Impact of Bt cotton on the immune system and histology of the midgut of the fall armyworm Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae)

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Despite the efficiency of transgenic plants expressing Bacillus thuringiensis (Bt) toxins as insecticides against several lepidopterans, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) is one species that presents low susceptibility to most Bt crops. This study investigated the effects of the Cry1Ac toxin expressed by Bt cotton in the midgut of S. frugiperda and its effects on the humoral and cellular immune responses. Three hypotheses were proposed and tested with contributing factors for the natural tolerance of S. frugiperda: (i) midgut regenerative cells are activated by the Cry1Ac toxin, and thus renew the epithelial cells damaged by the protein, (ii) Cry1Ac increased production of nitric oxide or phenoloxidase in the hemolymph, and (iii) there are qualitative and quantitative variations in the hemocyte levels of S. frugiperda. Caterpillars were reared using Bt cotton (Acala 90B) and non-Bt isolines (Acala 90), from the first to the fourth instar. The Bt cotton promoted elongation of the epithelial cells in the midgut of S. frugiperda caterpillars. Hence, evidence only supported the hypothesised increase of phenoloxidase (ii) and qualitative and quantitative differences in hemocyte levels (iii) in insects that were fed with Bt and non-Bt cotton. These parameters seem to explain the low susceptibility of S. frugiperda to Cry1Ac toxin and they are a viable set of responses for the evaluation of other xenobiotic factors.

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Figures

  • Cross-sectional view of the midgut of fourth instar S. frugiperda stained with Toluidine blue. (A, B) caterpillars fed with non-Bt cotton; (C, D) caterpillars fed with Bt cotton. Abbreviations and symbols: Cc, columnar cells; Cm, circular muscle; Gc, goblet cells; L, Lumen; Lm, longitudinal muscle; M, microvilli; n, nucleus; arrow head, peritrophic matrix; arros, regenerative cells. This figure is published in colour in the online version.

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  • Total count (average ± S.E.) of hemocytes in fourth instar S. frugiperda caterpillars fed with non-Bt and Bt cotton. Different letters indicate significant differences by Tukey’s HSD test (α=0.05, n = 10 larvae/treatment).

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  • Phenoloxidase activity [optical density (OD) at 492 nm] in the hemolymph of S. frugiperda caterpillars fed with non-Bt and Bt cotton as a function of time (sample size: N=10 larvae).

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