SacB-SacR Gene Cassette As the Negative Selection Marker to Suppress Agrobacterium Overgrowth in Agrobacterium-Mediated Plant Transformation

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dc.contributor.authorLiu, Yimingen
dc.contributor.authorMiao, Jiaminen
dc.contributor.authorTraore, Syen
dc.contributor.authorKong, Danyuen
dc.contributor.authorLiu, Yien
dc.contributor.authorZhang, Xunzhongen
dc.contributor.authorNimchuk, Zachary L.en
dc.contributor.authorLiu, Zongrangen
dc.contributor.authorZhao, Bingyu Y.en
dc.contributor.departmentSchool of Plant and Environmental Sciencesen
dc.date.accessioned2017-01-17T19:01:47Zen
dc.date.available2017-01-17T19:01:47Zen
dc.date.issued2016en
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
dc.description.versionPublished versionen
dc.format.extent70 - ? page(s)en
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.3389/fmolb.2016.00070en
dc.identifier.urihttp://hdl.handle.net/10919/74363en
dc.identifier.volume3en
dc.language.isoenen
dc.relation.urihttp://www.ncbi.nlm.nih.gov/pubmed/27833912en
dc.rightsIn Copyrighten
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en
dc.subjectAgrobacterium overgrowthen
dc.subjectCRISPR-Cas9en
dc.subjectNicotiana benthamianaen
dc.subjectSacB-SacR gene cassetteen
dc.subjectplant transformationen
dc.titleSacB-SacR Gene Cassette As the Negative Selection Marker to Suppress Agrobacterium Overgrowth in Agrobacterium-Mediated Plant Transformationen
dc.title.serialFrontiers in Molecular Biosciencesen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten
pubs.organisational-group/Virginia Techen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciencesen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences/CALS T&R Facultyen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences/Crop & Soil Environmental Scienceen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences/Horticultureen
pubs.organisational-group/Virginia Tech/All T&R Facultyen

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