Enhancement of somatic embryogenesis and plant regeneration in Japanese red pine (Pinus densiflora)

The highest proliferation rate (9.8-fold) of embryogenic suspensor mass (ESM) was obtained from half-strength Litvay (A1/2LM, Litvay et al. 1985) medium supplemented with 3.42 mM l-glutamine, while the lowest rate was noted when 0.84 mM l-glutamine (0.6-fold) was added to the medium. The highest growth ratio with brassinolide (BL) was observed for 1.0 mu M (2.3-fold, line 05-21) and 0.05 mu M (2.9-fold, line 06-22). However, in the ESM lines 05-21 and 06-22, high ESM growth rates (2.3-fold, line 05-21 and 2.1-fold, line 06-3) were seen without BL when compared with 1.0 mu M (05-21) or 0.05 mu M (06-22). BL-supplemented medium has been shown to have diverse, genotype-specific effects on the degree of ESM proliferation. For somatic embryo maturation with 0.05 % activated charcoal (AC), the highest number (798 g(-1) FW) of cotyledonary somatic embryos (line 06-29) were obtained on a maturation medium supplemented with AC. The influence of light-emitting diode (LED) sources on the germination of somatic embryos (four genotypes) in this species was studied and was strongly genotype-dependent. Germination of somatic embryos from ESM line 05-3 was strongly inhibited by fluorescent, and red + blue light, while lines 05-12, 05-29, and 05-37 showed a similar germination frequency for the five LED sources, where red light most stimulated somatic embryo germination.