Production of marker-free transgenic rice expressing tissue-specific Bt gene |
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Authors: | Chengxiang Qiu Jatinder Singh Sangha Fengshun Song Zhiyun Zhou Ao Yin Keyu Gu Dongsheng Tian Jianbo Yang Zhongchao Yin |
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Institution: | (1) Temasek Life Sciences Laboratory, National University of Singapore, Singapore, 117604, Singapore;(2) Rice Research Institute, Anhui Academy of Agricultural Sciences, Hefei, 230031, China;(3) Present address: Department of Environmental Sciences, Nova Scotia Agricultural College, P.O. Box 550, Truro, NS, B2N 5E3, Canada |
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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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