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Production of marker-free transgenic rice expressing tissue-specific Bt gene

文献类型： 外文期刊

作者： Qiu, Chengxiang 1 ; Sangha, Jatinder Singh 1 ; Song, Fengshun 2 ; Zhou, Zhiyun 1 ; Yin, Ao 1 ; Gu, Keyu 1 ; Tian, Dong 1 ;

作者机构： 1.Natl Univ Singapore, Temasek Life Sci Lab, Singapore 117604, Singapore

2.Anhui Acad Agr Sci, Rice Res Inst, Hefei 230031, Peoples R China

关键词： Bt rice;Cry1Ab/Ac;Marker-free transformation;Cre/loxP;Tissue-specific promoter;Rice leaffolder

期刊名称：PLANT CELL REPORTS （ 影响因子：4.57； 五年影响因子：4.463 ）

ISSN： 0721-7714

年卷期： 2010 年 29 卷 10 期

页码：

收录情况： SCI

摘要： The hybrid Bacillus thuringiensis (Bt) delta-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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