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.
The study introduces a CRISPR/Cas9‑based restoration system (CiRBS) that reactivates a disabled luciferase reporter (LUC40Ins26bp) in transgenic Arabidopsis, enabling long‑term single‑cell bioluminescence monitoring. Restoration occurs within 24 h after particle‑bombardment‑mediated CRISPR delivery, with ~7 % of cells regaining luminescence and most restored cells carrying a single correctly edited chromosome, facilitating reliable analysis of cellular gene‑expression heterogeneity.
The study used CRISPR/Cas9 to create rice lines with one to three tandem copies of the OsMADS18 gene and confirmed copy-number through high‑throughput qPCR. Incremental increases in OsMADS18 copy number produced proportional rises in transcript levels and corresponding enhancements in leaf blade and culm length, showing that gene dosage can be leveraged to fine‑tune agronomic traits.
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 authors adapted OpenPlant kit CRISPR/Cas9 tools to enable multiplex gRNA expression from a single transcript using tRNA sequences in the liverwort Marchantia polymorpha, markedly enhancing editing efficiency and scalability. They coupled this vector system with a simplified, optimized thallus transformation protocol, providing a rapid and versatile platform for generating CRISPR/Cas9 mutants and advancing functional genomics in this model species.
The study investigated how molecular factors determine the developmental fate of axillary buds (AXB) in Fragaria vesca, distinguishing between stolon formation and branch crown development. By combining phenotypic analysis of AXB development with RNA‑seq of undifferentiated buds across three genotypes, the authors identified FveBRC1 as a key regulator, and confirmed its role using CRISPR/Cas9‑generated brc1 mutants. These findings enhance understanding of AXB fate control and its impact on strawberry fruit yield.
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 demonstrates the implementation of CRISPR/Cas9-mediated targeted mutagenesis in the orphan crop grain amaranth (Amaranthus hypochondriacus) by editing genes of the betalain biosynthesis pathway using the CasCADE modular cloning system. It addresses the bottleneck of lacking efficient stable transformation and regeneration protocols for non‑model crops, providing a reproducible workflow for climate‑resilient breeding.