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 study compares transcriptional, proteomic, and metabolomic responses of wild‑type Arabidopsis and a cyp71A27 mutant to a plant‑growth‑promoting Pseudomonas fluorescens strain and a pathogenic Burkholderia glumeae strain, revealing distinct reprogramming and an unexpected signaling role for the non‑canonical P450 CYP71A27. Mutant analysis showed that loss of CYP71A27 alters gene and protein regulation, especially during interaction with the PGP bacterium, while having limited impact on root metabolites and exudates.
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 surveyed vegetative water use and life‑history traits across Arabidopsis thaliana ecotypes in both controlled and outdoor environments to assess how climatic history shapes water‑use strategies. Trait‑climate correlations and genome‑wide association analyses uncovered that ecotypes from warmer regions exhibit higher water use, and identified MYB59 as a key gene whose temperature‑linked alleles affect water consumption, a finding validated using myb59 mutants. These results indicate that temperature‑driven adaptive differentiation partly explains intraspecific water‑use variation.
The study investigates how miR394 influences flowering time in Arabidopsis thaliana by combining transcriptomic profiling of mir394a mir394b double mutants with histological analysis of reporter lines. Bioinformatic analysis identified a novel lncRNA overlapping MIR394B (named MIRAST), and differential promoter activity of MIR394A and MIR394B suggests miR394 fine‑tunes flower development through transcription factor and chromatin remodeler regulation.