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Schranz, M. E.

Publications and source records attributed to Schranz, M. E..

At least 19 recordsLinked to original sources

Enemies, more than sex, shape butterfly post-mating odor evolution

Not all odors influencing mating behavior evolve as sex pheromones. Female butterflies post-mating odors have been considered species-specific anti-aphrodisiac pheromones shaped by sexual selection but may also serve broader ecological roles shaped by natural selection. Males transfer odors to females that repel rivals, yet the widespread use of these compounds across phyla makes them targets for eavesdropping, such as by phoretic egg parasitoids. We show that in cabbage white butterflies (Pieris spp.), these odors are highly variable and attract parasitoids, deter predators, and influence oviposition. Using gas chromatography and electroantennography, we demonstrate that odor emission and perception lack species-specificity: compounds once thought unique to P. brassicae and P. rapae are shared across Pieridae. In P. napi, odor variation among populations correlates with parasitoid pressure, but not with latitude, genetic distance, or mating frequency, suggesting ecological rather than sexual drivers. In P. brassicae, CRISPR/Cas9 disruption of odor perception alters oviposition and increases susceptibility to parasitism. Moreover, these odors render females unpalatable to birds. Together, our results show that post-mating odors in Pieris butterflies may act as aposematic signals. We provide evidence that these signals evolve under multiple selective pressures, balancing deterrence of mates and predators, parasitoid avoidance, and host-plant interactions. These findings suggest that chemical signals should be viewed as integrating ecological and reproductive pressures, rather than being interpreted solely through the lens of sexual communication.

ecology↗

Systematic mass-spectrometry-guided metabolic fingerprinting elucidates diversity of specialized metabolites across the Brassicaceae

O_LIPlants produce diverse bouquets of specialized metabolites (SMs), yet only a fraction of the vast phytochemical space has been explored to date. Comparative analysis of SM profiles can reveal hotspots of biochemical novelty, while systematic profiling across taxonomic levels does presently not cover large plant families. C_LIO_LITo study core and accessory SM profiles in the Brassicaceae plant family, we fingerprinted 14 species by Liquid-Chromatography Mass-Spectrometry (LCMS/MS). We develop standardized experimental and computational workflows integrating in silico annotation tools to study consensus compound class and substructure distributions of SMs. Furthermore, we investigate the congruence of chemotaxonomy and species phylogeny across an extended panel of 17 species. C_LIO_LIUnique metabolite profiles were outstanding in Camelina sativa, Capsella rubella, and B. vulgaris, with the largest unique terpenoid profile annotated in C. sativa, accounting for 33.5% and 55.6% in positive and negative ionization mode, respectively. Substructure motifs were found to overlap with compound class predictions, highlighted for triterpenoids in Camelinodae. Furthermore, dual-tissue chemotaxonomic clustering resembled relationships of Brassica subgenomes across tissues. C_LIO_LIWe anticipate that our systematic approach can serve as a blueprint for investigating biochemical diversity in other plant lineages and can boost the characterization of plant natural product pathways. C_LI

plant biology↗

Metabolic fingerprinting of 17 Brassicaceae species across three tissues

Plants produce the most diverse blends of specialized metabolites on earth. Natural products derived from plants are valuable resources for drug development, food chemistry, and crop resistance breeding. Phenotypes of specialized metabolite profiles can be captured by untargeted mass-spectrometry across species phylogeny, tissues, and genotypes. Here, we collected metabolic fingerprints of 17 Brassicaceae species across three tissues (paired leaf and root; flower) using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in positive and negative ionization mode. Corresponding metadata has been refined for reuse according to ReDU guidelines, and for integration with public genomic and transcriptomic data. Standardization of in vitro growth conditions, and data processing workflows enables integration of acquired raw and processed data across platforms for single- and multi-omics analysis. Further, the inclusion of tissue-specific metabolic profiles across ploidy levels, as well as across crop species and wild relatives, makes this dataset a valuable resource for natural product discovery.

plant biology↗

KCFtools: Rapid alignment-free method for introgression screening and GWAS using k-mer profiles

MotivationIn the era of multiple genome references, researchers often align sequencing reads against distinct assemblies or even multiple references simultaneously. This enables applications such as the detection of introgressed segments or highly variable genomic regions, which are especially prevalent in large-genome crop species such as lettuce or wheat. However, these applications come at the cost of increased computational burden, inconsistencies in mapping methods, and reduced reproducibility across studies. To address these limitations, we developed KCFtools, a Java-based toolkit that identifies the presence and absence of k-mers in non-overlapping genomic or transcriptomic windows by comparing query and reference genomes. This alignment-free approach enables the efficient computation of an identity score for each window, thereby facilitating robust detection of introgressed or variable regions across genomes. ResultsWe systematically evaluated the performance and accuracy of the k-mer-based method implemented in KCFtools, benchmarking it against conventional SNP-based introgression detection pipelines. Our results demonstrate that KCFtools effectively captures introgressed segments and structurally diverse regions, even in species with fragmented or highly divergent reference genomes. In addition, we extended KCFtools to generate genotype matrices from k-mer variation tables. These matrices are compatible with Genome-Wide Association Studies (GWAS) software and allow the identification of loci associated with phenotypic traits. We showcase the utility of this approach by detecting known and novel associations for downy mildew resistance in lettuce, underscoring the pipelines potential for high-resolution, reference-agnostic population genetic analysis. Availabilityhttps://github.com/sivasubramanics/kcftools Contactc.s.sivasubramani@gmail.com

bioinformatics↗

The Lettuce Expression Browser: from lab to LEB

Lettuce (Lactuca sativa L.) is an economically important leafy vegetable within the Asteraceae family, cultivated worldwide across diverse agricultural systems. Recent advances in genomic and transcriptomic resources have positioned lettuce as a promising model system for functional genomics in the Asteraceae. However, currently available gene expression datasets lack comprehensive tissue-specific resolution, primarily focus on a single cultivar and are not visualised in an interpretable manner, limiting their utility for broader genetic and physiological studies. To bridge this gap, we developed the Lettuce Expression Browser (LEB), a publicly available platform providing high-resolution gene expression maps across various organs, tissues and developmental stages in both cultivated and wild lettuce species. The LEB integrates transcriptomic data from finely dissected seedlings, shoot tissues at various developmental stages and seedlings subjected to abiotic stresses (salt and far-red), visualised using the ggPlantmap R package. This platform offers an intuitive interface for exploring gene expression patterns and serves as a valuable resource for those studying lettuce development, stress responses, and evolutionary genomics. The LEB is hosted on the LettuceKnow Web Portal (https://lettuce.bioinformatics.nl) and can be expanded to include additional datasets, enhancing its role as a key tool for lettuce research and crop improvement. Significance statementThe Lettuce Expression Browser (LEB) provides the first high-resolution gene expression atlas for both cultivated and wild lettuce species. This open-access resource enables detailed exploration of gene activity across development stages and stress conditions, advancing functional genomics in the Asteraceae family.

plant biology↗

Scarcity of flowering plants on Sedum roofs limits pollinator diversity

Green roofs can provide suitable habitats for pollinating insects in urban areas. Pollinators are a large and diverse group with high economic and ecological relevance to society. Yet their populations are in decline, highlighting the need for new habitats such as green roofs. An understanding of how local and landscape factors of green roofs shape pollinator communities is crucial to optimize the design and development of green roofs. This study aimed to identify how the surrounding landscape and green roof characteristics influence pollinator diversity and abundance. Pollinators - such as bees, wasps, hoverflies and butterflies - were sampled using pan traps and hand netting on 25 green roofs that were categorized into three types based on functionality and vegetation structure: sedum roofs, nature roofs, and roof gardens. To assess pollinator communities, we examined species richness, abundance, and functional diversity--defined here as the range of ecological roles or traits. Roof characteristics (e.g., flower diversity, microrelief, vegetation cover, roof height, size, and age) and landscape factors (e.g. percentage of surrounding green, distance to surrounding green) were analysed in relation to these diversity metrics. We show that a continuous supply of flowers throughout the year, flower abundance and the type of green roof significantly influence pollinator diversity and abundance. Sedum roofs supported lower pollinator diversity compared to the other roof types. Furthermore, our models indicated a decrease in pollinator diversity on higher green roofs, and when honeybee hives are present, while the presence of bee hotels increased pollinator abundance. The study highlights the value of nature roofs and roof gardens, compared to sedum roofs for supporting pollinator diversity. This is linked to the importance of constant forage availability, especially in early spring, which is absent on most sedum roofs. The results from this study offer guidelines to green roof designers on how to design pollinator friendly roofs in urban areas. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=171 SRC="FIGDIR/small/676189v1_ufig1.gif" ALT="Figure 1"> View larger version (34K): org.highwire.dtl.DTLVardef@dfb4caorg.highwire.dtl.DTLVardef@d6478eorg.highwire.dtl.DTLVardef@1f3e725org.highwire.dtl.DTLVardef@1cc4ef0_HPS_FORMAT_FIGEXP M_FIG C_FIG Pollinator species: Macropis europea (Hymenoptera: Melittidae), Helophilus pendulus (Diptera: Syrhidae) and Polyommatus icarus (Lepidoptera: Lycaenidae). All three species were observed on green roofs during the authors fieldwork. Image created using biorender.com and personal drawings. HighlightsO_LIBiodiverse roofs and roof gardens support higher pollinator diversity compared to sedum roofs. C_LIO_LIA continuous supply of flowers throughout the year, along with a higher abundance of flowers, are key factors influencing pollinator diversity and abundance on green roofs. C_LIO_LIRoof height affects the diversity and abundance of different pollinator groups in distinct ways, a novel finding that needs further investigation. C_LIO_LIWe provide guidelines for green roof designers, emphasizing the importance of selecting roof types and designs that ensure constant flower availability. C_LI

ecology↗

MoGAAAP: A modular Snakemake workflow for automated genome assembly and annotation with quality assessment

With the current speed of sequencing, there is a desire for standardised and automated genome assembly and annotation to produce high-quality genomes as input for comparative (pan)genomics. Therefore, we created a convenience pipeline using existing tools that creates annotated genome assemblies from HiFi (and optionally ultra-long ONT and/or Hi-C) reads for a set of related accessions as well as a related reference genome. Our pipeline is species-agnostic and generates an extensive quality assessment report that can be used for manual filtering and refinement of the assembly and annotation. It includes statistics for individual completeness and contamination assessments as well as a concise pangenome view. The pipeline is implemented in Snakemake and available with a GPLv3 license at GitHub under github.com/dirkjanvw/MoGAAAP and at Zenodo under doi.org/10.5281/zenodo.14833021.

bioinformatics↗

Phenotypic similarity of NAD(P)-Malic Enzymes in Tomato: Unveiling Patterns of Convergent and Parallel Evolution

The Solanum lycopersicum (tomato) genome encodes seven malic enzymes (MEs): three cytosolic NADP-ME isoforms (SlNADP-ME1, -2, and -3), two plastidic NADP-ME proteins (SlNADP-ME4a and -4b), and two mitochondrial NAD-dependent subunits (SlNAD-ME1 and -2) of a heteromeric enzyme. Except for SlNADP-ME3, which is almost exclusively expressed in the root, all the genes are active in the major tissues of flowering plants (leaf, stem, flower, and root), with SlNADP-ME2 and -4b transcripts accumulating at relative high levels. During the cell expansion and ripening phases of fruit development, SlNADP-ME3 and -4b expression increased in the pericarp and seeds while SlNADP-ME2 remained at a high level. In green fruit, SlNADP-ME3 and -4b are down-regulated by ethylene, with SlNAD-ME1 and -2 being up-regulated. A correlation between SlNADP-ME4b accumulation and NADP-ME activity was observed from the early immature green to the mature green fruit stages, linking this plastidic isoform to starch and lipid biosynthesis. SlNADP-ME1 and -4a expression increase with temperature, suggesting involvement in defense mechanisms and supported by cis-elements composition in their promoters. Interesting, SlNADP-ME3 biochemical properties and its accumulation in seeds is part of an inherited and long-conserved genetic lineage in Angiosperms including Arabidopsis NADP-ME1, involved in normal seed germination. By analyzing the phylogeny and synteny of SlNAD(P)-ME genes and determining the biochemical properties of the recombinant proteins, we seek functional similarities to Arabidopsis NAD(P)-ME members. Our findings enhance the understanding of malate metabolism in tomatoes, which could inform strategies to improve fruit quality. Additionally, they prompt a re-evaluation of pre-existing notions regarding the functional orthology of enzymes based on phylogenetic relationships. SIGNIFICANCE STATEMENTEnzyme functional orthology among distant plant species cannot be inferred through phylogenetic analysis because of the large independent rearrangements of genomes during evolution. A comprehensive characterization of the tomato NAD(P)-ME family and their comparison with members from an unrelated plant species, such as Arabidopsis, allowed us to evaluate the impact of selective pressures on orthologous genes. This approach reveals that the phenotypic similarity of an entire enzyme family in different species is achieved through diverse evolutionary events such as ancestry, parallelism, and genetic convergence.

plant biology↗

Phylogenomic synteny analysis tracks conserved ancient polyploid-derived triplicated genomic blocks across Asteraceae genomes

The Asteraceae (Compositae) is the largest flowering plant family, ubiquitous in most terrestrial communities, and morphologically hyper-diverse. An ancient whole genome triplication (paleo-hexaploidization) occurred at approximately the same time as the evolutionary innovation and adaptive radiation of the family during the middle Eocene. Despite its importance, the genomic contents arising from this triplication have yet to be tracked in context of the Asteraceae genome evolution. We applied a synteny oriented phylogenomic analysis of 21 Asterales genomes and to study the paleo-hexaploidization and its consequences to gene, trait, and genome evolution. We identified 15 ancestral linkage groups (ALGs) that date back to the common diploid ancestor of all Asteraceae. Each of these groups was triplicated, resulting in 45 genomic blocks (3x15), which serve as the foundation for cross-family analyses. We demonstrate the complex evolutionary dynamics of the 45 genomic blocks across the Asteraceae phylogeny. We found that modern genomes are genetic mosaics of three progenitor genomes by extensive genomic exchange, chromosomal shuffling and gene fractionation. 157 genes retained three paleo-hexaploid derived syntenic paralogs across most Asteraceae species. Transcription factors (TFs) and auxin-related genes are significantly overrepresented in the conserved triplets, and expression of the paleo-hexaploidy paralogs is spatiotemporally differentiated. These genes are involved in the development of floral capitulum, a remarkable morphological innovation of the family. The discovery of conserved triplicated genes can direct further study to understand the evolutionary innovation, and the synteny-phylogenomic framework and ALGs provide a comparative framework to characterize newly sequenced Asteraceae genomes.

genomics↗

Identifying Agronomic, Nutrition and Leaf Vein Quantitative Trait Loci in the Orphan Crop Gynandropsis gynandra

A sustainable food supply is vital for addressing the challenges of a growing global population and climate change. Gynandropsis gynandra, a nutritious C4 orphan crop native to Africa and Asia, thrives in low-input agricultural systems, making it a valuable candidate for future food security. This crop also serves as a model for studying C4 photosynthesis. However, research on its genetic improvement is limited. In this study, two mapping populations from diverse parental lines were used to identify Quantitative Trait Loci (QTL) linked to agronomically relevant traits like plant height, leaf area, flowering time, nutritional content, and photosynthesis. Fifteen QTL were identified, with two affecting both leaf size and flowering time across populations, which can be applied for marker assisted selection for crop improvement. Additionally, QTL linked to C4 photosynthesis provide insights into the genetic mechanisms behind this pathway. Overall, this research enhances the potential of G. gynandra as a climate-resilient crop. One sentence summaryLeveraging natural variation in Gynandropsis gynandra to identify QTL associated with important traits.

plant biology↗

S-RNase Evolution in Self-Incompatibility: Phylogenomic Insights into Synteny with S-Like Genes

S-RNases are essential in the gametophytic self-incompatibility (GSI) system of many flowering plants, where they act as stylar-S determinants. Despite their significance, the syntenic genomic origin and evolutionary trajectory of S-RNase genes in eudicots have remained largely unclear. Here, we performed large-scale phylogenetic and microsynteny network analyses of RNase T2 genes across 130 angiosperm genomes, encompassing 35 orders and 56 families. Based on the connections observed in the synteny network, particularly the persisting synteny block in none-SI Cucurbitaceae species, we infer that well-characterized S-RNase genes (Class III-A RNase genes) were duplicated and evolved from Class I S-like RNase genes, possibly as a result of the gamma triplication event shared by core eudicots. Additionally, we identified frequent lineage-specific gene transpositions of S-RNase genes across diverse angiosperm lineages, including Rosaceae, Solanaceae, and Rutaceae families, accompanied by a significant increase in transposable element (TE) activity near these genes. Our findings delineate the precise genomic origin and evolutionary path of eudicot S-RNase genes, enhancing our understanding of the evolution of the S-RNase-based GSI system.

evolutionary biology↗

Lactuca super-pangenome reduces bias towards reference genes in lettuce research

Breeding of lettuce (Lactuca sativa L.), the most important leafy vegetable worldwide, for enhanced disease resistance and resilience relies on multiple wild relatives to provide the necessary genetic diversity. In this study, we constructed a super-pangenome based on four Lactuca species (representing the primary, secondary and tertiary gene pools) and comprising 474 accessions. We include 68 newly sequenced accessions to improve cultivar coverage and add important foundational breeding lines. With the super-pangenome we find substantial presence/absence variation (PAV) and copy-number variation (CNV). Functional enrichment analyses of core and variable genes show that transcriptional regulators are conserved whereas disease resistance genes are variable. PAV-genome-wide association studies (GWAS) and CNV-GWAS are largely congruent with single-nucleotide polymorphism (SNP)-GWAS. Importantly, they also identify several major novel quantitative trait loci (QTL) for resistance against Bremia lactucae in variable regions not present in the reference lettuce genome. The usability of the super-pangenome is demonstrated by identifying the likely origin of non-reference resistance loci from the wild relatives Lactuca serriola, Lactuca saligna and Lactuca virosa. The provided methodology and data provide a strong basis for research into PAVs, CNVs and other variation underlying important biological traits of lettuce and other crops.

plant biology↗

Oryza genome evolution through a tetraploid lens

Oryza is remarkable genus - with two domesticated (i.e. Asian and African rice) and 25 diploid and tetraploid wild species, 11 extant genome types, and a [~]3.4-fold genome size variation - that possesses a virtually untapped reservoir of genes that can be used for crop improvement. Here we unveil and interrogate 11 new chromosome-level assemblies of nine tetraploid and two diploid wild Oryza species in the context of [~]15 million years of evolution of the genus. We show that the core Oryza (sub)genome across all genome types is only [~]200 Mb and largely syntenic, while the remaining nuclear fractions, spanning [~]80-600 Mb, are intermingled, extremely plastic and rapidly evolving. For the halophyte O. coarctata, we show that - despite the detection of gene fractionation in the subgenomes - homoeologous genes are expressed at higher levels in one subgenome over the other in a mosaic form, thereby showing subgenome equivalence. The integration of these 11 new ultra-high quality reference genomes with our previously published genome data sets provide a nearly complete picture of the consequences of natural and artificial selection across the evolutionary history of Oryza. This in turn opens the door to unlock their genetic potential for future crop improvement and neodomestication.

genomics↗

Expanding the Triangle of U: The genome assembly of Hirschfeldia incana provides insights into chromosomal evolution, phylogenomics and high photosynthesis-related traits

The Brassiceae tribe encompasses many economically important crops and exhibits high intraspecific and interspecific phenotypic variation. After a shared whole-genome triplication (WGT) event (Br-, [~]15.9 million years ago), different lineages and species underwent differential chromosomal rearrangements (diploidization) leading to diverse patterns of gene retention and loss (fractionation). Lineage diversification and genomic changes contributed to an array of divergence in morphology, biochemistry, and physiology underlying photosynthesis-related traits. The C3 species Hirschfeldia incana is studied as it displays high photosynthetic rates under high-light conditions. We present an improved chromosome-level genome assembly for H. incana (Nijmegen, v2.0) using nanopore and chromosome conformation capture (Hi-C) technologies, with 409Mb in size and an N50 of 52Mb (a 10x improvement over the previously published scaffold-level v1.0 assembly). The updated assembly and annotation allowed to investigate the WGT history of H. incana in a comparative phylogenomic framework from the Brassiceae ancestral genomic blocks and related diploidized crops. Hirschfeldia incana (x=7) shares extensive genome collinearity with Raphanus sativus (x=9). These two species share some commonalities with Brassica rapa and B. oleracea (A genome, x=10 and C genome, x=9, respectively) and other similarities with B. nigra (B genome, x=8). Phylogenetic analysis revealed that H. incana and R. sativus form a monophyletic clade in between the Brassica A/C and B genomes. We postulate that H. incana and R. sativus genomes are results of reciprocal hybridization combinations of the Brassica A/C and B genome types. Our results might explain the discrepancy observed in published studies regarding phylogenetic placement of H. incana and R. sativus in relation to the "Triangle of U" species. Expression analysis of WGT retained gene copies revealed sub-genome expression divergence, likely due to neo- or sub-functionalization. Finally, we highlighted genes associated with physio-biochemical-anatomical adaptive changes observed in H. incana which likely facilitate its high-photosynthesis traits under high light.

plant biology↗

Maroon rice genomic diversity reflects 350 years of colonial history

Maroons in Suriname and French Guiana descend from enslaved Africans who escaped the plantations during colonial times. Maroon farmers still cultivate a large rice diversity, their oldest staple crop. Maroon oral history and written records by colonial authorities provide contrasting perspectives on the origins of Maroon rice. Here, we integrated genomic ancestry analyses of 136 newly sequenced Maroon rice varieties with ethnobotanical and archival research to reconstruct the historical contexts associated with the introduction of rice varieties to the Guianas. We found that a large subset traces to West Africa, linked to the transatlantic slave trade (c.1530-1825). Maroons obtained other varieties from indentured laborers from Java (1890 onwards), USA rice breeders (1932 onwards), and Hmong refugees from the Vietnam War (1991). Furthermore, we found rice types never documented before, indicating Maroon farmers selected from crosses. Overall, our results demonstrate that the Maroon farming system prioritizes maintenance of a high stock diversity, which we posit reflects the expertise they inherited from their (African) ancestors. Ignored by agricultural modernization initiatives, Maroon farmers today are important custodians of a unique cultural heritage. Moreover, the genomic findings underline many Maroon stories about their past. This study hence demonstrates the power of cross-disciplinary crop research to reconstruct aspects of the human past for which historical records may be biased or incomplete. We anticipate that a similar study approach can be applied to other heirloom crops of (Indigenous) communities that may have preserved their history on their farms to reconstruct, acknowledge and honor the past.

genomics↗

The dimorphic diaspore model Aethionema arabicum (Brassicaceae): Distinct molecular and morphological control of responses to parental and germination temperatures

Plants in habitats with unpredictable conditions are often characterized by diversifying their bet-hedging strategies that ensure fitness over a wider range of variable environmental factors. A striking example is the diaspore (seed and fruit) heteromorphism that evolved to maximize species survival in Aethionema arabicum (Brassicaceae) in which external and endogenous triggers allow the production of two distinct diaspores on the same plant. Using this dimorphic diaspore model, we identified contrasting molecular, biophysical, and ecophysiological mechanisms in the germination responses to different temperatures of the mucilaginous seeds (M+ seed morphs), the dispersed indehiscent fruits (IND fruit morphs), and the bare non-mucilaginous M- seeds obtained by pericarp (fruit coat) removal from IND fruits. Large-scale comparative transcriptome and hormone analyses of M+ seeds, IND fruits, and M- seeds provided comprehensive datasets for their distinct thermal responses. Morph-specific differences in co-expressed gene modules in seeds, as well as seed and pericarp hormone contents identified a role of the IND pericarp in imposing coat dormancy by generating hypoxia affecting ABA sensitivity. This involved expression of morph-specific transcription factors, hypoxia response and cell wall-remodeling genes, as well as altered abscisic acid (ABA) metabolism, transport, and signaling. Parental temperature affected ABA contents and ABA-related gene expression and altered IND pericarp biomechanical properties. Elucidating the molecular framework underlying the diaspore heteromorphism can provide insight into developmental responses to globally changing temperatures. IN A NUTSHELL BackgroundHeteromorphic diaspores (fruits and seeds) are an adaptive bet-hedging strategy to ensure survival in spatiotemporally variable environments. The stone cress Aethionema arabicum, an annual plant native to semi-arid habitats in Anatolia (Turkey), one of the worlds hotspots of biodiversity. It is a close relative of Arabidopsis, rapeseed, cabbage and other Brassica crops, but in contrast to these Ae. arabicum disperses two distinct diaspores from the same plant. These dimorphic diaspores are the mucilaginous seeds (dispersed by pod shatter) and indehiscent fruits (dispersed by abscission). The wing-like pericarp (fruit coat) of the single-seeded indehiscent fruit allows wind dispersal over large distances. The amounts and ratios of the dimorphic diaspores are variable and depend on the environmental conditions. The dimorphic diaspores differ in morphology, dormancy and germination properties and thereby make Ae. arabicum an excellent model for the comparative investigation of the underpinning molecular mechanisms. QuestionWe asked how temperature during fruit and seed formation and during seed germination affect dormancy release and germination speed, and how the morphology, hormonal regulation, and the expression of genes differ between the dimorphic diaspores. FindingsLarge-scale comparative transcriptome and hormone analyses of the mucilaginous seeds and the indehiscent fruits, as well as the seeds artificially extracted from indehiscent fruits by pericarp (fruit coat) removal, provided comprehensive datasets for their distinct thermal responses. Material obtained from plants grown at different temperatures during reproduction was imbibed at different temperatures for germination. This altered the abscisic acid (ABA) metabolism and the pericarp biomechanical properties. Diaspore-specific differences in response to distinct imbibition temperatures identified distinct gene expression patterns in seeds, distinct seed and pericarp hormone contents, and a role of the pericarp in generating hypoxia inside the fruit and imposing coat dormancy. This revealed distinct combinations of specific transcription factors, hypoxia responses and cell wall-remodeling genes, as well as altered signaling pathway genes. Next stepsOur large-scale comparative transcriptome datasets are easily and publicly accessible via the Aethionema arabicum web portal (https://plantcode.cup.uni-freiburg.de/aetar_db/index.php). We plan to expand this by future work on seedlings derived from the dimorphic diaspores, by comparing different Ae. arabicum genotypes, and by studying responses to specific stresses. Understanding the molecular basis of this fascinating example of developmental diversity and plasticity and its regulation by temperature is expected to add insight how plants respond to changing environmental conditions.

plant biology↗

Natural gene variation in Cannabis sativa unveils a key region of cannabinoid synthase enzymes

Cannabinoids are well-known specialised metabolites from the plant Cannabis sativa L. (cannabis). They exhibit various therapeutical to intoxicating psychoactive effects and have potential for medicinal applications. Among the enzymes involved in cannabinoid biosynthesis, cannabinoid oxidocyclases such as the tetrahydrocannabinolic acid (THCA) synthase play a key role in determining cannabis chemotype. To improve our understanding of cannabinoid oxidocyclase structure-function relationship, we proposed a new approach to targeted mutagenesis. By reviewing cannabis natural variation, three cannabinoid oxidocyclase mutations (S355N, CONF, G376R) associated to atypical plant chemotypes were selected. In-vitro characterization of THCA synthase mutants demonstrated these mutations significantly impact enzyme activity, correlating with the associated chemotype: S355N nearly inactivated the THCA synthase, CONF impaired CBGA metabolization and altered product specificity, while G376R drastically reduced enzyme activity and altered product specificity. In-silico docking experiments permitted to model the successive steps of THCA synthase substrate metabolization, revealing that the three mutations hamper substrate binding. Collectively, our results demonstrated how plant diversity can be leveraged to guide enzyme targeted mutagenesis, highlighted a key region of cannabinoid oxidocyclases, and permitted the establishment of a new model of the THCA synthase catalytic mechanism. This provides new insights into enzyme function, which can ultimately help developing medicinal cannabis cultivars and cannabinoid biotechnological production.

molecular biology↗

Genome assembly and analysis of Lactuca virosa: implications for lettuce breeding

Lettuce (Lactuca sativa L.) is a leafy vegetable crop with ongoing breeding efforts related to quality, resilience, and innovative production systems. Genetic variation of important traits in close relatives is necessary to meet lettuce breeding goals. Lactuca virosa (2x=2n=18), a wild relative assigned to the tertiary lettuce gene pool, has a much larger genome (3.7 Gbp) than Lactuca sativa (2.5 Gbp). It has been used in interspecific crosses and is a donor to modern crisphead lettuce cultivars. Here, we present a de novo reference assembly of L. virosa with high continuity and complete gene space. This assembly facilitated comparisons to the genome of L. sativa and to that of the wild species L. saligna, a representative of the secondary lettuce gene pool. To assess the diversity in gene content, we classified the genes of the three Lactuca species as core, accessory and unique. In addition, we identified three interspecific chromosomal inversions compared to L. sativa, which each may cause recombination suppression and thus hamper future introgression breeding. Using three-way comparisons in both reference-based and reference-free manners, we show that the proliferation of long-terminal repeat elements has driven the genome expansion of L. virosa. Further, we performed a genome-wide comparison of immune genes, nucleotide-binding leucine-rich repeat, and receptor-like kinases among Lactuca spp. and indicate the evolutionary patterns and mechanisms behind their expansions. These genome analyses greatly facilitate the understanding of genetic variation in L. virosa, which is beneficial for the breeding of improved lettuce varieties.

plant biology↗