Agrobacterium tumefaciens-mediated transformation of leek (Allium porrum) and garlic (Allium sativum)

Abstract

Transgenic leek (Allium porrum) and garlic (Allium sativum) plants have been recovered by the selective culturing of immature leek and garlic embryos via Agrobacterium-mediated transformation using a method similar to that described by Eady et al. (Plant Cell Rep 19:376-381, 2000) for onion transformation. This method involved the use of a binary vector containing the m-gfp-ER reporter gene and nptII selectable marker, and followed the protocol developed previously for the transformation of onions with only minor modifications pertaining to the post-transformation selection procedure which was simplified to have just a single selection regime. Transgenic cultures were selected for their ability to express the m-gfp-ER reporter gene and grown in the presence of geneticin (20 mg/l). The presence of transgenes in the genome of the plants was confirmed using TAIL-PCR and Southern analysis. This is the first report of leek and "true seed" garlic transformation. It now makes possible the integration of useful agronomic and quality traits into these crops.

Cloning and expression analysis of PpSUT2 encoding a sucrose transporter in pear

“…The pCXDG-PpSUT2 recombinant vector was transformed into Agrobacterium cells (strain EHA4404) using the freeze-thaw method and selected on LB plates containing 50 µg/mL rifamycin and 50 µg/mL kanamycin (Eady et al, 2005). The transformed colonies were further confirmed by PCR and grown in LB liquid medium containing 50 µg/mL rifamycin and 50 µg/mL kanamycin for 36 h. The primer sequences were P2r: 5'-TTACCCAAAATGAAAACCCGTTG-3' and P2f: 5'-GATGGCGGGGAGGACGGACT-3'.…”

Advances in Agrobacterium-mediated plant transformation with enphasys on soybean

“…GFP has been used as a reporter system for identifying transformation events in Arabidopsis thaliana (Haseloff et al, 1997;Ueda et al, 2003;Chang et al, 2005), apple (Maximova et al, 1998), rice (Vain et al, 1993), sugarcane, maize, lettuce, tobacco (Elliott et al, 1999), soybean (Ponappa et al, 1999), oat (Kaeppler et al, 2000), onion (Eady et al, 2000), wheat (Jordan, 2000), leek and garlic (Eady et al, 2005). The GFP reporter system has also been used for identifying successful plastid transformation events in potato (Sidorov et al, 1999).…”

Development of an Efficient Agrobacterium-Mediated Transformation System and Production of Herbicide-Resistant Transgenic Plants in Garlic (Allium sativum L.)

“…More recently, calli derived from different explant sources including roots as the most easily available explant and an Agrobacterium-mediated procedure were used to produce agronomically useful transgenic garlic plants resistant to beet armyworm (Spodoptera exigua), a polyphagous insect (Zheng et al, 2004). In addition, the Agrobacteriummediated transformation of garlic was also reported using embryos (Eady et al, 2005) and immature leaf materials (Kenel et al, 2010). These recent garlic transformation methods are still inefficient, although more transgenic events were obtained.…”