111 Successful in Vitro Micropropagation of Purported Blight-resistant Chestnuts (Castanea spp.)

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  • 1 Univ. of Nebraska-Lincoln, Dept. of Horticulture, Lincoln, NE 68583-0724

The American Chestnut Foundation (ACF) has conducted a breeding program aimed at developing blight-resistant chestnut trees exhibiting the phenotype of American Chestnut (Castanea dentata). We developed a protocol for in vitro micropropagation and multiplication of candidate blight-resistant plants from the ACF breeding program. The protocol included forcing dormant shoots to budbreak, culture establishment, shoot multiplication, inducing a functional root system on the microcuttings produced by this system and establishment of autotrophic plants. Because Castanea spp. is recalcitrant to rooting, a unique bilayer method of rooting was developed. The unique bilayer consisted of a clear basal medium of 50% DKW and 50% WPM (Long and Preece), with a continuous level of 0.01 mg IBA/L and 0.2 mg BA/L. The clear basal medium was over-laid with an opaque layer. Rooting response occurred for 27 of the 31 genotypes at various frequencies. Rooted plantlets were planted in 50% peat: 50% perlite in order to become autotrophic and acclimated. Acclimated trees were planted in 10″ × 2″ Deepots® and placed in the greenhouse. These trees exhibited a very vigorous functional root system. Acclimated trees were hardened off, placed in cold storage (≈4-5 °C) for 5 months. All trees placed in cold storage broke dormancy for spring growth and ≈100 trees were sent to ACF for planting into field trials.

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