Establishment of a High-Efficiency Hairy Root Transformation System for Forsythia suspensa and Enhancement of Transformation Efficiency by Developmental Regulators
This project encompasses sequencing data from a comprehensive study to establish and optimize a genetic transformation system for hairy roots in the medicinal plant Forsythia suspensa. Part I: Vector Construction The study employed a suite of engineered plant binary vectors designed for Agrobacterium-mediated transformation, all featuring the RUBY visual reporter gene (producing anthocyanin pigment). Key constructs include: 1. pCAMBIA1300-RUBY series: Derived from pCAMBIA1301-35S-EGFP by replacing the EGFP gene with RUBY. Variants were created by inserting different developmental regulator (DR) genes to drive tissue-specific expression. 2. pHSE401-RUBY: Constructed by inserting the RUBY gene into the binary vector pHSE401. 3. pHSE401-RUBY-FsMPsgRNA1-2: A CRISPR/Cas9 vector derived from pHSE401-RUBY, carrying the FsMP-sgRNA1-2 cassette for targeted genome editing. All plasmid constructs were sequenced (e.g., by Illumina MiSeq) to verify their integrity. Part II: Functional Validation in Transgenic Hairy Roots The project includes sequencing data generated to validate genetic modifications in transgenic Forsythia suspensa hairy roots: 1. Hi-TOM sequencing data: Targeted deep sequencing of edited genomic regions for precise detection of CRISPR/Cas9-induced mutations and editing efficiency analysis. 2. Sanger sequencing data: Validation of specific mutations and transgenic integration events in selected hairy root clones. 3. Vector sequencing data: Confirmation of T-DNA regions and expression cassettes used in the transformations. These integrated datasets document the entire workflow from vector engineering to functional assessment, supporting the findings in the associated manuscript titled "Establishment of a High-Efficiency Hairy Root Transformation System for Forsythia suspensa and Enhancement of Transformation Efficiency by Developmental Regulators".