biostudies-arrayexpressChristine FaulknerArabidopsis thalianahttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-16776We generated two independent genetic tools that can induce callose deposition and plasmodesmal closure following estradiol treatment: LexA::Icals3m and LexA::PLUG. We harvested mature leaf tissue at 12h, 24h, 48h and 72h post induction and extracted total RNAbiostudies-arrayexpressSample Collection - Leaf material was collected from 5 week old plants and frozen in liquid nitrogen following dissection.Nucleic Acid Extraction - Total RNA was extracted from leaf tissue using the Qiagen Plant RNeasy mini kit according to the manufacturers instructions. Samples were given rigorous treatment with Turbo DNase was undertaken to remove genomic DNA contamination.Sample Treatment - Leaves were painted with either a mock treatment (water + 0.001% DMSO) or estradiol (20uM + 0.001% DMSO).Library Construction - Library construction was outsourced to Novogene. Libraries were constructed for paired end reads, using oligo-dT enrichment.Sequencing - Library sequencing was outsourced. Libraries were sequenced by Novogene using RNA-seq using Illumina NovaSeq 6000.Growth Protocol - Arabidopsis thaliana plants were grown on MS plates for 5 weeks at 10 h light and 14h dark at 22 °C.OrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesSequence Alignment - Library quality was accessed with Fastqc v0.11.8. Libraries were trimmed using Trim_galore v0.5.0 and Cutadapt v1.7. Libraries were aligned to the Arabidopsis genome using Hisat2 v2.1.0.Data Transformation - SAMtools was used to convert files for input into StringTie, which was used calculate read counts and create a matrix of counts. Raw read counts were used in differential gene expression analysis using DESeq2 (v.1.38.346) in R studio (v.4.3.3).MetabolomicsUnknownTranscriptomicsGenomicsProteomicsIllumina NovaSeq 6000Plant cells are connected to their neighbors via plasmodesmata facilitating the exchange of nutrients and signaling molecules. During immune responses, plasmodesmata close, but how this contributes towards a full immune response is unknown. To investigate this, we developed two transgenic lines with which we could induce plasmodesmal closure independently of immune elicitors, using the over-active CALLOSE SYNTHASE3 allele icals3m and the C-terminus of PDLP1 to drive callose deposition at plasmodesmata. Induction of plasmodesmal closure increased the expression of stress responsive genes, salicylic acid accumulation and resistance to Pseudomonas syringae DC3000. More homogeneous plasmodesmal closure using icals3m also led to the accumulation of starch and sugars, decreased leaf growth, as well as hypersusceptibility to Botrytis cinerea . Based on the profile of responses, we conclude that plasmodesmal closure itself activates stress signaling, raising questions of what signals mediate this response and whether these responses occur in all circumstances when plasmodesmata close.RNA-seq of coding RNAArabidopsis thalianaPlasmodesmatal closure elicits stress responsesEstee E. Tee, Andrew Breakspear, Diana Papp, Hannah R. Thomas, Catherine Walker, Annalisa Bellandi, Christine FaulknerChristine FaulknerfalsePlasmodesmal closure triggers stress responsesWe generated two independent genetic tools that can induce callose deposition and plasmodesmal closure following estradiol treatment: LexA::Icals3m and LexA::PLUG. We harvested mature leaf tissue at 12h, 24h, 48h and 72h post induction and extracted total RNA2026-03-31T00:00:00Z2026-03-31T01:03:46.768Z2026-03-18T16:01:18.683ZE-MTAB-16776ERP190971EFO_0002944EFO_0004170EFO_0003789EFO_0004917EFO_0005518EFO_0003816EFO_0003738EFO_0004184EFO_000396910.1101/2024.05.08.593115