Abscisic acid and mannitol promote early development, maturation and storage protein accumulation in somatic embryos of interior spruce

Low levels of mannitol (2–6%) promoted the formation of globular embryos in embryogenic cultures of interior spruce ( Picea glauca engelmanni complex). However, these concentrations of mannitol were inhibitory to the formation of cotyledonary embryos. A short (1 week) pulse of mannitol in combination with abscisic acid doubled the production of late cotyledonary somatic embryos compared with the standard abscisic acid treatment. Higher levels of mannitol (13 and 20%) were required to inhibit precocious germination of spruce somatic embryos. These concentrations of mannitol promoted the accumulation of storage proteins during cotyledon maturation, but were not as effective as abscisic acid. Furthermore, 13 and 20% mannitol treatments did not substitute for abscisic acid in promoting the formation of cotyledonary embryos. Pre-treatment of late cotyledonary embryos with mannitol (13–25%) did not increase the frequency of germination compared with germination in non-treated embryos (approximately 10% germinated) although dehydration with high relative humidity treatment increased germination to 83%.