The Relationship of Apple Embryos and Their Cotyledons to Maturity, Dormancy, and the Potential to Form Shoots in Vitro

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Abstract
Embryos extracted from developing apple fruit were cultured in vitro. Immature embryos (up to 4 weeks post anthesis), cultured without their testa and endosperm, produced only callus tissue. Six and a half-week-old embryos produced multiple shoots and the percentage of embryos producing shoots rather than callus tissue increased with time. Ten and a half-week-old embryos produced the highest number of shoots per embryo, after which a decrease in multiple shoot formation was observed with mature (14 weeks post anthesis) embryos producing 1 shoot and 1 root per embryo. Simultaneously, embryonic-axis free cotyledon cultures were established from embryos of various ages. Adventitious shoots developed from cotyledon explants, and the number of multiple shoots formed, increased with time (June-September). Cotyledons extracted periodically from mature fruit stored at 3°C continued to produce multiple shoots in culture, but this gradually decreased with time (15-120 days). After 6 months in cold storage, cotyledon explants failed to develop any shoots.

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