8 results on '"Masani MY"'
Search Results
2. Production of polyhydroxybutyrate in oil palm (Elaeis guineensis Jacq.) mediated by microprojectile bombardment of PHB biosynthesis genes into embryogenic calli.
- Author
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Parveez GK, Bahariah B, Ayub NH, Masani MY, Rasid OA, Tarmizi AH, and Ishak Z
- Abstract
Biodegradable plastics, mainly polyhydroxybutyrate (PHB), which are traditionally produced by bacterial cells, have been produced in the cells of more than 15 plant species. Since the production of biodegradable plastics and the synthesis of oil in plants share the same substrate, acetyl-coenzyme A (acetyl-CoA), producing PHB in oil bearing crops, such as oil palm, will be advantageous. In this study, three bacterial genes, bktB, phaB, and phaC, which are required for the synthesis of PHB and selectable marker gene, bar, for herbicide Basta resistant, were transformed into embryogenic calli. A number of transformed embryogenic lines resistant to herbicide Basta were obtained and were later regenerated to produce few hundred plantlets. Molecular analyses, including polymerase chain reaction (PCR), Southern blot, and real-time PCR have demonstrated stable integration and expression of the transgenes in the oil palm genome. HPLC and Nile blue A staining analyses confirmed the synthesis of PHB in some of the plantlets.
- Published
- 2015
- Full Text
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3. Biotechnology of oil palm: strategies towards manipulation of lipid content and composition.
- Author
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Parveez GK, Rasid OA, Masani MY, and Sambanthamurthi R
- Subjects
- Arecaceae enzymology, Lipids biosynthesis, Palm Oil, Plants, Genetically Modified, Promoter Regions, Genetic genetics, Arecaceae genetics, Biotechnology methods, Lipids analysis, Plant Oils chemistry
- Abstract
Oil palm is a major economic crop for Malaysia. The major challenges faced by the industry are labor shortage, availability of arable land and unstable commodity price. This has caused the industry to diversify its applications into higher value products besides increasing its yield. While conventional breeding has its limitations, biotechnology was identified as one of the tools for overcoming the above challenges. Research on biotechnology of oil palm began more than two decades ago leveraging a multidisciplinary approach involving biochemical studies, gene and promoter isolation, transformation vector construction and finally genetic transformation to produce the targeted products. The main target of oil palm biotechnology research is to increase oleic acid in the mesocarp. Other targets are stearic acid, palmitoleic acid, ricinoleic acid, lycopene (carotenoid) and biodegradable plastics. Significant achievements were reported for the biochemical studies, isolation of useful oil palm genes and characterization of important promoters. A large number of transformation constructs for various targeted products were successfully produced using the isolated oil palm genes and promoters. Finally transformation of these constructs into oil palm embryogenic calli was carried out while the regeneration of transgenic oil palm harboring the useful genes is in progress.
- Published
- 2015
- Full Text
- View/download PDF
4. Efficient transformation of oil palm protoplasts by PEG-mediated transfection and DNA microinjection.
- Author
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Masani MY, Noll GA, Parveez GK, Sambanthamurthi R, and Prüfer D
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- Palm Oil, Plants, Genetically Modified cytology, Transfection methods, Transformation, Genetic genetics, Microinjections methods, Plant Oils metabolism, Plants, Genetically Modified genetics, Protoplasts metabolism
- Abstract
Background: Genetic engineering remains a major challenge in oil palm (Elaeis guineensis) because particle bombardment and Agrobacterium-mediated transformation are laborious and/or inefficient in this species, often producing chimeric plants and escapes. Protoplasts are beneficial as a starting material for genetic engineering because they are totipotent, and chimeras are avoided by regenerating transgenic plants from single cells. Novel approaches for the transformation of oil palm protoplasts could therefore offer a new and efficient strategy for the development of transgenic oil palm plants., Methodology/principal Findings: We recently achieved the regeneration of healthy and fertile oil palms from protoplasts. Therefore, we focused on the development of a reliable PEG-mediated transformation protocol for oil palm protoplasts by establishing and validating optimal heat shock conditions, concentrations of DNA, PEG and magnesium chloride, and the transfection procedure. We also investigated the transformation of oil palm protoplasts by DNA microinjection and successfully regenerated transgenic microcalli expressing green fluorescent protein as a visible marker to determine the efficiency of transformation., Conclusions/significance: We have established the first successful protocols for the transformation of oil palm protoplasts by PEG-mediated transfection and DNA microinjection. These novel protocols allow the rapid and efficient generation of non-chimeric transgenic callus and represent a significant milestone in the use of protoplasts as a starting material for the development of genetically-engineered oil palm plants.
- Published
- 2014
- Full Text
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5. Regeneration of viable oil palm plants from protoplasts by optimizing media components, growth regulators and cultivation procedures.
- Author
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Masani MY, Noll G, Parveez GK, Sambanthamurthi R, and Prüfer D
- Subjects
- Arecaceae growth & development, Cell Division, Palm Oil, Plant Oils, Protoplasts cytology, Protoplasts drug effects, Regeneration, Sepharose, Arecaceae physiology, Culture Media, Plant Growth Regulators pharmacology, Plant Somatic Embryogenesis Techniques methods, Protoplasts physiology
- Abstract
Oil palm protoplasts are suitable as a starting material for the production of oil palm plants with new traits using approaches such as somatic hybridization, but attempts to regenerate viable plants from protoplasts have failed thus far. Here we demonstrate, for the first time, the regeneration of viable plants from protoplasts isolated from cell suspension cultures. We achieved a protoplast yield of 1.14×10(6) per gram fresh weight with a viability of 82% by incubating the callus in a digestion solution comprising 2% cellulase, 1% pectinase, 0.5% cellulase onuzuka R10, 0.1% pectolyase Y23, 3% KCl, 0.5% CaCl2 and 3.6% mannitol. The regeneration of protoplasts into viable plants required media optimization, the inclusion of plant growth regulators and the correct culture technique. Microcalli derived from protoplasts were obtained by establishing agarose bead cultures using Y3A medium supplemented with 10μM naphthalene acetic acid, 2μM 2,4-dichlorophenoxyacetic acid, 2μM indole-3-butyric acid, 2μM gibberellic acid and 2μM 2-γ-dimethylallylaminopurine. Small plantlets were regenerated from microcalli by somatic embryogenesis after successive subculturing steps in medium with limiting amounts of growth regulators supplemented with 200mg/l ascorbic acid., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)
- Published
- 2013
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6. Transformation of oil palm using Agrobacterium tumefaciens.
- Author
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Izawati AM, Parveez GK, and Masani MY
- Subjects
- Acetophenones pharmacology, Herbicides pharmacology, Palm Oil, Plant Oils, Plants, Genetically Modified, Plasmids genetics, Promoter Regions, Genetic, Transformation, Genetic, Ubiquitin genetics, Zea mays genetics, Agrobacterium tumefaciens genetics, Aminobutyrates pharmacology, Biolistics methods, Cocos genetics, Gene Transfer Techniques, Herbicide Resistance genetics
- Abstract
Transgenic oil palm (Elaeis guineensis Jacq.) plantlets are regenerated after Agrobacterium tumefaciens-mediated transformation of embryogenic calli derived from young leaves of oil palm. The calli are transformed with an Agrobacterium strain, LBA4404, harboring the plasmid pUBA, which carries a selectable marker gene (bar) for resistance to the herbicide Basta and is driven by a maize ubiquitin promoter. Modifications of the transformation method, treatment of the target tissues using acetosyringone, exposure to a plasmolysis medium, and physical injury via biolistics are applied. The main reasons for such modifications are to activate the bacterial virulence system and, subsequently, to increase the transformation efficiency. Transgenic oil palm cells are selected and regenerated on a medium containing herbicide Basta. Molecular analyses revealed the presence and integration of the introduced bar gene into the genome of the transformants.
- Published
- 2012
- Full Text
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7. Construction of phosphomannose isomerase (PMI) transformation vectors and evaluation of the effectiveness of vectors in tobacco (Nicotiana tabacum L).
- Author
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Bahariah B, Parveez GK, Masani MY, and Khalid N
- Abstract
Unlabelled: Phosphomannose isomerase (pmi) gene isolated from Escherichia coli allows transgenic plants carrying it to convert mannose-6- phosphate (from mannose), a carbon source that could not be naturally utilized by plants into fructose-6-phosphate which can be utilized by plants as a carbon source. This conversion ability provides energy source to allow the transformed cells to survive on the medium containing mannose. In this study, four transformation vectors carrying the pmi gene alone or in combination with the β-glucuronidase (gusA) gene were constructed and driven by either the maize ubiquitin (Ubi1) or the cauliflower mosaic virus (CaMV35S) promoter. Restriction digestion, PCR amplification and sequencing were carried out to ensure sequence integrity and orientation. Tobacco was used as a model system to study the effectiveness of the constructs and selection system. PMI11G and pMI3G, which carry gusA gene, were used to study the gene transient expression in tobacco. PMI3 construct, which only carries the pmi gene driven by CaMV35S promoter, was stably transformed into tobacco using biolistics after selection on 30 g 1(-1) mannose without sucrose. Transgenic plants were verified using PCR analysis., Abbreviations: PMI/pmi - Phosphomannose isomerase, Ubi1 - Maize ubiquitin promoter, CaMV35S - Cauliflower mosaic virus 35S promoter, gusA - β-glucuronidase GUS reporter gene.
- Published
- 2012
- Full Text
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8. Construction of PHB and PHBV multiple-gene vectors driven by an oil palm leaf-specific promoter.
- Author
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Masani MY, Parveez GK, Izawati AM, Lan CP, and Siti Nor Akmar A
- Subjects
- Acetyl-CoA C-Acyltransferase genetics, Acyltransferases genetics, Alcohol Oxidoreductases genetics, Arabidopsis genetics, Arecaceae genetics, Artificial Gene Fusion, Cupriavidus necator enzymology, Escherichia coli genetics, Genetic Engineering, Plant Leaves genetics, Plant Leaves metabolism, Promoter Regions, Genetic genetics, Threonine Dehydratase genetics, Arecaceae metabolism, Cupriavidus necator genetics, Gene Expression Regulation, Plant, Genetic Vectors genetics, Hydroxybutyrates metabolism, Polyesters metabolism
- Abstract
One of the targets in oil palm genetic engineering programme is the production of polyhydroxybutyrate (PHB) and polyhydroxybutyrate-co-valerate (PHBV) in the oil palm leaf tissues. Production of PHB requires the use of phbA (beta-ketothiolase type A), phbB (acetoacetyl-CoA reductase) and phbC (PHB synthase) genes of Ralstonia eutropha, whereas bktB (beta-ketothiolase type B), phbB, phbC genes of R. eutropha and tdcB (threonine dehydratase) gene of Escherichia coli were used for PHBV production. Each of these genes was fused with a transit peptide (Tp) of oil palm acyl-carrier-protein (ACP) gene, driven by an oil palm leaf-specific promoter (LSP1) to genetically engineer the PHB/PHBV pathway to the plastids of the leaf tissues. In total, four transformation vectors, designated pLSP15 (PHB) and pLSP20 (PHBV), and pLSP13 (PHB) and pLSP23 (PHBV), were constructed for transformation in Arabidopsis thaliana and oil palm, respectively. The phosphinothricin acetyltransferase gene (bar) driven by CaMV35S promoter in pLSP15 and pLSP20, and ubiquitin promoter in pLSP13 and pLSP23 were used as the plant selectable markers. Matrix attachment region of tobacco (RB7MAR) was also included in the vectors to stabilize the transgene expression and to minimize silencing due to positional effect. Restriction digestion, PCR amplification and/or sequencing were carried out to ensure sequence integrity and orientation.
- Published
- 2009
- Full Text
- View/download PDF
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