Glutathione and abscisic acid supplementation influences somatic embryo maturation and hormone endogenous levels during somatic embryogenesis in Podocarpus lambertii Klotzsch ex Endl.

Prince Rupprecht’s larch (Larix principis-rupprechtii Mayr) is a native conifer in North China with great economic and ecological values. Somatic embryogenesis (SE) is a powerful tool for the mass clonal propagation in plants. In this study, we described a high-efficiency SE system via indirect pathways and investigated the effect of genotype, culture conditions and phytohormones on SE. Immature zygotic embryos (IZEs) of L. principis-rupprechtii Mayr were used as explant materials. In the induction stage, embryogenic tissues (ETs) were induced on mLV medium supplemented with 2.0 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg L−1 6-benzylaminopurine (6-BA). The initiation frequencies showed significant differences (p < 0.05) among 20 genotypes of open-pollinated mother trees with the highest induction frequency reaching 30%. For tissue proliferation, proliferation in liquid medium was more efficient compared with proliferation in semi-solid medium, providing a multiplication rate of 3.12 in an 8-day subculture period. As a necessary exogenous plant growth regulator (PGR) for somatic embryo maturation in conifers, abscisic acid (ABA) was optimized at 16 mg L−1 in this system. Next, an orthogonal test on osmotic pressure factors showed 50 g L−1 sucrose, 7 g L−1 phytagel and 75 g L−1 polyethylene glycol (PEG) was the optimal combination for somatic embryo maturation in L. principis-rupprechtii Mayr. Moreover, the dispersion culture method provided a more efficient somatic embryo maturation, up to 545 per gram of fresh weight (FW). Finally, 2 g L−1 of active charcoal (AC) was found to increase the somatic embryo germination rate to 63.46%. The improved protocol of SE will serve as a foundation for establishing mass propagation and genetic transformation of L. principis-rupprechtii Mayr.

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Somatic embryo maturation, germination, and soil establishment of plants of black and white spruce (Picea mariana and Picea glauca)

Canadian Journal of Botany ◽

10.1139/b90-326 ◽

1990 ◽

Vol 68 (12) ◽

pp. 2583-2589 ◽

Cited By ~ 33

Author(s):

S. M. Attree ◽

T. E. Tautorus ◽

D. I. Dunstan ◽

L. C. Fowke

Keyword(s):

Abscisic Acid ◽

Somatic Embryo ◽

Picea Mariana ◽

Picea Glauca ◽

Black Spruce ◽

White Spruce ◽

Embryo Maturation ◽

Black And White ◽

Somatic Embryo Maturation

Somatic embryo maturation, germination, and soil establishment frequencies were compared for two conifer species, white and black spruce (Picea glauca and Picea mariana). The comparison of the two species regenerated and established in soil under the same conditions showed black spruce to be the most responsive. Shorter exposure times to 32 μM abscisic acid were not as effective as maturation on a medium containing 16 μM abscisic acid for 28 days. This gave similar maturation frequencies for the two species (6–8%), and germination frequencies of 64% for white spruce and over 73% for black spruce. Over 1800 black and white spruce plantlets were recovered, and more than 400 were transferred from in vitro to nonsterile conditions. Sixty percent (160) of the black spruce plantlets survived transfer and continued to grow vigorously. By comparison only 18% (29) of the white spruce plantlets survived, and half of these rapidly produced dormant buds and underwent no further shoot growth. White spruce plants that did not produce dormant buds grew vigorously. These results indicate that there are large differences in the ability of these closely related species to respond to plantlet establishment following regeneration from somatic embryos, and that black spruce is highly responsive to micropropagation by this method. Key words: Picea glauca, Picea mariana, somatic embryogenesis, maturation, germination, soil establishment.

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