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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 15, No. 4, 2012, pp. 1-10
Bioline Code: ej12032
Full paper language: English
Document type: Short Communication
Document available free of charge

Electronic Journal of Biotechnology, Vol. 15, No. 4, 2012, pp. 1-10

 en Development of pollen mediated activation tagging system for Phalaenopsis check for this species in other resources and Doritaenopsis check for this species in other resources
Tsay, Hsin-Sheng; Ho, Hui-Min; Gupta, Sushim K.; Wang, Chang-Sheng; Chen, Pei-Tsu & Chen, Emily Chin-Fun

Abstract

Abstract In the present study, a novel plant transformation system for Doritaenopsis check for this species in other resources and Phalaenopsis check for this species in other resources has been developed. The pollen-mediated activation tagging system was established by artificial pollination. The pollens, co-cultured with Agrobacterium tumefaciens check for this species in other resources strain EHA105 harbouring an activation tagging vector (pTAG-8), were used for pollination. In order to optimize the transformation efficiency, several factors (concentration of A. tumefaciens, concentration of acetosyringone during co-cultivation and the duration of co-cultivation) known to influence Agrobacterium-mediated DNA transfer were examined. A concentration of 0.5-1 x 108 CFU/ml for A. tumefaciens, 0.1 mM acetosyringone, and 6 hrs of co-culture period were found to be the optimal condition for high transformation efficiency. Integration of T-DNA into the genome of putative transgenic plants was confirmed by PCR and DNA blot analyses. Single copy of the transgene was observed in all transgenic plants analyzed. Most of the transgenic plants had a morphologically normal phenotype and the overall capsule formation efficiency was similar to control plant. Our results showed a new approach of genetic transformation in orchids and this method can be employed for genetic improvement of the orchids.

Keywords
activation tagging; Agrobacterium tumefaciens; Doritaenopsis cultivars; Phalaenopsis amabilis; transformation

 
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