caa a22 4500 463235673 CHVBK 20180405153256.0 cr unu---uuuuu 170326e20070101xx s 000 0 eng 10.1007/s11240-006-9170-7 doi (NATIONALLICENCE)springer-10.1007/s11240-006-9170-7 Plate flooding as an alternative Agrobacterium -mediated transformation method for American chestnut somatic embryos [Elektronische Daten] [Ronald Rothrock, Linda Polin-McGuigan, Andrew Newhouse, William Powell, Charles Maynard] In an attempt to improve Agrobacterium-mediated transformation frequency of American chestnut somatic embryos, a novel method of inoculation/co-cultivation was developed. Plate flooding is a simple method where the Agrobacterium inoculum is poured onto the embryos while they remain on multiplication medium. This method tested the hypothesis that wounding tissues prior to co-cultivation was unnecessary or counterproductive. Two clones, WB296 and P1-1, were tested for differences in transformation efficiency as measured by the number of transformed embryogenic cell lines per Petri dish, the total number of transformed cell lines (embryos plus callus) and percentage of transformants that remained embryogenic. Plate flooding using clone WB296 produced significantly more transformed embryo cell lines and had a higher percentage of transformants remain embryogenic. The number of total transformed cell lines (embryos plus callus) was the same as obtained by other methods (desiccation, blot dry, sand abrasion, sonication and vacuum infiltration). With clone P1-1 there were no significant differences among the inoculation/co-cultivation treatments tested. Polymerase chain reaction and Southern hybridizations confirmed that the transgene of interest had been stably integrated into both American chestnut clones. Whole plants were regenerated from clone P1-1. Springer Science+Business Media B.V., 2006 BAR nationallicence Castanea dentata nationallicence Genetic engineering nationallicence GFP nationallicence Oxalate oxidase nationallicence OxO nationallicence PEM nationallicence Sonication nationallicence Transgenic nationallicence WB296 : American chestnut somatic embryo clone WB296-10A-2 nationallicence BAR : Bialaphos (and PPT)-resistance nationallicence GFP : Green fluorescent protein nationallicence OxO : Oxalate oxidase nationallicence P1-1 : American chestnut somatic embryo clone Pond1-1 nationallicence PEM : Pro-embryogenic mass nationallicence PPT : Phosphinothricin nationallicence T-embryos : Transformed embryogenic cell lines nationallicence T-events : Transformed cell lines (embryos plus callus) nationallicence Rothrock Ronald Institute for Sustainable and Renewable Resources, Institute for Advanced Learning and Research, Danville, VA, USA aut Polin-McGuigan Linda Faculty of Forest and Natural Resources Management, SUNY College of Environmental Science and Forestry, Syracuse, NY, USA aut Newhouse Andrew Faculty of Environmental and Forest Biology, SUNY College of Environmental Science and Forestry, Syracuse, NY, USA aut Powell William Faculty of Environmental and Forest Biology, SUNY College of Environmental Science and Forestry, Syracuse, NY, USA aut Maynard Charles Faculty of Forest and Natural Resources Management, SUNY College of Environmental Science and Forestry, Syracuse, NY, USA aut Plant Cell, Tissue and Organ Culture Springer Netherlands 88/1(2007-01-01), 93-99 0167-6857 88:1<93 2007 88 11240 https://doi.org/10.1007/s11240-006-9170-7 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11240-006-9170-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Rothrock Ronald Institute for Sustainable and Renewable Resources, Institute for Advanced Learning and Research, Danville, VA, USA aut NATIONALLICENCE 700 1- Polin-McGuigan Linda Faculty of Forest and Natural Resources Management, SUNY College of Environmental Science and Forestry, Syracuse, NY, USA aut NATIONALLICENCE 700 1- Newhouse Andrew Faculty of Environmental and Forest Biology, SUNY College of Environmental Science and Forestry, Syracuse, NY, USA aut NATIONALLICENCE 700 1- Powell William Faculty of Environmental and Forest Biology, SUNY College of Environmental Science and Forestry, Syracuse, NY, USA aut NATIONALLICENCE 700 1- Maynard Charles Faculty of Forest and Natural Resources Management, SUNY College of Environmental Science and Forestry, Syracuse, NY, USA aut NATIONALLICENCE 773 0- Plant Cell, Tissue and Organ Culture Springer Netherlands 88/1(2007-01-01), 93-99 0167-6857 88:1<93 2007 88 11240 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer