Overproduction of alfalfa glutamine synthetase in transgenic tobacco plants

Eckes, Peter; Schmitt, Petra M.; Daub, Winfried; Wengenmayer, Friedrich

doi:10.1007/bf02464891

Cited by 96 publications

(48 citation statements)

References 24 publications

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“…In this system we made use of A. tumefaciens for gene delivery, and the bar gene, which confers resistance to the herbicide Basta (PPT), as the selectable marker. PPT is an inhibitor of Gin synthase in plants (Eckes et al, 1989;Krieg et al, 1990) and is the active ingredient in the nonselective herbicide Basta. The bar gene encodes the enzyme PAT, which catalyzes the conversion of PPT into a nontoxic acetylated product (De Block et al, 1987).…”

Section: Discussionmentioning

confidence: 99%

Agrobacterium-Mediated Transformation of Subterranean Clover (Trifolium subterraneum L.)

et al. 1994

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We have developed a rapid and reproducible transformation system for subterranean clover (Trifolium subterraneum 1.) using Agrobacterium tumefaciens-mediated gene delivery. Hypocotyl segments from seeds that had been allowed to imbibe were used as explants, and regeneration was achieved via organogenesis. Clucose and acetosyringone were required in the co-cultivation medium for efficient gene transfer. DNA constructs containing four genes encoding the enzymes phosphinothricin acetyl transferase, 8-glucuronidase (GUS), neomycin phosphotransferase, and an a-amylase inhibitor were used to transform subterranean clover. lransgenic shoots were selected on a medium containing 50 mg/L of phosphinothricin. Four commercial cultivars of subterranean clover (representing all three subspecies) have been successfully transformed. Southern analysis revealed the integration of 1-DNA into the subterranean clover genome. l h e expression of the introduced genes has been confirmed by enzyme assays and northern blot analyses. Transformed plants grown in the glasshouse showed resistance to the herbicide Basta at applications equal to or higher than rates recommended for killing subterranean clover in field conditions. In plants grown from the selfed seeds of the primary transformants, the newly acquired gene encoding CUS segregated as a dominant Mendelian trait. ~ Subterranean clover (Trifolium subterraneum L., subclover) is native to the Mediterranean regions and is grown as a pasture legume mainly in Australia and the countries around the Mediterranean. To a lesser extent, it is also grown in the Americas, Africa, New Zealand, and Japan (Johnstone and McLean, 1987). In Australia, subterranean clover is the major pasture legume and is grown on more than 16 million ha of mainly acidic and infertile lands. As a pasture crop and because of its ability to fix nitrogen, subterranean clover is a major contributor to the productivity of the meat, wool, dairy, and wheat industries in Australia (Johnstone and McLean, 1987). It also has the potential to be grown in other temperate regions of the world. A number of grain and pasture legumes are now amenable to gene transfer by genetic engineering using the Agrobacte-rium-mediated gene delivery system. These include soybean (Hinchee et al.

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Section: Discussionmentioning

confidence: 99%

Agrobacterium-Mediated Transformation of Subterranean Clover (Trifolium subterraneum L.)

et al. 1994

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“…PPT is a potent inhibitor of Gln synthase in plants (Eckes et al, 1989;Krieg et al, 1990) and is available commercially as a nonselective herbicide called Basta. The bar gene, which confers resistance to PPT, encodes the enzyme PAT, which catalyzes the conversion of PPT to a nontoxic acetylated product (De Block et al, 1987).…”

Section: Dlscusslonmentioning

confidence: 99%

Transformation and Regeneration of Two Cultivars of Pea (Pisum sativum L.)

et al. 1993

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The pea (Pisum sativum) is an important grain legume crop plant that has gained worldwide economic importance as a source of protein for animal and human nutrition. In addition, it has well-defined genetics, and it has been commonly used as a model plant for research in plant physiology and biochemistry. The productivity and value of peas could be greatly increased by the introduction of stably inherited traits such as pest and disease resistance, herbicide resistance, and improved protein quality. These traits are not available in natural populations of near relatives of cultivated peas, but current advances in plant genetic engineering provide a potentially powerful tool for achieving these goals by another means.The prerequisites for the transfer of foreign genes into any plant species by genetic engineering are an efficient gene delivery system, such as Agrobacterium-mediated DNA transfer, an effective selectable marker for transformed cells, and the ability to regenerate mature, fertile, transgenic plants from transformed tissue in culture.Regeneration via embryogenesis or organogenesis has been described for a variety of pea explants, e.g. from immature leaflets (Mroginski and Kartha, 1981;Rubluo et al., 1984), from cotyledonary node (Jackson and Hobbs, 1990), from hypocotyls (Nielsen et al., 1991), from embryos (Kysely et * Corresponding author; fax 61-6-246-5000. 75 1 Natali and Cavallini, 1987;Tetu et al., 1990), from various organs of seedlings (Malmberg, 1979;Hussey and Gunn, 1984;Ezhova et al., 1985), and from protoplast cultures (Jacobsen and Kysely, 1984; Puonti-Kaerlas and Eriksson, 1988; Lehminger-Mertens and Jacobsen, 1989). Agrobacterium-mediated transfonnation of various pea explants has also been reported, e.g. stem explants (Lulsdorf et al., 199 l), embryonic axis and epicotyl segments (Filippone and Lurquin, 1989;Puonti-Kaerlas et al., 1989), nodus explants (De Kathen and Jacobsen, 1990; Nauerby et al., 1991), and root explants and protoplasts (Schaerer and Pilet, 1991). Tumors were induced in young pea plants by wild-type Agrobacterium . However, no mature transgenic pea plants were regenerated from any of the above transformation systems. The only report to date of stable transformation of peas and the production of mature, flowering, transgenic pea plants is by Puonti-Kaerlas et al. (1990), who achieved regeneration by organogenesis via callus formation using a gene encoding hygromycin phosphotransferase as a selectable marker.In this paper we report the development of a routine, reliable transformation and regeneration system for peas. The procedure has been used to introduce herbicide resistance and the expression of an antibiotic resistance gene into two cultivars of peas using an Agrobacterium tumefaciens-mediated delivery system. Integration of the two traits was stable, and their frequency in the first generation progeny followed the Mendelian pattem. MATERIALS AND METHODS Plant Material and Transformation ProcedurePea (Pisum sativum L.) cv Greenfeast and cv Rondo were grown in t...

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“…Several attempts have been made to modulate the levels of the GS enzyme in different plants using genetic engineering tools, with rather mixed outcomes (Eckes et al 1989;Hemon et al 1990;Hirel et al 1992Hirel et al , 1997Temple et al 1993Temple et al , 1994Temple et al , 1998Su et al 1995;Vincent et al 1997;Brugiere et al 1999;Gallardo et al 1999;Limami et al 1999;Ortega et al 2001;Fuentes et al 2001;Oliveira et al 2002;Carvalho et al 2003;Fei et al 2003;Fu et al 2003;Harrison et al 2003). Tobacco plants transformed with different GS 1 genes driven by the CaMV 35S promoter have shown an increase in GS activity and GS 1 polypeptide level in the leaves (Eckes et al 1989;Hemon et al 1990;Hirel et al 1992Hirel et al , 1997Temple et al 1993;Fuentes et al 2001;Oliveira et al 2002). These plants showed little change in growth or in leaf protein content under optimal growth conditions.…”

Section: Introductionmentioning

confidence: 99%

Biochemical and molecular characterization of transgenic Lotus japonicus plants constitutively over-expressing a cytosolic glutamine synthetase gene

Ortega

Temple

Bagga

et al. 2004

Planta

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Higher plants assimilate nitrogen in the form of ammonia through the concerted activity of glutamine synthetase (GS) and glutamate synthase (GOGAT). The GS enzyme is either located in the cytoplasm (GS 1 ) or in the chloroplast (GS 2 ). To understand how modulation of GS activity affects plant performance, Lotus japonicus L. plants were transformed with an alfalfa GS 1 gene driven by the CaMV 35S promoter. The transformants showed increased GS activity and an increase in GS 1 polypeptide level in all the organs tested. GS was analyzed by non-denaturing gel electrophoresis and ion-exchange chromatography. The results showed the presence of multiple GS isoenzymes in the different organs and the presence of a novel isoform in the transgenic plants. The distribution of GS in the different organs was analyzed by immunohistochemical localization. GS was localized in the mesophyll cells of the leaves and in the vasculature of the stem and roots of the transformants. Our results consistently showed higher soluble protein concentration, higher chlorophyll content and a higher biomass accumulation in the transgenic plants. The total amino acid content in the leaves and stems of the transgenic plants was 22-24% more than in the tissues of the non-transformed plants. The relative abundance of individual amino acid was similar except for aspartate/asparagine and proline, which were higher in the transformants.

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Overproduction of alfalfa glutamine synthetase in transgenic tobacco plants

Cited by 96 publications

References 24 publications

Agrobacterium-Mediated Transformation of Subterranean Clover (Trifolium subterraneum L.)

Agrobacterium-Mediated Transformation of Subterranean Clover (Trifolium subterraneum L.)

Transformation and Regeneration of Two Cultivars of Pea (Pisum sativum L.)

Biochemical and molecular characterization of transgenic Lotus japonicus plants constitutively over-expressing a cytosolic glutamine synthetase gene

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