A protocol for Agrobacterium-mediated transformation of banana with a rice chitinase gene.
A rice chitinase gene (RCC2) multiplied in Agrobacterium strain (EHA 101), was simultaneously introduced into single buds of in vitro grown banana cultivar, Rastali (AAB). Plasmid pBI333-EN4-RCC2 contained a hygromycin phosphotransferase gene (hptII) as the selectable marker and gusA gene as a repor...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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United Arab Emirates University
2009
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| _version_ | 1825945112759238656 |
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| author | Sreeramanan, S. Mahmood, Maziah Xavier, R. |
| author_facet | Sreeramanan, S. Mahmood, Maziah Xavier, R. |
| author_sort | Sreeramanan, S. |
| collection | UPM |
| description | A rice chitinase gene (RCC2) multiplied in Agrobacterium strain (EHA 101), was simultaneously introduced into single buds of in vitro grown banana cultivar, Rastali (AAB). Plasmid pBI333-EN4-RCC2 contained a hygromycin phosphotransferase gene (hptII) as the selectable marker and gusA gene as a reporter marker to identify the transformants.. Single buds derived from multiple bud clumps (Mbcs), were the target explants for transformation. Transformation frequency based on hygromycin selection (25 mg L-1) was higher, although no positive transformant was confirmed based on PCR and Southern blot analyses.. Stable gusA gene expression was detectable in transformed single buds, multiple bud clumps, shoots, leaves and roots derived from hygromycin selection at 50 mg L-1 ). An assay was performed to identify the minimum concentration of two antibiotics most effective against Agrobacterium EHA 101. Protein assay showed an increase in chitinase enzyme activity of transformed plantlets. The Agrobacterium-mediated transformation protocol reported here is suitable for future selection of banana meristem tissues resistant to fungal disease. |
| first_indexed | 2024-03-06T07:26:26Z |
| format | Article |
| id | upm.eprints-12797 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T07:26:26Z |
| publishDate | 2009 |
| publisher | United Arab Emirates University |
| record_format | dspace |
| spelling | upm.eprints-127972011-12-15T02:05:15Z http://psasir.upm.edu.my/id/eprint/12797/ A protocol for Agrobacterium-mediated transformation of banana with a rice chitinase gene. Sreeramanan, S. Mahmood, Maziah Xavier, R. A rice chitinase gene (RCC2) multiplied in Agrobacterium strain (EHA 101), was simultaneously introduced into single buds of in vitro grown banana cultivar, Rastali (AAB). Plasmid pBI333-EN4-RCC2 contained a hygromycin phosphotransferase gene (hptII) as the selectable marker and gusA gene as a reporter marker to identify the transformants.. Single buds derived from multiple bud clumps (Mbcs), were the target explants for transformation. Transformation frequency based on hygromycin selection (25 mg L-1) was higher, although no positive transformant was confirmed based on PCR and Southern blot analyses.. Stable gusA gene expression was detectable in transformed single buds, multiple bud clumps, shoots, leaves and roots derived from hygromycin selection at 50 mg L-1 ). An assay was performed to identify the minimum concentration of two antibiotics most effective against Agrobacterium EHA 101. Protein assay showed an increase in chitinase enzyme activity of transformed plantlets. The Agrobacterium-mediated transformation protocol reported here is suitable for future selection of banana meristem tissues resistant to fungal disease. United Arab Emirates University 2009 Article PeerReviewed Sreeramanan, S. and Mahmood, Maziah and Xavier, R. (2009) A protocol for Agrobacterium-mediated transformation of banana with a rice chitinase gene. Emirates Journal of Food and Agriculture, 21 (2). pp. 18-33. ISSN 2079-052X http://ejfa.info/article/view/5161/2632 Agrobacterium Plant-microbe relationships - Genetic aspects. Plant genetic transformation. English |
| spellingShingle | Agrobacterium Plant-microbe relationships - Genetic aspects. Plant genetic transformation. Sreeramanan, S. Mahmood, Maziah Xavier, R. A protocol for Agrobacterium-mediated transformation of banana with a rice chitinase gene. |
| title | A protocol for Agrobacterium-mediated transformation of banana with a rice chitinase gene. |
| title_full | A protocol for Agrobacterium-mediated transformation of banana with a rice chitinase gene. |
| title_fullStr | A protocol for Agrobacterium-mediated transformation of banana with a rice chitinase gene. |
| title_full_unstemmed | A protocol for Agrobacterium-mediated transformation of banana with a rice chitinase gene. |
| title_short | A protocol for Agrobacterium-mediated transformation of banana with a rice chitinase gene. |
| title_sort | protocol for agrobacterium mediated transformation of banana with a rice chitinase gene |
| topic | Agrobacterium Plant-microbe relationships - Genetic aspects. Plant genetic transformation. |
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