Evaluation of Off-target Effects of Anti-Miridae Transgenic dsAllim and Transgenic dsAsfar Cottons on Harmonia axyridis

doi:10.16409/j.cnki.2095-039x.2025.09.023

Abstract

Abstract: Off-target effects are a major drawback of RNAi, and for transgenic dsRNA crops using RNAi strategy for pest control, the off-target effects of dsRNA on non-target organisms (NTOs) are an important indicator for their safety evaluation. In this study, Harmonia axyridis was selected as NTOs representing the natural enemy insect group, and dsRNA was introduced into NTOs by feeding. The relationship between the efficiency of dsRNA uptake and feeding days was explored by qRT-PCR. The off-target effects of transgenic dsAsfar cotton and transgenic dsAllim cotton were evaluated by RNA-Seq combined with quantitative analysis for validation. The uptake efficiency of dsRNA by H. axyridis larvae was not directly related to the duration of dsRNA feeding, and the uptake efficiency of dsRNA by adults was significantly higher than that in larvae. In addition, neither dsAsfar nor dsAllim expressed in the two transgenic dsRNA cotton plants had a significant effect on the expression of homologous genes in H. axyridis. The number of down-regulated differentially expressed genes (DEGs) was 2 and 8 in H. axyridis larvae and adults feeding on dsAsfar, and 3 and 26 on dsAllim, respectively. KEGG and GO enrichment analyses of H. axyridis larvae fed on dsAllim revealed that DEGs were significantly enriched in KEGG pathways related to muscle development and GO terms associated with cuticle structural components, with all 33 DEGs showing upregulated expression. The results of the KEGG and GO enrichment analyses for dsAllim in adults, as well as for dsAsfar in both adults and larvae, showed that DEGs were not significantly enriched in the corresponding KEGG pathways and GO entries of far and lim genes. Overall, the results of this study provide valid support for the safety of transgenic dsRNA cotton.

Key words: RNA interference, transgenic cotton, off-target effects, Harmonia axyridis, transcriptome

CLC Number:

S476.2

CAO Danyang, YAO Haiqin, CHEN Lizhen, MA Weihua. Evaluation of Off-target Effects of Anti-Miridae Transgenic dsAllim and Transgenic dsAsfar Cottons on Harmonia axyridis[J]. Chinese Journal of Biological Control, 2026, 42(2): 412-424.

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