Article (Scientific journals)
Agrobacterium-mediated transformation of friable embryogenic calli and regeneration of transgenic cassava.
Bull, S. E.; Owiti, J. A.; Niklaus, M. et al.
2009In Nature Protocols, 4 (12), p. 1845-54
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Keywords :
Genetic Engineering/methods; Glucuronidase/analysis; Manihot/embryology/genetics/physiology; Plants, Genetically Modified/physiology; Regeneration; Rhizobium/genetics; Tissue Culture Techniques; Transformation, Genetic
Abstract :
[en] Agrobacterium-mediated transformation of friable embryogenic calli (FEC) is the most widely used method to generate transgenic cassava plants. However, this approach has proven to be time-consuming and can lead to changes in the morphology and quality of FEC, influencing regeneration capacity and plant health. Here we present a comprehensive, reliable and improved protocol, taking approximately 6 months, that optimizes Agrobacterium-mediated transformation of FEC from cassava model cultivar TMS60444. We cocultivate the FEC with Agrobacterium directly on the propagation medium and adopt the extensive use of plastic mesh for easy and frequent transfer of material to new media. This minimizes stress to the FEC cultures and permits a finely balanced control of nutrients, hormones and antibiotics. A stepwise increase in antibiotic concentration for selection is also used after cocultivation with Agrobacterium to mature the transformed FEC before regeneration. The detailed information given here for each step should enable successful implementation of this technology in other laboratories, including those being established in developing countries where cassava is a staple crop.
Disciplines :
Biotechnology
Author, co-author :
Bull, S. E.
Owiti, J. A.
Niklaus, M.
Beeching, J. R.
Gruissem, W.
Vanderschuren, Hervé  ;  Université de Liège - ULiège > Sciences agronomiques > Biologie végétale
Language :
English
Title :
Agrobacterium-mediated transformation of friable embryogenic calli and regeneration of transgenic cassava.
Publication date :
2009
Journal title :
Nature Protocols
ISSN :
1754-2189
eISSN :
1750-2799
Publisher :
Nature Publishing Group, United Kingdom
Volume :
4
Issue :
12
Pages :
1845-54
Peer reviewed :
Peer Reviewed verified by ORBi
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