Rice Genomics and Genetics 2024, Vol.15, No.4, 164-177 http://cropscipublisher.com/index.php/rgg 173 QTL-seq approach has been used to identify novel QTLs controlling grain weight and nutrient content, which can be exploited for biofortification (Bommisetty et al., 2020). Genomic tools have also been employed to improve grain quality traits, such as texture, flavor, and processing quality. Studies have utilized next-generation sequencing and GWAS to uncover genetic loci associated with these traits. The integration of high-throughput SNP analysis and genomic selection has further refined the breeding of rice varieties with superior grain quality (Xu et al., 2021). 7 Future Prospects and Challenges 7.1 Emerging genomic technologies The field of Oryza genomics is rapidly evolving with the advent of new genomic technologies. Emerging technologies such as CRISPR/Cas9 and its variants (e.g., CRISPR/Cpf1 and base editors) have revolutionized the way we can edit the rice genome, enabling precise and efficient modifications for crop improvement. These technologies hold great promise for accelerating breeding programs by introducing desirable traits such as disease resistance, stress tolerance, and improved nutritional quality (Mishra et al., 2018). Another significant development is the construction of high-quality reference genomes and pan-genomes, which provide a comprehensive understanding of genetic diversity and facilitate the identification of novel genes for crop improvement (Huang et al., 2021). Figure 5 Agrobacterium-mediated transformation of CLCWR by using scutellum tissue of embryos in mature seeds (Adopted from Xiang et al., 2022) Image caption: A: Embryogenic calli induced on callus induction medium containing 2.5 mg/L 2,4-D for 10 days. (B) Calli subcultured on callus induction medium containing 2.5 mg/L 2,4-D for 12 days; C: Subcultured calli co-cultivated with Agrobacterium on co-cultivation medium for 3 days; D: Infected calli were screened by Hygromycin. The co-cultured calli were transferred to the selection medium supplemented with 2.5 mg/L 2,4-D, 30 mg/L Hygromycin B, and 400 mg/L carbenicillin for 3 rounds of 15 days each; E: Shoot regeneration from transformed calli. The Hygromycin-resistant calli were regenerated on the regeneration medium containing 2 mg/L ZT and 0.1 mg/L NAA for 30 days; F: Roots were induced on the root-induction medium containing 0.135 mg/L NAA for 14 days; G: Transgenic plant after transplanting to the soil for 3 months. (H-L) GUS staining of transgenic calli and plant. The calli co-cultivated with Agrobacterium; H: selected by Hygromycin B (I) and regenerated (J), also the leaves (K) and roots (L) of the transgenic plant (left was non-transgenic and right was transgenic) were stained using GUS staining solution and observed under a stereomicroscope. Scale bars, 1 cm (A-E), 5 cm (F,G), and 2 mm (H-L) (Adopted from Xiang et al., 2022)
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