Bt Research 2024, Vol.15, No.5, 240-247 http://microbescipublisher.com/index.php/bt 241 involve bioassays and biochemical analyses. These methods typically include concentration-response bioassays where purified Bt toxins are administered to target insect populations to observe their effects on insect survival, growth, and reproduction. For instance, bioassays have been used to assess the potential occurrence of hormesis induced by Bt toxins in both susceptible and resistant insect populations (Campos et al., 2019; Deng et al., 2019). Biochemical studies focus on understanding the structural and functional aspects of Bt toxins, such as the formation of pores in insect gut cells, which is crucial for their insecticidal activity. Techniques like crystallography and mutagenesis have been employed to elucidate the pre-pore and pore structures of Bt toxins, providing insights into their mode of action (Torres et al., 2023). Figure 1 Comparison between experimental and AF2-predicted structures (Adopted from Torres et al., 2023) Image caption: (A) Comparison of experimental and AF2-predicted α-helical stretches (helices α1 to α7) in domain I for Cry1Aa and Cry4Ba; (B,C) overlay of experimental (cylinders) and AF2 structures (ribbon) of trimeric domain I in Cry4Ba (top and side views) (B) and for monomeric domain I of Cry1Aa (C); (D) Cry4Ba monomer including domains I-III (cyan: experimental; red: AF2); (E) same as (D) without domain I, removed for clarity (Adopted from Torres et al., 2023) 2.2 Limitations of experimental methods Despite their utility, traditional experimental methods have several limitations. One significant drawback is the time and resource-intensive nature of bioassays and biochemical studies. These methods often require extensive laboratory work and can be slow in generating results. Moreover, they may not always accurately predict field performance due to the controlled conditions under which they are conducted. For example, while bioassays can
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