A Robust Agrobacterium-Mediated Genetic Transformation Protocol for East African Coastal Farmer-Preferred Cassava Landraces
Journal Title: IOSR Journal of Biotechnology and Biochemistry (IOSR-JBB - Year 2018, Vol 4, Issue 4
Abstract
Cassava (Manihot esculenta) is the third biggest source of carbohydrates after rice and maize, feeding almost a billion people daily in the tropics. Recently it has been proposed as one of the more ideal biomass producers for biofuel production due to its high biomass yield and low input needs. A robust reproducible genetic transformation protocol is needed for germplasm improvement or functional genomics studies in cassava. In the present study we report, for the first time the establishment of a reproducible Agrobacterium-mediated transformation protocol for cassava landraces Mkombozi and Albert preferred by farmers in the coastal region of east Africa. Friable embryogenic calli (FEC) were produced using axillary buds and infected using Agrobacterium strain LBA4404 harboring the binary vector pCAMBIA2301. About 32–40 transgenic plants per 1 ml settled cell volume (SCV) were regenerated on selective medium. Histochemical GUS assays confirmed the expression of GUS gene in transformed calli, somatic embryos and transgenic plants. The presence and expression of the gusA gene were confirmed by PCR and RT-PCR analysis of transgenic plants.
Authors and Affiliations
Joshua Njiru, Evans Nyaboga
Keywords
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- EP ID EP415124
- DOI 10.9790/264X-0404013646.
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