In vitro plantlet propagation and efficiency of virus elimination from ornamentals by sterile culture.

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Simple methods for preparing sterile medium without autoclaving and inoculating explains without the laminar air-flo cabinet using direct application of chlorine disinfectants were developed. The sterile medium could be prepared without autoclaving by immediately incorporating sodium hypochlorite solution or sodium dichloroisocyanurate solution (active chlorine, 0.005-0.01%) into me medium. Residual chlorine in the medium was effective for maintaining sterility of medium. By incorporating chlorine disinfectant plastic vessels and bags mat can not be sterilized by autoclaving could be utilized as simple culture vessels for micropropagation of orchids and other omamentals.Spraying the surface of a medium and the whole of explants and plantlets with sodium hypochlorite solution or sodium dichloroisocyanurate solution (active chlorine, 0.005-0.05%) after inoculating them on the medium, proved to be feasibl an practical for inoculating procedure without the laminar air-flow cabinet in the open air under non-sterile conditions. Solution concentrations of the incorporated and sprayed disinfectants did not appear to be toxic to the orchid explains chrysanthemum and carnation plantlets etc. These techniques which are easy to prepare medium and to inoculate explants and transplanting plantlets without special equipment can be applied to various procedures of horticultural plants in vitro.The efficiency of vims elimination by simple shoot tip cultures and simple in vitro cultures was investigated using, Inacase of Cymbidium, the frequency of CyMV elimination tended to increase with increasing subcultures of PLBs. In PLBs obtained flora the simple cultures using flower stalk sections and leaf sections of Phalaenopsis, all the PLBs were CyMV-fiee by dot immnobinding assay (DIBA). These results indicate that the methods developed in this study can also be utilized as a novel method for in vitro propagation of virus-free plants.