Journal of Tea Science Research, 2024, Vol.14, No.3, 160-168 http://hortherbpublisher.com/index.php/jtsr 163 pathways. This can lead to a more robust and rapid response to pathogen attacks (Veillet et al., 2020; Zaidi et al., 2020; Wang et al., 2022). Furthermore, CRISPR/Cas9 can be employed in a way that does not leave any foreign DNA in the plant genome, addressing public concerns about genetically modified organisms (GMOs). This is particularly important for crops like tea, which are consumed directly (Chandrasekaran et al., 2016; Chen et al., 2019). 4.3 Case studies: successful implementations Some case studies have demonstrated the successful application of CRISPR/Cas9 in cultivating disease-resistant plants, serving as models for tea plant improvement. Researchers used CRISPR/Cas9 technology to target the eIF4E gene, developing virus-resistant cucumber plants. The resulting plants exhibited immunity to multiple viruses without any transgenic elements, highlighting the potential for similar strategies in tea plants (Figure 2) (Chandrasekaran et al., 2016). CRISPR/Cas9 has also been used to confer resistance to fungal pathogens in crops such as rice and tomatoes by editing susceptibility genes. These improvements significantly reduced disease incidence and increased crop yields (Figure 3) (Borrelli et al., 2018; Paul et al., 2021). Figure 2 Gene editing of eIF4E mediated by CRISPR/Cas9 in transgenic cucumber plants (Adopted from Chandrasekaran et al., 2016) Image caption: (A) Schematic representation of the cucumber eIF4E genomic map and the sgRNA1 and sgRNA2 target sites (red arrows). The target sequence is shown in red letters together with the restriction site (underlined), and the protospacer adjacent motif (PAM) is marked in bold underlined letters. The black arrows indicate the primers flanking the target sites used to detect the mutations. (B) Restriction analysis of T0 polymerase chain reaction (PCR) fragments of CEC-1, CEC1-4 and CEC2-5. (C) Alignment of nine colony sequences from the undigested fragment of line 1 with the wild-type (wt) genome sequence. DNA deletions are shown by red dashes and deletion sizes (nucleotides) are marked on the right side of the sequence (Adopted from Chandrasekaran et al., 2016)
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