The study functionally characterizes a conserved structured RNA motif (45ABC) in Arabidopsis RBP45 pre‑mRNAs, showing that its sequence and pairing elements mediate a negative auto‑ and cross‑regulatory feedback loop through alternative splicing that produces unproductive isoforms and reduces RBP45 expression. Transcriptome‑wide splicing analysis and phenotypic assessment of rbp45 mutants reveal that RBP45B plays a dominant role and that proper regulation of this motif is essential for root growth and flowering time.
Spatiotemporal regulation of arbuscular mycorrhizal symbiosis at cellular resolution
Authors: Chancellor, T., Ferreras-Garrucho, G., Akmakjian, G. Z., Montero, H., Bowden, S. L., Hope, M., Wallington, E., Bhattacharya, S., Korfhage, C., Bailey-Serres, J., Paszkowski, U.
The study applied dual-species spatial transcriptomics at single-cell resolution to map plant and fungal gene activity in rice roots colonized by Rhizophagus irregularis, revealing transcriptional heterogeneity among morphologically similar arbuscules. By pioneering an AM-inducible TRAP-seq using stage‑specific promoters, the authors uncovered stage‑specific reprogramming of nutrient transporters and defence genes, indicating dynamic regulation of nutrient exchange and arbuscule lifecycle.
The study identifies two diel regulatory modules that coordinate plant cuticle formation: the LRB‑phyB‑PIF4 pathway suppresses wax biosynthesis during daylight, while the COP1‑CFLAP1 pathway promotes cutin accumulation at night. Degradation of phyB and CFLAP1 via specific E3 ubiquitin ligases modulates the activity of transcription factors PIF4 and BDG1 to ensure timely cuticle assembly.
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 shows that high ambient temperature triggers extensive changes in ROS homeostasis in Arabidopsis seedlings, with H2O2 balance being essential for thermomorphogenic hypocotyl elongation. PIF4 directly activates catalase genes CAT2 and CAT3 to regulate H2O2 levels, forming a PIF4‑CAT‑H2O2 module that operates alongside the PIF4‑auxin pathway, while elevated H2O2 feeds back to reduce PIF4 protein abundance.
The study applied spatial transcriptomics to map the transcriptional landscape of wheat (Triticum aestivum) inflorescences during spikelet development, revealing two distinct regions—a RAMOSA2‑active primordium and an ALOG1‑expressing boundary. Developmental assays showed that spikelets arise from meristematic zones accompanied by vascular rachis formation, identifying key regulators that could be targeted to improve spikelet number and yield.
An optimized workflow was developed to apply the Xenium in situ sequencing platform to formalin‑fixed paraffin‑embedded (FFPE) sections of Medicago truncatula roots and nodules, incorporating customized tissue preparation, probe design, and imaging to overcome plant‑specific challenges such as cell wall autofluorescence. The protocol was validated across nodule developmental stages using both a 50‑gene panel for mature cell identity and an expanded 480‑gene panel covering multiple cell types, providing a scalable high‑resolution spatial transcriptomics method adaptable to other plant systems.
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 applied the STOmics spatial transcriptomics platform to map gene expression at subcellular resolution in developing wheat (Triticum aestivum) seeds during grain filling, analyzing over four million transcripts. Eight functional cellular groups were identified, including four distinct endosperm clusters with radial expression patterns and novel marker genes, and subgenome‑biased expression was observed among specific paralogs. These results highlight spatial transcriptomics as a powerful tool for uncovering tissue‑specific and polyploid‑specific gene regulation in seeds.
Spatial and single-cell transcriptomics capture two distinct cell states in plant immunity
Authors: Hu, Y., Schaefer, R., Rendleman, M., Slattery, A., Cramer, A., Nahiyan, A., Breitweiser, L., Shah, M., Kaehler, E., Yao, C., Bowling, A., Crow, J., May, G., Tabor, G., Thatcher, S., Uppalapati, S. R., Muppirala, U., Deschamps, S.
The study combined spatial transcriptomics and single-nuclei RNA sequencing to map soybean (Glycine max) responses to Asian soybean rust caused by Phakopsora pachyrhizi, revealing two distinct host cell states: pathogen‑occupied regions and adjacent non‑infected regions that show heightened defense gene expression. Gene co‑expression network analysis identified a key immune‑related module active in the stressed cells, highlighting a cell‑non‑autonomous defense mechanism.