International Journal of Molecular Evolution and Biodiversity, 2025, Vol.15, No.2, 84-98 http://ecoevopublisher.com/index.php/ijmeb 90 5.4 Integrated application of biotechnology in drought-resistant breeding Genetic engineering technology has opened up new avenues for the precise improvement of drought-resistant traits. By targeted introduction of functional genes related to the ABA signaling pathway or osmotic regulation (Ashraf, 2010), this technology breaks through the limitations of traditional breeding. Compared with conventional hybridization methods, genetic engineering not only simplifies the multi-generation screening process, but also ensures the stable inheritance of target traits in offspring (Cao et al., 2022), showing its important application value in drought-resistant breeding. At the same time, the CRISPR/Cas9 gene editing system has shown broad application prospects in regulating drought-responsive genes due to its high targeting and editing efficiency. By precisely knocking out or activating regulatory elements or coding regions, this technology can effectively enhance the adaptability of plants under water stress conditions (Ashraf, 2010). More importantly, CRISPR/Cas9 is not only an independent improvement tool, but its flexibility also makes it easy to integrate with traditional breeding methods, QTL positioning and genomic selection (GS) technology to build a multi-dimensional synergistic drought-resistant breeding system (Figure 1) (Cao et al., 2022). With the continuous optimization and in-depth application of these biotechnologies, rapeseed drought-resistant breeding is gradually moving towards a new stage of precision, efficiency and controllability. Figure 1 Drought resistance testing of p35S::BnA.JAZ5plants at the five-leaf stage (Adopted from Cao et al., 2022) Image caption: (A) Drought resistance assay of five overexpressing lines. (B) Survival rates of five overexpressing lines after 3-day recovery from drought stress treatment. WT, accession K407. OE-1, OE-2, OE-3, OE-4 and OE-5, p35S::BnA.JAZ5 plants. Asterisks indicate significant differences between wild-type and p35S::BnA.JAZ5 lines (P< 0.05, Student’s t-test). Scale bar, 10 cm (Adopted from Cao et al., 2022)
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