Biology subjects

Heinicke, S.

Publications and source records attributed to Heinicke, S..

13 recordsLinked to original sources

bioRxiv2026

North American bird occupancy dynamics attributed to climate and land use change

Evidence is accumulating that global change is altering species distributions. Yet, detailed knowledge is missing about the relative and joint contribution of different drivers to observed species responses. Here, we implemented an impact attribution framework based on counterfactual simulations to assess the impact of climate and land use change on occupancy dynamics of North American breeding birds. We used a Bayesian framework to fit process-explicit dynamic occupancy models to long-term survey data for 159 species from 1995 to 2019, and quantified predictive performance using spatial and temporal cross-validation. We then assessed the relative importance and effect direction of climate and land use change while accounting for model predictive accuracy. Results indicate that climate change negatively affected 90 % of the species and land use change negatively impacted 96 %. Climate change emerged as more important than land use change for driving changes in occupancy across species. Remarkably, the effects of both drivers were mostly antagonistic rather than acting additively or synergistically. Climate was the most important driver for bird communities in the western USA, while land use change dominated in the southeast, and combined climate and land use change in the northeast. Our analysis demonstrates that recent changes in North American bird distributions are shaped by multiple global change drivers acting in concert. The effect of recent climate and land use change were mostly antagonistic, and thus trends in bird occupancy dynamics could not be understood by studying the impact of those drivers in isolation. By disentangling the effects of climate and land use change on biodiversity trends, impact attribution approaches can improve our understanding of global change impacts and can support conservation planning and more accurate and realistic projections of biodiversity response to global change.

ecology↗

bioRxiv2026

Birds protected in Europe, overlooked in Africa: conservation synergies between African primates and wintering Afro-Palearctic migratory birds

Primates are frequent targets of international conservation efforts, due to their anthropological significance and function as biodiversity conservation flagship species. Afro-Palearctic migratory birds receive considerable conservation attention in their breeding habitats, but much less in their wintering areas. Good wintering habitat conditions are, however, crucial for long-term population viability. Integrating complementary conservation efforts for these taxa may provide substantial co-benefits. Here, we evaluate the spatial, ecological, and conservation congruence between 248 Afro-Palearctic migratory bird and 213 African primate species to identify potential synergies in intercontinental conservation efforts. The wintering areas of 218 migratory bird species overlap with primate ranges. Up to 125 Afro-Palearctic migratory bird species co-occur with at least one primate species locally and highest regional cross-taxon richness occurs in forest-savanna ecotones in West Africa and eastern DRC. Agriculture and hunting threaten both taxa (88-97% of primate and 59-70% of bird species). Several conservation interventions have demonstrated effectiveness for both taxa, particularly habitat protection and hunting reduction. We propose three policy priorities: strengthening cross-taxa monitoring within existing primate research infrastructure, improving the conservation evidence base in the African context, and creating financing synergies that allow conservation investments to simultaneously benefit both taxa.

ecology↗

bioRxiv2026

Discovery of a Plant Pictet-Spenglerase with R-Stereoselectivity

The class-defining monoterpenoid indole alkaloid (MIA) scaffold strictosidine is generated by condensation of tryptamine and secologanin by the Pictet-Spenglerase strictosidine synthase (STR). All previously characterized STR orthologs are strictly 3S-stereoselective. Here, we report that the medicinal plant species Pogonopus speciosus (Rubiaceae) accumulates the 3R epimer vincosidic acid produced by a an ortholog of STR. This ortholog, named here Epi-STR, exclusively produces the 3R configuration and is capable of accepting both secologanic acid and the methylester secologanin as aldehyde substrates. Using comparative phylogenetic and structural analyses, we determine the amino acid residues that confer stereoselectivity. Through rational design of reciprocal amino acid substitutions, we achieved switches in stereoselectivity in a canonical STR and in Epi-STR. The stereoselectivity of the engineered mutants is also dependent on the identity of the aldehyde substrate. Notably, previous and extensive engineering efforts have failed to switch the stereo-selectivity of STR. Therefore, this discovery now allows cost-effective epimer-pure access to the R-epimer, and offers mechanistic insights into the enzyme stereoselectivity of this important reaction. This work also highlights the importance of phytochemical analyses of poorly described plant species.

biochemistry↗

bioRxiv2025

Conserved early steps of stemmadenine biosynthesis

Stemmadenine acetate is a pivotal intermediate in the production of pharmacologically active monoterpene indole alkaloids. Here, we identify orthologs of stemmadenine acetate pathway genes (SGD, GS, GO, Redox1, Redox2, SAT) from Tabernanthe iboga. We characterize these enzymes in vitro, and additionally, we reconstitute stemmadenine acetate biosynthesis in Nicotiana benthamiana, comparing the formation of intermediates and shunt products that are produced when previously characterized orthologs from the related plant Catharanthus roseus are used. Ortholog pairs are catalytically indistinguishable, except in the case of GS. Surprisingly, the T. iboga ortholog catalyzes formation of an alternate stereoisomer 19Z-geissoschizine, seeding a low-flux Z-series in heterologous reconstitution systems in vitro and in planta. We additionally characterize the major shunt products that arose during reconstitution of stemmadenine acetate biosynthesis. We show that the substrate promiscuity of Redox2 results in formation of the shunt products 16(R/S)-isositsirikines, hampering pathway flux and yields. Additionally, we show that stemmadenine can be oxidized by endogenous N. benthamiana enzymes, leading to the shunt product condylocarpine. Nevertheless, we could produce stemmadenine at a 6 mg yield from 19E-geissoschizine by heterologous expression in N. benthamiana. Overall, we highlight the prospects for milligram production of important biosynthetic intermediates in N. benthamiana.

biochemistry↗

bioRxiv2025

Discovery of iridoid cyclase completes the iridoid pathway in asterids

Iridoids are specialized monoterpenes ancestral to asterid flowering plants (Albach et al, 2001; Stull et al, 2018). Iridoids play key roles in plant defense and are also essential precursors for pharmacologically important alkaloids (Dobler et al, 2011; Eisner, 1964). The biosynthesis of all iridoids involves the cyclization of a reactive enol intermediate. While this cyclization occurs spontaneously at low yields, it has long been hypothesized that a dedicated enzyme is involved in this process (Geu-Flores et al, 2012; Lichman et al, 2019b). Here, we report the discovery of asterid iridoid cyclases (ICYC). We show that these enzymes catalyze cyclization of the reactive intermediate to form the two major iridoid stereoisomers found in plants. Our work uncovers the last missing key step in the otherwise well-characterized iridoid biosynthesis pathway in asterids. This discovery unlocks the possibility to generate previously inaccessible iridoid stereoisomers, which will enable metabolic engineering for the sustainable production of valuable iridoid and iridoid-derived compounds.

plant biology↗

bioRxiv2025

Single-cell metabolome and RNA-seq multiplexing on single plant cells

Plants produce valuable natural products used for a wide variety of industrial applications. Since these molecules have important applications in a variety of industrial sectors, there is enormous interest in elucidating the biosynthetic pathways that are responsible for the production of these compounds. Identification of the genes that comprise these biosynthetic pathways has been enabled by gene-to-metabolite networks that are generated from transcriptomic and metabolomic datasets. Recent advances in both single-cell RNA-seq (scRNA-seq) and single-cell mass spectrometry metabolomics (scMS) have enabled the measurement of either gene expression or metabolite levels in individual cells. However, these individual datasets can only be used to indirectly correlate gene expression levels with metabolite concentrations at the single cell level. Here we demonstrate that both scRNA-seq and scMS can be applied to the same plant cell, thereby enabling direct comparisons between gene expression and metabolite levels. This multiplexing approach reveals both qualitative and quantitative correlations between metabolite levels and biosynthetic gene expression in individual cells. This integrated approach sheds light on the underlying processes driving complex plant biosynthesis.

plant biology↗

bioRxiv2025

It runs in the family: Discovery of enzymes in the oleuropein pathway in Olive (Olea europaea) by comparative transcriptomics

Olive (Olea europaea L.) is one of the most important crop trees, with olive oil being a key ingredient of the Mediterranean diet. Oleuropein, an oleoside-type secoiridoid, is the major determinant of flavor and quality of olive oil. Iridoid biosynthesis has been elucidated in Catharanthus roseus, which produces secologanin-type secoiridoids, but iridoid biosynthesis in other species remains unresolved. In this work, we sequenced RNA from olive fruit mesocarp of six commercial olive cultivars with varying oleuropein content, during maturation and ripening. Using this data we discovered three polyphenol oxidases with oleuropein synthase (OS) activity, a novel oleoside-11-methyl ester glucosyl transferase (OMEGT) synthesizing a potential intermediate in the route, and a 7-epi-loganic acid O-methyltransferase (7eLAMT). Interestingly, integrating transcriptomics data from 15 plant species from three iridoid-producing plant orders (Lamiales, Gentianales, and Cornales), and tissue expression panels from Jasminum sambac and Fraxinus excelsior, we discovered two 2-oxoglutarate dependent dioxygenases (named 7eLAS) that synthesize 7-epi-loganic acid; in contrast C. roseus 7-deoxy-loganic acid hydroxylase (7DLH), a known bottleneck in MIA production, is a cytochrome p450. This comparative co-expression method, which combines guilt by association and comparative transcriptomics approaches, can successfully leverage big datasets for untargeted discovery of enzymes. Key FindingsO_LIExpression of genes involved in iridoid biosynthesis, from the early MEP pathway to the last step of oleuropein biosynthesis, decreases during olive fruit maturation. C_LIO_LIWe discovered an oxoglutarate dependent dioxygenase, 7-epi-loganic acid synthase (7eLAS), catalyzing the stereoselective oxidation of 7-deoxy-loganic acid to 7-epi-loganic acid, in a reaction analogous to C. roseus 7-deoxy-loganic acid hydroxylase (7DLH), a cytochrome p450. C_LIO_LIWe report a 7-epi-loganic acid O-methyltransferase (7eLAMT) orthologous to Catharanthus roseus loganic acid O-methyltransferase and found a novel oleoside-11-methyl ester glucosyl transferase (OMEGT) synthesizing 7-{beta}-1-D-glucopyranosyl-oleoside-11-methyl ester, a potential intermediate in the oleuropein biosynthesis route. C_LIO_LIWe discovered three olive polyphenol oxidases that have oleuropein synthase (OS) activity, catalyzing the conversion of ligstroside to oleuropein. C_LI

biochemistry↗

bioRxiv2024

Single-cell mass spectrometry reveals heterogeneous triterpenic acid accumulation in apple callus-derived cells

The use of plant cell cultures for large scale production of natural compounds, although promising, has been hindered by their genetic instability and heterogeneity. Here, we show how single cell mass spectrometry can be used to characterize the natural product profile of a callus culture at a highly resolved level. We identify and quantify triterpenic acids in a population of callus cells derived from Annurca apple (Malus pumila Miller cv Annurca) leaf. The analysis demonstrated that a high degree of metabolic heterogeneity exists in the cell population, with the levels of detected metabolites varying significantly across the callus cells. This metabolic heterogeneity was underpinned by variable expression levels of key biosynthetic genes in the single cells. The application of an abiotic stress, near ultraviolet radiation (NUV), to the callus culture resulted in increased levels of triterpenic acids. Single cell mass spectrometry analysis revealed that after treatment, a larger percentage of callus cells produced detectable amounts of these metabolites, ultimately resulting in a more homogeneous production of the metabolites. Furthermore, it showed that intracellular concentrations of ursolic acid derivatives can reach more than 100 mM. Single cell mass spectrometry analyses provide a starting foundation for understanding the molecular mechanisms responsible for metabolic heterogeneity in plant cell cultures, which could in turn facilitate efforts to improve these cell cultures for commercial purposes.

plant biology↗

bioRxiv2024

Independent evolution of ipecac alkaloid biosynthesis

Ipecac alkaloids are medicinal monoterpenoid-derived tetrahydroisoquinoline alkaloids found in two distantly related plants: Carapichea ipecacuanha (Gentianales) and Alangium salviifolium (Cornales). We have elucidated ipecac alkaloid biosynthesis in both species, conclusively demonstrating that biosynthesis of the structurally complex ipecac alkaloid protoemetine has evolved independently. We show that although protoemetine biosynthesis proceeds via the same chemical logic in both species, each plant uses a distinct monoterpene precursor. Moreover, we provide evidence that both plants initiate ipecac biosynthesis by a non-enzymatic Pictet-Spengler reaction, and we elucidate the biosynthetic fate of both the 1R and 1S stereoisomers that are produced in this non-stereoselective reaction. Phylogenetic analyses clearly show independent pathway evolution through parallel and convergently evolved enzymes. This work provides insight into how nature can capitalize on highly reactive starting substrates, the manner in which multi-step pathways can arise, and also lays the foundation for metabolic engineering of these important medicinal compounds.

plant biology↗

bioRxiv2024

Streamlined screening platforms lead to the discovery of pachysiphine synthase from Tabernanthe iboga

Plant-specialized metabolism is largely driven by the oxidative tailoring of key chemical scaffolds catalyzed by cytochrome P450 (CYP450s) enzymes. The monoterpene indole alkaloids tabersonine and pseudo-tabersonine, found in the medicinal plant Tabernanthe iboga, are extensively modified by oxidative reactions. Here we developed a streamlined screening strategy to screen the activity of T. iboga CYP450s in Nicotiana benthamiana. Using multigene constructs encoding the biosynthesis of tabersonine and pseudo-tabersonine scaffolds, we set out to uncover the CYP450s responsible for oxidative transformations of these scaffolds. Our approach identified two T. iboga cytochrome P450 enzymes: pachysiphine synthase (PS) and 16-hydroxy-tabersonine synthase (T16H). These enzymes catalyze an epoxidation and site-specific hydroxylation of tabersonine to produce pachysiphine and 16-OH-tabersonine, respectively. We further demonstrated that these genes produced the expected products when expressed in Catharanthus roseus flowers. This work provides new insights into the biosynthetic pathways of MIAs and underscores the utility of N. benthamiana and C. roseus as platforms for the functional characterization of plant enzymes.

biochemistry↗

bioRxiv2024

Quantitative single cell mass spectrometry reveals the dynamics of plant natural product biosynthesis

Plants produce an extraordinary array of complex natural products (specialized metabolites). Since the biosynthetic genes that are responsible for synthesis of these molecules are often localized to rare or distinct cell types, recently developed single cell RNA-sequencing (scRNA-seq) approaches have tremendous potential to resolve these complex pathways. In contrast, detection, identification, and quantification of metabolites in single cells has remained challenging. Here, we report a robust method for single cell mass spectrometry in which we rigorously characterize and quantify the concentrations of four classes of natural products in individual cells of leaf, root, and petal of the medicinal plant Catharanthus roseus. These single cell mass spectrometry datasets reveal information about the biosynthetic processes that cannot be determined from the corresponding scRNA-seq data alone, providing a highly resolved picture of natural product biosynthesis at cell-specific resolution.

biochemistry↗

bioRxiv2023

Threat of mining to African great apes

The rapid growth of clean energy technologies is driving a rising demand for critical minerals. In 2022 at the UN Biodiversity Conference (COP 15), seven major economies formed an alliance to enhance the sustainability of mining these essential decarbonization minerals. However, there is a scarcity of studies assessing the threat of mining to global biodiversity. By integrating a global mining dataset with ape density distribution estimates, we explored the potential negative impact of industrial mining on African great apes. Our findings reveal that up to one-third of Africas great ape population faces mining-related risks. This is especially pronounced in West Africa, where numerous mining areas overlap with fragmented ape habitats, often occurring in high-density ape regions. For 97% of mining areas, no ape survey data are available, underscoring the importance of increased accessibility to environmental data within the mining sector to facilitate research into the complex interactions between mining, climate, biodiversity and sustainability. TeaserMining for clean energy minerals could put one-third of Africas ape population at risk.

ecology↗

bioRxiv2022

Global analysis of the yeast knock-out phenome

Genome-wide phenotypic screens in the budding yeast Saccharomyces cerevisiae have produced the largest, richest and most systematic phenotypic description of any organism. Such an achievement was enabled by the development of highly scalable phenotypic assays and construction of the yeast knock-out (YKO) collection, comprising ~5,000 isogenic strains each deleted for exactly one open reading frame. Systematic screening of the YKO collection led to ~500 publications describing ~14,500 phenotypes capturing nearly every aspect of yeast biology. Yet, integrative analyses of this rich data source have been virtually impossible due to the lack of a central repository and consistent meta-data annotations. Here, we describe the aggregation, harmonization and analysis of all published phenotypic screens of the YKO collection, which we refer to as the Yeast Phenome (www.yeastphenome.org). To demonstrate the power of data integration and illustrate how much it facilitates the generation of testable hypotheses, we present three discoveries uniquely enabled by Yeast Phenome. First, we use the variation in the number of phenotypes per gene to identify tryptophan homeostasis as a central point of vulnerability to a wide range of chemical compounds, including FDA-approved drugs. Second, using phenotypic profiles as a tool for predicting gene function, we identify and validate the role of YHR045W as a novel regulator of ergosterol biosynthesis and DNA damage response, and YGL117W as a new member of the aromatic amino acid biosynthesis pathway. Finally, we describe a surprising exponential relationship between phenotypic similarity and intergenic distance in both yeast and human genomes. This relationship, which stretches as far as 380 kb in yeast and 100 Mb in humans, suggests that gene positions are optimized for function to a much greater extent than appreciated previously. Overall, we show that Yeast Phenome enables systematic enquiries into the nature of gene-gene and gene-phenotype relationships and is an important new resource for systems biology.

systems biology↗