Efficient octopine Ti plasmid-derived vectors for<i>Agrobacterium</i>-mediated gene transfer to plants

Deblaere, Rolf; Bytebier, Benny; Greve, Henri De; Deboeck, Francine; Schell, Jeff; Montagu, Marc Van; Leemans, J.

doi:10.1093/nar/13.13.4777

Cited by 684 publications

(382 citation statements)

References 32 publications

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“…The resulting plasmid, pASGRl ( Fig. l), was introduced into Agrobacterium tumefaciens C58C1 Rif harboring a Ti plasmid pGV2260 (Deblaere et al, 1985) by electroporation using E. coli Pulser (Bio-Rad Laboratories, Tokyo, Japan). This bacterium was used to transform leaf discs of Nicotiana tabactrm SR1 (Horsh et al, 1985).…”

Section: Materials a N D Methodsmentioning

confidence: 99%

Decrease in Activity of Glutathione Reductase Enhances Paraquat Sensitivity in Transgenic Nicotiana tabacum

et al. 1995

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Section: Materials a N D Methodsmentioning

confidence: 99%

Decrease in Activity of Glutathione Reductase Enhances Paraquat Sensitivity in Transgenic Nicotiana tabacum

et al. 1995

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“…The constructs generated were used to transform the C58C1 (pGV2260) Agrobacterium tumefaciens strain (Deblaere et al, 1985) and delivered into leaf cells of tobacco (Nicotiana benthamiana) by Agrobacterium infiltration as described previously (Voinnet et al, 2003), together with the C58C1 (pCH32) Agrobacterium strain to avoid gene silencing. Basically, Agrobacterium strains were resuspended in 10 mM MgCl 2 , 10 mM MES, pH 5.6, and 200 mM acetosyringone.…”

Section: Agroinfiltration Analysismentioning

confidence: 99%

“…The PYL8/RCAR3 clone was excised from the pGEM-T vector using SacI-XbaI double digestion and subcloned into the doubly digested pBIN121 vector. The T-DNA region of the pBIN121-PYL8/RCAR3 construct was transferred to Agrobacterium C58C1 (pGV2260; Deblaere et al, 1985) by electroporation. Arabidopsis plants (Col-0 ecotype) were transformed by the floral dip method (Clough and Bent, 1998).…”

Section: Vector Construction and Plant Transformationmentioning

confidence: 99%

The Nuclear Interactor PYL8/RCAR3 ofFagus sylvaticaFsPP2C1 Is a Positive Regulator of Abscisic Acid Signaling in Seeds and Stress

et al. 2009

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The functional protein phosphatase type 2C from beechnut (Fagus sylvatica; FsPP2C1) was a negative regulator of abscisic acid (ABA) signaling in seeds. In this report, to get deeper insight on FsPP2C1 function, we aim to identify PP2C-interacting partners. Two closely related members (PYL8/RCAR3 and PYL7/RCAR2) of the Arabidopsis (Arabidopsis thaliana) BetV I family were shown to bind FsPP2C1 in a yeast two-hybrid screening and in an ABA-independent manner. By transient expression of FsPP2C1 and PYL8/RCAR3 in epidermal onion (Allium cepa) cells and agroinfiltration in tobacco (Nicotiana benthamiana) as green fluorescent protein fusion proteins, we obtained evidence supporting the subcellular localization of both proteins mainly in the nucleus and in both the cytosol and the nucleus, respectively. The in planta interaction of both proteins in tobacco cells by bimolecular fluorescence complementation assays resulted in a specific nuclear colocalization of this interaction. Constitutive overexpression of PYL8/RCAR3 confers ABA hypersensitivity in Arabidopsis seeds and, consequently, an enhanced degree of seed dormancy. Additionally, transgenic 35S:PYL8/RCAR3 plants are unable to germinate under low concentrations of mannitol, NaCl, or paclobutrazol, which are not inhibiting conditions to the wild type. In vegetative tissues, Arabidopsis PYL8/RCAR3 transgenic plants show ABA-resistant drought response and a strong inhibition of early root growth. These phenotypes are strengthened at the molecular level with the enhanced induction of several ABA response genes. Both seed and vegetative phenotypes of Arabidopsis 35S:PYL8/RCAR3 plants are opposite those of 35S: FsPP2C1 plants. Finally, double transgenic plants confirm the role of PYL8/RCAR3 by antagonizing FsPP2C1 function and demonstrating that PYL8/RCAR3 positively regulates ABA signaling during germination and abiotic stress responses.

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“…Agrobacteriurn tumefaciens strain C58C1 containing pGV2260 (Deblaere et al, 1985) was cultivated in YEB medium (Vervliet et al, 1975).…”

Section: Plants Bacterial Strains and Mediamentioning

confidence: 99%

Reduction of the cytosolic fructose‐1,6‐bisphosphatase in transgenic potato plants limits photosynthetic sucrose biosynthesis with no impact on plant growth and tuber yield

et al. 1996

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SummarySucrose produced in source leaves is the predominant carbon source for developing sink tissues in most higher plants. Consequently the rate of sucrose synthesis is likely to be important for sink development and final crop yield. Two sucrose biosynthetic enzymes are believed to possess regulatory properties with respect to the rate of sucrose synthesis: (i) cytosolic FBPase and (ii) sucrose phosphate synthase. To study the impact of reduced photosynthetic sucrose biosynthesis on plant growth and crop yield a cDNA clone encoding cytosolic FBPase was isolated from a potato leaf cDNA library and used for antisansa experiments in transgenic potato plants. The cDNA clone cy-F1, containing an open reading frame of 1020 bp highly homologous (85%) to other known sequences of plant cytosolic FBPases, was cloned in reversed orientation between the 35S CaMV promoter and the octopine synthase polyadenylation signal. Out of 75 independent transformants five transgenic lines having 9 to 55 % of the wild-type FBPase activity were chosen for further analysis.A 45% reduction of the cytosolic FBPasa activity did not cause any measurable change in metabolite concentrations, growth behaviour or photosynthetic parameters of the transgenic plants. Inhibition of cytosolic FBPase activity below 20% of the wild-type activity led to an accumulation of 3-PGA, triose-phosphates and fructose-l,6-bisphosphete in source leaves. This resulted in a reduced light-saturated rate of assimilation measured via gas exchange and a decreased photosynthetic rate under conditions of the leaf disc electrode with saturating light and CO2. Measuring photosynthetic carbon fluxes by labelling leaf discs with 14C02 revealed a 53-65% reduction of sucrose synthesis whereas starch synthesis decreased only by 18-24%. The flux into the anionic and cationic

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Efficient octopine Ti plasmid-derived vectors forAgrobacterium-mediated gene transfer to plants

Cited by 684 publications

References 32 publications

Decrease in Activity of Glutathione Reductase Enhances Paraquat Sensitivity in Transgenic Nicotiana tabacum

Decrease in Activity of Glutathione Reductase Enhances Paraquat Sensitivity in Transgenic Nicotiana tabacum

The Nuclear Interactor PYL8/RCAR3 ofFagus sylvaticaFsPP2C1 Is a Positive Regulator of Abscisic Acid Signaling in Seeds and Stress

Reduction of the cytosolic fructose‐1,6‐bisphosphatase in transgenic potato plants limits photosynthetic sucrose biosynthesis with no impact on plant growth and tuber yield

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