Propagation of Lirlodendron hybrids via somatic embryogenesis

Propagation of hybrids between the Chinese tuliptree (Liriodendron chinense) and the North American yellow-poplar (Liriodendron tulipiferea) could be greatly accelerated with a highly productive somatic embryogenesis system. Flowers were collected from a single Chinese tuliptree and the anthers used for controlled pollinations of 4 yellow-poplar mother trees. Aggregates of samaras resulting from the pollinations were harvested 8 weeks post-pollination. Following surface disinfestation, samaras were dissected and embryos and endosperm were cultured together on a semisolid induction medium containing 9.0 M 2,4-dichlorophenoxyacetic acid and 1.1 M benzyladenine. Following 2–3 months on induction medium, an average of 15.6 percent of the explants produced either somatic embryos or proembryogenic masses. Compared to pure yellow-poplar embryogenic cultures, putative hybrid cultures tended not to maintain growth as proembryogenic masses while exposed to auxin, instead proliferating via repetitive embryogenesis as globular-stage embryos. Four to six weeks following transfer of globular embryos to basal medium, mature embryos were produced from the putative hybrid lines. Mature embryos germinated following transfer to basal medium lacking casein hydrolysate. Plantlets survived transfer to potting mix and acclimatization to greenhouse conditions. Verification of the hybrid nature of the embryogenic lines and somatic embryo-derived plantlets was accomplished by Southern hybridization analysis with a species-specific DNA marker.Abbreviations BA benzyladenine - 2,4-d 2,4-dichlorophenoxyacetic acid - PEM proembryogenic mass - CH casein hydrolysate - RFLP restricted fragment length polymorphism - CTAB hexadecyltrimethylammonium bromide - TBE Tris-borate-EDTA - SDS sodium dodecylsufate - SSC sodium citrate/chloride