toward endogenous sinks beneficial for a biosynthetic chassis. Establishing metabolite-modified fruit chassis demonstrates efficient combinatorial editing of entire branches of plant specialized metabolism, facilitating engineering of heterologous terpenes of industrial interest in tomato fruit.

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.

Obtaining tomato plants with firm and intact fruit is one of the main goals in tomato breeding programs. Achieving these goals through conventional breeding is time-consuming and can lead to the loss of unwanted traits. In other hand, consumers are concerned about the presence of transgenic elements in plants acquired through RNA interference. The use of CRISPR/Cas9 technology has made it possible to overcome the above-mentioned shortcomings. In this study, the {beta}-D-N-acetylhexosaminidase ({beta}-hex) gene, which is involved in tomato fruit ripening, was knocked out using CRISPR/Cas9. In the resulting mutant plant genome, an indel mutation was found in exons 1 and 2 of the {beta}-hex gene. Plants with a mutation in their genome were observed to have increased fruit firmness and shelf life compared to control plants without affecting fruit quality.

In plant breeding and genetics, recent advances in high-throughput phenotyping are beginning to meet the growing demand for large-scale, high-quality phenotypic data that emerged after the development of next-generation sequencing technologies. Recent developments in phenomics have been incorporated into almond breeding programs, facilitating the large-scale acquisition of quantitative phenotypes and the dissection of the genetic architecture underlying morphological and quality-related traits. The implementation of a high-throughput phenotyping platform integrating RGB and hyperspectral imaging with genotyping using the 60K almond SNP array enabled the large-scale characterization of almond populations and the identification of 567 robust marker-trait associations across 66 traits. These analyses revealed two major genomic hotspots on chromosomes 2 and 5 associated with morphological and quality-related traits. These regions harbored biologically relevant candidate genes, including genes associated with OVATE family proteins, brassinosteroid signaling, protein ubiquitination, and acyl-CoA metabolism, as well as other regulators of organ growth, cell proliferation, hormone signaling, and seed development. Furthermore, a novel candidate gene encoding a COMT-like O-methyltransferase involved in lignin biosynthesis was identified and proposed to contribute to shell hardness, a major genetically controlled trait in almond. Together, these findings demonstrate the potential of integrating high-throughput phenomics and genomics to dissect complex traits, identify candidate genes, and accelerate genomics-informed breeding in almond.

Understanding how plants respond to high temperature is critical under global warming. Metabolite markers can provide insights into stress-responsive mechanisms and help guide strategies to maintain crop quality. However, heat-associated metabolite markers in grape berries remain poorly defined, particularly at the green stage, a critical phase of berry development during which early metabolic perturbations can influence subsequent ripening and ultimately determine berry composition and quality. Here, we applied berry-scale heat treatments of eight durations of two major wine cultivars, Cabernet Sauvignon and Merlot. Untargeted LC-MS profiling revealed both conserved and cultivar-dependent responses to heat. Based on these patterns, three time points were selected for targeted GC-MS analysis, and subsequent statistical analyses identified robust "cultivar-common heat markers": glycine decreased, whereas galactinol increased consistently across time points and cultivars. "Cultivar-dependent heat markers" were identified: xylose, lyxose, citrulline, quinic acid, and glutamine, that consistently distinguished CS and Merlot fruits under heat stress. Notably, xylose, lyxose, citrulline, and quinic acid also differentiated the two cultivars under ambient conditions, underscoring their potential as stable cultivar-discriminating metabolites. Together, these results reveal dynamic metabolic remodeling in grape berries under heat stress, particularly in amino acid, nitrogen, central carbon metabolism, raffinose family oligosaccharides pathway and the glutathione-ascorbate cycle.

Tomato (Solanum lycopersicum) is one of the most important agricultural crops and serves as a model system for fleshy fruit biology. However, domestication bottlenecks have led to limited genetic diversity for further crop improvement. Wild relatives represent a rich reservoir of phenotypic diversity, yet the molecular basis underlying such phenotypic diversity remains largely unknown. As gene expression variation is a major driver of phenotypic diversity, we performed genome-wide allele-specific expression analyses in F1 hybrids between cultivated tomato and three wild relatives spanning a range of evolutionary distances, across three fruit tissues and multiple developmental stages. Our analysis generated a multi-species map of cis - and trans-regulatory variation, revealing a predominant role of cis-regulatory effects in expression divergence across species, tissues, and stages. The majority of cis effects were tissue- and/or stage-specific, underscoring the importance of tissue context in regulatory variation. Regulatory mechanisms and inheritance patterns shifted with evolutionary distance, with more distantly related species showing increased cis-regulatory contributions. Finally, we identified extensive regulatory divergence in biosynthetic pathways related to fruit nutritional value and flavor quality, including carotenoids, flavonoids, alkaloids, sugars, and volatiles. This study presents a high-resolution map of regulatory variation underlying tomato fruit development and provides evolutionary insights into the regulation of fruit nutrition and flavor traits.

As extreme heat events increase in frequency and intensity worldwide, understanding how woody perennial crops respond to higher maximum temperatures is critical. Perennials face distinct challenges, persisting across many seasons under increasingly variable and extreme conditions, and heat waves threaten the viability of wine grape cultivars through impacts on yield, wine quality, and long-term vine health. To test whether irrigation practices before and during heat waves affect grapevine carbon (C) storage and health, we experimentally manipulated irrigation regimes surrounding heat waves from 2019-2021 in a commercial Cabernet Sauvignon vineyard in the Lodi AVA of Californias Central Valley. Vine physiological traits and yield were measured throughout, and whole-vine nonstructural carbohydrate (NSC) concentrations were quantified after three growing seasons. Although lower supplemental irrigation reduced photosynthesis, stomatal conductance, and fruit yield, whole-vine NSCs did not differ significantly in any perennial organ by the experiments end, indicating that reproductive output and final NSC status responded to irrigation on different timescales. These results suggest that moderate supplemental irrigation during heat events is sufficient to mitigate negative impacts on yield and quality while supporting recovery of NSC reserves, though longer-term monitoring is needed to confirm that this short-term resilience persists.

Here, we present a comprehensive multiomics analysis of anthocyanin biosynthesis in Rubus armeniacus, known for its dark fruits. A phased genome sequence of the tetraploid blackberry was generated, achieving an N50 of 34 Mb with an assembly size of 1.2 Gbp based on Oxford Nanopore Technology sequencing (ONT). The BUSCO score for the total assembly shows a high completeness of 99.1%. The assembly was separated into 4 pseudohaplophases, with the pseudohaplophase A representing the R. armeniacus genome in 7 chromosome scale contigs, with an N50 of 46 Mbp and 98.8% conserved BUSCO genes. A total of 118,183 protein coding genes were annotated within the genome assembly and all relevant genes encoding enzymes and transcriptional regulators of the anthocyanin biosynthesis pathway were identified within each pseudohaplophase. To further understand the underlying cause of dark pigmentation, the gene expression was analysed during different stages of berry development revealing a strong induction of anthocyanin biosynthesis genes including the anthocyanin activating subgroup 6 MYB transcriptions during the berry ripening process. Further, a quantification of cyanidin-3-O-glucoside in methanolic berry extract, utilizing a UHPLC-HRAM-MS analysis, revealed an approximately 500-fold increase of cyanidin-3-O-glucoside from green to black fruit, indicating that dark pigmentation in R. armeniacus results from high anthocyanin accumulation. Significance statementThis study provides a multiomics analysis of the dark pigmentation of Rubus armeniacus, including a high quality phased assembly and an in-depth analysis of the anthocyanin biosynthesis pathway. A transcriptional and metabolomic analysis revealed that dark berry pigmentation is caused by a high accumulation of cyanidin-3-O-glucoside during fruit ripening.

Potato crop is highly vulnerable to abiotic stresses like salinity and low nutrient availability. Rapid identification of stress-resilient genotypes is therefore essential for breeding, yet conventional phenotyping is often slow, space-demanding and expensive. We present LOCOPOTS -- a LOw-COst high-throughput screening platform for in vitro POTatoes under abiotic Stress -- which combines individual in vitro plant culture, low-cost RGB imaging and machine-learning-based automatic segmentation using a trained model of a convolutional neural network, based on U-Net architecture. LOCOPOTS enabled the automated extraction of growth, colour, and vegetation-index traits and demonstrated robust performance across independent phenotyping rounds. We screened 30 potato varieties under control, low-nutrient and saltinity conditions, identifying contrasting growth and physiological responses. Integrated traits such as final area and height, Area_AUC and height_AUC, together with GLI, Chol, cive and chlorophyll fluorescence parameters, discriminated genotype performance under stress. Metabolic profiling further revealed genotype-specific reprogramming in carbon and nitrogen metabolism under low nutrition and salt stress, including changes in fructose, myo-inositol, {beta}-aminobutyric acid, {gamma}-aminobutyric acid, proline, and certain polyamines, identifying them as specific chemical biomarkers of plant stress responses. LOCOPOTS provides a scalable, affordable and space-efficient platform for early screening of potato genetic diversity and identification of candidate traits associated with stress resilience.