Transgenic superroots of Lotus corniculatus can be regenerated from superroot-derived leaves following Agrobacterium-mediated transformation

FOX-superroots of Lotus corniculatus, overexpressing Arabidopsis full-length cDNA, show stable variations in morphological traits

“…Transgenic calli were selected on MS medium containing hygromycin. Hygromycin-resistant calli were induced to undergo caulogenesis and transgenic plants obtained from regenerated shoots as described by Tanaka et al (2008). We have generated, in approximately 50 transformation experiments, 162 independent putative SR transgenic callus lines of which 153 regenerated plants.…”

“…The transformation of SR was performed as described by Tanaka et al (2008). For the production of target tissue, 4-week-old SR cultures were transferred to light under stationary conditions.…”

“…These features are unique to root cultures and are now stably expressed since the culture was isolated more than 13 years ago (1997). Therefore, SR are unique and promising experimental system for developmental studies, such as root formation, regeneration or nodulation in vitro (Tanaka et al, 2008;Jian et al, 2009). The application of FOX hunting to L. corniculatus would supplement existing techniques in mutagenesis and gene discovery in legumes, such as targeted EMS and fast neutron mutagenesis in soybean (Carroll et al, 1985;Men et al, 2002), L. japonicus (Wopereis et al, 2000;Hoffmann et al, 2007) and Medicago truncata (Penmetsa and Cook, 1997).…”

Gene Transfer in Legumes

“…Tanaka et al (2008) obtained transgenic L. corniculatus from Superroot-derived leaves via A. tumefaciens-mediated transformation. Transgenicity of kanamycin-resistant calli was verified by GUS staining, but transformation efficiency was low and the process from gene transfer to PCR identification took 6 months.…”

The National BioResource Project (NBRP) Lotus and Glycine in Japan

“…These characteristics are still maintained 14 years after the discovery of SR (Akashi et al 1998, 2003). SRs can be used in physiological research as well as in functional analysis of genes using A. tumefaciens (Tanaka et al 2008) or A. rhizogenes -mediated transformation (Jian et al 2009). Himuro et al (2011) developed 130 Arabidopsis full-length cDNA overexpressor (FOX)-superroot lines using the FOX hunting system.…”