Optimization of Brassica napus (canola) explant regeneration for genetic transformation

Improved Agrobacterium-mediated transformation and high efficiency of root formation from hypocotyl meristem of spring Brassica napus ‘Precocity’ cultivar

“…Upon increasing 6-BA concentration, callus regeneration in the explants gradually declined. This result was similar to that previously described by Maheshwari et al (2011), which showed that increasing the concentration of BA in medium containing 2,4-D, decreased callus regeneration. We also analyzed combinations containing different concentrations of 2,4-D (Figure 2A and B).…”

“…Explants were cultured on MS medium containing various combinations of cytokinin (6-BA) and auxin (2,4-D). A previous study showed that the combination of a higher concentration of auxin and a lower concentration of cytokinin was more efficient for induction of callus formation (Maheshwari et al, 2011). Similarly, results from the present study demonstrated that callus regeneration was minimal in the absence of 6-BA.…”

“…Regeneration depends on many factors, including plant cultivar type, age of explants, media composition, environmental conditions, and hormonal combinations (Bhalla and Smith, 1998;Zhang et al, 2005;Maheshwari et al, 2011). In addition, various types of tissues have been reported to be useful for Agrobacterium-mediated transformation in B. napus, such as cotyledons, stems (Fry et al, 1987;Pua et al, 1987), roots, and hypocotyls (Zhang and Bhalla, 2004;Moghaieb et al, 2006;Mashayekhi et al, 2008).…”

The Use of Herbicides in Biotech Oilseed Rape Cultivation and in Generation of Transgenic Homozygous Plants of Winter Oilseed Rape (Brassica napus L.)

“…Due to its economical importance, Brassica napus L. was among the first genetically transformed crops in the history of genetically transformed plants (Maheshwari et al, 2011). Various physical techniques of gene transfer to oilseed rape have been tested, including microprojectile bombardment (Chen & Beversdorf, 1994;Fukuoka et al, 1998), electroporation (Chapel & Glimelius, 1990), microinjection (Jones-Velleneunes et al, 1995) and PEG-mediated DNA uptake (Golz et al, 1990).…”

“…However, Agrobacterium tumefaciens-mediated method is the preferred choice since it is an easy and effective way to transfer foreign genes to plant cell. (Cardoza and Stewart, 2003;Friedt & Snowdon, 2009;Jonoubi et al, 2005;Moghaieb et al, 2006;Maheshwari et al, 2011;Sharma et al, 2005;Takahata et al, 2005). Many factors have been investigated to elaborate transformation protocols, including oilseed rape varieties, Agrobacterium tumefaciens and Agrobacterium rhizogenes strains, binary vectors, type of explant (leaf pieces, cotyledons, hypocotyls, stem segments etc.)…”