Limits...
Evaluation on the effectiveness of 2-deoxyglucose-6-phosphate phosphatase (DOG(R)1) gene as a selectable marker for oil palm (Elaeis guineensis Jacq.) embryogenic calli transformation mediated by Agrobacterium tumefaciens.

Izawati AM, Masani MY, Ismanizan I, Parveez GK - Front Plant Sci (2015)

Bottom Line: The plantlets were later transferred into soil and grown in a biosafety screenhouse.PCR and subsequently Southern blot analyses were carried out to confirm the integration of the transgene in the plantlets.This result demonstrates the potential of using combination of DOG(R)1 gene and 2-DOG for regenerating transgenic oil palm.

View Article: PubMed Central - PubMed

Affiliation: Advanced Biotechnology and Breeding Centre, Malaysian Palm Oil Board Selangor, Malaysia.

ABSTRACT
DOG(R)1, which encodes 2-deoxyglucose-6-phosphate phosphatase, has been used as a selectable marker gene to produce transgenic plants. In this study, a transformation vector, pBIDOG, which contains the DOG(R)1 gene, was transformed into oil palm embryogenic calli (EC) mediated by Agrobacterium tumefaciens strain LBA4404. Transformed EC were exposed to 400 mg l(-1) 2-deoxyglucose (2-DOG) as the selection agent. 2-DOG resistant tissues were regenerated into whole plantlets on various regeneration media containing the same concentration of 2-DOG. The plantlets were later transferred into soil and grown in a biosafety screenhouse. PCR and subsequently Southern blot analyses were carried out to confirm the integration of the transgene in the plantlets. A transformation efficiency of about 1.0% was obtained using DOG(R)1 gene into the genome of oil palm. This result demonstrates the potential of using combination of DOG(R)1 gene and 2-DOG for regenerating transgenic oil palm.

No MeSH data available.


Related in: MedlinePlus

Protocol for Agrobacterium-mediated transformation and regeneration of transgenic oil palm selected on 2-DOG medium.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4585222&req=5

Figure 2: Protocol for Agrobacterium-mediated transformation and regeneration of transgenic oil palm selected on 2-DOG medium.

Mentions: The simplified schematic protocol for Agrobacterium-mediated transformation and regeneration of transgenic oil palm selected on 2-deoxyglucose medium is summarized in Figure 2. In this study, suspension EC, were chosen as the starting material for oil palm transformation as they are easily regenerated into plantlets (Tarmizi et al., 2008). Embryogenic cultures were also reported to be a good target tissue as they provided a source of dividing cells that were recognized as the most competent cells for genetic transformation of pine species, cassava, sweet potato, rice, and garlic (Trontin et al., 2007; Bull et al., 2009; Zang et al., 2009; Rahman et al., 2010; Alvarez and Ordás, 2013; Lagunes-Fortiz et al., 2013). The use of highly regenerative tissue as target for transformation often results in the regeneration of a large number of independently transformed lines.


Evaluation on the effectiveness of 2-deoxyglucose-6-phosphate phosphatase (DOG(R)1) gene as a selectable marker for oil palm (Elaeis guineensis Jacq.) embryogenic calli transformation mediated by Agrobacterium tumefaciens.

Izawati AM, Masani MY, Ismanizan I, Parveez GK - Front Plant Sci (2015)

Protocol for Agrobacterium-mediated transformation and regeneration of transgenic oil palm selected on 2-DOG medium.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4585222&req=5

Figure 2: Protocol for Agrobacterium-mediated transformation and regeneration of transgenic oil palm selected on 2-DOG medium.
Mentions: The simplified schematic protocol for Agrobacterium-mediated transformation and regeneration of transgenic oil palm selected on 2-deoxyglucose medium is summarized in Figure 2. In this study, suspension EC, were chosen as the starting material for oil palm transformation as they are easily regenerated into plantlets (Tarmizi et al., 2008). Embryogenic cultures were also reported to be a good target tissue as they provided a source of dividing cells that were recognized as the most competent cells for genetic transformation of pine species, cassava, sweet potato, rice, and garlic (Trontin et al., 2007; Bull et al., 2009; Zang et al., 2009; Rahman et al., 2010; Alvarez and Ordás, 2013; Lagunes-Fortiz et al., 2013). The use of highly regenerative tissue as target for transformation often results in the regeneration of a large number of independently transformed lines.

Bottom Line: The plantlets were later transferred into soil and grown in a biosafety screenhouse.PCR and subsequently Southern blot analyses were carried out to confirm the integration of the transgene in the plantlets.This result demonstrates the potential of using combination of DOG(R)1 gene and 2-DOG for regenerating transgenic oil palm.

View Article: PubMed Central - PubMed

Affiliation: Advanced Biotechnology and Breeding Centre, Malaysian Palm Oil Board Selangor, Malaysia.

ABSTRACT
DOG(R)1, which encodes 2-deoxyglucose-6-phosphate phosphatase, has been used as a selectable marker gene to produce transgenic plants. In this study, a transformation vector, pBIDOG, which contains the DOG(R)1 gene, was transformed into oil palm embryogenic calli (EC) mediated by Agrobacterium tumefaciens strain LBA4404. Transformed EC were exposed to 400 mg l(-1) 2-deoxyglucose (2-DOG) as the selection agent. 2-DOG resistant tissues were regenerated into whole plantlets on various regeneration media containing the same concentration of 2-DOG. The plantlets were later transferred into soil and grown in a biosafety screenhouse. PCR and subsequently Southern blot analyses were carried out to confirm the integration of the transgene in the plantlets. A transformation efficiency of about 1.0% was obtained using DOG(R)1 gene into the genome of oil palm. This result demonstrates the potential of using combination of DOG(R)1 gene and 2-DOG for regenerating transgenic oil palm.

No MeSH data available.


Related in: MedlinePlus