caa a22 4500 46790006X CHVBK 20180406152815.0 cr unu---uuuuu 170328e20060601xx s 000 0 eng 10.1007/s00299-006-0114-9 doi (NATIONALLICENCE)springer-10.1007/s00299-006-0114-9 A highly efficient in vitro plant regeneration system and Agrobacterium -mediated transformation in Plumbago zeylanica [Elektronische Daten] [Xiaoping Wei, Xiaoping Gou, Tong Yuan, Scott Russell] Plumbago zeylanica is a unique model for studying flowering plant gametogenesis, heterospermy, and preferential fertilization, yet understanding the control of related molecular mechanisms is impossible without efficient and reproducible regeneration and stable genetic transformation. We found three key factors for enhancing successful regeneration: (1) tissue source of explants, (2) combination and concentration of growth regulators, and (3) culture conditions. The highest frequency of shoot regeneration was achieved using hypocotyl segments cultured on MS basal medium supplemented with BA 2.0mg/l, NAA 0.75mg/l, adenine 50mg/l and 10% (v/v) coconut milk under subdued light at 25±2°C; under these conditions, each hypocotyl segment produced over 30 shoots, arising primarily through direct organogenesis after 3 weeks of culture. Regenerated shoots rooted easily on half-strength basal MS medium and were successfully established in the greenhouse. Using this tissue culture protocol, reporter gene GUS under the constitutive CaMV 35S promoter was introduced into P. zeylanica cells of petiole, cotyledon and hypocotyl with A. tumefaciens strains AGL1 and LBA4404. Transient expression was observed in all recipient tissues. Stable transgenic calli originating from petiole were obtained. Springer-Verlag, 2006 Agrobacterium tumefaciens nationallicence Gene transformation nationallicence Growth regulators nationallicence Organogenesis nationallicence Plumbago zeylanica nationallicence Regeneration nationallicence AD : adenine free base nationallicence AS : acetosyringone nationallicence BA : 6-benzylaminopurine nationallicence CM : coconut milk nationallicence GUS : β-glucuronidase nationallicence IAA : indole-3-acetic acid nationallicence IBA : indole-3-butyric acid nationallicence MS medium : Murashige and Skoog (1962) medium nationallicence NAA : 1-naphthaleneacetic acid nationallicence NPTII : neomycin phosphotransferase nationallicence PGR : plant growth regulator nationallicence SEM : scanning electron microscopy nationallicence X-Gluc : 5-bromo-4-chloro-3-indolyl-β-d-glucuronide nationallicence ZT : zeatin nationallicence Wei Xiaoping Department of Botany and Microbiology, University of Oklahoma, 73019, Norman, OK, USA aut Gou Xiaoping Department of Botany and Microbiology, University of Oklahoma, 73019, Norman, OK, USA aut Yuan Tong Department of Botany and Microbiology, University of Oklahoma, 73019, Norman, OK, USA aut Russell Scott Department of Botany and Microbiology, University of Oklahoma, 73019, Norman, OK, USA aut Plant Cell Reports Springer-Verlag 25/6(2006-06-01), 513-521 0721-7714 25:6<513 2006 25 299 https://doi.org/10.1007/s00299-006-0114-9 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00299-006-0114-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wei Xiaoping Department of Botany and Microbiology, University of Oklahoma, 73019, Norman, OK, USA aut NATIONALLICENCE 700 1- Gou Xiaoping Department of Botany and Microbiology, University of Oklahoma, 73019, Norman, OK, USA aut NATIONALLICENCE 700 1- Yuan Tong Department of Botany and Microbiology, University of Oklahoma, 73019, Norman, OK, USA aut NATIONALLICENCE 700 1- Russell Scott Department of Botany and Microbiology, University of Oklahoma, 73019, Norman, OK, USA aut NATIONALLICENCE 773 0- Plant Cell Reports Springer-Verlag 25/6(2006-06-01), 513-521 0721-7714 25:6<513 2006 25 299 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer