Cryopreservation of yellow-poplar and sweetgum embryogenic cultures

Cryopreservation has become anessential tool for operational application offorest tree embryogenic cultures, due to thelong evaluation periods needed for treesregenerated from these cultures. Fiveyellow-poplar (Liriodendron tulipifera)and seven sweetgum (Liquidambar spp.)embryogenic culture lines werestored in liquid nitrogen for 48 hours, afterwhich they were thawed and tested for regrowthand ability to produce somatic seedlings.Combinations of two sorbitol pretreatments andthree dimethylsulfoxide (DMSO) cryoprotectantlevels were evaluated for their impact onrecovery following cryogenic storage. The bestresults were obtained with 0.4 M sorbitol and5% DMSO, which provided 100% recovery.Somatic seedlings were regenerated from allculture lines and treatments, except for atransgenic sweetgum line.     

Light quality treatments enhance somatic seedling production in three southern pine species

Embryogenic cultures of loblolly pine (Pinus taeda L.), slash pine (Pinus elliottii Engelm.), longleaf pine (Pinus palustris Mill.) and slash pine x longleaf pine hybrids were initiated from immature seeds on an initiation medium containing 13.57 microM 2,4-dichlorophenoxyacetic acid and 2.22 microM benzylaminopurine. Embryogenic cultures proliferated and somatic embryos developed, matured and germinated following a modified protocol and media originally developed for radiata pine (Pinus radiata D. Don.) somatic seedling production. A discrete, light-sensitive pre-germination stage and a later germination (radicle emergence) stage were identified by the differential response of somatic embryos to light of different wavelengths. Different light quality treatments were applied during the pre-germination and germination steps, using cool white fluorescent bulbs or light-emitting diodes (LEDs), or both. In general, red wavelengths provided by LEDs during these steps resulted in higher frequencies of somatic embryo germination (up to 64%) and conversion (up to 50%), longer tap roots and more first-order lateral roots than the standard cool white fluorescent treatments or treatment with blue wavelengths from LEDs. In addition, exposure to red light allowed germination of somatic embryos of some clones that failed to produce germinants under fluorescent light. Germination and conversion were further enhanced by sequential application of cool white fluorescent light and red light, resulting in up to 100% germination and conversion in one experiment. Longleaf pine somatic embryos were especially responsive to the light quality treatments, resulting in the first report of somatic seedling production for this species.