Transgenic Research

, Volume 14, Issue 3, pp 261–272 | Cite as

Two different Bacillus thuringiensis toxin genes confer resistance to beet armyworm (Spodoptera exigua Hübner) in transgenic Bt-shallots (Allium cepa L.)

  • Si-Jun Zheng
  • Betty Henken
  • Ruud A. de Maagd
  • Agus Purwito
  • Frans A. Krens
  • Chris Kik


Agrobacterium-mediated genetic transformation was applied to produce beet armyworm (Spodoptera exigua Hübner) resistant tropical shallots (Allium cepa L. group Aggregatum). A cry1Ca or a H04 hybrid gene from Bacillus thuringiensis, driven by the chrysanthemum ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (Rubisco SSU) promoter, along with the hygromycin phosphotransferase gene (hpt) driven by the CaMV 35S promoter, was employed for genetic transformation. An average transformation frequency of 3.68% was obtained from two shallot cultivars, Tropix and Kuning. After transfer of the in vitro plants to the greenhouse 69% of the cry1Ca and 39% of the H04 transgenic shallots survived the first half year. After one year of cultivation in the greenhouse the remaining cry1Ca and H04 transgenic plants grew vigorously and had a normal bulb formation, although the cry1Ca transgenic plants (and controls) had darker green leaves compared to their H04 counterparts. Standard PCR, adaptor ligation PCR and Southern analyses confirmed the integration of T-DNA into the shallot genome. Northern blot and ELISA analyses revealed expression of the cry1Ca or H04 gene in the transgenic plants. The amount of Cry1Ca expressed in transgenic plants was higher than the expression levels of H04 (0.39 vs. 0.16% of the total soluble leaf proteins, respectively). There was a good correlation between protein expression and beet armyworm resistance. Cry1Ca or H04 gene expression of at least 0.22 or 0.08% of the total soluble protein in shallot leaves was sufficient to give a complete resistance against beet armyworm. This confirms earlier observations that the H04 toxin is more toxic to S. exigua than the Cry1Ca toxin. The results from this study suggest that the cry1Ca and H04 transgenic shallots developed could be used for introducing resistance to beet armyworm in (sub) tropical shallot.


Agrobacterium tumefaciens Allium cepa beet armyworm Bt resistance cry1Ca hybrid H04 transformation 



adaptor ligation PCR


Bacillus thuringiensis


a synthetic gene from Bacillus thuringiensis




a hybrid gene from B. thuringiensis


hygromycin phosphotransferase gene


T-DNA left border


T-DNA right border


β-glucuronidase gene


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Copyright information

© Springer 2005

Authors and Affiliations

  • Si-Jun Zheng
    • 1
    • 3
  • Betty Henken
    • 1
  • Ruud A. de Maagd
    • 1
  • Agus Purwito
    • 2
  • Frans A. Krens
    • 1
  • Chris Kik
    • 1
  1. 1.Plant Research InternationalWageningen University and Research CenterWageningenThe Netherlands
  2. 2.Research Centre for BiotechnologyBogor Agricultural UniversityBogorIndonesia
  3. 3.Laboratory of EntomologyWageningen UniversityWageningenThe Netherlands

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