Herbicide-resistant transgenic creeping bentgrass plants obtained by electroporation using an altered buffer

Asano, Yasuhiro; Ito, Yasuyuki; Fukami, Masanobu; Sugiura, Kiyoyuki; Fujiie, Azusa

doi:10.1007/s002990050518

Cited by 19 publications

(13 citation statements)

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“…There is an increasing demand for improving the quality and stress resistance of creeping bentgrass via biotechnology approach. The majority of grass including creeping bentgrass now can be transformed by direct methods such as silicon-carbide fibermediated transformation, micro-projectile bombardment and protoplast transformation (either using PEG or by electroporation) (Ha et al, 1992;Wang et al, 1992;Zhong et al, 1993;Spangenberg et al, 1995;Asano et al, 1998, Bettany et al, 1998Dalton et al, 1998;Altpeter and Xu, 2000;Zhang et al, 2003;Goldman et al, 2004;Li and Qu et al, 2004). Previous reports have suggested that Agrobacterium tumefaciens-mediated transformation may offer a better alternative than the direct method for delivery of transgenes into plants.…”

Section: Introductionmentioning

confidence: 95%

Production of transgenic creeping bentgrass Agrostis stolonifera var. palustris plants by Agrobacterium tumefaciens-mediated transformation using hygromycin selection

Han

Chen

Bian

et al. 2005

Plant Cell Tiss Organ Cult

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A protocol was developed for Agrobacterium tumefaciens-mediated transformation of creeping bentgrass [Agrostis stolonifera L. var. palustris (Huds) Farw]. The transformation was performed using the vector pCAMBIA 1301 which contains the reporter (uidA) gene and the selectable marker hygromycin phosphotransferase (hph) gene. Embryogenic calli initiated from mature seeds were infected with A. tumefaciens strain EHA105 followed by hygromycin selection. Effects of some variables influencing efficiency of T-DNA delivery and selection were examined, and the protocol was successfully employed in the production of transgenic creeping bentgrass. Southern blot analysis demonstrated the integration of a single or two copies of foreign gene, and RT-PCR confirmed the transcription of the genes.

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Section: Introductionmentioning

confidence: 95%

Production of transgenic creeping bentgrass Agrostis stolonifera var. palustris plants by Agrobacterium tumefaciens-mediated transformation using hygromycin selection

Han

Chen

Bian

et al. 2005

Plant Cell Tiss Organ Cult

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“…For the T 11 generation, we extracted DNA using the CTAB method for PCR and Southern blotting. The PCR reaction procedure described by Asano et al (1998) was used in our study. A Biometra T1 Thermocycler was used for amplification.…”

Section: Molecular Tests To Transgenic Progeniesmentioning

confidence: 99%

Transformation of upland rice with the bar gene and selection for resistance to the herbicide Basta

et al. 2015

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In this study, Chinese upland rice cultivars including Handao297 and Handao502 were transformed with the anti-herbicide bar gene in order to achieve efficient weed control. PCR amplification, Southern blot analysis and field Basta resistance identification in early T 0 , T 1 , and late generation T 6 and T 11 -T 13 illustrated that the exogenous bar gene had been integrated into the genome of the upland rice varieties with stable expression in successive generations. Different transformants had a different gene copy numbers, which was not related to Basta resistance. Some of the transgenic lines showed gene silencing, whereas multiple copies of the bar gene were observed in other late generation transgenic lines. Basta resistance can thus be inherited stably among generations, and an optimal Basta concentration and selective pressure are suitable for resistance segregation and breeding of homozygous progenies with high resistance. After three generations of successive Basta resistance selection in 207 T 0 plants, we obtained 555 T 3 lines. Comprehensive identification and selection for Basta resistance, drought resistance, yield, and other agronomic traits in nine generations yielded two promising transgenic lines, HD297T-31 and HD297T-523. Although both varieties were resistant to 0.1 % Basta, HD297T-31 possessed high (100 %) resistance to Basta while HD297T-523 showed moderate (95 %) resistance. Moreover, during the reproductive stage, the stems, sheaths, leaves, and panicles of HD297T-31 were immune-highly resistant to highly-moderately resistant to 0.5 % Basta, whereas, they were highly resistantmoderately resistant to 1 % Basta. HD297T-523 was immune-highly resistant and immune-highly resistant to 0.5 and 1 % Basta respectively. The plant and stem percentage sensitive to 0.5 % Basta was lower than 1 and 3 % respectively. Since HD297T-31 showed isozygosity, it could be used as a new variety in production.

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“…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. The herbicide-resistance transformants are expected to be useful for reducing the cost and labor for eliminating weeds (Hoshino and Mii.…”

Section: Introductionmentioning

confidence: 98%

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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Herbicide-resistant transgenic creeping bentgrass plants obtained by electroporation using an altered buffer

Cited by 19 publications

References 0 publications

Production of transgenic creeping bentgrass Agrostis stolonifera var. palustris plants by Agrobacterium tumefaciens-mediated transformation using hygromycin selection

Production of transgenic creeping bentgrass Agrostis stolonifera var. palustris plants by Agrobacterium tumefaciens-mediated transformation using hygromycin selection

Transformation of upland rice with the bar gene and selection for resistance to the herbicide Basta

Production of bialaphos-resistant Nierembergia repens by electroporation

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