Skip to Content
Merck
  • 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
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Calcium chloride dihydrate, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0%
Sigma-Aldrich
D-(+)-Glucose, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.5%
Sigma-Aldrich
Pyridoxine hydrochloride, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture
Sigma-Aldrich
L-Aspartic acid, from non-animal source, meets EP, USP testing specifications, suitable for cell culture, 98.5-101.0%
Sigma-Aldrich
L-Proline, from non-animal source, meets EP, USP testing specifications, suitable for cell culture
Sigma-Aldrich
1-Naphthaleneacetic acid, suitable for plant cell culture, BioReagent, ≥95%, crystalline
Sigma-Aldrich
Rifampicin, ≥95% (HPLC), powder or crystals
Sigma-Aldrich
Potassium chloride, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99.0%
Sigma-Aldrich
6-Benzylaminopurine, suitable for plant cell culture
Sigma-Aldrich
Kinetin, suitable for plant cell culture, BioReagent, amorphous powder
Sigma-Aldrich
Thiamine hydrochloride, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture
Sigma-Aldrich
Sodium chloride, BioXtra, ≥99.5% (AT)
Sigma-Aldrich
3′,5′-Dimethoxy-4′-hydroxyacetophenone, 97%
Sigma-Aldrich
Manganese(II) sulfate monohydrate, BioReagent, suitable for plant cell culture, suitable for cell culture
Sigma-Aldrich
Boric acid, BioReagent, for molecular biology, suitable for cell culture, suitable for plant cell culture, ≥99.5%
Sigma-Aldrich
L-Arginine, from non-animal source, meets EP, USP testing specifications, suitable for cell culture, 98.5-101.0%
Sigma-Aldrich
L-Glutamine, meets USP testing specifications, suitable for cell culture, 99.0-101.0%, from non-animal source
Sigma-Aldrich
Potassium phosphate monobasic, powder, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0%
Sigma-Aldrich
Magnesium chloride, anhydrous, ≥98%
Sigma-Aldrich
Sodium phosphate monobasic, BioPerformance Certified, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0% (titration)