Bioscience Methods 2024, Vol.15, No.4, 149-161 http://bioscipublisher.com/index.php/bm 153 (Liu et al., 2021). The development of multiplex genome editing strategies, such as BREEDIT, further accelerates the improvement of complex traits by targeting multiple genes simultaneously (Lorenzo et al., 2022). Figure 2 Exploring photosynthetic pathways for enhanced crop yield and future improvement strategies (Adapted from Sharwood et al., 2022) Image caption: The key photosynthetic components-light reactions, CO2 fixation, and photorespiration that contribute to crop yield. Recent gene technology advancements have identified several targets for improving these processes. Enhancements include increasing electron transport for the Calvin cycle by modifying Cyt b6f levels, optimizing Rubisco activity through catalytic alterations, and engineering thermotolerant Rubisco activase for better performance at elevated temperatures. Ambitious strategies like introducing CO2-concentrating mechanisms (CCMs) such as the C4 CCM, carboxysomes, or pyrenoids into crops are also explored. Additional opportunities involve improving CO2 diffusion, Calvin cycle flux, sugar signaling, and altering the photorespiratory pathway to boost plant productivity. These innovations present potential pathways for future-proofing crop productivity in response to environmental challenges (Adapted from Sharwood et al., 2022) 4.4 Transgenic approaches and their successes Transgenic approaches have been employed to introduce foreign genes into maize to enhance photosynthetic efficiency and yield. These approaches have led to the development of maize varieties with improved stress tolerance, higher yield, and better photosynthetic performance. For example, the integration of CRISPR/Cas9 with transgenic techniques has facilitated the creation of novel maize germplasms with enhanced traits (Wang et al., 2022). The use of haploid-inducer mediated genome editing has also accelerated the breeding process by generating homozygous pure lines with desired traits within two generations (Wang et al., 2019). 4.5 Challenges and future directions in photosynthesis enhancement Despite the advancements in genetic and biotechnological approaches, several challenges remain in enhancing photosynthesis in maize. The complex genetic architecture of photosynthetic traits and the interaction of multiple small-effect genes pose significant hurdles. Additionally, the acclimation of the photosynthetic machinery to fluctuating environments needs to be better understood to identify relevant genetic variations (Bezouw et al., 2019). Future research should focus on optimizing gene editing and transformation systems, exploring novel
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