24/10/2025
Vol12, Iss9 The global citrus industry is threatened by biotic and abiotic stresses, particularly Huanglongbing (HLB), prompting extensive research efforts to mitigate these challenges. Recent progress in biotechnology offers promising solutions. Transgene-free gene editing, in particular, presents a viable alternative to conventional breeding. This study introduces an optimized Agrobacterium-mediated transient expression system for Cas9 and gRNAs, incorporating treatments on Agrobacterium and citrus explants along with strategic chemical applications to significantly improve transgene-free gene editing efficiency. Through technologies such as gene editing and molecular breeding, researchers are able to develop superior varieties with enhanced disease resistance, higher yields, and broader adaptability, providing robust scientific and technological support for the sustainable development of citrus industry. Moreover, this methodology holds potential for extension to other economically significant heterozygous perennial crops, providing a versatile tool for crop improvement. https://doi.org/10.1093/hr/uhaf153
