Genetic transformation of the oilseed crop Crambe abyssinica
Engineering oilseed crops for industrial purposes requires a suitable crop that does not outcross to any food oilseed crop, thus eliminating problems of gene flow. Crambe abyssinica is such a dedicated crop as it does not hybridize with any existing food oilseed crops. However, lack of regeneration and transformation protocols has limited the use of C. abyssinica in genetic manipulation studies. In this study, efficient regeneration and transformation protocols for Crambe have been developed. Hypocotyls of C. abyssinica cv. Galactica were incubated on a Murashige and Skoog medium supplemented with various plant growth regulators (PGRs). Among the different PGR combinations tested, 10 μM thidiazuron and 2.7 μM α-naphthaleneacetic acid promoted highest frequency of regeneration, up to 60%. Among six Agrobacterium stains evaluated, each harbouring the cloning vector containing the neomycin phosphotransferase (nptII) and β-glucuronidase (gus) genes. EHA101 and AGL-1 yielded the highest transformation frequencies of 1.3 and 2.1%, respectively. Putative transgenic lines were recovered, and confirmed as transgenic by Southern blot analysis. Subsequently, Agrobacterium-mediated transformation of hypocotyls of cv. Galactica with constructs harbouring the wax synthase and fatty acid reductase genes have also successfully recovered confirmed transgenic plants carrying these transgenes.
KeywordsCrambe abyssinica Regeneration Agrobacterium transformation
We wish to thank Dr. Robert van Loo for providing the seeds of Glactica, Dr. Ed Cahoon for providing the vector pMS9, Dr. John Dyer for providing the pKan-Wax-FAR vector, and Prof. Sten Stymne for his support to the work, SIDA (Swedish Research Link), Dr. Anders Carlsson and Prof. Bangquan Huang for providing financial support to Xiaofeng Yan. This work is part of ICON, a European Commission FP7 project and E.C. is greatly acknowledged for its financial contribution.
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