Background Development of precise genome editing strategies is a prerequisite for producing edited plants that can aid in the study of gene function and help understand the genetic traits in a cultivar. Citrus embryogenic cell cultures can be used to rapidly produce a large population of genome edited transformed citrus lines. The ability to introduce specific mutations in the genome of these cells using two constructs (pC-PDS1 and pC-PDS2) was evaluated in this study. Results Citrus sinensis ‘EV2’ embryogenic cell cultures are amenable to Agrobacterium-mediated CRISPR/Cas9-based genome editing. Guide RNAs (gRNAs) targeting two locations in the phytoene desaturase (PDS) gene were either driven by the Arabidopsis U6–26 promoter (pC-PDS1) or assembled as a Csy4 array under the control of the CmYLCV promoter (pC-PDS2). All transgenic embryos were completely albino and no variegated phenotype was observed. We evaluated 12 lines from each construct in this study and the majority contain either insertion (1–2 bp), substitution (1 bp), or deletion (1–3 bp) mutations that occurred close to the protospacer adjacent motif. Conclusions Both the pC-PDS1 and pC-PDS2 could successfully edit the citrus embryogenic cell cultures. However, the editing efficiency was dependent on the gRNA, confirming that the selection of a proper gRNA is essential for successful genome editing using the CRISPR/Cas9 technique. Also, utilization of embryogenic cell cultures offers another option for successful genome editing in citrus.

Citrus cultivation is challenging due to the plethora of abiotic and biotic stresses faced by the crop. In recent years, production has been severely affected by diseases such as citrus canker and huanglongbing (HLB). Disease management is hampered as there is no field resistance to these diseases in any of the important commercially planted varieties. Traditionally, conventional breeding approaches have been applied for the improvement of the susceptible cultivars; however, this technique is laborious and time consuming. Genetic transformation of citrus allows for the rapid integration of novel genes into the plant's genome to develop disease-resistant transgenic plants. Therefore, efforts have been made to utilize genetic engineering tools to develop genetically modified citrus that are resistant to citrus canker and HLB. This review summarizes the major achievements made in the development of citrus canker and HLB tolerance using transgenic technologies.

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