Agrobacterium-Mediated Transformation Of Dendrobium Sonia-28

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Date
2012-06
Authors
Julkifle, Advina Lizah
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Publisher
Universiti Sains Malaysia
Abstract
Effective propagation of protocorm-like bodies (PLBs) of Dendrobium sonia-28 was achieved by using micropropagation technique. The highest PLBs proliferation within 21 days was observed in semi-solid media over liquid culture. Half strength of Murashige and Skoog (MS) media produced the highest PLBs growth rate percentage compared to full and double strength MS media in semi-solid and liquid cultures. In sucrose treatment, the highest PLBs growth rate percentage was in 10 g.L-1 sucrose. In the conditions of absence of sucrose (0 g.L-1) or excessive of sucrose (30 g.L-1) had proven detrimental for the PLBs growth and development. PLBs growth rate percentage was the highest (14.45%) in media devoid of exogenous 6-Benzylaminopurine (BAP) or Naphthalene acetic acid (NAA). However, the addition of BAP and NAA in combinations of BAP (4.44 or 8.88 μM) and NAA (8.88 μM) resulted in increased PLBs growth to 14% compared to when added separately. The development of PLBs was monitored at cellular level based on histology analysis. Secondary PLBs were directly formed from the actively dividing meristematic cells of the epidermal layer of primary tissue without any intermediate tissue development. Based on scanning electron microscope (SEM) observation, the surface structure of PLBs was found suitable for facilitating Agrobacterium infection during transformation process. The interaction of Agrobacterium to plant tissue was evaluated based on Agrobacterium motility and attachment competency. Agrobacterium motility respond was the highest (1.5 unit) toward large size (3-4mm) PLBs regardless of wounding conditions.
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Keywords
Agrobacterium-mediated transformation , of dendrobium sonia-28
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