Advances in Tissue Culture-Free Genetic Engineering and Genome Editing of Peanut.

Plant transformation and genome editing are pivotal in advancing peanut biotechnology, yet traditional tissue culture-dependent methods are hindered by lengthy protocols, genotype dependency, and somaclonal variation. CRISPR/Cas technologies have revolutionized breeding by enabling precise, multiplex genome editing to improve traits such as disease resistance, allergen reduction, seed quality, and yield. However, variable transformation efficiencies and chimerism remain challenges. This review examines emerging tissue culture-independent techniques such as nanoparticle-based delivery, viral vectors, pollen magnetofection, pollen tube injection, node injection, and vacuum infiltration that offer rapid, cost-effective gene transfer. It also highlights the integration of high-throughput screening, robust selection markers, and automation, including robotics and advanced imaging, to refine transformation pipelines. These methodological breakthroughs promise to overcome current limitations and accelerate the development of improved peanut cultivars for sustainable agriculture.