Synergism of Bacillus thuringiensis Toxin Cry1Ac by a Fragment of Toxin-Binding Polycalin from Plutella xylostella
文献类型： 外文期刊
作者： Zhu, Qing 1 ; Gao, Meijing 1 ; Lu, Lina 1 ; Liu, Xianjin 1 ;
作者机构： 1.Jiangsu Acad Agr Sci, Inst Food Safety & Nutr, Minist Agr,State Key Lab Breeding,Key Lab Food Qu, Key Lab Control Technol & Stand Agroprod Safety &, Nanjing 210014, Peoples R China
关键词： Plutella xylostella; polycalin; Cry1Ac; synergism; larvae; Sf9 cells; cytotoxicity
期刊名称：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY （ 2020影响因子：5.279； 五年影响因子：5.269 ）
ISSN： 0021-8561
年卷期： 2021 年 69 卷 40 期
收录情况： SCI
摘要： The continued success of pest control using insecticidal crystal (Cry) proteins from Bacillus thuringiensis (Bt) in transgenic plants was threatened by the evolution of resistance. Previous studies suggested that polycalin from Plutella xylostella could bind to Cry1Ac toxin as a potential receptor. In this study, a fragment of P. xylostella polycalin (Pxpolycalinf, G2209-A2942) containing a carboxyl-terminal GPI-anchored signal peptide was cloned and expressed. Purified Pxpolycalinf retained the binding ability to Cry1Ac and synergized Cry1Ac toxicity to the third larvae of P. xylostella in bioassays. Moreover, the polyclonal antibody of Pxpolycalinf decreased the Cry1Ac activity after being fed together with normal food. Further, the ELISA results showed the concentration-dependent binding of Pxpolycalinf to P. xylostella brush border membrane vesicles (BBMV). Spodoptera frugiperda 9 (Sf9) cells expressing Pxpolycalinf were not susceptive to Cry1Ac, whereas Pxpolycalinf increased Cry1Ac cytotoxicity to Sf9 cells expressing P. xylostella ATP-dependent binding cassette transporter C2 (PxABCC2). Immunolocalization presented the binding of Pxpolycalinf to the Sf9 cell membrane, and ELISA showed the concentration-dependent binding of Pxpolycalinf to Sf9 cell extraction. These results here provide the first evidence that a fragment of P. xylostella polycalin, a potential receptor of Cry1Ac, synergizes Cry1Ac toxicity to P. xylostella larvae and Sf9 cells expressing PxABCC2.
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