Field Crop 2025, Vol.8, No.2, 72-81 http://cropscipublisher.com/index.php/fc 76 greatly reduced. These two fatty acids are not very stable and are prone to problems during processing. So overall, the edited oil is indeed more stable and of better quality. 4.2 Comparison of CRISPR-modified rapeseed with conventional high-oleic varieties The high-oleic rapeseed on the market now has better performance after being modified by CRISPR technology. After the oleic acid content is increased, the antioxidant property of the oil is significantly enhanced, which is particularly suitable for use in the industrial field (Jiang et al., 2017). However, traditional varieties also have their own advantages, such as being more stable in certain processing techniques. From a health perspective, the cardiovascular benefits of this type of modified oil have been confirmed (Shi et al., 2022), and after the PUFA and erucic acid content is reduced, it is more convenient for both consumption and industrial application (Liu et al., 2022; Sandgrind et al., 2023). In terms of economic benefits, this type of rapeseed variety is indeed very promising. Due to the improved quality, the market price may be higher (Shi et al., 2022). The processing link can also save a lot of costs. After all, the unnecessary fatty acids are reduced, and the refining efficiency is naturally improved (Jiang et al., 2017). The environmental advantages are also obvious. Compared with traditional breeding methods, CRISPR technology is cleaner and safer (Tian et al., 2022). Although some people were concerned about the accuracy of gene editing in the early days, subsequent studies have shown that the risk of off-target effects can be completely controlled (Zhang et al., 2019; Karunarathna et al., 2020). Overall, this technology has indeed brought new opportunities to rapeseed breeding. Not only has the quality of oil products been significantly improved, but the entire industry chain from planting to processing can benefit. As market demand changes, this precisely improved rapeseed variety should become more and more popular. Figure 3 The general workflow of CRISPR/Cas9-based gene editing in rapeseed (Adopted from Tian et al., 2022) Image caption: (A) Select genes for editing according to target traits. The traits that B. napus genes edited by CRISPR/Cas9 are involved in are shown here; (B) perform single or multiple copy knockout according to function differentiation-related information among homologous copies; (C) design sgRNAs for selected target genes using online and offline tools (sgRNAs are usually designed in exon or functional domain, which can lead to a greater probability of functional mutation); (D) CRISPR/Cas9 vector with the sgRNA coding sequence construction and transformation; (E) select genetic transformation object (the representative receptor materials successfully used for transformation for CRISPR/Cas9 vector are shown on the right); (F) deliver CRISPR/Cas9 vector by co-culturing Agrobacterium with transfection object in the culture medium; (G,H) differentiate callus and regenerate plants by inducing reagents in the culture medium, and identify positive plants by resistance or fluorescence; (I) identify target and/or off-target editing by (a) polymerase chain reaction restricted enzyme (PCR-RE) and/or (b) sequencing; (J) build target gene-edited rapeseed lines for phenotyping (Adopted from Tian et al., 2022)
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