A Two-Stage Culture Method for Zygotic Embryos Effectively Overcomes Constraints Imposed by Hypocotyl and Epicotyl Seed Dormancy in Paeonia ostii 'Fengdan'

Abstract

The effect of the exogenous hormone and light quality on breaking hypocotyl and epicotyl dormancy was studied. The results showed that the greatest percentage of hypocotyl dormancy breaking was observed with the Murashige and Skoog (MS) medium supplemented with or without 1.0 mg.L-1 gibberellin 3 (GA(3)), while ABA and endosperm greatly inhibited hypocotyl dormancy breaking. This suggests that hypocotyl dormancy of the Paeonia ostii 'Fengdan' embryo could be easily overcome by removing constraints of the surrounding endosperm, and ABA may be one of the constraint factors contained in the endosperm. The percentage of epicotyl dormancy breaking was also greatly affected by the concentration of 6-benzylaminopurine (BA) and GA(3). Compared to BA by itself, adding GA(3) to the medium containing BA highly enhanced epicotyl dormancy breaking, with the greatest percentage of epicotyl dormancy breaking in MS medium supplemented with both 0.5 mg.L-1 BA and 0.5-1.0 mg.L-1 GA(3). The percentage of hypocotyl and epicotyl dormancy breaking was also affected by light and its quality. Red light-emitting diodes (LEDs) had the same effect as a dark condition on the hypocotyl dormancy breaking, while blue LEDs and a combination of red and blue LEDs had a negative effect on the hypocotyl dormancy breaking. Unexpectedly, blue LEDs greatly enhanced, whereas red LEDs inhibited, epicotyl dormancy breaking. Conclusively, a two-stage culture method was recommended for breaking the hypocotyl and epicotyl dormancy: hypocotyl dormancy was broken first using the MS medium without any plant growth regulators in the dark (25 degrees C), and epicotyl dormancy was subsequently broken with the MS medium supplemented with both 1.0 mg.L-1 GA(3) and 0.5 mg.L-1 BA under blue light.