Establishment of an Efficient Micropropagation System in Anthurium Hybrids Through In Vitro Callogenesis and Suspension Culture

Abstract

The development of new Anthurium cultivars relies on efficient micropropagation, which is highly dependent on genotypes, to provide enough young plants for cultivation. Microbial contamination is the critical factor for successful initiation of aseptic culture of Anthurium cultivars. The aims of this study were to investigate surface disinfection with either sodium hypochlorite or a fungicide Plant Preservative Mixture^TM (PPM), and supplemental PPM in the medium to evaluate their effect on reducing contamination and tissue browning. Next, a liquid suspension culture system was developed to regenerate the adventitious shoots efficiently in a short time for subsequent establishment of young plantlets for transplanting. Disinfecting newly developed leaves of ‘Kaohsiung No. 1’ with 0.6% sodium hypochlorite was more effective than other disinfection treatments even though only 5% of leaf explants produced callus. However, disinfecting the leaves of ‘Kaohsiung No. 2’ with 0.15% sodium hypochlorite was more effective than other treatments, with a callus induction rate up to 100%. No callus was induced on ‘Orange Hot’ at any hypochlorite concentration. Disinfection using PPM could reduce the contamination rate. By supplementing 0.01% PPM to the culture medium, the callus induction rate of ‘Kaohsiung No. 2’ was up to 55%, and the browning rate was lower than the control with hypochlorite disinfection. However, surface disinfection using 25% and 50% PPM did not lead to any callus formation in any of the three cultivars. Calluses of A. ‘Kaohsiung No. 2’ induced after 60 days of culture were transferred onto liquid suspension after 60 days for further proliferation of adventitious shoots and subsequent plantlet regeneration. Young plantlets could be successfully transplanted in peat moss and later flowered within 16 months.