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  • Efficient Agrobacterium-mediated Transformation of The Elite-Indica Rice Variety Komboka.

Efficient Agrobacterium-mediated Transformation of The Elite-Indica Rice Variety Komboka.

Bio-protocol (2021-03-05)
Van T Luu, Melissa Stiebner, Paula Emmerich Maldonado, Sandra Valdés, Didier Marín, Gerardo Delgado, Virginia Laluz, Lin-Bo Wu, Paul Chavarriaga, Joe Tohme, Inez H Slamet-Loedin, Wolf B Frommer
要旨

Genetic transformation is crucial for both investigating gene functions and for engineering of crops to introduce new traits. Rice (Oryza sativa L.) is an important model in plant research, since it is the staple food for more than half of the world's population. As a result, numerous transformation methods have been developed for both indica and japonica rice. Since breeders continuously develop new rice varieties, transformation protocols have to be adapted for each new variety. Here we provide an optimized transformation protocol with detailed tips and tricks for a new African variety Komboka using immature embryos. In Komboka, we obtained an apparent transformation rate of up to 48% for GUS/GFP reporter gene constructs using this optimized protocol. This protocol is also applicable for use with other elite indica rice varieties.

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D-(+)-グルコース, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.5%
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L-グルタミン, meets USP testing specifications, suitable for cell culture, 99.0-101.0%, from non-animal source
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リファンピシン, ≥95% (HPLC), powder or crystals
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L-プロリン, from non-animal source, meets EP, USP testing specifications, suitable for cell culture
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塩化カルシウム 二水和物, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0%
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塩化ナトリウム, BioXtra, ≥99.5% (AT)
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6-ベンジルアミノプリン, suitable for plant cell culture
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