Leibniz Institute of Plant Genetics and Crop Plant Research

Aralia elata is closely related to Panax ginseng and contains high levels of saponins and other medicinal compounds. Successful A. elata micropropagation is commercially significant; however, the genomic stability of tissue culture-derived regenerants is unclear. In this study, callus-derived regenerated A. elata plants were obtained, and their cytogenomic constitutions were assessed. Using RepeatExplorer, pre-labeled oligonucleotide probes (PLOPs) were developed with newly mined tandem repeats from < 1× NGS whole-genome short reads, fluorescence in situ hybridization (FISH) was performed using six repeat probes, including three universal PLOPs, and genomic DNA content was estimated using flow cytometry. Regenerated A. elata plants (50) exhibited consistent ploidy, repeat distribution, and genome sizes compared with those exhibited by the mother plant. Six repeat probes were detected using FISH. Tandem repeat AeTR49 was identified as an excellent cytogenetic marker for homologous chromosomes, and AeTR161 and AeTR178 were localized in the centromeric and telomeric sections, respectively. Genomic DNA content (2C) was estimated at 2.46 ± 0.04 pg in the mother plant and 2.41 ± 0.05 pg in regenerated plants, with no significant variations in genome size or chromosome length. These results demonstrate that cytogenomics can be used to effectively evaluate chromosome-level genomic stability in regenerated A. elata plants. Supplementary Information The online version contains supplementary material available at 10.1038/s41598-024-75004-0.

WHIRLY1 belongs to a family of plant‐specific transcription factors capable of binding DNA or RNA in all three plant cell compartments that contain genetic materials. In Arabidopsis thaliana , WHIRLY1 has been studied at the later stages of plant development, including flowering and leaf senescence, as well as in biotic and abiotic stress responses. In this study, WHIRLY1 knockout mutants of A. thaliana were prepared by CRISPR/Cas9‐mediated genome editing to investigate the role of WHIRLY1 during early seedling development. The loss‐of‐function of WHIRLY1 in 5‐day‐old seedlings did not cause differences in the phenotype and the photosynthetic performance of the emerging cotyledons compared with the wild type. Nevertheless, comparative RNA sequencing analysis revealed that the knockout of WHIRLY1 affected the expression of a small but specific set of genes during this critical phase of development. About 110 genes were found to be significantly deregulated in the knockout mutant, wherein several genes involved in the early steps of aliphatic glucosinolate (GSL) biosynthesis were suppressed compared with wild‐type plants. The downregulation of these genes in WHIRLY1 knockout lines led to decreased GSL contents in seedlings and in seeds. Since GSL catabolism mediated by myrosinases was not altered during seed‐to‐seedling transition, the results suggest that AtWHIRLY1 plays a major role in modulation of aliphatic GSL biosynthesis during early seedling development. In addition, phylogenetic analysis revealed a coincidence between the evolution of methionine‐derived aliphatic GSLs and the addition of a new WHIRLY in core families of the plant order Brassicales.

A single reference genome does not fully capture species diversity. By contrast, a pangenome incorporates multiple genomes to capture the entire set of nonredundant genes in a given species, along with its genome diversity. New sequencing technologies enable researchers to produce multiple high-quality genome sequences and catalog diverse genetic variations with better precision. Pangenomic studies have detected structural variants in plant genomes, dissected the genetic architecture of agronomic traits, and helped unravel molecular underpinnings and evolutionary origins of plant phenotypes. The pangenome concept has further evolved into a so-called superpangenome that includes wild relatives within a genus or clade and shifted to graph-based reference systems. Nevertheless, building pangenomes and representing complex structural variants remain challenging in many crops. Standardized computing pipelines and common data structures are needed to compare and interpret pangenomes. The growing body of plant pangenomics data requires new algorithms, huge data storage capacity, and training to help researchers and breeders take advantage of newly discovered genes and genetic variants.

Chromatin modeling enables the characterization of chromatin architecture at a resolution so far unachievable with experimental techniques. Polymer models fill our knowledge gap on a wide range of structures, from chromatin loops to nuclear compartments. Many physical properties already known for polymers can thus explain the dynamics of chromatin. With molecular simulations, it is possible to probe an ensemble of conformations, which attest to the variability observed in individual cells and the general behavior of a population of cells. In this review, we describe universal characteristics of polymers that chromatin carries. We introduce how these characteristics can be assessed with polymer simulations while also addressing specific aspects of chromatin and its environment. Finally, we give examples of plant chromatin models that, despite their paucity, augur well for the future of polymer simulations to plant chromosome biology.

Predicting phenotypes from a combination of genetic and environmental factors is a grand challenge of modern biology. Slight improvements in this area have the potential to save lives, improve food and fuel security, permit better care of the planet, and create other positive outcomes. In 2022 and 2023 the first open-to-the-public Genomes to Fields (G2F) initiative Genotype by Environment (GxE) prediction competition was held using a large dataset including genomic variation, phenotype and weather measurements and field management notes, gathered by the project over nine years. The competition attracted registrants from around the world with representation from academic, government, industry, and non-profit institutions as well as unaffiliated. These participants came from diverse disciplines include plant science, animal science, breeding, statistics, computational biology and others. Some participants had no formal genetics or plant-related training, and some were just beginning their graduate education. The teams applied varied methods and strategies, providing a wealth of modeling knowledge based on a common dataset. The winner’s strategy involved two models combining machine learning and traditional breeding tools: one model emphasized environment using features extracted by Random Forest, Ridge Regression and Least-squares, and one focused on genetics. Other high-performing teams’ methods included quantitative genetics, machine learning/deep learning, mechanistic models, and model ensembles. The dataset factors used, such as genetics; weather; and management data, were also diverse, demonstrating that no single model or strategy is far superior to all others within the context of this competition.

The remobilization of stored assimilates from stems to seeds plays a pivotal role in augmenting barley yield, particularly under water stress conditions. This study examines the molecular mechanisms underlying stem reserve utilization by conducting a comparative analysis of the proteome and metabolome across three barley contrasting genotypes: Yousef, Morocco, and PBYT17. Evaluations were performed at 21 and 28 days after anthesis (DAA) under both water stress and control conditions. The results indicate that the Yousef genotype exhibits superior remobilization of stem reserves, thereby demonstrating its potential to thrive even in adverse environmental conditions. Utilizing advanced quantitative proteomics and targeted metabolomics, this investigation identified a significant number of metabolites and proteins exhibiting differential accumulation across the genotypes. Specifically, 17 metabolites and 1580 proteins were catalogued, highlighting the intricate biochemical responses to water stress. Noteworthy enzymes such as sucrose synthase, inositol monophosphatase 3, and galactokinase were found to be closely associated with remobilization efficiency. In the drought-tolerant genotype, these enzymes maintained stable levels, in stark contrast to the decline observed in the susceptible genotype. This stability is crucial for promoting seed development through ascorbic acid synthesis and for mitigating oxidative stress, which is exacerbated by drought conditions. The elevated levels of certain metabolites, including glucose 6-phosphate, and UDP-glucose, in the drought-tolerant genotype suggest a robust mechanism for maintaining signalling molecules for carbon availability, which is then instrumental in regulating plant growth and seed size development. The findings from this study strongly imply that the drought-tolerant genotype, through enhanced antioxidant capacity, can effectively produce energy-rich storage compounds, thereby optimizing carbon allocation under water stress. Such insights are invaluable for future breeding strategies aimed at improving barley resilience in the face of climate variability.

Nucleotide-binding leucine-rich repeat (NLR) disease resistance genes typically confer resistance against races of a single pathogen. Here, we report that Yr87/Lr85, an NLR gene from Aegilops sharonensis and Aegilops longissima, confers resistance against both P. striiformis tritici (Pst) and Puccinia triticina (Pt) that cause stripe and leaf rust, respectively. Yr87/Lr85 confers resistance against Pst and Pt in wheat introgression as well as transgenic lines. Comparative analysis of Yr87/Lr85 and the cloned Triticeae NLR disease resistance genes shows that Yr87/Lr85 contains two distinct LRR domains and that the gene is only found in Ae. sharonensis and Ae. longissima. Allele mining and phylogenetic analysis indicate multiple events of Yr87/Lr85 gene flow between the two species and presence/absence variation explaining the majority of resistance to wheat leaf rust in both species. The confinement of Yr87/Lr85 to Ae. sharonensis and Ae. longissima and the resistance in wheat against Pst and Pt highlight the potential of these species as valuable sources of disease resistance genes for wheat improvement.

Pangenomes are collections of annotated genome sequences of multiple individuals of a species¹. The structural variants uncovered by these datasets are a major asset to genetic analysis in crop plants². Here we report a pangenome of barley comprising long-read sequence assemblies of 76 wild and domesticated genomes and short-read sequence data of 1,315 genotypes. An expanded catalogue of sequence variation in the crop includes structurally complex loci that are rich in gene copy number variation. To demonstrate the utility of the pangenome, we focus on four loci involved in disease resistance, plant architecture, nutrient release and trichome development. Novel allelic variation at a powdery mildew resistance locus and population-specific copy number gains in a regulator of vegetative branching were found. Expansion of a family of starch-cleaving enzymes in elite malting barleys was linked to shifts in enzymatic activity in micro-malting trials. Deletion of an enhancer motif is likely to change the developmental trajectory of the hairy appendages on barley grains. Our findings indicate that allelic diversity at structurally complex loci may have helped crop plants to adapt to new selective regimes in agricultural ecosystems.

The genomes of many plants, animals, and fungi frequently comprise dispensable B chromosomes that rely upon various chromosomal drive mechanisms to counteract the tendency of non-essential genetic elements to be purged over time. The B chromosome of rye – a model system for nearly a century – undergoes targeted nondisjunction during first pollen mitosis, favouring segregation into the generative nucleus, thus increasing their numbers over generations. However, the genetic mechanisms underlying this process are poorly understood. Here, using a newly-assembled, ~430 Mb-long rye B chromosome pseudomolecule, we identify five candidate genes whose role as trans-acting moderators of the chromosomal drive is supported by karyotyping, chromosome drive analysis and comparative RNA-seq. Among them, we identify DCR28, coding a microtubule-associated protein related to cell division, and detect this gene also in the B chromosome of Aegilops speltoides. The DCR28 gene family is neo-functionalised and serially-duplicated with 15 B chromosome-located copies that are uniquely highly expressed in the first pollen mitosis of rye.

Agriculture is confronted with several challenges such as climate change, the loss of biodiversity and stagnating productivity. The massive increasing amount of data and new digital technologies promise to overcome them, but they necessitate careful data integration and data management to make them usable. The FAIRagro consortium is part of the National Research Data Infrastructure (NFDI) in Germany and will develop FAIR compliant infrastructure services for the agrosystems science community, which will be integrated in the existing research data infrastructure service landscape. Here we present the initial steps of designing and implementing the FAIRagro middleware infrastructure to connect existing data infrastructures. The middleware will feature services for the seamless data integration across diverse infrastructures. Data and metadata are streamlined for research in agrosystems science by downstream processing in the central FAIRagro Search and Inventory Portal and the data integration and analysis workflow system “SciWIn”.

Seed quality is the set of physical, genetic, and physiological characteristics, reflecting the overall germination potential. Maintaining an optimal seed quality is essential for agriculture and seed banks to preserve genetic diversity. Compared to conventional methods (e.g., germination tests), non-invasive approaches allow a more sustainable and rapid evaluation of seed quality but this is limited by high costs. The measurement of ultra-weak photon emission (UPE) and delayed fluorescence (DL), defined as biological phenomena potentially related to the physiological status of living systems, may represent a suitable approach to estimate seed quality. To test this hypothesis, seeds of five agriculturally relevant legume species (Phaseolus vulgaris L., Lathyrus sativus L., Cicer arietinum L., Pisum sativum L., and Vicia faba L.), stored at different conditions (room temperature or -18 °C) for several years, were analysed using a LIANA© prototype to collect data regarding DL and UPE occurring after UV excitation. The obtained data were integrated with germination parameters which underline species-specific behaviours in response to storage conditions. The prediction models show variable efficiency in classifying seeds based on germination which underline species-dependent links between photon emission and seed quality. Therefore, these measurements represent novel, non-invasive, and rapid approaches to evaluate seed quality.

In most studied eukaryotes, chromosomes are monocentric, with centromere activity confined to a single region. However, the rush family (Juncaceae) includes species with both monocentric (Juncus) and holocentric (Luzula) chromosomes, where centromere activity is distributed along the entire chromosome length. Here, we combine chromosome-scale genome assembly, epigenetic analysis, immuno-FISH and super-resolution microscopy to study the transition to holocentricity in Luzula sylvatica. We report repeat-based holocentromeres with an irregular distribution of features along the chromosomes. Luzula sylvatica holocentromeres are predominantly associated with two satellite DNA repeats (Lusy1 and Lusy2), while CENH3 also binds satellite-free gene-poor regions. Comparative repeat analysis suggests that Lusy1 plays a crucial role in centromere function across most Luzula species. Furthermore, synteny analysis between L. sylvatica (n = 6) and Juncus effusus (n = 21) suggests that holocentric chromosomes in Luzula could have arisen from chromosome fusions of ancestral monocentric chromosomes, accompanied by the expansion of CENH3-associated satellite repeats.

The proteinaceous synaptonemal complex (SC) structure forms between meiotic homologous chromosomes. Its central region (CR) consists of transverse filament and central element proteins, in Arabidopsis ZYP1 and SCEP1/SCEP2, respectively. We describe a novel CR protein in Arabidopsis. SCEP3 spatiotemporally overlaps with other CR components and is conserved in plants. In scep3 , SC formation, crossover (CO) assurance (minimum one CO per chromosome pair), CO interference (limited closely-spaced CO) and heterochiasmy (male/female CO rate difference) vanish while genome-wide and particularly female CO increase. Compared with other CR proteins, SCEP3 is also critical for some synapsis-independent CO. SCEP3 interacts with ZYP1 but loads onto recombination intermediates independent of other CR proteins. We propose SCEP3’s loading onto recombination intermediates may stabilize and/or recruit further factors such as ZYP1 to a subset of these intermediates designated to form CO. Hence, SCEP3 interlinks SC and CO formation, being structurally likely the plant ortholog of yeast Ecm11.

Faba bean is a globally adapted legume protein crop with a high yield potential. Currently, yield variation across environments limits more widespread cultivation and the underlying genetics remain unknown. Here, we identify major QTL for faba bean yield and yield stability. We genotype the ProFaba diversity panel with high resolution and carry out coordinated multi-year/location trials across Europe. Based on these data, we identify more than one hundred loci associated with mean performance and stability for 14 complex traits, including yield. Furthermore, we introduce a method for integrating environmental data in the analysis of trait stability, which enables prediction of performance in untested environments. Our work offers insights into the genetics underlying traits and trait stability in faba bean and provides the approaches and resources needed to drive future protein crop improvement.

In eukaryotes, topologically associating domains (TADs) organize the genome into functional compartments. While TAD-like structures are common in mammals and many plants, they are challenging to detect in Arabidopsis thaliana. Here, we demonstrate that Arabidopsis PDS5 proteins play a negative role in TAD-like domain formation. Through Hi-C analysis, we show that mutations in PDS5 genes lead to the widespread emergence of enhanced TAD-like domains throughout the Arabidopsis genome, excluding pericentromeric regions. These domains exhibit increased chromatin insulation and enhanced chromatin interactions, without significant changes in gene expression or histone modifications. Our results suggest that PDS5 proteins are key regulators of genome architecture, influencing 3D chromatin organization independently of transcriptional activity. This study provides insights into the unique chromatin structure of Arabidopsis and the broader mechanisms governing plant genome folding.

Seeds represent essential stages of the plant life cycle: embryogenesis, the intermittent quiescence phase and germination. Each stage has its own physiological requirements, genetic program and environmental challenges. Consequently, the effects of developmental and environmental hypoxia can vary from detrimental to beneficial. Past and recent evidence shows how low-oxygen signalling and metabolic adaptations to hypoxia affect seed development and germination. Here, we review the recent literature on seed biology in relation to hypoxia research, and present our perspective on key challenges and opportunities for future investigations.

Winter faba beans exhibit significant yield advantages over spring cultivars and hold promise for enhancing local protein production and agricultural sustainability. However, the threat of winter kill limits wider cultivation and the genetics of faba bean winter hardiness remain unresolved. Here, we develop a highly improved faba bean reference genome and combine this with resequencing and phenotyping of winter and spring faba bean accessions to identify genetic determinants of winter hardiness. Genome-wide association analysis of frost tolerance traits identifies a major winter hardiness locus where the most strongly associated variant explains the vast majority of phenotypic variation and accurately differentiates between winter and spring types. Furthermore, we identify additional signals within the winter faba bean gene pool that pave the way for further improvement of winter hardiness. Our work provides improved genomic resources and resolves the genetics of a key agronomic trait in a global protein crop to facilitate future breeding efforts.

In plants, L‐serine (Ser) biosynthesis occurs through various pathways and is highly dependent on the atmospheric CO2 concentration, especially in C3 species, due to the association of the Glycolate Pathway of Ser Biosynthesis (GPSB) with photorespiration. Characterization of a second plant Ser pathway, the Phosphorylated Pathway of Ser Biosynthesis (PPSB), revealed that it is at the crossroads of carbon, nitrogen, and sulphur metabolism. The PPSB comprises three sequential reactions catalysed by 3‐phosphoglycerate dehydrogenase (PGDH), 3‐phosphoSer aminotransferase (PSAT) and 3‐phosphoSer phosphatase (PSP). PPSB was overexpressed in plants exhibiting two different modes of photosynthesis: Arabidopsis (C3 metabolism), and maize (C4 metabolism), under ambient (aCO2) and elevated (eCO2) CO2 growth conditions. Overexpression in Arabidopsis of the PGDH1 gene alone or PGDH1, PSAT1 and PSP1 in combination increased the Ser levels but also the essential amino acids threonine (aCO2), isoleucine, leucine, lysine, phenylalanine, threonine and methionine (eCO2) compared to the wild‐type. These increases translated into higher protein levels. Likewise, starch levels were also increased in the PPSB‐overexpressing lines. In maize, PPSB‐deficient lines were obtained by targeting PSP1 using Cas9 endonuclease. We concluded that the expression of PPSB in maize male gametophyte is required for viable pollen development. Maize lines overexpressing the AtPGDH1 gene only displayed higher protein levels but not starch at both aCO2 and eCO2 conditions, this translated into a significant rise in the nitrogen/carbon ratio. These results suggest that metabolic engineering of PPSB in crops could enhance nitrogen content, particularly under upcoming eCO2 conditions where the activity of GPSB is limited.

Main conclusion Rice exudation patterns changed in response to P deficiency. Higher exudation rates were associated with lower biomass production. Total carboxylate exudation rates mostly decreased under P-limiting conditions. Abstract Within the rhizosphere, root exudates are believed to play an important role in plant phosphorus (P) acquisition. This could be particularly beneficial in upland rice production where P is often limited. However, knowledge gaps remain on how P deficiency shapes quality and quantity of root exudation in upland rice genotypes. We therefore investigated growth, plant P uptake, and root exudation patterns of two rice genotypes differing in P efficiency in semi-hydroponics at two P levels (low P = 1 µM, adequate P = 100 µM). Root exudates were collected hydroponically 28 and 40 days after germination to analyze total carbon (C), carbohydrates, amino acids, phenolic compounds spectrophotometrically and carboxylates using a targeted LC–MS approach. Despite their reported role in P solubilization, we observed that carboxylate exudation rates per unit root surface area were not increased under P deficiency. In contrast, exudation rates of total C, carbohydrates, amino acids and phenolics were mostly enhanced in response to low P supply. Overall, higher exudation rates were associated with lower biomass production in the P-inefficient genotype Nerica4, whereas the larger root system with lower C investment (per unit root surface area) in root exudates of the P-efficient DJ123 allowed for better plant growth under P deficiency. Our results reveal new insights into genotype-specific resource allocation in rice under P-limiting conditions that warrant follow-up research including more genotypes.