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Silencing a cotton bollworm P450 monooxygenase gene by plant-mediated RNAi impairs larval tolerance of gossypol

Abstract

We identify a cytochrome P450 gene (CYP6AE14) from cotton bollworm (Helicoverpa armigera), which permits this herbivore to tolerate otherwise inhibitory concentrations of the cotton metabolite, gossypol. CYP6AE14 is highly expressed in the midgut and its expression correlates with larval growth when gossypol is included in the diet. When larvae are fed plant material expressing double-stranded RNA (dsRNA) specific to CYP6AE14, levels of this transcript in the midgut decrease and larval growth is retarded. Both effects are more dramatic in the presence of gossypol. As a glutathione-S-transferase gene (GST1) is silenced in GST1 dsRNA–expressing plants, feeding insects plant material expressing dsRNA may be a general strategy to trigger RNA interference and could find applications in entomological research and field control of insect pests.

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Figure 1: Net weight increases of larvae reared on a gossypol-supplemented diet.
Figure 2: Expression pattern of CYP6AE14.
Figure 3: CYP6AE14 suppression in larvae fed leaves expressing CYP6AE14 dsRNA.
Figure 4: Enhanced effect of gossypol on larval growth fed on dsRNA leaves.
Figure 5: Suppression of GST1 by dsRNA-producing plants.
Figure 6: Suppression of CYP6AE14 by dcl2 dcl3 dcl4 triple mutant plants expressing dsCYP6AE14.

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Acknowledgements

We are grateful to Z. Xie, Bin Luo and Yuqian Jiang for experimental assistance and David Nelson for P450 nomenclature. This research was supported by The National Science Foundation of China (30421001), The Chinese Academy of Sciences (KSCX2-SW-329), and The Ministry of Science and Technology of China (2007CB108800). The A. thaliana dcl2 dcl3 dcl4 triple mutant, generated from dcl2-1 (SALK 064627), dcl3-1 (SALK 005512) and dcl4-2 (GABI160G05), and rosette leaves of the A. thaliana dcl2 dcl3 dcl4 triple mutant, generated from dcl2-1 (SALK 064627), dcl3-1 (SALK 005512) and dcl4-2 (GABI160G05), was provided by Z. Xie.

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X.-Y.C. and Y.-B.M. designed the research; Y.-B.M. performed most of the experiments; Y.-B.M. and J.-W.W. designed the dsRNA constructs; W.-J.C., L.-J.W., G.-J.H., X.-Y.T. did some of the RNA analysis experiments; Y.-B.M., X.-Y.C. and Y.-P.H. wrote the manuscript.

Corresponding author

Correspondence to Xiao-Ya Chen.

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Mao, YB., Cai, WJ., Wang, JW. et al. Silencing a cotton bollworm P450 monooxygenase gene by plant-mediated RNAi impairs larval tolerance of gossypol. Nat Biotechnol 25, 1307–1313 (2007). https://doi.org/10.1038/nbt1352

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