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 combined ecometabolomics of root exudates with fungal community profiling to assess how abiotic (soil moisture, temperature legacy) and biotic (microbial inoculum, plant density) treatments shape metabolite diversity and fungal assemblages in Guarea guidonia seedlings. While soil microbial legacy and moisture drove metabolite diversity, antimicrobial treatments altered metabolite composition, and fungal community structure was linked to metabolite profiles, revealing metabolite‑fungal associations as early indicators of plant response to disturbance.
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 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 plant roots promote water infiltration through dry soil layers using dye tracing in model soil microcosms. Results indicate that dissolved root exudates, possibly by altering surface tension, are the primary drivers of infiltration, with root architecture also contributing. These insights suggest that root traits influencing exudation and structure could improve drought resistance in crops.
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 examined how altering ethylene biosynthesis (ACO1) or perception (etr1.1) in a hybrid poplar (P. tremula × P. tremuloides T89) influences the assembly of root and shoot fungal and bacterial communities, using amplicon sequencing and confocal microscopy. Ethylene modulation had limited impact on the sterile plant metabolome but triggered distinct primary and secondary metabolic changes in microbe‑colonized plants, correlating with reduced fungal colonisation of shoots and increased root fungal colonisation, while arbuscular mycorrhizal fungi and bacterial communities were largely unchanged.
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.