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dc.contributor.authorLiu, Yen_US
dc.contributor.authorMiao, Jen_US
dc.contributor.authorTraore, Sen_US
dc.contributor.authorKong, Den_US
dc.contributor.authorLiu, Yen_US
dc.contributor.authorZhang, Xen_US
dc.contributor.authorNimchuk, ZLen_US
dc.contributor.authorLiu, Zen_US
dc.contributor.authorZhao, Ben_US
dc.coverage.spatialSwitzerlanden_US
dc.date.accessioned2017-01-17T19:01:47Z
dc.date.available2017-01-17T19:01:47Z
dc.date.issued2016en_US
dc.identifier.urihttp://hdl.handle.net/10919/74363
dc.description.abstractAgrobacterium overgrowth is a common problem in Agrobacterium-mediated plant transformation. To suppress the Agrobacterium overgrowth, various antibiotics have been used during plant tissue culture steps. The antibiotics are expensive and may adversely affect plant cell differentiation and reduce plant transformation efficiency. The SacB-SacR proteins are toxic to most Agrobacterium tumefaciens strains when they are grown on culture medium supplemented with sucrose. Therefore, SacB-SacR genes can be used as negative selection markers to suppress the overgrowth of A. tumefaciens in the plant tissue culture process. We generated a mutant A. tumefaciens strain GV2260 (recA-SacB/R) that has the SacB-SacR cassette inserted into the bacterial genome at the recA gene locus. The mutant Agrobacterium strain is sensitive to sucrose but maintains its ability to transform plant cells in both transient and stable transformation assays. We demonstrated that the mutant strain GV2260 (recA-SacB/R) can be inhibited by sucrose that reduces the overgrowth of Agrobacterium and therefore improves the plant transformation efficiency. We employed GV2260 (recA-SacB/R) to generate stable transgenic N. benthamiana plants expressing a CRISPR-Cas9 for knocking out a WRKY transcription factor.en_US
dc.format.extent70 - ? page(s)en_US
dc.languageengen_US
dc.relation.urihttp://www.ncbi.nlm.nih.gov/pubmed/27833912en_US
dc.subjectAgrobacterium overgrowthen_US
dc.subjectCRISPR-Cas9en_US
dc.subjectNicotiana benthamianaen_US
dc.subjectSacB-SacR gene cassetteen_US
dc.subjectplant transformationen_US
dc.titleSacB-SacR Gene Cassette As the Negative Selection Marker to Suppress Agrobacterium Overgrowth in Agrobacterium-Mediated Plant Transformationen_US
dc.typeArticle - Refereed
dc.description.versionPublished online (Publication status)en_US
dc.title.serialFrontiers in Molecular Biosciencesen_US
dc.identifier.doihttps://doi.org/10.3389/fmolb.2016.00070
dc.identifier.volume3en_US
pubs.organisational-group/Virginia Tech
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences/CALS T&R Faculty
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences/Crop & Soil Environmental Science
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences/Horticulture
pubs.organisational-group/Virginia Tech/All T&R Faculty


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