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dc.contributor.authorPhlaetita, Wasana
dc.contributor.authorChin, Dong Poh
dc.contributor.authorNtui, Valentine Otang
dc.contributor.authorNakamura, Ikuo
dc.contributor.authorMii, Masahiro
dc.date.accessioned2021-03-30T07:51:58Z
dc.date.available2021-03-30T07:51:58Z
dc.date.issued2015
dc.identifier.citationPhlaetita, W., Chin, D. P., Otang, N. V., Nakamura, I., & Mii, M. (2015). High efficiency Agrobacterium-mediated transformation of Dendrobium orchid using protocorms as a target material. Plant Biotechnology, 32(4), 323–327. doi:10.5511/plantbiotechnology.15.0804a
dc.identifier.issn1342-4580
dc.identifier.issn1347-6114
dc.identifier.doi10.5511/plantbiotechnology.15.0804a
dc.identifier.urihttp://hdl.handle.net/10754/668390
dc.description.abstractDendrobium orchid is one of the most popular cut flower and potted plants. In this study, a protocol for efficient genetic transformation of D. nobile-type orchids was established by co-cultivating 21 day-old protocorms for 3 days with Agrobacterium tumefaciens strain EHA101 carrying pIG121Hm harboring β-glucuronidase (GUS) gene as reporter gene and hygromycin phosphotransferase (hpt) gene as selectable marker gene. After selection of the infected protocorms on New Dogashima (ND) medium containing 10 gl−1 maltose, 30 mgl−1 hygromycin and 20 mgl−1 meropenem for 3 months followed by the culture on hygromycin-free recovery medium for 1 month, secondary protocorm-like bodies (PLBs) produced on this medium were again transferred onto secondary selection (regeneration) medium. Plantlets were successfully regenerated from these secondary PLBs after the transfer. The highest transformation efficiency of 27.3% was obtained when protocomrs were inoculated with 10 times diluted Agrobacterium solution (OD600=0.1) for 300 min. Transformation of the selected plants was confirmed by GUS assay, PCR and Southern blot analysis. This protocol could be adopted to produce transgenic D. nobile-type orchids with various traits such as novel flower color and resistances to biotic and abiotic stresses.
dc.description.sponsorshipWe thank Mr. Nobuyuki Asai, Asai Daikeikan Co., Ltd. to kindly provide the seeds as plant materials in this research.
dc.publisherJapanese Society for Plant Cell and Molecular Biology
dc.rightsArchived with thanks to Japanese Society for Plant Cell and Molecular Biology
dc.subjectAgrobacterium tumefaciens
dc.subjectDendrobium nobile-type hybrid
dc.subjectGenetic transformation
dc.subjectOrchid
dc.subjectProtocorm
dc.titleHigh efficiency Agrobacterium-mediated transformation of Dendrobium orchid using protocorms as a target material
dc.typeArticle
dc.contributor.departmentBiological and Environmental Science and Engineering (BESE) Division
dc.contributor.departmentCenter for Desert Agriculture
dc.eprint.versionPost-print
dc.contributor.institutionGraduate School of Horticulture, Chiba University
dc.contributor.institutionCenter for Environment, Health and Field Sciences, Chiba University, Kashiwanoha 6-2-1 Kashiwa, Chiba 277-0882, Japan
dc.identifier.volume32
dc.identifier.issue4
dc.identifier.pages323-327
kaust.personNtui, Valentine Otang
dc.identifier.eid2-s2.0-84952641725


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