Appeal 2007-1026
Application 10/405,819
28. A method for producing cotyledonary pine embryos, the
method comprising the steps of:
(a) culturing pine embryonal suspensor masses on solid
maintenance medium that does not comprise a gibberellin;
(b) culturing the pine embryonal suspensor masses treated in
accordance with step (a) in liquid maintenance medium that
does not comprise a gibberellin;
(c) culturing the pine embryonal suspensor masses treated in
accordance with step (b) in a liquid maintenance medium
comprising at least one gibberellin, provided that the liquid
maintenance medium comprising a gibberellin does not
comprise abscisic acid; and
(d) culturing the pine embryonal suspensor masses treated in
accordance with step (c) on solid development medium, that
does not comprise a gibberellin, to yield cotyledonary pine
embryos.
THE PRIOR ART
Pullman describes “a method for reproducing coniferous trees by
somatic embryogenesis using plant tissue culture techniques in a multistage
culturing process.” (Abstract.) Four primary culturing stages are described,
each associated with a particular culture medium composition:
1) Initiation or induction. “A suitable explant, typically the fertilized
embryo excised from an immature seed, is first cultured on a medium that
induces multiple early stage proembryos.” (Abstract). The medium utilized
in this step is referred to as “initiation” or “induction” medium (col. 6, ll.
22-24; col. 13-14, Table 2).
3
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