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 engineers Type‑B response regulators to alter their transcriptional activity and cytokinin sensitivity, enabling precise modulation of cytokinin‑dependent traits. Using tissue‑specific promoters, the synthetic transcription factors were deployed in Arabidopsis thaliana to reliably increase or decrease lateral root numbers, demonstrating a modular platform for controlling developmental phenotypes.
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.
Imputation integrates single-cell and spatial gene expression data to resolve transcriptional networks in barley shoot meristem development
Authors: Demesa-Arevalo, E., Dorpholz, H., Vardanega, I., Maika, J. E., Pineda-Valentino, I., Eggels, S., Lautwein, T., Kohrer, K., Schnurbusch, T., von Korff, M., Usadel, B., Simon, R.
The study uses an imputation strategy that integrates deep single-cell RNA sequencing with spatial gene expression data to map transcriptional dynamics across barley inflorescence development at cellular resolution. By leveraging the BARVISTA web interface, the authors identify key transcriptional events in meristem founder cells, characterize complex branching mutants, and reconstruct spatio‑temporal trajectories of flower organogenesis, offering insights for targeted trait manipulation.
The study examines how the SnRK1 catalytic subunit KIN10 integrates carbon availability with root growth regulation in Arabidopsis thaliana. Loss of KIN10 reduces glucose‑induced inhibition of root elongation and triggers widespread transcriptional reprogramming of metabolic and hormonal pathways, notably affecting auxin and jasmonate signaling under sucrose supplementation. These findings highlight KIN10 as a central hub linking energy status to developmental and environmental cues in roots.
The study characterizes the tomato class B heat shock factor SlHSFB3a, revealing its age‑dependent expression in roots and its role in enhancing lateral root density by modulating auxin homeostasis. Overexpression of SlHSFB3a increases lateral root emergence, while CRISPR‑mediated knockouts produce the opposite phenotype, indicating that SlHSFB3a regulates auxin signaling through repression of auxin repressors and activation of the ARF7/LOB20 pathway.
A forward genetic screen in light-grown Arabidopsis seedlings identified the Evening Complex component ELF3 as a key inhibitor of phototropic hypocotyl bending under high red:far-red and blue light, acting upstream of PIF4/PIF5. ELF3 and its partner LUX also mediate circadian regulation of phototropism, and the orthologous ELF3 in Brachypodium distachyon influences phototropism in the opposite direction.
The study investigates the altered timing of the core circadian oscillator gene ELF3 in wheat compared to Arabidopsis, revealing that dawn-specific expression in wheat arises from repression by TOC1. An optimized computational model integrating experimental expression data and promoter architecture predicts that wheat’s circadian oscillator remains robust despite this shift, indicating flexibility in plant circadian network design.
The study tests whether the circadian clock component ELF3 shapes developmental trait heterogeneity, proposing that faster‑developing populations are more heterogeneous early but less so at maturity, whereas slower growers show the opposite pattern. Experiments with Arabidopsis elf3 and barley Hvelf3 mutants confirmed these predictions, showing ELF3 influences hypocotyl and bolting variability via maturation rate, and that smaller barley plants exhibit increased osmotic stress resilience, suggesting ELF3‑driven heterogeneity serves as a bet‑hedging strategy.