Bt Research 2024, Vol.15, No.2, 96-109 http://microbescipublisher.com/index.php/bt 106 In the context of IPM, Bt crops can be integrated with other pest control methods, such as biological control, crop rotation, and the use of refuges, to create a diversified and sustainable pest management strategy. The use of Bt crops can reduce the reliance on chemical insecticides, thereby minimizing the risk of secondary pest outbreaks and promoting the conservation of natural enemies (Anderson et al., 2019; Romeis et al., 2019). Additionally, the integration of Bt crops with other IPM practices can enhance the overall effectiveness and sustainability of pest management programs, ensuring long-term agricultural productivity and environmental health (Gassmann and Reisig, 2022). By leveraging the benefits of gene stacking strategies and integrating them with comprehensive IPM plans, farmers can achieve more sustainable and resilient agricultural systems. This holistic approach not only addresses the immediate challenges of pest control but also contributes to the broader goals of sustainable agriculture and environmental conservation. 9 Concluding Remarks The systematic review on gene stacking strategies to enhance the durability of Bt crops has highlighted several key findings. Transgene stacking, which involves the introduction of multiple genes into crop plants, has been shown to confer higher and more durable resistance to insects and diseases compared to single-gene technology. This approach has been particularly effective in managing resistance to Bacillus thuringiensis (Bt) toxins, which are crucial for pest control in transgenic crops. Additionally, the study has underscored the importance of combining resistance genes with other agricultural practices to slow down the evolution of virulent pathogen genotypes. The use of linker peptides, such as LP4/2A, has been identified as a reliable method for producing gene stacking in crops like maize and tobacco, enhancing both pest resistance and herbicide tolerance. Gene stacking is of paramount importance for the sustainability and effectiveness of Bt crops. By incorporating multiple resistance genes, gene stacking can significantly delay the development of resistance in pest populations, thereby extending the useful life of Bt crops. This strategy not only improves pest resistance but also enhances crop yield and reduces the need for chemical pesticides, contributing to more sustainable agricultural practices. Moreover, gene stacking can be combined with other resistance management strategies, such as crop rotation and the use of quantitative resistance genes, to further enhance the durability of Bt crops. The ability to stack genes for multiple traits, including herbicide tolerance and disease resistance, makes this approach highly versatile and beneficial for modern agriculture. Future research should focus on optimizing gene stacking techniques to overcome the current technical challenges associated with the expression and manipulation of multiple genes in plants. There is a need for more empirical studies to compare different gene deployment strategies and their long-term effects on pest resistance and crop yield. Additionally, exploring the potential of combining gene stacking with other innovative technologies, such as RNA interference (RNAi) and marker-assisted selection (MAS), could provide new avenues for enhancing crop resistance and sustainability. Research should also investigate the ecological impacts of gene stacking, including its effects on non-target organisms and overall biodiversity, to ensure that this approach contributes to environmentally friendly agricultural practices. Finally, developing more efficient and reliable methods for gene stacking, such as the use of advanced linker peptides, will be crucial for the successful implementation of this strategy in a wider range of crops. Acknowledgments Thank you to the peer reviewers for their suggestions on the manuscript. Conflict of Interest Disclosure The author affirms that this research was conducted without any commercial or financial relationships that could be construed as a potential conflict of interest.
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