Abstract
The hybrid Bacillus thuringiensis (Bt) δ-endotoxin gene Cry1Ab/Ac was used to develop a transgenic Bt rice (Oryza sativa L.) targeting lepidopteran insects of rice. Here, we show the production of a marker-free and tissue-specific expressing transgenic Bt rice line L24 using Agrobacterium-mediated transformation and a chemically regulated, Cre/loxP-mediated DNA recombination system. L24 carries a single copy of marker-free T-DNA that contains the Cry1Ab/Ac gene driven by a maize phosphoenolpyruvate carboxylase (PEPC) gene promoter. The marker-free T-DNA was integrated into the 3′ untranslated region of rice gene Os01g0154500 on the short arm of chromosome 1. Compared to the constitutive and non-specific expression of the P Actin1 :Cry1Ab/Ac:T Nos gene in the control Bt rice line T51-1, the P Pepc :Cry1Ab/Ac:T Nos gene was detected only in the leaf and stem tissues of L24. More importantly, compared to high levels of CRY1Ab/Ac proteins accumulated in T51-1 seeds, the CRY1Ab/Ac proteins were not detectable in L24 seeds by Western blot analysis. As demonstrated by insect bioassay, L24 provided similar level of resistance to rice leaffolder (Cnaphalocrocis medinalis) as T51-1. The marker-free transgenic line L24 can be used directly in rice breeding for insect resistance to lepidopteran insects where absence of Bt toxin protein in the seed is highly desirable.
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Acknowledgments
The authors thank Yunlu Fan for providing plasmid pFWZ16 and Qifa Zhang for providing Bt line T51-1. This work was supported by intramural research funds from Temasek Life Sciences Laboratory (Z.Y.). The authors declare that they have no conflict of interest.
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Communicated by K. Chong.
C. Qiu and J. S. Sangha contributed equally to this work and their last names appear in alphabetical order.
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Qiu, C., Sangha, J.S., Song, F. et al. Production of marker-free transgenic rice expressing tissue-specific Bt gene. Plant Cell Rep 29, 1097–1107 (2010). https://doi.org/10.1007/s00299-010-0893-x
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DOI: https://doi.org/10.1007/s00299-010-0893-x