Agrobacterium-mediated transformation of friable embryogenic calli and regeneration of transgenic cassava
- 3 December 2009
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Protocols
- Vol. 4 (12), 1845-1854
- https://doi.org/10.1038/nprot.2009.208
Abstract
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 ∼ 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.This publication has 23 references indexed in Scilit:
- Recent trends in global production and utilization of bio-ethanol fuelApplied Energy, 2009
- Where Next for Genome Sequencing?Science, 2006
- Cassava breeding: opportunities and challengesPlant Molecular Biology, 2004
- Development and application of transgenic technologies in cassavaPublished by Springer Science and Business Media LLC ,2004
- Efficient production of transgenic plants by Agrobacterium-mediated transformation of cassava (Manihot esculenta Crantz)Euphytica, 2001
- Efficient production of transgenic cassava using negative and positive selection.Transgenic Research, 2000
- Regeneration of transgenic cassava plants ( Manihot esculenta Crantz) through Agrobacterium -mediated transformation of embryogenic suspension culturesPlant Cell Reports, 1998
- Regeneration of transgenic cassava plants (Manihot esculenta Crantz) from microbombarded embryogenic suspension culturesNature Biotechnology, 1996
- Genetic transformation of cassava (Manihot esculenta Crantz)Nature Biotechnology, 1996
- Development of friable embryogenic callus and embryogenic suspension culture systems in cassava (Manihot esculenta Crantz)Nature Biotechnology, 1996