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Gamma linolenic acid (GLA; C18:3Δ6,9,12 cis), also known as γ-Linolenic acid, is an important essential fatty acid precursor for the synthesis of very long chain polyunsaturated fatty acids and important pathways involved in human health. GLA is synthesized from linoleic acid (LA; C18:2Δ9,12 cis) by endoplasmic reticulum associated Δ6-desaturase activity. Currently sources of GLA are limited to a small number of plant species with poor agronomic properties, and therefore an economical and abundant commercial source of GLA in an existing crop is highly desirable. To this end, the seed oil of a high LA cultivated species of safflower (Carthamus tinctorius) was modified by transformation with Δ6-desaturase from Saprolegnia diclina resulting in levels exceeding 70% (v/v) of GLA. Levels around 50% (v/v) of GLA in seed oil was achieved when Δ12-/Δ6-desaturases from Mortierella alpina was over-expressed in safflower cultivars with either a high LA or high oleic (OA; C18:1Δ9 cis) background. The differences in the overall levels of GLA suggest the accumulation of the novel fatty acid was not limited by a lack of incorporation into the triacylgylcerol backbone (>66% GLA achieved), or correlated with gene dosage (GLA levels independent of gene copy number), but rather reflected the differences in Δ6-desaturase activity from the two sources. To date, these represent the highest accumulation levels of a newly introduced fatty acid in a transgenic crop. Events from these studies have been propagated and recently received FDA approval for commercialization as Sonova™400.

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A protocol is described for the in vitro production of plantlets from embryonic explants of eastern white cedar (Thuja occidentalis L.). Bud induction was optimal when embryonic explants were cultured for 20-25 days on half-strength Quoirin and Le Poivre mineral salts containing equimolar concentrations (10(-6) M) of N(6)-benzyladenine and 2-isopentyl adenine. Bud development was achieved in phytohormone-free medium in the presence of activated charcoal. Maximal shoot elongation occurred on half-strength Quoirin and Le Poivre salts, whereas shoot multiplication was optimal on half-strength Bornman's MCM salts in the presence of cytokinin. Hardened shoots, dipped in commercial rooting powder containing indole-3-butyric acid, rooted optimally in mist under non-sterile greenhouse conditions. Both rooting and subsequent plantlet growth was best when Redi-Earth((R)) was used as a substrate. Over 250 plantlets per embryo can be produced annually by this technique.

A protocol is described for plantlet regeneration using embryonic explants of Juniperus cedrus Webb & Berth.An average of 6 adventitious buds were induced on whole excised embryos cultured for 15 days on Quoirin and LePoivre (QP) half-strength medium supplemented with 5 laM N6-benzyladenine. For bud development, explants were transferred to phytohormone-free 1/2 QP medium. For shoot elongation, explants were cultured on 1/2 QP with 0.05% activated charcoal and 2% sucrose. Adventitious shoots were rooted successfully in peat-vermiculiteperlite (1:1:1) moistened with 1/4 QP containing 1% sucrose and 5 laM a-napthaleneacetic acid, pH 5.0. Axillary shoots elongated spontaneously in culture from leaf axils.