Somatic embryogenesis in American chestnut
1991
Merkle, S.A. | Wiecko, A.T. | Watson-Pauley, B.A.
Cultures were initiated from developing ovules and excised embryos of American chestnut (Castanea dentata (Marsh.) Borkh.), collected from five source trees on three dates during early and middle stages of fruit development. Explants were cultured initially on semisolid induction medium containing 0.25 mg/L benzyladenine and either 6 mg/L naphthaleneacetic acid or 4 mg/L 2,4-dichlorophenoxyacetic acid for 1 or 2 weeks. Then they were either transferred to hormone-free medium or medium with 0.25 mg/L benzyladenine or maintained on the original induction media. Ovules collected from three of the five trees 6 or 7 weeks postanthesis produced embryogenic cultures. Those pulsed for 1 or 2 weeks on auxin-containing medium prior to transfer to media without auxin produced multiple somatic embryos directly from the radicle end of the zygotic embryo. Cultures maintained on auxin-supplemented media initially produced proembryogenic masses, which formed globular and heart-stage embryos as they aged. Transfer of clusters of somatic embryos from auxin-supplemented media to hormone-free medium promoted maturation of embryos to the cotyledon stage.
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