Transgenic fertile Scoparia dulcis L., a folk medicinal plant, conferred with a herbicide-resistant trait using an Ri binary vector

Yamazaki, Mami; Son, Lin; Hayashi, Toshimitsu; Morita, Naokata; Asamizu, Tetsuya; Mourakoshi, Isamu; Saito, Kazuki

doi:10.1007/s002990050024

Cited by 7 publications

(11 citation statements)

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“…So far, there is only one report of production of transgenic herbicide-resistant S. dulcis plants using Agrobacterium rhizogenes (Yamazaki et al 1996), and there are no reports available on Agrobacterium tumefaciens-mediated transformation for this valuable medicinal plant. Establishment of an efficient transformation method may facilitate its improvement in terms of the accumulation levels of secondary metabolites.…”

Section: Introductionmentioning

confidence: 99%

Highly efficient production of transgenic Scoparia dulcis L. mediated by Agrobacterium tumefaciens: plant regeneration via shoot organogenesis

Mahender

Abbagani

Zhang

2011

Plant Biotechnol Rep

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Efficient Agrobacterium-mediated genetic transformation of Scoparia dulcis L. was developed using Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pCAMBIA1301 with b-glucuronidase (GUS) (uidA) and hygromycin phosphotransferase (hpt) genes. Two-day precultured leaf segments of in vitro shoot culture were found to be suitable for cocultivation with the Agrobacterium strain, and acetosyringone was able to promote the transformation process. After selection on shoot organogenesis medium with appropriate concentrations of hygromycin and carbenicillin, adventitious shoots were developed on elongation medium by twice subculturing under the same selection scheme. The elongated hygromycin-resistant shoots were subsequently rooted on the MS medium supplemented with 1 mg l -1 indole-3-butyric acid and 15 mg l -1 hygromycin. Successful transformation was confirmed by PCR analysis using uidA-and hpt-specific primers and monitored by histochemical assay for b-GUS activity during shoot organogenesis. Integration of hpt gene into the genome of transgenic plants was also verified by Southern blot analysis. High transformation efficiency at a rate of 54.6% with an average of 3.9 ± 0.39 transgenic plantlets per explant was achieved in the present transformation system. It took only 2-3 months from seed germination to positive transformants transplanted to soil. Therefore, an efficient and fast genetic transformation system was developed for S. dulcis using an Agrobacteriummediated approach and plant regeneration via shoot organogenesis, which provides a useful platform for future genetic engineering studies in this medicinally important plant.

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Section: Introductionmentioning

confidence: 99%

Highly efficient production of transgenic Scoparia dulcis L. mediated by Agrobacterium tumefaciens: plant regeneration via shoot organogenesis

Mahender

Abbagani

Zhang

2011

Plant Biotechnol Rep

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“…The bar gene has been used for plant transformation studies as a selection marker (De Block et al 1987;Spencer et al 1990;Saito et al 1992;Russell et al 1993;Anzai et al 1996;Hoshino and Mii 1998) and a target gene of interest for conferring herbicide resistance to crops such as tobacco (De Block et al 1987), potato (De Block 1988, Brassica (De Block et al 1989), Atropa (Saito et al 1992), Scoparia (Yamazaki et al 1996), Antirrhinum majus (Hoshino and Mii 1998), carrot (Chen and Punja 2002), rice (Dekeyser et al 1989), Phaseolus (Russell et al 1993), orchids (Anzai et al 1996;Knapp et al 2000), lawn grasses (Hartman et al1994;Asano et al 1998), and jute (Ghosh et al 2002). In our study, most nontransformed colonies turned brown by adding bialaphos into the culture medium, and the transformed colonies were efficiently selected in the selection process.…”

Section: Discussionmentioning

confidence: 99%

“…For introducing bialaphos-based herbicide resistance, pARK22 (Yamazaki et al 1996;Asano et al 1998), which consists of EcoRI-HindIII fragment from pARK5 (Saito et al 1992) and Escherichia coli vector pUC19, which contains the phosphinothricin acetyltransferase (bar) gene (Murakami et al 1986) driven by the CaMV 35s promoter and flanked by nopaline synthase terminator, were used.…”

Section: Plasmid Dnamentioning

confidence: 99%

“…Recently, many herbicide-resistant transformants were produced in Nicotiana tabacum (De Block et al 1987), potato (De Block 1988), Brassica (De Block et al 1989), Atropa belladonna (Saito et al 1992), Scoparia dulcis (Yamazaki et al 1996), Antirrhinum majus (Hoshino and Mii. 1998), carrot (Chen and Punja 2002), and Oryza sativa (Osakabe et al 2005) by Agrobacterium-mediated transformation, and O. sativa (Dekeyser et al 1989), Phaseolus (Russell et al 1993), orchids (Anzai et al 1996;Knapp et al 2000), grasses (Asano et al 1998;Gao et al 2006), jute (Ghosh et al 2002), maize (Howe et al2002), and Allium sativum (Park et al 2002) by direct gene-transfer methods such as particle bombardment and electroporation.…”

Section: Introductionmentioning

confidence: 99%

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Production of bialaphos-resistant Nierembergia repens by electroporation

Shizukawa

Mii

2008

Plant Biotechnol Rep

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Transgenic plants with the herbicide-resistance gene (bar gene) were obtained via organogenesis from isolated mesophyll protoplasts of Nierembergia repens after applying electroporation. Transient b-glucuronidase (GUS) activity of electroporated protoplasts assayed 2 days after applying an electric pulse showed that optimum condition (transient GUS activity 319 pmol 4 MU/mg per min and plating efficiency 2.43%) for electroporation was 0.5 kV/cm in field strength and 100 lF in capacitance. The protoplasts electroporated with the bar gene at this condition initiated formation of microcolonies on medium after 2 weeks. After 4 weeks of culture, equal volume of fresh 1/2-strength Murashige and Skoog (MS) medium containing 0.2 mg/l bialaphos was added for selection of transformed colonies. After 6 weeks of culture, growing colonies were transferred onto regeneration medium containing 1.0 mg/l bialaphos, on which they formed adventitious shoots 1-2 months after electroporation. The adventitious shoots rooted easily after transfer onto MS medium with bialaphos lacking plant-growth regulators. Transformation of these regenerants with the bar gene was confirmed by Southern analysis. Some of the transformants showed strong resistance to the application of bialaphos solution at 10.0 mg/l.

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“…This technique depends on the fact that the T-DNA derived from a Ti plasmid can be mobilized in traps by vir gene products of the Ri plasmid. We have been exploring the Ri binary vector system for the genetic transformation of pharmaceutically important plants [3][4][5][6][7][8][9] . In most cases, doubly transformed tissues integrated with both T-DNAs derived from an Ri plasmid and a binary vector were obtained in a high frequency without any selection of transformed cells by the traits conferred by binary vectors.…”

Section: Introductionmentioning

confidence: 99%

Transformation of Perilla frutescens var. crispa Using an Agrobacterium-Ri Binary Vector System.

Yamazaki

Kobayashi

Saito

1997

Plant Biotechnology

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Transgenic fertile Scoparia dulcis L., a folk medicinal plant, conferred with a herbicide-resistant trait using an Ri binary vector

Cited by 7 publications

References 0 publications

Highly efficient production of transgenic Scoparia dulcis L. mediated by Agrobacterium tumefaciens: plant regeneration via shoot organogenesis

Highly efficient production of transgenic Scoparia dulcis L. mediated by Agrobacterium tumefaciens: plant regeneration via shoot organogenesis

Production of bialaphos-resistant Nierembergia repens by electroporation

Transformation of Perilla frutescens var. crispa Using an Agrobacterium-Ri Binary Vector System.

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