Agrobacterium-mediated genetic transformation of economically important oilseed rape cultivars

Abstract

Oilseed rape is considered relatively recalcitrant for genetic modification. This work was performed to establish conditions for effective transformation and regeneration of commercially used cultivars Campino, Haydn, Heros, Hunter and Topas (Brassica napus L.). Cotyledonary petioles and hypocotyls (obtained from the seedlings grown under dark conditions) as a source of explants were used. Our experiments revealed that a lower selection pressure in combination with postponing of selection by 14 days after co-cultivation resulted in recovery of transgenic plants from all cultivars. Transformants were obtained with efficiency from 1 to 5.5%. Histochemically GUS-positive plants were analysed by PCR using primers corresponding to the internal fragments of gus and nptII genes. The transgenic nature was confirmed by Southern blot analyses using a specific nptII probe. This work enriches the list of oilseed rape cultivars available for genetic modification.

CRISPR-Cas9 Targeted Mutagenesis Leads to Simultaneous Modification of Different Homoeologous Gene Copies in Polyploid Oilseed Rape (Brassica napus)

“…Compared with other rapeseed cultivars, cv Haydn has an elevated shatter resistance (data not shown). We had chosen this genotype for our experiments because of its high transformation efficiency (Boszoradova et al, 2011). However, cv Haydn is considered an old variety because it was released in the year 2000.…”

Targeting transgene to seed resulted in high rate of morphological abnormalities of Camelina transformants

“…Since aberrant transformants are undesired sideproducts of many experiments, they are rarely discussed in detail and their reasons are not further explored. There are many quite well described reasons for changes at DNA level (ranging from point mutations and methylation differences to transposon induction, gene amplification, chromosomal aberrations and ploidy level changes; Kaeppler et al 2000;Jain 2001;Bregitzer et al 2002;Boszorádová et al 2011Boszorádová et al , 2014. Numerous aspects of plant transformation are associated with the stress of the exposed tissues.…”

Transformation of sundew: pitfalls and promises

“…Production of chimeras seems to be a more frequent phenomenon than originally thought and it has been reported in several species (Berthomieu et al 1994;Boszoradova et al 2011;Petri et al 2011). Transgenic plants regenerated directly from shoot apical meristems that were targeted for transformation are always chimerical (Yookongkaew et al 2007;Yevtushenko and Misra 2010).…”