Qu-1: a transformation-and regeneration-amenable doubled haploid cell line with a reference genome sequence for genetic and functional studies in Populus.

Abstract

Plant suspension homogeneous cells are invaluable materials for investigating molecular mechanisms underlying various biological processes. In this study, we established and characterized a doubled haploid cell line from Populus simonii × P. nigra, designated as Qu-1. This cell line exhibited high viability and dispersibility under suspension culture conditions and retained the ability to regenerate into whole plants. K-mer analysis confirmed the homozygous genome of Qu-1, and a chromosome-level genome assembly was subsequently achieved by using PacBio sequencing. Additionally, we established an efficient transient transformation protocol using PEG-mediated protoplasts and a stable Agrobacterium-mediated transformation system for Qu-1. To explore the mutagenesis potential of this cell line, we treated the cell line with ethyl methanesulfonate (EMS) and performed genome resequencing to identify mutation sites. Overall, Qu-1 represents the first poplar cell line with a homozygous genome and is as a powerful tool for molecular biology research in woody plants.