Pine embryogenic tissue derived from immature zygotic embryos may consist of multiple genotypes due to simple polyembryony. To test this hypothesis, megagametophytes with intact zygotic embryos were cultured from immature loblolly pine (Pinus taeda L.) seeds of clone WV42 control pollinated with a 1:1:1 pollen mix of clones WV44, WV47, and WV48. Each pollen parent contained a marker allele at one or more of the following loci: aconitase, malic dehydrogenase, 6-phosphogluconate dehydrogenase, and shikimate dehydrogenase, allowing determination of the paternal parent. After two to four weeks in culture, embryogenic tissue derived from zygotic embryos extruded from megagametophytes was separated into individual embryos and sectors of embryogenic tissue. The paternal genotype of each resulting cell line was determined by starch gel electrophoresis. Three of thirty-six explants produced multiple cell lines with genotypic differences among the cell lines within each explant. Our results unequivocally show that it is possible to initiate embryogenic tissue from more than one zygotic embryo of a loblolly pine seed and that the resulting cell lines may be genetically different.Abbreviations: ACO -aconitase, MDH -malic dehydrogenase, SKDH -shikimate dehydrogenase, 6PGD -6-phosphogluconate dehydrogenase

Vegetative long-shoot buds, greenwood stems, and immature needles of 20year-old western larch (Larix occidentalis Nutt.) were cultured to induce multiple bud formation. Explants were collected year-round and cultured on a modified Schenk and Hildebrandt (SH) medium containing 6-benzyladenine (BA) at 0, 1, 5, 10, 50, or 100 µM. Multiple buds were produced on buds and stems with terminal meristems, but not on needles or stem sections. The induction of de novo buds and development of axillary buds required BA at 1 to 10 µM; higher concentrations of BA were less effective. More explants formed multiple buds on SH than on modified Murashige and Skoog (MS) media. Multiple buds formed on more buds and stems excised during the growing season than from dormant buds. Buds cultured on media containing gibberellin died within 6 weeks; auxin caused bud elongation but no multiple buds formed. Chemical names used: N-[(trichloromethyl)thio]-4-cyclohexene-1,2-dicarboximide (captan); 6-benzyladenine (BA); 1H-indole-3-butyric acid (IBA); 1H-indole-3-acetic acid (IAA); gibberellin (GA 4+7 ).

White pine embryos were grown on 4 different media with 6 different benzyladenine (BA) concentrations. Maximum adventitious shoot initiation and growth were obtained on a modified Lepoivre medium with 20/zM BA. Modified Schenk and Hildebrandt, Murashige and Skoog, and Gresshoff and Doy media were also tested. Shoot elongation was achieved on half-strength basal medium lacking growth regulators. Three rooting experiments involving indole-3-butyric acid (IBA), sucrose concentration, shoot orientation, IBA pulse length, and light or dark were carried out. Treatment of shoots in an upright position with 50/xM IBA for eight days followed by culture in a medium with 3% sucrose in the light produced the most rooting (50% at 3 months). Rooted shoots were transplanted to the greenhouse for further growth.

No abstract