Dihaploid Callus Production from Ulmus americana Anthers

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Abstract
Diphaploid (2x = 28) Ulmus americana callus was consistently produced from anthers. Dihaploidy was verified by chromosome counts. Specific media requirements for producing dihaploid callus were a high sucrose concentration of at least 3% and high levels of either auxin or auxin with cytokinin. Also, callus formation from anthers of four genotypes varied significantly, from 3 5% to 12 5%. During two seasons, 17 media and three incubation regimes (darkness, 16 h light/8 h dark, and transfer from dark to light after 2 wk) were tested for callus production. Anthers incubated under dark conditions produced a greater frequency of dihaploid callus than anthers cultured under the other two regimes. Pretreating anthers by centrifugation at 5 C did not increase callus formation. Although budlike structures formed from dihaploid callus in several instances, no further development occurred. This is the first report of dihaploid callus production from U. americana anthers.

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