Evaluation of combined root exudate and rhizosphere microbiota sampling approaches to elucidate plant-soil-microbe interaction
Authors: Escudero-Martinez, C., Browne, E. Y., Schwalm, H., Santangeli, M., Brown, M., Brown, L., Roberts, D. M., Duff, A. M., Morris, J., Hedley, P. E., Thorpe, P., Abbott, J. C., Brennan, F., Bulgarelli, D., George, T. S., Oburger, E.
The study benchmarked several sampling approaches for simultaneous profiling of root exudates and rhizosphere microbiota in soil-grown barley, revealing consistent exudate chemistry across methods but variation in root morphology and nitrogen exudation. High‑throughput amplicon sequencing and quantitative PCR showed protocol‑specific impacts on microbial composition, yet most rhizosphere-enriched microbes were captured by all approaches. The authors conclude that different protocols provide comparable integrated data, though methodological differences must be aligned with experimental objectives.
A large-scale proteomic study in Arabidopsis thaliana identified over 32,000 isoform-specific peptides, confirming that alternative splicing, particularly intron retention, produces translated protein isoforms. Integrated proteogenomic analysis, SUPPA classification, and AlphaFold modeling revealed structural impacts and a non-linear regulation of transcript and protein abundance, with mutant phenotypes linking splicing to growth, chlorophyll content, and anthocyanin accumulation.
The study profiled root transcriptomes of Arabidopsis wild type and etr1 gain-of-function (etr1-3) and loss-of-function (etr1-7) mutants under ethylene or ACC treatment, identifying 4,522 ethylene‑responsive transcripts, including 553 that depend on ETR1 activity. ETR1‑dependent genes encompassed ethylene biosynthesis enzymes (ACO2, ACO3) and transcription factors, whose expression was further examined in an ein3eil1 background, revealing that both ETR1 and EIN3/EIL1 pathways regulate parts of the network controlling root hair proliferation and lateral root formation.
The study shows that the SnRK1 catalytic subunit KIN10 directs tissue-specific growth‑defense programs in Arabidopsis thaliana by reshaping transcriptomes. kin10 knockout mutants exhibit altered root transcription, reduced root growth, and weakened defense against Pseudomonas syringae, whereas KIN10 overexpression activates shoot defense pathways, increasing ROS and salicylic acid signaling at the cost of growth.
The study used phospho‑proteomics to uncover rapid phosphorylation changes in Arabidopsis seedlings upon light or sucrose exposure, identifying RS41 as a hyperphosphorylated SR protein. By creating single and higher‑order mutants of four RS genes, the authors demonstrated that these RS proteins are essential for photomorphogenic development and regulate light‑dependent alternative splicing, with loss of all four causing sterility.
The study investigated how Arabidopsis thaliana SR protein kinases (AtSRPKs) regulate alternative RNA splicing by using chemical inhibitors of SRPK activity. Inhibition with SPHINX31 and SRPIN340 caused reduced root growth and loss of root hairs, accompanied by widespread changes in splicing and phosphorylation of genes linked to root development and other cellular processes. Multi‑omics analysis (transcriptomics and phosphoproteomics) revealed that AtSRPKs modulate diverse splicing factors and affect the splicing landscape of numerous pathways.
The study used TurboID-based proximity labeling coupled with mass spectrometry to map the Arabidopsis alternative splicing machinery centered on ACINUS, PININ, and SR45, identifying 298 high-confidence components and revealing that splicing is tightly linked to transcription and other RNA processing steps. Bioinformatic and genetic analyses, including O-glycosylation double mutants, demonstrated both conserved and plant‑specific regulatory networks and highlighted the role of sugar modifications in modulating splicing.
The study investigates the role of the Arabidopsis transcription factor AtMYB93 in sulfur (S) signaling and root development, revealing that AtMYB93 mutants exhibit altered expression of S transport and metabolism genes and increased shoot S levels, while tomato plants overexpressing SlMYB93 show reduced shoot S. Transcriptomic profiling, elemental analysis, and promoter activity assays indicate that AtMYB93 contributes to root responses to S deprivation, though functional redundancy masks clear phenotypic effects on lateral and adventitious root formation.
The study investigates the Arabidopsis ribosomal protein RPS6A and its role in auxin‑related root growth, revealing that rps6a mutants display shortened primary roots, fewer lateral roots, and defective vasculature that are not rescued by exogenous auxin. Cell biological observations and global transcriptome profiling show weakened auxin signaling and reduced levels of PIN auxin transporters in the mutant, indicating a non‑canonical function of the ribosomal subunit in auxin pathways.
The study demonstrates that abscisic acid (ABA) accumulates in darkness to suppress cotyledon opening during seedling deetiolation, and that light exposure lifts this repression, enabling cotyledon aperture. Genome‑wide transcriptional and alternative‑splicing changes accompany this process, and the light‑dependent regulation requires the splicing factors RS40 and RS41, whose activity is repressed in the dark.