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Straube, H.

Publications and source records attributed to Straube, H..

5 recordsLinked to original sources

Deoxyribonucleotide dephosphorylation by VENOSA4 supports organellar genome replication in plants

The replication of the three genomes in plant cells during germination is a complex process, which requires a high degree of coordination between the genome-containing compartments: the nucleus, mitochondria, and chloroplasts. The first committed step for the de novo synthesis of deoxynucleoside triphosphates (dNTPs), the building blocks for DNA replication, occurs exclusively in the cytosol. A major unresolved question is how an adequate supply of dNTPs for the organelles is achieved. Here, we show that VENOSA4 (VEN4), a dNTP triphosphohydrolase, is critical for this process in Arabidopsis thaliana. Using isotope feeding combined with mass spectrometry analysis, we demonstrate that VEN4 converts most de novo-synthesized dNTPs to deoxynucleosides (dNs). Preventing this conversion by VEN4 mutation strongly diminishes both cpDNA and mtDNA amounts, but these can be partially rescued by the application of exogenous dNs. Hence, the dNTP catabolic activity of VEN4 ensures that sufficient DNA precursors in form of dNs reach the chloroplasts and the mitochondria to be salvaged there to dNTPs for DNA synthesis. This seemingly counterintuitive coupling of de novo synthesis, dNTP hydrolysis, and salvage may provide a mechanism to control DNA precursor allocation between cellular compartments through selective transport and metabolic trapping.

plant biology↗

Exploring L-tyrosine and L-DOPA biosynthesis in faba bean (Vicia faba L.)

Background and AimsL-DOPA is an important pharmaceutical that accumulates to high levels in the legume faba bean (Vicia faba). L-DOPA is likely derived from L-tyrosine but the responsible enzyme (L-tyrosine oxidase) remains unknown. Availability of L-tyrosine may be a key factor controlling L-DOPA accumulation. In legumes, L-tyrosine is supplied via either a plastidial TyrA enzyme (ADH) or a deregulated cytosolic homolog (PDH). This study aimed at identifying L-tyrosine oxidase and TyrA genes from faba bean. MethodsWe used gene-to-metabolite correlations and homology-based searches to select fifteen L-tyrosine oxidase candidates, which were tested in yeast and in the model plant Nicotiana benthamiana. We also used isotopically labeled L-tyrosine to measure biosynthetic activity in different faba bean tissues and to test an alternative biosynthetic hypothesis. Three faba bean TyrA genes were inferred by homology and assayed in N. benthamiana by co-expression with a known L-tyrosine oxidase, CYP76AD6. Key ResultsNone of the L-tyrosine oxidase candidates produced L-DOPA upon heterologous expression. Feeding experiments showed a lack of correlation between L-DOPA accumulation and biosynthetic capacity. Feeding studies also disproved an alternative route to L-DOPA by oxidation of 4-hydroxyphenylpyruvate. Of the TyrA genes, two were able to increase L-tyrosine levels in N. benthamiana 2-3-fold (VfADH and VfPDH), and one of them was able to boost the levels of L-DOPA derivatives up to 6-fold (VfADH). ConclusionsThe faba bean L-tyrosine oxidase remains unidentified, with a possible transport of L-DOPA across tissues likely having confounded our correlation-based selection strategies. In N. benthamiana, both VfADH and VfPDH can increase the levels of L-tyrosine, while VfADH can further boost the levels of L-DOPA derivatives. Our work delivers a strategy to boost the provision of L-tyrosine in N. benthamiana and provides valuable insights in the search for the elusive L-tyrosine oxidase from faba bean.

plant biology↗

Climate-driven in-situ trait variation in an annual ruderal grass across Europe

O_LIPlant functional traits link environmental conditions to plant performance and adaptation. Growing evidence shows that in-situ intraspecific trait variation can be as important as differences between species, yet large intraspecific datasets measured in situ are rare. While most studies focused on plant morphological traits, contents of elemental nutrients in the seeds received much less attention so far. C_LIO_LIWe conducted a large-scale in-situ study of the widespread ruderal grass Hordeum murinum. We sampled 2070 individuals from 207 populations from a large part of the native range in Europe and Northern Africa. In-situ, we measured seed ripening phenology and morphological traits, and analyzed the content of elemental nutrients in the seeds. C_LIO_LIWe found that Hordeum murinum grew larger, produced seeds later, and had heavier seeds in colder and wetter areas. Plants in denser vegetation were taller with heavier seeds but formed fewer spikes. Seed nutrient content generally declined with seed weight and was mainly driven by climate. Soil conditions had only minor effects on plant traits and seed nutrients. Population identity explained much of the variation, indicating a possible genetic component. C_LIO_LISynthesis: Our findings provide a comprehensive view of Hordeum murinum responding to environmental gradients across its European distribution. Climatic variables, especially temperature, are key drivers for reproductive success and seed nutrient content, while local environments, such as biotic pressures, are more critical for growth-related traits. These patterns indicate that H. murinum modulates its growth and reproductive investment along environmental gradients, balancing phenology, stress tolerance, and limited competitive capacity. C_LI

ecology↗

A fungal endophyte-generated nucleoside signal regulates host cell death and promotes root colonization

The intracellular colonization of plant roots by the beneficial fungal endophyte Serendipita indica follows a biphasic strategy. After an early biotrophic phase, the interaction transitions to a host cell death phase restricted to the epidermal and cortex layers of the root. Host cell death contributes to the successful accommodation of the fungus during the beneficial interaction in Arabidopsis thaliana. How host cell death is initiated and controlled is largely unknown. Here we show that two fungal enzymes, the ecto-5-nucleotidase SiE5NT and the nuclease SiNucA, act synergistically in the plant apoplast at the onset of cell death to produce deoxyadenosine (dAdo), a potent cell death inducer in animal systems. The uptake of extracellular dAdo, but not the structurally related adenosine (Ado), activates a previously undescribed cell death mechanism in A. thaliana. Mutation of the equilibrative nucleoside transporter ENT3 in A. thaliana results in resistance to cell death triggered by extracellular dAdo and reduced fungal-mediated cell death during root colonization. A library screen of A. thaliana T-DNA insertion lines identified a toll/interleukin-1 receptor nucleotide-binding leucine-rich repeat (TIR-NLR) protein as an additional intracellular component in dAdo-triggered cell death. Mutation of this previously uncharacterised TIR-NLR, which we have named ISI (induced by S. indica), affects host cell death, fungal colonization and growth promotion, suggesting a key role in the regulation of root cell death and plant-microbe interaction. Our data show that the combined activity of two fungal apoplastic enzymes leads to the production of a metabolite that, upon uptake, triggers TIR-NLR-modulated plant cell death, providing a link to immunometabolism in plants. Short summaryEfficient intraradical colonization by the beneficial fungal endophyte Serendipita indica requires restricted host cell death. How this symbiotic host cell death is initiated and controlled is largely unknown. Here we show that two fungal enzymes, the ecto-5-nucleotidase SiE5NT and the nuclease SiNucA, act synergistically in the apoplast at the onset of cell death to produce deoxyadenosine (dAdo), a potent cell death inducer in animal systems. Uptake of extracellular dAdo activates a previously undescribed cell death mechanism in plants. Mutation of the A. thaliana equilibrative nucleoside transporter ENT3 leads to resistance to cell death triggered by uptake of extracellular dAdo and to reduced fungal-mediated cell death during colonization. A library screen of A. thaliana T-DNA insertion lines identified a TIR-NLR protein as an additional intracellular component in dAdo-triggered cell death, providing a link to immunometabolism in plants. In a nutshellRegulated host cell death is part of the plant defense strategy against pathogens, but it is also involved in the accommodation of certain beneficial microbes in the roots. We have identified extracellular metabolites and intracellular metabolic signals that contribute to colonization by beneficial root fungal endophytes and uncovered a conserved cell death mechanism likely co-opted for establishing plant-endophyte symbiosis.

plant biology↗

An inosine triphosphate pyrophosphatase safeguards nucleic acids from aberrant purine nucleotides and prevents a constitutive salicylic acid response

- In plants, inosine is enzymatically introduced in some tRNAs but not in other RNAs or DNA. Nonetheless, our data show that RNA and DNA from Arabidopsis thaliana contain (deoxy)inosine, probably derived from non-enzymatic adenosine deamination in nucleic acids and usage of (deoxy)inosine triphosphate (dITP and ITP) during nucleic acid synthesis. - We combined biochemical approaches, sample preparation and LC-MS, as well as RNA-Seq to characterize a plant INOSINE TRIPHOSPHATE PYROPHOSPHATASE (ITPA) from Arabidopsis thaliana, which is conserved in many organisms, and investigated the sources of deaminated purine nucleotides in plants. - ITPA dephosphorylates deaminated nucleoside di- and triphosphates to the respective monophosphates. ITPA loss-of-function causes inosine di- and triphosphate accumulation in vivo and an elevated (deoxy)inosine content in DNA and RNA, as well as salicylic acid (SA) accumulation, early senescence and upregulation of transcripts associated with immunity and senescence. Cadmium-induced oxidative stress leads to more ITP in the wildtype, and this effect is enhanced in itpa mutants, suggesting that ITP originates from ATP deamination. - ITPA is part of a molecular protection system, preventing accumulation of (d)ITP, its usage for nucleic acid synthesis, and probably nucleic acid stress leading to SA accumulation, stress gene induction and early senescence.

plant biology↗