Use of a temporary immersion bioreactor system for the sustainable production of thapsigargin in shoot cultures of <i>Thapsia garganica</i>

Abstract Background Thapsigargin and nortrilobolide are sesquiterpene lactones found in the Mediterranean plant Thapsia garganica L. Thapsigargin is a potent inhibitor of the sarco/endoplasmic reticulum calcium ATPase pump, inducing apoptosis in mammalian cells. This mechanism has been used to develop a thapsigargin-based cancer drug first by GenSpera and later Inspyr Therapeutics (Westlake Village, California). However, a stable production of thapsigargin is not established. Results In vitro regeneration from leaf explants, shoot multiplication and rooting of T. garganica was obtained along with the production of thapsigargins in temporary immersion bioreactors (TIBs). Thapsigargin production was enhanced using reduced nutrient supply in combination with methyl jasmonate elicitation treatments. Shoots grown in vitro were able to produce 0.34% and 2.1% dry weight of thapsigargin and nortrilobolide, respectively, while leaves and stems of wild T. garganica plants contain only between 0.1 and 0.5% of thapsigargin and below detectable levels of nortrilobolide. In addition, a real-time reverse transcription PCR (qRT-PCR) study was performed to study the regulatory role of the biosynthetic genes HMG-CoA reductase (HMGR), farnesyl diphosphate synthase (FPPS), epikunzeaol synthase (TgTPS2) and the cytochrome P450 (TgCYP76AE2) of stem, leaf and callus tissues. Nadi staining showed that the thapsigargins are located in secretory ducts within these tissues. Conclusions Shoot regeneration, rooting and biomass growth from leaf explants of T. garganica were achieved, together with a high yield in vitro production of thapsigargin in TIBs

Coastal Research Seen Through an Early Career Lens—A Perspective on Barriers to Interdisciplinarity in Norway

The value of interdisciplinarity for solving complex coastal problems is widely recognized. Many early career researchers (ECRs) therefore actively seek this type of collaboration through choice or necessity, for professional development or project funding. However, establishing and conducting interdisciplinary research collaborations as an ECR has many challenges. Here, we identify these challenges through the lens of ECRs working in different disciplines on a common ecosystem, the Norwegian Skagerrak coast. The most densely populated coastline in Norway, the Skagerrak coast, is experiencing a multitude of anthropogenic stressors including fishing, aquaculture, eutrophication, climate change, land runoff, development, and invasive species. The Skagerrak coastline has also been the focus of environmental science research for decades, much of which aims to inform management of these stressors. The region provides a fantastic opportunity for interdisciplinary collaboration, both within and beyond the environmental sciences. This perspective article identifies the barriers ECRs in Norway face in establishing interdisciplinary and collaborative research to inform management of coastal ecosystems, along with their root causes. We believe our discussion will be of broad interest to all research institutions who employ or educate ECRs (in Norway and worldwide), and to those who develop funding mechanisms for ECRs and interdisciplinary research.publishedVersio

Assessing specialized metabolite diversity in the cosmopolitan plant genus Euphorbia l.

Coevolutionary theory suggests that an arms race between plants and herbivores yields increased plant specialized metabolite diversity and the geographic mosaic theory of coevolution predicts that coevolutionary interactions vary across geographic scales. Consequently, plant specialized metabolite diversity is expected to be highest in coevolutionary hotspots, geographic regions, which exhibit strong reciprocal selection on the interacting species. Despite being well-established theoretical frameworks, technical limitations have precluded rigorous hypothesis testing. Here we aim at understanding how geographic separation over evolutionary time may have impacted chemical differentiation in the cosmopolitan plant genus Euphorbia. We use a combination of state-of-the-art computational mass spectral metabolomics tools together with cell-based high-throughput immunomodulatory testing. Our results show significant differences in specialized metabolite diversity across geographically separated phylogenetic clades. Chemical structural diversity of the highly toxic Euphorbia diterpenoids is significantly reduced in species native to the Americas, compared to Afro-Eurasia. The localization of these compounds to young stems and roots suggest a possible ecological relevance in herbivory defense. This is further supported by reduced immunomodulatory activity in the American subclade as well as herbivore distribution patterns. We conclude that computational mass spectrometric metabolomics coupled with relevant ecological data provide a strong tool for exploring plant specialized metabolite diversity in a chemo-evolutionary framework

Travel Tales of a Worldwide Weed: Genomic Signatures of Plantago major L. Reveal Distinct Genotypic Groups With Links to Colonial Trade Routes

Retracing pathways of historical species introductions is fundamental to understanding the factors involved in the successful colonization and spread, centuries after a species’ establishment in an introduced range. Numerous plants have been introduced to regions outside their native ranges both intentionally and accidentally by European voyagers and early colonists making transoceanic journeys; however, records are scarce to document this. We use genotyping-by-sequencing and genotype-likelihood methods on the selfing, global weed, Plantago major, collected from 50 populations worldwide to investigate how patterns of genomic diversity are distributed among populations of this global weed. Although genomic differentiation among populations is found to be low, we identify six unique genotype groups showing very little sign of admixture and low degree of outcrossing among them. We show that genotype groups are latitudinally restricted, and that more than one successful genotype colonized and spread into the introduced ranges. With the exception of New Zealand, only one genotype group is present in the Southern Hemisphere. Three of the most prevalent genotypes present in the native Eurasian range gave rise to introduced populations in the Americas, Africa, Australia, and New Zealand, which could lend support to the hypothesis that P. major was unknowlingly dispersed by early European colonists. Dispersal of multiple successful genotypes is a likely reason for success. Genomic signatures and phylogeographic methods can provide new perspectives on the drivers behind the historic introductions and the successful colonization of introduced species, contributing to our understanding of the role of genomic variation for successful establishment of introduced taxa.publishedVersio

Travel Tales of a Worldwide Weed: Genomic Signatures of Plantago major L. Reveal Distinct Genotypic Groups With Links to Colonial Trade Routes

Retracing pathways of historical species introductions is fundamental to understanding the factors involved in the successful colonization and spread, centuries after a species’ establishment in an introduced range. Numerous plants have been introduced to regions outside their native ranges both intentionally and accidentally by European voyagers and early colonists making transoceanic journeys; however, records are scarce to document this. We use genotyping-by-sequencing and genotype-likelihood methods on the selfing, global weed, Plantago major, collected from 50 populations worldwide to investigate how patterns of genomic diversity are distributed among populations of this global weed. Although genomic differentiation among populations is found to be low, we identify six unique genotype groups showing very little sign of admixture and low degree of outcrossing among them. We show that genotype groups are latitudinally restricted, and that more than one successful genotype colonized and spread into the introduced ranges. With the exception of New Zealand, only one genotype group is present in the Southern Hemisphere. Three of the most prevalent genotypes present in the native Eurasian range gave rise to introduced populations in the Americas, Africa, Australia, and New Zealand, which could lend support to the hypothesis that P. major was unknowlingly dispersed by early European colonists. Dispersal of multiple successful genotypes is a likely reason for success. Genomic signatures and phylogeographic methods can provide new perspectives on the drivers behind the historic introductions and the successful colonization of introduced species, contributing to our understanding of the role of genomic variation for successful establishment of introduced taxa.info:eu-repo/semantics/publishedVersio

Thapsigargins and induced chemical defence in Thapsia garganica

Thapsigargin and related compounds are produced by Thapsia garganica L. (Apiaceae) and are thought to be a defence compound against herbivory. Thapsigargin inhibits the sarco-endoplasmic reticulum Ca2+-ATPase (SERCA) in both vertebrates and invertebrates. This activity is responsible for its potent toxicity, as well as the potential use to treat solid tumours. However, the ecological role and regulation of thapsigargin are not well understood, and the aim of this study was to investigate if thapsigargin biosynthesis was responsive to leaf damage. To test the response to potential leaf damage during a herbivory, greenhouse plants were subjected to clipping to mimic the physical damage. Unclipped versus clipped plants were sampled for chemical analysis and the gene expression for the two known thapsigargin biosynthetic genes (TgTPS2 and TgCYP76AE2) was investigated. Data obtained by LC–ESI–MS/MS were used to perform molecular networking to identify chemical constituents related to thapsigargin and its biosynthesis. The results show a significant change in a plant’s chemical profile after mimicking an herbivory event. Both the chemical analysis and gene expression data show that T. garganica plants can induce the biosynthesis of this class of defence compounds at the site of an attack. Thapsigargins are clearly the dominant defence compounds in these plants, and they seem to be produced through a common biosynthetic pathway with little diversity. This likely means that T. garganica has a relatively simple response to herbivory, as opposed to many other plant species that have been shown to have complex metabolite responses to herbivory