Molecular Plant Breeding 2025, Vol.16, No.3, 156-164 http://genbreedpublisher.com/index.php/mpb 160 5.2 Targeting disease resistance genes Through CRISPR/Cas9 editing, genes that play a key role in pathogen recognition and defense responses can enhance the disease resistance of sugarcane. Editing the resistance (R) gene can enhance the recognition ability of sugarcane against the effector of pathogenic bacteria, thereby strengthening its immune response ability. This method. CRISPR/Cas9 can edit the R gene and is also applicable to other crops, which can endow crops with resistance to various pathogens (Arora and Narula, 2017; Krishna et al., 2023). Researchers have utilized CRISPR/Cas9 to target the specific gene, the R gene, in sugarcane that is involved in pathogen recognition and defense responses, in order to enhance the susceptibility of sugarcane to diseases. As a result, the frequency of sugarcane being infected with diseases will be reduced, allowing farmers to use less spray and grow healthier sugarcane. This gene editing technology helps control other crops such as beans and rice, enhancing the resistance of these plants to bacteria, fungi and viruses (Wang et al., 2021; Kumar et al., 2024). The precise editing of these genes has made the new sugarcane varieties more stress-resistant and have higher yields. 5.3 Edit the stress response pathways CRISPR/Cas9 can help sugarcane cope with environmental stress during growth by modifying the genes that control the stress response. These genes, transcription factors and kinases play an important role in the process by which plants cope with adverse conditions. For example, by editing the genes in the ABA signaling pathway, CRISPR/Cas9 can enhance the water-saving capacity of sugarcane, thereby enhancing the drought resistance of sugarcane (Farhat et al., 2019; Ahmar et al., 2023). Similarly, the genes in the salicylic acid (SA) and jasmonic acid (JA) pathways were edited to enhance the disease and pest resistance of sugarcane (Kumar et al., 2023). CRISPR/Cas9 can simultaneously edit multiple genes and is also more likely to regulate complex stress response systems. Sugarcane is polyploid, and each gene in sugarcane can have multiple copies. CRISPR can target multiple genes simultaneously, which is particularly beneficial for sugarcane. CRISPR/Cas9 modifying different genes of the same pathway can more effectively enhance the stress resistance of sugarcane, enabling it to grow stronger and have a higher yield under harsh conditions, and also laying the foundation for cultivating better sugarcane varieties (Hussin et al., 2022; Mir et al., 2022). 6 Challenges and Future Perspectives 6.1 Complexity and off-target issues of the sugarcane genome The genome of sugarcane is very complex because it is polyploid. Polyploid leads to multiple copies of genes, so it is difficult to achieve targeted modifications without affecting the copies of other genes, and it may accidentally change the wrong part of the DNA. When using CRISPR/Cas9, it may also affect areas that should not be changed. These unwanted changes may bring about new problems or reduce the ability of plants to adapt to the environment (Mohan, 2016; Oz et al., 2021; Hussin et al., 2022). Researchers are also striving to improve the CRISPR/Cas9 system, studying a newer and more precise version of the Cas9 enzyme and designing better guide RNA to help the system locate the correct position (Oz et al., 2021; Hussin et al., 2022). Combining multi-omics tools such as genomics, transcriptomics and proteomics can help researchers better understand the genome of pitaya, which is conducive to selecting better target genes and conducting more precise gene editing (Figure 2) (Oz et al., 2021; Hussin et al., 2022). 6.2 Management challenges and public acceptance Sugarcane is one of the genome-edited crops. The main challenge in genome-edited crops is the relevant laws and regulations. Many countries have strict management regulations on genetically modified organisms (Gmos), and genome-edited plants usually also face scrutiny. The presence of genome editing in plants has raised concerns among regulatory agencies, leading to restrictions or delays in the application of some technologies in agriculture (Krishna et al., 2023; Kumar et al., 2024). Some genome editing methods, such as ribonucleoprotein technology, do not introduce exogenous DNA into plants, which can reduce some regulatory challenges (Arora and Narula, 2017; Krishna et al., 2023).
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