Arabidopsis lines with modified ascorbate concentrations reveal a link between ascorbate and auxin biosynthesis
Authors: Fenech, M., Zulian, V., Moya-Cuevas, J., Arnaud, D., Morilla, I., Smirnoff, N., Botella, M. A., Stepanova, A. N., Alonso, J. M., Martin-Pizarro, C., Amorim-Silva, V.
The study used Arabidopsis thaliana mutants with low (vtc2, vtc4) and high (vtc2/OE-VTC2) ascorbate levels to examine how ascorbate concentration affects gene expression and cellular homeostasis. Transcriptomic analysis revealed that altered ascorbate levels modulate defense and stress pathways, and that TAA1/TAR2‑mediated auxin biosynthesis is required for coping with elevated ascorbate in a light‑dependent manner.
The authors adapted OpenPlant kit CRISPR/Cas9 tools to enable multiplex gRNA expression from a single transcript using tRNA sequences in the liverwort Marchantia polymorpha, markedly enhancing editing efficiency and scalability. They coupled this vector system with a simplified, optimized thallus transformation protocol, providing a rapid and versatile platform for generating CRISPR/Cas9 mutants and advancing functional genomics in this model species.
The study used comparative transcriptomics across Erysimum species to identify two 2‑oxoglutarate‑dependent dioxygenases, CARD5 and CARD6, responsible for the 14β‑ and 21‑hydroxylation steps in cardenolide biosynthesis in Erysimum cheiranthoides. Knockout mutants lacking these genes accumulated pathway intermediates, and transient expression in Nicotiana benthamiana confirmed their enzymatic functions, while structural modeling pinpointed residues linked to neofunctionalization.
Comparative transcriptomics uncovers plant and fungal genetic determinants of mycorrhizal compatibility
Authors: Marques-Galvez, J. E., de Freitas Pereira, M., Nehls, U., Ruytinx, J., Barry, K., Peter, M., Martin, F., Grigoriev, I. V., Veneault-Fourrey, C., Kohler, A.
The study used comparative and de‑novo transcriptomic analyses in poplar to uncover plant and fungal gene regulons that govern ectomycorrhizal (ECM) compatibility, distinguishing general fungal‑sensing responses from ECM‑specific pathways. Key findings include modulation of jasmonic acid‑related defenses, coordinated regulation of secretory and cell‑wall remodeling genes, and dynamic expression of the Common Symbiosis Pathway during early and mature symbiosis stages.
High radiosensitivity in the conifer Norway spruce (Picea abies) due to lesscomprehensive mobilisation of protection and repair responses compared to the radiotolerant Arabidopsis thaliana
Authors: Bhattacharjee, P., Blagojevic, D., Lee, Y., Gillard, G. B., Gronvold, L., Hvidsten, T. R., Sandve, S. R., Lind, O. C., Salbu, B., Brede, D. A., Olsen, J. E.
The study compared early protective, repair, and stress responses to chronic gamma irradiation in the radiosensitive conifer Norway spruce (Picea abies) and the radiotolerant Arabidopsis thaliana. Norway spruce exhibited growth inhibition, mitochondrial damage, and higher DNA damage at low dose rates, while Arabidopsis maintained growth, showed minimal organelle damage, and activated DNA repair and antioxidant genes even at the lowest dose rates. Transcriptomic analysis revealed that the tolerant species mounts a robust transcriptional response at low doses, whereas the sensitive species only responds at much higher doses.
The study used comparative transcriptomics to examine how Fusarium oxysporum isolates with different lifestyles on angiosperms regulate effector genes during infection of the non‑vascular liverwort Marchantia polymorpha. Core effector genes on fast core chromosomes are actively expressed in the bryophyte host, while lineage‑specific effectors linked to angiosperm pathogenicity are silent, and disruption of a compatibility‑associated core effector alters the expression of other core effectors, highlighting conserved fungal gene networks across plant lineages.
The study investigated how molecular factors determine the developmental fate of axillary buds (AXB) in Fragaria vesca, distinguishing between stolon formation and branch crown development. By combining phenotypic analysis of AXB development with RNA‑seq of undifferentiated buds across three genotypes, the authors identified FveBRC1 as a key regulator, and confirmed its role using CRISPR/Cas9‑generated brc1 mutants. These findings enhance understanding of AXB fate control and its impact on strawberry fruit yield.
The study applied a CRISPR/Cas9 multiplex guide RNA strategy to delete entire open reading frames of four reproductive genes in Arabidopsis thaliana, achieving homozygous deletions already in the T1 generation with rates of 8.3–30%. Deletion efficiencies correlated with DeepSpCas9 prediction scores, and phenotypic analyses revealed unexpected effects of residual gene fragments on fertilization and seed development.
The study generated a high-quality genome assembly for Victoria cruziana and used comparative transcriptomics to identify anthocyanin biosynthesis genes and their transcriptional regulators that are differentially expressed between white and light pinkish flower stages. Differential expression of structural genes (VcrF3H, VcrF35H, VcrDFR, VcrANS, VcrarGST) and transcription factors (VcrMYB123, VcrMYB-SG6_a, VcrMYB-SG6_b, VcrTT8, VcrTTG1) correlates with the observed flower color change.
The study demonstrates the implementation of CRISPR/Cas9-mediated targeted mutagenesis in the orphan crop grain amaranth (Amaranthus hypochondriacus) by editing genes of the betalain biosynthesis pathway using the CasCADE modular cloning system. It addresses the bottleneck of lacking efficient stable transformation and regeneration protocols for non‑model crops, providing a reproducible workflow for climate‑resilient breeding.