Large-scale plantlets conversion from cotyledonary somatic embryos of Kalopanax septemlobus tree using bioreactor cultures

A suitable bioreactor system for large scale embryo-to-plantlets conversion of Kalopanax septemlobus was established. In temporary immersion with net (TIN) bioreactor, 85% of embryos successfully produced plantlets whereas in continuous immersion with net (CIN) bioreactor, only conversion rate of 29.3% was obtained. Embryos cultured in TIN bioreactor produced more vigorous plantlets in terms of fresh weight, height, root length, roots and leaves quantity. In CIN bioreactor, Kalopanax plantlets showed high malondialdehyde (MDA) content and increased activities of reactive oxygen species (ROS)-processing enzymes, such as ascorbate peroxidase (APX) and glutathione reductase (GR) indicating the occurrence of oxidative stress. However, superoxide dismutase (SOD) and catalase (CAT) showed similar activities in plantlets grown in different bioreactors. Kalopanax plantlets grown in both TIN and CIN bioreactors were harvested and transferred to greenhouse for their acclimatization. Plantlets grown in CIN bioreactor exhibited low survival rate (75.8%) compared to those grown in TIN bioreactor (100%). MDA content decreased with progression of acclimatization indicating a decrease in oxidative stress. However, MDA level in CIN derived plantlets was higher than TIN derived plantlets. In TIN derived plantlets, an increase in SOD and GR activities were observed after 1 week and thereafter decreased. CAT activity decreased while APX activity started to increase after 1 week of acclimatization. The results indicated that Kalopanax plantlets were able to overcome oxidative stress mainly through SOD activity. However, levels of antioxidant enzyme activities were higher in CIN derived plantlets than TIN derived plantlets. Kalopanax plantlets obtained from TIN bioreactor performed better during the acclimatization phase and showed higher survival rate than material obtained on CIN bioreactor or conventional culture systems.