Abstract
An efficient method of axenic seed propagation of large numbers of Cypripedium reginae Walt, was developed as a means of supplying seedlings for a restoration attempt in the northeastern United States. Seedlings were grown on ¼ strength Murashige and Skoog basal salts supplemented with 100 ml/l coconut milk and solidified with 7g/l agar. The development of Cypripedium reginae was studied with both light and scanning electron microscopy. Dose response experiments of seed germination versus exposure time to sodium hypochlorite indicate germination and developmental progress was greatest at longer exposure times. Experiments on seed germination subsequent to exposure to increased concentrations of sodium hypochlorite (0%, 0.125%, 0.25%, and 0.5%) indicate germination and developmental progress was greatest at higher concentrations. Earliest germination was seen at 14 days after initial plating of seed. Stage 5 seedlings were present 27 days after initial plating and in greatest proportion in seeds exposed to longer times in sodium hypochlorite. Micropropagation experiments showed a success rate of 32% with an indication of difficulty in cutting the seedlings to allow for both shoot and root development. Approximately 10,000 seedlings have been successfully germinated and have developed both healthy roots and shoots.
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