Direct gene delivery into isolated microspores of rapeseed ( Brassica napus L.) and the production of fertile transgenic plants

Haploids and doubled haploids in Brassica spp. for genetic and genomic research

“…A highly efficient doubled haploidy protocol is required for any transformation protocols, making Brassica species ideal candidates. Different microspore transformation methods have been attempted in Brassica including Agrobacterium tumefaciens (Pechan 1989;Dormann et al 2001), particle bombardment (Fukuoka et al 1998;Nehlin et al 2000), microinjection (Jones-Villeneuve et al 1995), and electroporation (Guerche et al 1987;Jardinaud et al 1993). A recent paper combined particle bombardment and Agrobacterium-mediated transformation in B. napus microspores (Abdollahi et al 2009a).…”

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

“…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.…”

“…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).…”

Identification and evaluation of intervarietal substitution lines of rapeseed (Brassica napus L.) with donor segments affecting the diploidization rate of isolated microspores

“…In addition, microspores, MDEs and DH plants are being used for transformation studies. Different approaches have been reported, using Agrobacterium tumefaciens (Dormann et al 2001;Cegielska-Taras et al 2008), particle gun bombardment (Fukuoka et al 1998;Chen and Beversdorf 1994;Nehlin et al 2000), or microinjection (Guerche et al 1987;Jardinaud et al 1993;Huang 1992).…”