Somatic embryo and root regeneration from quince leaves cultured in ventilated vessels or under different oxygen and carbon dioxide levels

Abstract

The influence of the gaseous composition of the atmosphere inside culturing vessels on somatic embryogenesis and on adventitious root formation was investigated in the quince clone (Cydonia ablonga Mill.) BA29. Leaves taken from in vitro-grown shoots were cultured in glass Petri dishes and exposed to ventilation with atmospheric air (flow rate 25 ml min−1) for 0, 5, 10, 20, and 40 d. Twenty days of ventilation reduced the frequency of embryogenic leaves and a further decrease was observed after 40d of treatment. Conversely, adventitious root formation in the ventilated dishes was never different from the untreated cultures. In a second test, leaves were incubated in atmospheres containing different levels of oxygen (0, 5.0, 10.0, and 21.0%) or carbon dioxide (0, 0.04, 0.15, 1.5, and 3.0%). Anoxia conditions almost completely inhibited somatic embryo and adventitious root formation, but without compromising callus formation and explant viability. In contrast, embryo and root regeneration occurred even in totally CO2-free atmosphere. Oxygen seemed to influence somatic embryogenesis according to a quadratic response; a similar relationship was also observed for root regeneration. Instead, no clear trend could be inferred between embryo or root regeneration and CO2 levels. Furthermore, in dishes flushed with gas mixtures containing oxygen or carbon dioxide somatic embryo formation was almost always lower than in confined dishes. A different result was observed for root regeneration, since the number of roots was never lower than in the control and increased appreciably with 3.0% CO2. These results demonstrate that atmosphere composition of the culture head-space can influence somatic embryogenesis in quince. The finding that both vessel ventilation and atmosphere replacement with different gas mixtures reduced somatic embryo formation suggests that gaseous compounds, different from O2 an CO2, present in the gaseous environment may promote embryogenesis in this species.