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Ari Indrianto
Graduate school of Biotechnology and Faculty of Biology, Universitas Gajah Mada

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Improvement of Genetic Transformation Efficiency in Vanda tricolor Orchid Using Acetosyringone Rindang Dwiyani; Azis Purwantoro; Ari Indrianto; Endang Semiarti
Annales Bogorienses Vol. 14 No. 2 (2010): Annales Bogorienses
Publisher : National Research and Innovation Agency (BRIN)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ann.bogor.2010.v14.n2.27-32

Abstract

Vanda tricolor Lindl. var. suavis is an Indonesian wild orchid which is now extremely rare in nature due to its habitat destruction. Development of an appropriate method for improving Vanda orchid through genetic modification could be valuable for horticulture and, indirectly, also for conservation. In this research, a method of Agrobacterium-mediated transformation of two V. tricolor obtained from Salak Mount, West Java and Merapi Mount, Yogyakarta in Indonesia protocorms was improved using acetosyringone (AS). Concentrations of 0 and 25 ppm AS were used in transformation of pG35S binary vector containing kanamycin resistance geneinto V. tricolor protocorms. The result showed that 25 ppm AS was required on inoculation with Agrobacterium solution, without AS on cocultivation. Five weeks after treatment on the 300 ppm kanamicyn containing medium, green protocorms were obtained, that was 11.01% for V. tricolor from Salak Mount with pre-culture treatment prior to inoculation, 9.39% for V. tricolor from Merapi Mount with pre-culture treatment prior to inoculation, and 1.37% for V. tricolor from Merapi Mount without pre-culture treatment prior to inoculation. The best condition to set high efficiency of transformation is pre-culture protocorms prior inoculation, soaking protocorm on 25 ppm AS for 30 minutes, then co-cultivate its on AS-free callus induction medium.
Co-Authors Azis Purwantoro Endang Semiarti Rindang Dwiyani