Peer Review #3 of "A multi-generational risk assessment of Cry1F on the non-target soil organism Folsomia candida (Collembola) based on whole transcriptome profiling (v0.1)"

Abstract

The Bacillus thuringiensis (Bt) toxin Cry1F has been used to develop insect-resistant genetically engineered (IRGE) crops. There has been great interest in evaluating its potential risk to non-target organisms (NTOs). However, the majority of previous risk assessments only examined one generation of NTOs using several physiological indicators, which cannot comprehensively detect some potential sub-lethal effects at the molecular level. In this study, we conducted a laboratory-based, multi-generational risk assessment of Cry1F for the collembolan Folsomia candida, an important representative of soil arthropods in terms of survival, reproduction, and differentially expressed genes (DEGs) identified from whole transcriptome profiles. Our results demonstrated that Cry1F was continuously ingested by collembolans over three consecutive generations, but it did not affect the survival or reproduction of F. candida. There were no significant differences in the global gene expression between F. candida-fed diets with and without Cry1F, and no consistent co-expressed DEGs over three generations. In addition, Cry1F did not obviously alter the expression profiles of seven sensitive biological markers. Our composite data indicates that Cry1F had no long-term harmful effects on collembolan F. candida.