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Optimization Of Biolistic Transformation Of Yellow Watermelon Cotyledons Using Gene For Î’-Glucuronidase As Visual Marker.

UNSPECIFIED (2007) Optimization Of Biolistic Transformation Of Yellow Watermelon Cotyledons Using Gene For Î’-Glucuronidase As Visual Marker. In: APAC (Asia Pacific Conference on Plant Tissue Culture), 17 - 20 June 2007, PWTC, Kuala Lumpur, Malaysia. (Unpublished)

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Abstract

Regeneration system of watermelon in vitro has been in place for over a decade and has paved the way for studies involving genetic improvement through Agrobacterium-mediated and biolistics-mediated transformation. In this study, cotyledon sections from yellow watermelon were successfully regenerated in MS medium containing 20 µM of BAP (100%) with multiple shoots from the axillary meristems. The rooting system was achieved using half-strength of MS medium supplemented with 2.5 µM IBA at 88%. Parameters for biolistic were optimized using β-gluruconidase (GUS) as the reporter gene. The plasmid pRQ6 was used as microprojectile containing both gusA and hph gene to optimize biolistic parameters that included helium pressure (650, 900, 1100psi), target distance (6, 9, 12cm), explant ages (4 and 5 days old) and osmotic treatment using sorbitol and mannitol at 0.2, 0.4, 0.6M concentrations. The optimized biolistic parameters for cotyledon explant were at 1100 helium pressure, 9cm target distance, 4-day-old explants and osmotic treatment for 24 h on medium supplemented with 0.6 M mannitol prior to bombardment. These parameters have been used to obtain stable transformed explants for subsequent experiments.

Item Type:Conference or Workshop Item (Poster)
Subjects:S Agriculture > SB Plant culture
Divisions:Others
ID Code:9246
Deposited By: Ms. Zaidah Rahmat
Last Modified:17 Aug 2009 10:14

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