Somatic embryogenesis (SE) in Norway spruce (Picea abies). (top left) Dissected seed with zygotic embryo used for initiation of SE; (top right) proliferating culture of early stages of somatic embryos (Pro Embryogenic Masses; PEMs); (bottom left) culture of mature somatic embryos in bioreactors; (bottom right) germinated somatic embryos. (Top pictures are photos by Sofie Johansson and bottom pictures are photos by Izabela Dobrowolska)
Different genotypes of embryogenic cell lines of Norway spruce of are kept in the SE lab and are available for gene testing experiments. These cell lines are all potential elite clones derived from the operational spruce breeding program run by the Forestry Research Institute of Sweden (Skogforsk).The cell lines have been tested for plant regeneration capabilities and the somatic embryos have been shown to mature, germinate and form plants when cultured with standard protocols.
SE Fluidics system
The SE fluidics system can be used for harvest of somatic embryos or dispersion, separation and singulation of other biomaterials.
Automated fluidics system for harvest of mature somatic embryos: dispersion, separation/singulation, imaging-selection and deposition/positioning
The SE fluidics system is primarily used to study development of mature somatic embryos of Norway spruce. Correlation between embryo morphology and plant formation help guide selection of ‘good’ embryos for plant production.
The bioreactors can be used for somatic embryogenesis multiplication, maturation and germination as well as micropropagation.
The SE lab has a custom-made biolistic gene transfer apparatus for particle-mediated transfer of foreign DNA into particularly somatic embryos of spruce or pine but also works for other species e.g. Arabidopsis, Eucalyptus, onion etc. The device works well with most cell lines and has been extensively used in the past in other labs for studying gene function in conifers. So far, several different SE cell lines in the SE lab have been transformed with a reporter gene construct and stably transformed plants have been developed.
The SE lab is also using and developing the Agrobacterium mediated transformation technology and has successfully generated transformed spruce seedlings and plants.
Embryogenic cell lines used in research programs and cell lines from the breeding program are important to maintain in a viable state for future uses. Decline of vitality and maturation capacity is often the result of continuous in vitro culture; it is thus important to preserve the cell lines under long term storage conditions that preserve viability. For this, we use cryopreservation techniques and instrumentation that allows us to store the embryogenic cell lines for extended times without damage to the viability of the somatic embryos.
The Genomics facility has designed and created transformation vectors (pBGWSPA-UBI and pBGWSXA-GUS) to enable easy cloning of UPSC candidate genes and for use with the SE-lab's biolistic transformation systems. In addition new vectors (pSBIIIAGUSSXA and pSBIIIAUBISPA) has been created for the Agrobacterium-based transformation system.
- The SE-lab provides a standard transformation work-package for research purpose only which includes transformation (by biolistic device or Agrobacterium), generation of transgenic SE cultures and transgenic plants.
- It does NOT include design and preparation of the construct for transformation.
- The transgenic SE cultures can be cryopreserved as an additional service.
To place an order for spruce transformation
Request form for Spruce transformation
Sofie Johansson (Research engineer and Lab manager)