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Abstract Detail


Economic Botany: Applied Plant Biology

Ho, Hui-Min [1], Chen, Chin-Fun [2], Tsay, Hsin-Sheng [2].

A novel method of transformation of orchid Doritaenopsis by pollen co-cultured with Agrobacterium tumefaciens.

In the present study, a novel plant transformation technique for orchid Doritaenopsis has been developed. Gene transfer was affected by pollination of flowers with pollen co-cultured with Agrobacterium tumefaciens strains EHA105 harboring plasmids pCAMBIA1301 or pCAMBIA1302, and strain EHA105 with an activation tagging vector (pTAG-8). The construct pCAMBIA1302 had GFP and pigment biosynthetic gene DFR (dihydroflavonol 4-reductase), while pCAMBIA1301 had £]-glucuronidase (GUS) and F3H (flavanone 3-hydroxylase) genes. Putative transformants (1016 nos) of Doritaenopsis cultivars SPH33 - Dtps.Sinica Sunday were obtained using Agrobacterium tumefaciens of 1*108 CFU/ml without acetosyringone and a coculture period of 12 h. Concentrations of acetosyringone (AS), Agrobacterium and co-cultivation period between pollen and Agrobacterium, before pollination had significant influences on capsule rate and number of transformants obtained among three cultivars of Doritaenopsis. In Doritaenopsis cultivars SPH33 - Dtps.Sinica Sunday, green fluorescent protein (GFP) activity could be seen in 4.89% of transformant seeds. Presence of hygromycin phosphotransferase (hpt) and GUS genes by polymerase chain reaction (PCR), and DFR gene by PCR-Southern in selected transformants was confirmed.


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1 - Chaoyang University of Technology, Graduate Institute of Biotechnology, 168, Jifong East Road, Wufong Township, Taichung County, 41349, Taiwan
2 - Chaoyang University of Technology, Graduate Institute of Biotechnology, 168 Jifong East Road, Wufong Township, Taichung County, 41349, Taiwan
3 - Chaoyang University of Technology, Graduate Institute of Biotechnology, 168 Jifong East Road, Wufong Township, Taichung County, 41349, Taiwan

Keywords:
Agrobacterium tumefaciens
orchid.

Presentation Type: Oral Paper:Papers for Topics
Session: 59
Location: 157/Law
Date: Wednesday, July 30th, 2008
Time: 10:15 AM
Number: 59001
Abstract ID:86


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