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

Abstract

A two-component cloning system to transfer foreign DNA into plants was derived from the octopine Ti plasmid pTiB6S3. pGV2260 is a non-oncogenic Ti plasmid from which the T-region is deleted and substituted by pBR322. pGV831 is a streptomycin-resistant pBR325 derivative that contains a kanamycin resistance marker gene for plant cells and a site for cloning foreign genes between the 25-bp border sequences of the octopine T-region. Conjugative transfer of pGV831 derivatives to Agrobacterium and cointegration by homologous recombination between the pBR322 sequences present on pGV831 and pGV2260, can be obtained in a single step. Strains carrying the resulting cointegrated plasmids transfer and integrate T-DNA into the genome of tobacco protoplasts, and transformed tobacco calli are readily selected as resistant to kanamycin. Intact plants containing the entire DNA region between the T-DNA borders have been regenerated from such clones. In view of these properties we present pGV831 and its derivatives as vectors for efficient integration of foreign genes into plants.

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

“…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).…”

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

“…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.…”

“…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).…”

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

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