Utilization of Microspore-Derived Embryos

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

“…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.)…”

“…or other techniques. So far transformation attempts have shown very limited success, despite using selection system based on npt II gene, conferring resistance to the mildly acting antibiotic kanamycin (Grison et al, 1996;Poulsen, 1996;Takahata et al, 2005;Wang Y.P. et al, 2005).…”

“…Although both oilseed rape microspores and MDEs characterise high regeneration potential (Fig. 3) (Cegielska et al, 2002, Friedt andZarhloul 2005;Takahata et al, 2005), only MDEs were able to regenerate post Agrobacterium inoculation (Fig. 4).…”

“…Doubled haploids, obtained by tissue cultures of microspores and microspore-derived embryos (MDEs), show uniformity and complete homozygosity, thus they are particularly useful for breeding programmes and have been extensively used to produce homozygous breeding lines and cultivars of oilseed rape (Cegielska et al, 2002;Friedt and Zarhloul, 2005;Takahata et al, 2005). DH plants can also be efficiently used in basic studies, for example in genetic analysis and genetic transformation (Cegielska-Taras et al, 2008;Fukuoka et al, 1998;Huang, 1992;Jardinaud et al, 1993;Kazan et al, 1997;Nehlin et al, 2000;Oelck et al, 1991;Swanson & Erickson, 1989;Takahata et al, 2005;Troczyńska et al, 2003). It is postulated that in the process of genetic transformation, the gene introduced into the haploid genome, in this case MDE and subsequent chromosome doubling (diploidization), it will give rise to homozygous transgenic oilseed rape (Dorman et al, 1998;Huang, 1992).…”

Improvement of microspore culture method for multiple samples in Brassica

“…In genus Brassica, since successful isolated microspore culture of B. napus was reported by Lichter (1982), a large amount of research has been carried out to improve this technique and to expand it other Brassica species and allied genera (Ferrie and Keller 2004, Palmer and Keller 1999, Takahata 1997. Microspore culture of Brassicas has several advantages; 1) the techniques of microspore culture are simple and easier than those of anther culture, 2) a high yield of embryos can be obtained within a short period of 2-3 weeks, and 3) this technique offers not only the fastest route to homozygosity, but it also provides several valuable tools for genetic engineering and a model system of developmental research because of its single-cell system (Takahata et al 2005).…”