Thai Journal of Botany
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Title: Regeneration and transformation of Vanda lilacina Teijsm.& Binn.
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Pages: 41 - 48
Year: 2553
Year No.: 2
Volume: 1   Show All Articles
Abstract:           The objectives of this research were to improve the efficiency of regeneration and establish optimal condition for transformation of Vanda lilacina Teijsm & Binn. The new Dogashima (ND) medium supplemented with 1% potato extract was suitable for seed germination. Experiments were performed to determine the effect of plant growth regulators on the number of shoots. It was found that the shoot number per protocorm was 6 when cultured on ND medium supplemented with1 mg/l napthalene acetic acid and 5 mg/l benzylaminopurine. The protocorm regeneration was completely inhibited by hygromycin at 9 mg/l. Agrobacterium tumefaciens strain LBA 4404 (pCAMBIA1305.1) was used to establish a transformation protocol. The plasmid pCAMBIA containing antisense ACC oxidase,beta- glucuronidase (GUS) and hygromycin resistant (hpt) genes, was used. Co-cultivation of the protocorms with Agrobacterium LBA 4404 led to numerous positive GUS staining. Successful transformation of antisense ACC oxidase into V. lilacina was verified by GUS assay. The GUS assay revealed the GUS activity while PCR method indicated the integration of DNA.
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