Biology subjects

Yokoyama, R.

Publications and source records attributed to Yokoyama, R..

7 recordsLinked to original sources

bioRxiv2026

The proteotoxicity of azetidine 2-carboxylic acid is associated with reactive oxygen species accumulation

Plants make diverse metabolites to outcompete neighboring organisms for space and resources. Some of these toxic metabolites broadly disrupt conserved molecular mechanisms, such as protein biosynthesis. Nonproteogenic amino acids (NPAAs) are a structurally diverse class of metabolites that interfere with protein biosynthesis. The proline (Pro) analog azetidine-2-carboxylic acid (Aze) inhibits plant growth through misincorporation during protein biosynthesis. However, it is unknown if a cascade of downstream stress responses is triggered following Aze misincorporation. Here, we investigate the morphological and stress responses in Arabidopsis grown on Aze. Investigation of root morphological responses show not only reduced root growth, but increased root branching following growth on Aze. Altered root morphology is coupled with a reduced gravitropic response. Aboveground organs were also affected by Aze, including reduced chlorophyll content, reduced photosynthetic efficiency, and increased anthocyanin content. We then tested whether Aze induces reactive oxygen species (ROS) accumulation using multiple approaches and observed both immediate and sustained accumulation of general ROS and H2O2 following treatment with Aze. When plants were grown on Aze supplemented with Pro, ROS levels were restored to normal levels, suggesting that reducing misincorporation events results in less downstream stress responses. In summary, we find that following Aze treatment a cascade of downstream stress responses is induced that exacerbates the effects of toxic NPAAs. This study sheds light on the mechanism of action of NPAAs and provides information on the downstream consequences of translational errors.

plant biology↗

bioRxiv2026

Enhanced phenylalanine biosynthesis amplifies light-stress-driven phenylpropanoid production in Arabidopsis

Phenylpropanoids are L-phenylalanine (Phe)-derived specialized metabolites with crucial roles in plant stress adaptation. Among various abiotic stresses, high-light (HL) is a major threat that leads to oxidative damage and therefore upregulates the biosynthesis of antioxidant phenylpropanoids, including anthocyanins, in plants. Phenylpropanoid production is initiated by the conversion of Phe by phenylalanine ammonia-lyase (PAL) enzyme, which acts as the key gatekeeper controlling carbon influx from the Phe pool into phenylpropanoid biosynthesis. Despite the importance of Phe as a precursor for phenylpropanoid production, we have limited knowledge of how Phe precursor availability influences downstream pathways, particularly in the context of stress adaptation. To tackle this question, Arabidopsis thaliana wild-type and production of anthocyanin pigment 1 dominant (pap1D) mutant, a line genetically enhanced for anthocyanin production, were subjected to HL treatment, followed by proteome and metabolome analyses. Our multi-omics data indicated that Phe biosynthesis was insufficiently responsive to meet the increased precursor demand from the downstream phenylpropanoid pathway, likely making Phe availability a rate-limiting factor for HL-induced phenylpropanoid production. We then generated pap1D double mutants with Phe-overaccumulating mutants. Untargeted metabolomics revealed that enhanced Phe availability had little impact on phenylpropanoid accumulation under standard growth conditions but additively promoted the accumulation of specific phenylpropanoid intermediates and anthocyanin species under HL stress. This metabolic difference between light treatment and genotypes was correlated with the activation of PAL enzymatic activity. This study demonstrates that enhanced Phe biosynthesis amplifies HL-induced phenylpropanoid biosynthesis.

plant biology↗

bioRxiv2025

Parasitism-mediated horizontal transfer of a functional cytochrome P450 gene entails transposon colonization in newly gained introns

Plants produce a wide variety of specialized metabolites that are typically present only in specific lineages. However, some specialized metabolites are found sporadically across distantly related plant species. While the latter cases have been explained as outcomes of convergent evolution, the molecular mechanism behind such metabolic evolution has remained largely elusive. Here, we report that parasitic dodders belonging to the genus Cuscuta accumulate sesamin, and that this accumulation may be attributed to the acquisition of enzymatically active homologs of Sesamum indicum CYP81Q1, which encodes piperitol/sesamin synthase (PSS). Phylogenetic analysis of CYP81Q homologs in Cuscuta and Grammica subgenera supports a trajectory in which ancestral Cuscuta species acquired CYP81Q from an ancestral host plant of Lamiales through horizontal gene transfer (HGT), and that the gene has been maintained during the speciation of Cuscuta. The evolution of the CYP81Q genes was accompanied by sequential intron gains, which likely involved the colonization of transposons. Experiments involving C. campestris and S. indicum suggested that expression of the host CYP81Q gene could be induced by parasitism, and physical connection to the host plant allowed the transfer of genetic elements to Cuscuta. These data suggest that parasitism-mediated HGT contributed to the transfer of a gene encoding a key enzyme in specialized lignan metabolism to Cuscuta, and that the acquired metabolic gene underwent structural modification while retaining its enzymatic function in dodders.

plant biology↗

bioRxiv2025

Chromosome-scale Genome Assemblies of Two Allopolyploid Cuscuta Species Uncover Genomic Signatures of Parasitic Lifestyle and Polyploid Evolution

Dodders (Cuscuta spp.) is an obligated parasitic plant, which lost a large part of photosynthetic genes but gained host genes through parasitism-mediated horizontal gene transfer (HGT). Their migratory ecology across the world would contribute to complexity of the speciation via geographic isolation. Here we report the de novo genome assemblies of the two phylogenetically distinct dodders; C. chinensis (2n=4x=60) and C. campestris (2n=4x=60) that are classified into Clade B and Clade H of subgenus Grammica, respectively. Relatively low completeness of BUSCO genes ca.87% indicated progressive gene loss after evolution of parasitic lifestyle due to release from functional constraints as photosynthesis and organ development. Comparative genomics uncovered that both species are the genome size is completely different regardless of the same cytotypes and allopolyploid through the independent ancient hybridization between different parents. Various genomic rearrangements including 1) gene gain and loss events, 2) homoeologous recombination between two subgenomes and 3) the lineage-specific proliferation of transposable elements likely contribute to their genomic diversity and sexual isolation of the two lineages partly sharing their habitats. Our findings not only provide genomic basis to survey parental species for allopolyploidization but also help to understand unique speciation of parasitic dodders through these chromosomal natures.

genomics↗

bioRxiv2024

Arabidopsis 3-deoxy-D-arabino-heptulosonate 7-phosphate (DAHP) synthases of the shikimate pathway display both manganese- and cobalt-dependent activities

The plant shikimate pathway directs a significant portion of photosynthetically assimilated carbon into the downstream biosynthetic pathways of aromatic amino acids (AAA) and aromatic natural products. 3-Deoxy-D-arabino-heptulosonate 7-phosphate synthase (DHS) catalyzes the first step of the shikimate pathway, playing a critical role in controlling the carbon flux from central carbon metabolism into the AAA biosynthesis. Previous biochemical studies suggested the presence of manganese- and cobalt-dependent DHS enzymes (DHS-Mn and DHS-Co, respectively) in various plant species. Unlike well-studied DHS-Mn, however, the identity of DHS-Co is still unknown. Here, we show that all three DHS isoforms of Arabidopsis thaliana exhibit both DHS-Mn and DHS-Co activities in vitro. A phylogenetic analysis of various DHS orthologs and related sequences showed that Arabidopsis 3-deoxy-D-manno-octulosonate-8-phosphate synthase (KDOPS) proteins were closely related to microbial Type I DHSs. Despite their sequence similarity, these Arabidopsis KDOPS proteins showed no DHS activity. Meanwhile, optimization of the DHS assay conditions led to the successful detection of DHS-Co activity from Arabidopsis DHS recombinant proteins. Compared to DHS-Mn, DHS-Co activity displayed the same redox dependency but distinct optimal pH and cofactor sensitivity. Our work provides biochemical evidence that the DHS isoforms of Arabidopsis possess DHS-Co activity.

plant biology↗

bioRxiv2024

Effect of environmental conditions on seed germination and seedling growth in Cuscuta campestris

The dodder, Cuscuta, is an obligate parasitic plant that cannot survive without a host plant and causes major damage to crop yields. To know its growth characteristics before parasitism, we examined the effect of various environmental conditions on seed germination and seedling growth in Cuscuta campestris. As for the effect of light on germination, far-red light was rather preferable to red light and the reversible response of the seeds to red and far-red light was confirmed, implicating a phytochrome-mediated signaling opposite to that in many seed plants. Among amino acids, aspartic acid and alanine had a promotive effect and histidine had an inhibitory effect on germination. We further found that, in addition to gibberellic acid, methyl jasmonate was stimulatory to germination and shoot elongation. While 2,4-D extended the viability of trichomes around the root cap, kinetin induced the formation of scale leaves on the shoot and calli at the base of the shoot and the root tip. RT-PCR experiments confirmed that expression of a putative RbcS gene for photosynthesis showed no response to light but that of a Phytochrome A homolog was increased in the dark. Our results indicate that some of the molecular mechanisms to respond to light and hormone signals are uniquely modified in dodder seedlings, providing a clue for understanding the survival strategy of parasitic plants.

plant biology↗

bioRxiv2022

Environmental pH signals the release of monosaccharides from cell wall in coral symbiotic alga

Reef-building corals thrive in oligotrophic environments due to their possession of endosymbiotic algae. Confined to the low pH interior of the symbiosome within the cell, the algal symbiont provides the coral host with photosynthetically fixed carbon. However, it remains unknown how carbon is released from the algal symbiont for uptake by the host. Here we show, using cultured symbiotic dinoflagellate, Breviolum sp., that decreases in pH directly accelerates the release of monosaccharides, i.e., glucose and galactose, into the ambient environment. Under low pH conditions, the cell surface structures were deformed and genes related to cellulase were significantly upregulated in Breviolum. Importantly, the release of monosaccharides was suppressed by the cellulase inhibitor, glucopyranoside, linking the release of carbon to degradation of the agal cell wall. Our results suggest that the low pH signals the cellulase-mediated release of monosaccharides from the algal cell wall as an environmental response in coral reef ecosystems.

evolutionary biology↗