Appeal 2007-1026
Application 10/405,819
2) Multiplication. The proembryos are multiplied in a maintenance
medium (col. 7, ll. 29-36). “It appears now that the inclusion . . . of selected
active gibberellins and/or abscisic acid in the maintenance . . . media is also
beneficial for improvement of proembryo quality.” (Col. 8, ll. 4-8.)
3) Singulation. “The proembryos tend to form in tight clumps or
clusters which must first be singulated before going to the development
stage. This singulation is carried out in a series of liquid shake cultures
which . . . have exogenous abscisic acid as a necessary new hormone. . . . It
now appears to be beneficial to include . . . an active gibberellin in the
singulation medium.” (Col. 8, ll. 21-39.)
4) Development. “The early stage embryos may then be placed in or
on a late stage proembryo development culture in order to develop very
robust late stage proembryos having at least 100 cells. Culturing from this
point continues in a cotyledonary embryo development medium containing
an active gibberellin (GA) . . . . Preferably exogenous abscisic acid (ABA) is
also present . . . . After several weeks somatic embryos having the
appearance of zygotic embryos will have formed.” (Abstract.)
Example 1 in Pullman describes a culture method (col. 13, ll. 65-67).
“The embryonal-suspensor masses containing early stage proembryos are
transferred to a solid Stage II maintenance and multiplication medium
containing greatly reduced plant growth hormones and preferably a
somewhat raised osmotic level. . . . Low concentrations of a gibberellin
and/or abscisic acid are frequently beneficial at this stage of culture.”
(Col. 14, ll. 55-64.)
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