Bt Research 2025, Vol.16, No.5, 182-193 http://microbescipublisher.com/index.php/bt 191 metabolites in culture medium and host insects, it can be found which metabolites change significantly during infection and their function can be inferred. These may include secondary metabolites or signaling molecules that have not received previous attention and are of great value for elucidating the pathogenic mechanism of Bt. At the same time, the combination of transcriptomics and proteomics will improve the perspective of metabolic research. Systematic biological methods integrate multi-omics data and can reconstruct Bt's metabolic regulation network model. Using these models, we can simulate metabolic flow allocation and virulence factor expression of Bt in different environments, thereby predicting optimization strategies. 8.2 Transformation and improvement of metabolic pathway Bt pathogenicity and stability Based on an in-depth understanding of the relationship between Bt metabolism and virulence, we can try to further improve the pathogenic performance and environmental adaptability of Bt strains through metabolic engineering. A direct idea is to remove metabolic inhibition and improve toxin synthesis. By knocking out the global inhibitor gene in the Bt strain, it can continue to express toxins under nutritious conditions, rather than waiting for the nutrients to be exhausted before producing toxins. This type of transformation may allow Bt to release insecticide factors earlier when entering the insect body, speeding up the lethal process. Of course, attention should be paid to avoid the strains that have premature spore production during the fermentation period affects growth, and regulatory elements should be used to carefully control the knockout effect. Another idea is to introduce or strengthen specific metabolic pathways to improve the environmental stress resistance and survival rate of Bt. For example, Bt can be overexpressed by genetic engineering to overexpress key enzymes of the melanin synthesis pathway, thereby accumulating more melanin particles on the spore surface area and improving the anti-UV capability of the bacteria agent. 8.3 Application direction of green agriculture and sustainable development In the context of pursuing green agriculture and sustainable development, the metabolic research and application of Bt have ushered in new opportunities and challenges. On the one hand, it is expected to develop more efficient, environmentally friendly biopesticides through in-depth understanding of Bt metabolism to reduce dependence on chemical pesticides. On the other hand, the application of Bt is expanding from a single pesticide to a broader biotechnology field such as soil improvement and plant health. Studies have found that metabolites of some Bt strains can promote plant growth or inhibit plant pathogens. For example, certain volatile substances produced by Bt metabolism may induce insect resistance and improve crop self-resistance; for example, Bt is applied in combination with crop endophytes, and its metabolic activity can improve rhizosphere microbial community and thereby enhance plant nutrient absorption. These new application directions require us to break out of the traditional framework of "Bt is insecticide" and re-examine the function of Bt from the perspective of metabolic interaction. Acknowledgments We would like to thank Cuixi Biotechnology Institute for its funding and technical platform support for this research, and we would also like to thank the peer reviewer for its review opinions. Conflict of Interest Disclosure The authors affirm that this research was conducted without any commercial or financial relationships that could be construed as a potential conflict of interest. References Al-Harbi A., Lary S., Edwards M., Qusti S., Cockburn A., Poulsen M., and Gatehouse A., 2019, A proteomic-based approach to study underlying molecular responses of the small intestine of Wistar rats to genetically modified corn (MON810), Transgenic Research, 28: 479-498. https://doi.org/10.1007/s11248-019-00157-y Cao Z., Tan T., Jiang K., Mei S., Hou X., and Cai J., 2018, Complete genome sequence of Bacillus thuringiensis L-7601 a wild strain with high production of melanin, Journal of Biotechnology, 275: 40-43. https://doi.org/10.1016/j.jbiotec.2018.03.020 Chen H., Verplaetse E., Slamti L., and Lereclus D., 2022, Expression of the Bacillus thuringiensis vip3A insecticidal toxin gene is activated at the onset of stationary phase by VipR an autoregulated transcription factor, Microbiology Spectrum, 10(4): e01205-22. https://doi.org/10.1128/spectrum.01205-22
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