Agrobacterium-mediated transformation of Hevea anther calli and their regeneration into plantlets


Citation

Coomber S., . and Jones H., . and Charlwood B. V., . and Arokiaraj P., . and Hafsah J., . Agrobacterium-mediated transformation of Hevea anther calli and their regeneration into plantlets. pp. 77-87. ISSN 0127-7065

Abstract

A gene transfer system for Hevea brasiliensis was established with Agrobacterium tumefaciens GV2260 (p35SGUSINT) and LBA4404 (pAL4404/pMON9793). In this system H. brasiliensis anther-derived calli were transformed with vectors harbouring the b-glucuronidase (gus) gene and the neomycin phosphotransferase (nptII) gene. The success of gene transfer was determined by histochemical staining and fluorometric assay for b-glucuronidase activity and enzyme linked immunosorbent assay for detecting neomycin phosphotransferase II protein levels. These independent assays all showed a several-fold increase compared to control values in enzyme activity and protein levels in extracts from transformed calli and embryoids of H. brasiliensis. The presence of the gus gene in transformed plantlets was confirmed by using the polymerase chain reaction protocol as performed on DNA isolated from transformed calli and embryoids.


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Abstract

A gene transfer system for Hevea brasiliensis was established with Agrobacterium tumefaciens GV2260 (p35SGUSINT) and LBA4404 (pAL4404/pMON9793). In this system H. brasiliensis anther-derived calli were transformed with vectors harbouring the b-glucuronidase (gus) gene and the neomycin phosphotransferase (nptII) gene. The success of gene transfer was determined by histochemical staining and fluorometric assay for b-glucuronidase activity and enzyme linked immunosorbent assay for detecting neomycin phosphotransferase II protein levels. These independent assays all showed a several-fold increase compared to control values in enzyme activity and protein levels in extracts from transformed calli and embryoids of H. brasiliensis. The presence of the gus gene in transformed plantlets was confirmed by using the polymerase chain reaction protocol as performed on DNA isolated from transformed calli and embryoids.

Additional Metadata

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Item Type: Article
AGROVOC Term: Hevea
AGROVOC Term: Hevea brasiliensis
AGROVOC Term: Genetic transformation
AGROVOC Term: Agrobacterium
AGROVOC Term: Gene transfer
AGROVOC Term: Genes
AGROVOC Term: Plant tissues
AGROVOC Term: Isolation
AGROVOC Term: Enzyme activity
AGROVOC Term: Callus
Depositing User: Ms. Suzila Mohamad Kasim
Last Modified: 24 Apr 2025 06:28
URI: http://webagris.upm.edu.my/id/eprint/23361

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