title: Efficient transformation of oil palm protoplasts by PEG-mediated transfection and DNA microinjection.
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.
  
  
  RESULTS
  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
  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.
authors.affiliation: Advanced Biotechnology and Breeding Centre, Malaysian Palm Oil Board (MPOB), Kuala Lumpur, Malaysia.
authors.forename: Mat Yunus Abdul
authors.initials: MY
authors.lastname: Masani
authors.affiliation: Westfälische Wilhelms-Universität Münster, Institut für Biologie und Biotechnologie der Pflanzen, Münster, Germany.
authors.forename: Gundula A
authors.initials: GA
authors.lastname: Noll
authors.affiliation: Advanced Biotechnology and Breeding Centre, Malaysian Palm Oil Board (MPOB), Kuala Lumpur, Malaysia.
authors.forename: Ghulam Kadir Ahmad
authors.initials: GK
authors.lastname: Parveez
authors.affiliation: Advanced Biotechnology and Breeding Centre, Malaysian Palm Oil Board (MPOB), Kuala Lumpur, Malaysia.
authors.forename: Ravigadevi
authors.initials: R
authors.lastname: Sambanthamurthi
authors.affiliation: Westfälische Wilhelms-Universität Münster, Institut für Biologie und Biotechnologie der Pflanzen, Münster, Germany; Fraunhofer Institut für Molekularbiologie und Angewandte Ökologie, Münster, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Prüfer
mesh_terms: D008845:Microinjections / Q000379:methods*; D000073878:Palm Oil; D010938:Plant Oils / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000166:cytology / Q000235:genetics*; D011523:Protoplasts / Q000378:metabolism*; D014162:Transfection / Q000379:methods; D014170:Transformation, Genetic / Q000235:genetics
chemical_list: D010938:Plant Oils; D000073878:Palm Oil