title: Improved production of transgenic Dioscorea  zingiberensis (Dioscoreaceae) by Agrobacterium tumefaciens-mediated  transformation.
abstract: The establishment of high-efficiency Agrobacterium-mediated  transformation techniques could improve the production of Dioscorea  zingiberensis, a medicinal species with a high diosgenin content. We  co-cultivated embryogenic calli induced from mature seeds with A.  tumefaciens strain EHA105. A binary vector, pCAMBIA1381, which contains the gfp and hpt genes under the control of the ubiquitin promoter and  the CaMV 35S promoter, respectively, was used for transformation. Pre-culture,  basic medium, acetosyringone, and bacterial density were evaluated to establish  the most efficient protocol. The optimal conditions consisted of MS medium  without CaCl(2) for pre- and co-cultivation, three days  for pre-culture, addition of 200 μM AS, and an OD(600) of 0.5. The transgenic plants grown under selection  were confirmed by PCR analysis and Southern blot analysis. This protocol  produced transgenic D. zingiberensis plants in seven months, with a  transformation efficiency of 6%.
authors.affiliation: College of Life Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, China.
authors.forename: L
authors.initials: L
authors.lastname: Shi
authors.forename: J Q
authors.initials: JQ
authors.lastname: Fan
authors.forename: C G
authors.initials: CG
authors.lastname: Hu
authors.forename: J
authors.initials: J
authors.lastname: Luo
authors.forename: J L
authors.initials: JL
authors.lastname: Yao
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics*; D002228:Carbenicillin / Q000494:pharmacology; D002934:Cinnamates / Q000494:pharmacology; D027742:Dioscorea / Q000235:genetics*; D004144:Diosgenin / Q000378:metabolism; D004351:Drug Resistance; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D006921:Hygromycin B / Q000031:analogs & derivatives / Q000494:pharmacology; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D030821:Plants, Genetically Modified / Q000235:genetics; D010946:Plants, Medicinal / Q000235:genetics; D012639:Seeds / Q000166:cytology / Q000235:genetics / Q000502:physiology; D014170:Transformation, Genetic
chemical_list: D002934:Cinnamates; C485191:GFP10 Protein; D049452:Green Fluorescent Proteins; D006921:Hygromycin B; C026273:hygromycin A; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase; D002228:Carbenicillin; D004144:Diosgenin