Přehled o publikaci

Large-scale chromosome rearrangements, including deletions, inversions, and reciprocal translocations, have long been recognized as critical events in plant genome evolution and speciation. In era of CRISPR/Cas, the capacity to engineer such rearrangements provides unparalleled opportunities to investigate the genetic and phenotypic consequences of these genetic alternations. This study utilized CRISPR/Cas technology to induce large chromosome rearrangements in the genome of Cardamine hirsuta (Brassicaceae; n = 8), a model organism widely employed for investigating developmental processes, ecological interactions, and evolutionary adaptations. Agrobacterium-mediated floral dip transformation was utilized, with a transfer vector carrying SaCas9 under the control of an egg-cell-specific promoter, along with two highly specific gRNAs designed to target specific loci for chromosome rearrangement induction. PCR analysis, Sanger sequencing, and chromosome painting confirmed the presence of a 3-kb deletion and inversion on chromosome Ch8, as well as 0.1 to 2-Mb reciprocal translocation between chromosomes Ch6 and Ch8. Importantly, the heritability of these engineered chromosome rearrangements was confirmed across subsequent generations (T2 and T3). These findings highlight the potency of CRISPR/Cas tools in manipulating chromosomal structures in C. hirsuta genome, paving the way for future experiments aimed at reshaping crucifer karyotype structure and elucidating the profound consequences of such rearrangements.