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Somatic Embryogenesis and Spruce Transformation |
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The spruce somatic embryogenesis and transformation laboratory (the “SE lab”) is a new technical platform at UPSC. The primary focus of the activities at the SE lab is on somatic embryogenesis and transformation as a platform for testing of gene function in gymnosperm species using Norway spruce (Picea abies) as the model species. The laboratory functions as a service platform for all groups at UPSC that whish to extend their research and gene- testing into conifers. The laboratory is also working with basic aspects of the somatic embryo technology in order to enable this technology to be used for large scale clonal forestry applications. This not only involves work with conifers, but also with hardwood species such as Poplar and Eucalyptus.
Several embryogenic cell lines of Norway spruce are maintained and are available for gene testing experiments. These cell lines are all potential elite lines derived from the operational spruce breeding program run by the Forestry Research Institute of Sweden (Skogforsk).The cell lines have been tested and the somatic embryos have been shown to mature, germinate and form plants when cultured with standard protocols.
The SE lab has a custom made biolistic gene transfer apparatus for particle mediated transfer of foreign DNA into the somatic embryos. The device works well with most cell lines and has been extensively used in the past in other labs for studying gene function in gymnosperms. So far, several different SE cell lines in the SE lab have been transformed with a reporter gene construct and stably transformed plants are underway.
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| Agrobacterium transformation of gymnosperm somatic embryos is still not routine in most labs. The pictures show positive GUS staining in somatic embryos at different developmental stages; (left) proliferating cultures of somatic embryos and (right) mature somatic embryos. |
To date there are only a few reports available on success- ful transformation of spruce somatic embryos by Agrobacterium mediated transformation. The SE lab is also using and developing this transformation technology and has already generated Agrobacterium-transformed spruce seedlings.
Valuable embryogenic cell lines are important to maintain 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 in a safe long term storage conditions. For this, we use cryopreservation techniques and instrumentation that allows us to store the embryogenic cell lines under liquid nitrogen for extended times without damage to the cells. The SE lab is also involved in developing the protocols for these cryopreservation techniques.
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