Agrobacterium tumefaciens -mediated transformation of Robinia pseudoacacia

Igasaki, Tomohiro; Mohri, Takeshi; Ichikawa, Hiroaki; Shinohara, Kenji

doi:10.1007/s002990050754

Cited by 40 publications

(25 citation statements)

References 32 publications

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“…We established previously a simple and reliable procedure for the regeneration of transgenic Japanese broad-leaved trees (Mohri et al 1996(Mohri et al , 1997(Mohri et al , 1999Igasaki et al 2000). However, to our knowledge, no studies of the transformation of Japanese coniferous species have been reported.…”

Section: Resultsmentioning

confidence: 88%

Somatic embryogenesis and plant regeneration from immature zygotic embryos of Cryptomeria japonica D. Don

et al. 2003

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This report describes the successful plant regeneration via somatic embryogenesis from immature zygotic embryos of Cryptomeria japonica D. Don. For the induction of embryogenic tissue, we determined that the optimal medium contained N6-benzyladenine and 2,4-dichlorophenoxyacetic acid. Immature zygotic embryos that were collected at the end of June yielded embryogenic tissue at the highest frequency. Embryogenic tissues that had proliferated in liquid medium included small and loosely packed cells and elongating or elongated cells. We used ten cell lines to determine the optimal medium for the development of somatic embryos. Induced somatic embryos germinated with synchronous sprouting of cotyledons, hypocotyls and roots. Gibberellin A3 in the germination medium had a positive effect on both the elongation of hypocotyls and the survival of seedlings. The frequencies of induction and germination of somatic embryos differed among the cell lines examined. Most of the seedlings grew normally. This system of somatic embryogenesis required 4-5 months for the regeneration of C. japonica plantlets from immature zygotic embryos.

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Section: Resultsmentioning

confidence: 88%

Somatic embryogenesis and plant regeneration from immature zygotic embryos of Cryptomeria japonica D. Don

et al. 2003

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“…This observation is surprising because Agrobacterium strain EHA101 (or its derivative EHA105, or its oncogenic progenitor A281) is generally recognized as being superior in facilitating gene transfer to plant cells; for example, in leguminous species, such as soybean (Meurer et al 1998;Donaldson and Simmonds 2000), pea (Nadolska-Orczyk and Orczyk 2000) and peanut (Egnin et al 1998), but also in other crops, such as apple (De Bondt et al 1994), cabbage (Takasaki et al 1997), blueberry (Cao et al 1998) and wheat and barley (Guo et al 1998). However, the fact that this strain is not better than other strains, or even worse, has also been observed in Phaseolus vulgaris (Zhang et al 1997) and in other plants, such as cotton (Sunilkumar and Rathore 2001), rice (Hiei et al 1994(Hiei et al , 1997, kalanchoe (Jia et al 1989), black locust (Igasaki et al 2000) and hibiscus (Srivatanakul et al 2001). This underscores the importance of testing various A. tumefaciens strains for every species and genotype under study.…”

Section: Resultsmentioning

confidence: 88%

An optimized Agrobacterium -mediated transformation procedure for Phaseolus acutifolius A. Gray

et al. 2002

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To improve Agrobacterium tumefaciensmediated transformation of Phaseolus acutifolius, we examined the effect of different factors on T-DNA transfer by measuring transient expression levels of an introncontaining β-glucuronidase gene. Improved transformation frequencies were obtained with an A. tumefaciens strain carrying nopaline-type virulence genes and when calli were infected with Agrobacterium cells in the earlylog growth phase. Optimized co-cultivation was performed at 22°C under a 16/8-h (day/night) photoperiod in an acidic medium (pH 5.5) in the presence of 200 µM acetosyringone. By combining the best treatments, an efficient and reproducible transformation procedure was established for the P. acutifolius genotype NI576. Southern and immunoblot analyses confirmed the stable integration and expression of the transgenes in the primary transgenic plants and their progeny.

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“…Arrillaga and Merkle (1994) reported transient b-glucuronidase (GUS) expression after bombardment of black locust proembryogenic masses but no production of transgenic somatic embryos. Recently, phenotypically normal transgenic plants were obtained after infection of black locust stem and leaves with Agrobacterium tumefaciens (Igasaki et al 2000). In this work, we present a transformation protocol, based on SAAT (sonication-assisted Agrobacterium-mediated transformation, Trick and Finer 1997), to obtain herbicide-resistant transgenic black locust plants from cotyledon explants.…”

Section: Introductionmentioning

confidence: 99%

Regeneration of herbicide-tolerant black locust transgenic plants by SAAT

2004

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A protocol based on SAAT (sonication-assisted Agrobacterium-mediated transformation) has been developed to obtain herbicide-resistant transgenic black locust (Robinia pseudoacacia L.) plants. Cotyledon explants were co-cultivated with Agrobacterium AGL1 strain carrying the pTAB16 plasmid (bar and gusA genes). The effects of bacterial concentration (OD550 of 0.3, 0.6, 0.8) and method of infection (sonication vs immersion) on bacterial delivery were determined by assaying cotyledons for transient beta-glucuronidase expression 3 days after infection. SAAT increases transient expression efficiency especially at an OD550 of 0.6. After determining bacterial concentration and infection method, other factors affecting transformation efficiency, such as explant preconditioning and period of time before applying selection, were tested. From these experiments, the preferred protocol for black locust cotyledon transformation should include sonication of preconditioned cotyledons in AGL1 suspension, coculture for 3 days with 100 microM acetosyringone and transfer to selection medium with 4 mg/l phosphinothricin and 150 mg/l timentin. Of the initial explants, 2% produced at least one transgenic shoot. Genetic transformation was confirmed by Southern hybridization, chlorophenol red assay and herbicide tolerance of the regenerated plants.

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Agrobacterium tumefaciens -mediated transformation of Robinia pseudoacacia

Cited by 40 publications

References 32 publications

Somatic embryogenesis and plant regeneration from immature zygotic embryos of Cryptomeria japonica D. Don

Somatic embryogenesis and plant regeneration from immature zygotic embryos of Cryptomeria japonica D. Don

An optimized Agrobacterium -mediated transformation procedure for Phaseolus acutifolius A. Gray

Regeneration of herbicide-tolerant black locust transgenic plants by SAAT

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