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Insect-resistant chrysanthemum calluses by introduction of aBacillus thuringiensis crystal protein gene

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Abstract

A 3′-end truncated crystal protein gene, derived fromBacillus thuringiensis (Bt) subsp.aizawai 7.21, encoding the toxic fragment of the insecticidal proteincryIA(b), was constructed. The gene was inserted into a transformation vector, also carrying the neomycin phosphotransferase II (nptII) gene and the β-glucuronidase (gus) gene, and introduced in the oncogenicAgrobacterium tumerfaciens strain A281, harbouring the Ti-plasmid pTiBO542. The recombinantAgrobacterium strain was used to transform leaf explants of chrysanthemum (Dendranthema grandiflora) cultivar Parliament. The resulting tumours were kanamycin-resistant, exhibited β-glucuronidase activity and produced agropine and mannopine. In most tumours, all simultaneously transferred genes were expressed, owing to selection for the presence of both T-DNAs, but no correlation was found between the level of expression of the various genes. A bioassay was developed, in which larvae were fed with tumorous chrysanthemum tissue, in order to detect the effect of the transferred toxin gene on larval development. Using this bioassay with second instar larvae ofHeliothis virescens (tobacco budworm), 17 tumour lines were tested. Several of these lines proved to be strongly inhibitory to larval growth. These results indicate thatBt-based insect resistance might be used as a tool in reducing the amount of pesticides used in chrysanthemum culture.

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  1. Guy Honée

    Present address: Department of Phytopathology, Agricultural University, Binnenhaven 9, 6709 PD, Wageningen, Netherlands

Authors and Affiliations

  1. Department of Developmental Biology and Molecular Biology, DLO-Centre for Plant Breeding and Reproduction Research (CPRO-DLO), P.O. Box 16, 6700 AA, Wageningen, Netherlands

    Monique F. van Wordragen, Guy Honée & Hans J. M. Dons

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  1. Monique F. van Wordragen
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  2. Guy Honée
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van Wordragen, M.F., Honée, G. & Dons, H.J.M. Insect-resistant chrysanthemum calluses by introduction of aBacillus thuringiensis crystal protein gene. Transgenic Research 2, 170–180 (1993). https://doi.org/10.1007/BF01972611

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