Bt Research 2025, Vol.16, No.3, 95-102 http://microbescipublisher.com/index.php/bt 99 5.2 Environmental benefits: reduced chemical pesticide use, improved biodiversity The use of Bt cotton has significantly reduced chemical pesticides. Research has found that the frequency and dosage of pesticide spraying in Bt cotton fields are 25%~55% less than those in ordinary cotton fields, and in some areas, it even drops to less than 10%. After the reduction of pesticides, environmental pollution and health risks decline. It can also protect natural enemies and promote biodiversity (Chatla and SumanthKumar, 2017; Singh, 2017). Meanwhile, more effective pest control, increased output and income have also promoted the sustainable development of agriculture. 5.3 Challenges: secondary pest outbreaks, resistance development in pink bollworm Although Bt cotton has a good effect on major pests, some minor pests have begun to break out in local areas, such as stink bugs and armyworm (Bilal et al., 2012). What is more serious is that pink cotton bollworm developed resistance to Bt protein, resulting in a decline in efficacy and an increase in pesticide usage in some areas (Gutierrez et al., 2023) The illegal cultivation of Bt cotton and the inadequate implementation of sheltered areas have also made the resistance problem more prominent (Srinivas, 2002). 5.4 Adaptive strategies: refuge planting enforcement, stacking multiple Bt genes, farmer education To address these challenges, India has implemented a sanctuary policy. Farmers need to plant non-BT cotton beside Bt cotton fields to delay the development of resistance. Meanwhile, crops containing multiple Bt genes (such as Bollgard II) are widely used and can be effective against various pests. Farmer education and technology promotion have also been continuously strengthened, enhancing farmers' awareness of confrontational management and ecological protection (Rani and Selvaraj, 2009; Mukherjee et al., 2022). 5.5 Lessons learned for future applications The experience of Bt cotton in India shows that biotechnology can also bring about a win-win situation for the environment and the economy in small-scale farming systems. However, for long-term and stable development, it is also necessary to attach importance to resistance management and ecological risks, and ensure the active participation of farmers. In the future, supervision should be strengthened, multi-gene superimposition should be promoted, the sheltered area policy should be strictly implemented, and farmers' training and ecological monitoring should be intensified at the same time, so as to achieve the sustainable application of Bt technology (Mukherjee et al., 2022). 6 Future Perspectives 6.1 Next-generation Bt technologies (e.g., RNAi-Bt hybrids, CRISPR-engineered Bt toxins) Bt technology is entering a new stage of greater precision and diversity. In the future, RNA interference (RNAi) and Bt toxins can be combined to act on multiple targets of pests, improving the control effect and delaying resistance (Castagnola and Jurat-Fuentes, 2012; Valicente, 2014). Meanwhile, gene editing tools such as CRISPR can modify the Bt toxin gene, create more efficient and specific new proteins, expand the application scope, and reduce the impact on non-target organisms. 6.2 Role of bioinformatics and omics in designing more targeted Bt toxins Bioinformatics and omics technologies (such as genomics and proteomics) provide new tools for the design of Bt toxins. By using big data and molecular simulation, the binding characteristics of Bt protein and pest receptors can be predicted, thereby screening out more specific new toxins. These technologies can also monitor the changing trends of pest resistance genes, providing references for resistance management. 6.3 Prospects for combining Bt with microbial consortia or synthetic biology innovations The application of Bt in the future will no longer be limited to a single strain or toxin. It can form a consortium with other beneficial microorganisms, such as fungi or actinomycetes, to jointly control pests and pathogens (Valicente, 2014). The development of synthetic biology can also support the modular design of Bt toxins, superimposing multiple functions to develop safer and more versatile biopesticides.
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