Pan, Po-Hung , Chiang, Yu-Chung , Chou, Chang-Hung , Tsai, Chi-Chu .
The Development of a pollen-mediated gene transformation system for Spathoglottis plicata (Orchidaceae).
Traditionally, developing a plant regeneration system is a prerequisite for most of plant gene transformation. In present study, we show that the gene transformation of Spathoglottis plicata (Orchidaceae) can be achieved by pollen-mediated transformation which does not need to develop a regeneration system. Agrobacterium tumefaciens strain EHA 105 harboring a plasmid pCAMBIA 1302 which contains a reporter gene green fluorescent protein (GFP) and a hygromycin resistance gene controlling by CaMV 35S promoter and NOS terminator. Spathoglottis plicata pollinia was cultivated in pollen germination medium to germinate for 16 hours and germinated pollens were¬†cocultivated with EHA 105. The infected pollens were pollinated on the stigma of Spathoglottis plicata. After about two weeks later, those developing fruits were harvested. Developing embryos were observed under fluorescent microscope to detect the expression of GFP protein. About one month after pollination, fruits were matured and cultured on seed germination medium. The plantlets were transplanted on selection medium containing hygromycin to obtain transgenic plantlets. Those transgenic plants were confirmed by PCR and RT-PCR for GFP gene. These results show that gene transfer by pollen-mediated is an easy and efficient method for Spathoglottis plicata.
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1 - National Pingtung University of Science and Technology, Graduate Institute of Biotechnology
2 - Pingtung University of Science and Technology, Department of Life Sciences, Depart. Life Science, NPUST,, No. 1, Hseuhfu Rd. Neipu, Pingtung, 912, Taiwan
3 - China Medical University, Research Center for Biodiversity, Taichung, 404, Taiwan
4 - Kaohsiung District Agricultural Research and Extension Station, Crop Improvement Division, Pingtung, 900, Taiwan
Presentation Type: Poster:Posters for Sections
Location: Ball Room & Party Room/SUB
Date: Monday, July 28th, 2008
Time: 12:30 PM