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United States Department of Agriculture

Agricultural Research Service

Research Project: MANAGEMENT OF GENETIC RESOURCES FOR VITIS, PRUNUS, JUGLANS, FICUS, OLEA, PISTACIA, PUNICA, DIOSPYROS, ACTINIDIA, AND MORUS Title: Somatic embryogenesis in immature cotyledons of Manchurian ash (Fraxinus mandshurica Rupr.)

Authors
item Kong, Dong-Mei -
item Preece, John
item Shen, Hai-Long -

Submitted to: Plant Cell Tissue And Organ Culture
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: September 18, 2011
Publication Date: September 29, 2011
Citation: Kong, D. 2011. Somatic embryogenesis in immature cotyledons of Manchurian ash (Fraxinus mandshurica Rupr.). Plant Cell Tissue And Organ Culture. 108:485-492.

Interpretive Summary: Somatic embryogenesis was obtained from immature cotyledon explants that were cultured on half-strength Murashige and Skoog (MS) salts and vitamins with 5.4 uM naphthaleneacetic acid (NAA) and 0.2 uM thidiazuron (TDZ) plus a 4x4 factorial combination of 0,9.8, 34.6, or 49.2 uM indole-3-butyric acid (IBA) and 0, 8.9, 22.2, or 44.4 uM 6-benzyladenine (BA). The addition of 44.4 uM BA improved the percentage of cotyledon explants that produced somatic embryos to >20%, if 9.8 or 24.6 uM IBA was also present.Somatic embryogenesis was >30% when seeds were harvested on 31 July or 15 August. The addition of 50 or 70 g 1- sucrose enhanced embryo-genesis. Histological examination showed that somatic embryos originated from epidermis cells of zygotic embryos. A peak germination rate (69%) was attained when somatic embryos were desiccated for 10 min before they produced green cotyledons and elongated shoot tips. Of the germinated embryos from this desiccation treatment, 65.6% also grew roots and therefore converted into plants. Chilling somatic embryos at 4 degrees Celsius for 15 days resulted in the highest germination rate (69.4%), which was significantly higher than those without chilling treatment (27.6%). However<10% of the chilled germinated embryos formed roots and grew into plants. Plantlets from somatic embryos were transplanted into a 2 vermiculite: 1 sphagnum peat medium, where they had a survival rate of 80.8%, and had no morphological abnormalities.

Technical Abstract: Somatic embryogenesis was obtained from immature cotyledon explants that were cultured on half-strength Murashige and Skoog (MS) salts and vitamins with 5.4 uM naphthaleneacetic acid (NAA) and 0.2 uM thidiazuron (TDZ) plus a 4x4 factorial combination of 0,9.8, 34.6, or 49.2 uM indole-3-butyric acid (IBA) and 0, 8.9, 22.2, or 44.4 uM 6-benzyladenine (BA). The addition of 44.4 uM BA improved the percentage of cotyledon explants that produced somatic embryos to >20%, if 9.8 or 24.6 uM IBA was also present.Somatic embryogenesis was >30% when seeds were harvested on 31 July or 15 August. The addition of 50 or 70 g 1- sucrose enhanced embryo-genesis. Histological examination showed that somatic embryos originated from epidermis cells of zygotic embryos. A peak germination rate (69%) was attained when somatic embryos were desiccated for 10 min before they produced green cotyledons and elongated shoot tips. Of the germinated embryos from this desiccation treatment, 65.6% also grew roots and therefore converted into plants. Chilling somatic embryos at 4 degrees Celsius for 15 days resulted in the highest germination rate (69.4%), which was significantly higher than those without chilling treatment (27.6%). However<10% of the chilled germinated embryos formed roots and grew into plants. Plantlets from somatic embryos were transplanted into a 2 vermiculite: 1 sphagnum peat medium, where they had a survival rate of 80.8%, and had no morphological abnormalities.

Last Modified: 10/21/2014
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