Control of relative humidity and root-zone water content for acclimation of in vitro-propagated M9 apple rootstock plantlets

The present study aimed to evaluate the effects of controlling the relative humidity (RH) and water content of the root-zone on the survival rate and growth of in vitro-propagated virus-free M9 apple plantlets in closed-type plant production systems. In the first experiment, three RH regimes were applied to pre-acclimated (PA) and non-PA apple plantlets for 6 weeks after transplantation. In the second experiment, the apple plantlets were transplanted into several growth media, including a mixture of peat moss and perlite (PP), rock wool (RW), and urethane sponge (SP), and in a deep flow technique (DFT) system for controlled root zone water content under controlled RH. In the first experiment, pre-acclimation improved the survival rate by preventing the loss of leaf water potential and promoting antioxidant capacity during the acclimation period. However, no clear difference was found among the three RH regimes. The antioxidant capacity was increased at 2 weeks after transplantation, followed by root initiation. The leaf water potential, which decreased continuously until 3 weeks after transplanting, tended to remain constant after root initiation. These results suggested that pre-acclimation is necessary for the survival of in vitro-propagated apple plantlets, and that the underdeveloped roots of apple plantlets have restricted water absorption under controlled RH. In the second experiment, the survival rate of plantlets grown in PP at 6 weeks after transplantation was only 70% accompanied by an increase in antioxidant capacity, whereas the survival rates of plantlets grown in RW, SP, DFT, and DFT-PP (replanted to PP from DFT 4 weeks after transplantation) were 98, 96, 93.8, and 93.8%, respectively. Most of the growth parameters of the plantlets grown in DFT were the highest among the growth media at 6 weeks after transplantation. The results of the second experiment implied that the application of DFT for in vitro-propagated apple plantlets can reduce the problems caused by poor root architecture during acclimation.