Regeneration and Agrobacterium- mediated transformation of hop ( Humulus lupulus L.)

Abstract

An efficient procedure for direct organogenesis and regeneration of hop (Humulus lupulus L.) was established. For the first time Agrobacterium-mediated genetic transformation of hop (cv. "Tettnanger") was achieved. Shoot internodes from in vitro cultures were identified as the most suitable type of explant for regeneration. Using this type of explant, a shoot-inducing medium was developed that supported direct organogenesis of approximately 50% of the explants. Plantlets were successfully rooted and transferred to the greenhouse. Overall, in less than 6 months hop cultures propagated in vitro were regenerated to plants in the greenhouse. Agrobacterium-mediated genetic transformation was performed with the reporter gene GUS (beta-glucuronidase). The presence and function of transgenes in plants growing in the greenhouse was verified by PCR (polymerase chain reaction) and enzyme assay for GUS activity, respectively. We have obtained 21 transgenic plants from 1,440 explants initially transformed, yielding an overall transformation efficiency of 1.5%.

Effects of plant growth regulators and l-glutamic acid on shoot organogenesis in the halophyte Leymus chinensis (Trin.)

“…Monocotyledonous plants, in particular cereal plants, are naturally outside of the host range of A. tumefaciens, however, so the development of a system for the genetic transformation of monocotyledons has been slow (Hiei et al 1997). Nonetheless, successful transformations has been reported in some monocotyledonous plants, including Oryza (Hiei et al 1994), Hordeum (Tingay et al 1997), Humulus (Horlemann et al 2003) and Alstroemeria (Akutsu et al 2004). The bombardment of cells and tissues, another option for genetic improvement, has been attempted in many plant species (Iida et al 1991;Christou et al 1992;Batista et al 2008).…”

Genetic Transformation for Quality Improvement in Ornamental Climbers

“…Škof and Luthar ( 2005 ) reported the Agrobacteriummediated genetic transformation of H. Lupulus introducing gus reporter gene. Many other authors have also reported the effi cient transformation and regeneration system of H. lupulus (Horlemann et al 2003 ;Schwekendiek et al 2005 ;Batista et al 2008 ). Bombarded with the plasmid pSR5-2 ( gusA and nptII ), delivery into H. lupulus organogenic calli using the gene gun system was also reported by .…”

"Transgenic" metabolome of hop, some aspects of its development and prospects of utilization.

“…Tento první názorný pří-klad transgenního metabolomu u chmelu by měl být v budoucnu ná-sledován cílenějšími změnami podle toho, jak bude postupovat analýza genetické determinace biochemických drah účastnících se v syntéze chmelového metabolomu. Transgenní pletiva chmelu lze v současnosti získat klasickým způsobem při použití transformace bakteriemi A. tumefaciens (Oriniaková et al, 1996) a regenerace na selekčním médiu (Horlemann et al, 2003, Schwekendiek et al, 2007 nebo s použitím biolistické metody (Batista et al, 2008), i když v obou případech je oproti některým jiným rostlinným druhům efektivita transformace stále nízká (Horlemann et al, 2003). V obou přípa-dech lze úspěšně využít klasické monocistronické transformační vektory, tj.…”

“…This first illustrative example of transgenic metabolome in hop should be followed up with the targeted application of transgenosis in connection to the progress of our knowledge of biosynthesis pathways of hop metabolome. At present the transgenic hop tissues can be prepared either by a classical way using A. tumefaciens-mediated (Oriniaková et al, 1996) transformation and regeneration on selection media (Horlemann et al, 2003, Schwekendiek et al, 2007 or with the use of biolistic method (Batista et al, 2008), although in both cases the efficiency of hop transformation is still low in comparison with some other plant species (Horlemann et al, 2003). In both transformation methods it is possible to use classical monocistronic plant binary vectors i.e.…”