Plant Cell, Tissue and Organ Culture

, Volume 57, Issue 1, pp 47-55

First online:

Development and germination of American chestnut somatic embryos

  • Zizhuo XingAffiliated withDepartment of Microbiology and Immunology, State University of New York, Health Sciences Center
  • , William A. PowellAffiliated withFaculty of Environmental and Forest Biology, State University of New York, College of Environmental Science and Forestry
  • , Charles A. MaynardAffiliated withFaculty of Forestry, State University of New York, College of Environmental Science and Forestry

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American chestnut (Castanea dentata (Marsh.) Borkh.) plants were regenerated from developing ovules through somatic embryogenesis. On an initiation medium containing 18.18 μM 2,4-dichlorophenoxyacetic acid and 1.11 μM 6-benzyladenine (BA), 25 out of 1,576 ovules were induced to form proembryogenic masses (PEMs). These PEMs were cultivated on a development medium for 4 weeks. Individual somatic embryos were then grown on a maturation medium for at least one month, until shoot meristems and radicles were developed. Both development and maturation media consisted of Gamborg's B-5 basal medium, 0.5 μM BA, and 0.5 μM α-naphthaleneacetic acid, but the former contained 20 g l−1 sucrose and the later contained 60 g l−1 sucrose. A range of 86 to 586 embryos per gram PEMs was observed beyond the cotyledonary stage. These embryos then germinated, resulting in plantlets with a 3.3% conversion rate. An additional 6.3% of the mature embryos produced shoots, which could also result in plantlets by rooting of microcuttings. Proembryogenic masses that were established in continuous culture and maintained on initiation medium for 17 months retained regenerability, though the embryo yield decreased over time. Twenty plantlets were acclimatized and grown in potting mix in a greenhouse. The largest 6 were transplanted, along with seedling controls, into a nursery bed in 1997. As of July, 1999, 4 out of the 6 were surviving.

acclimatization Castanea dentata embryogenesis establishment plant regeneration