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Fractional disinfection procedure for Huperzia lucidulum explants

, : Fractional disinfection procedure for Huperzia lucidulum explants. Hortscience 36(3): 583-584, June

The shining club moss, Huperzia lucidulum, can be propagated in tissue culture, but it is difficult to obtain sterile explants. To facilitate surface sterlization of H. lucidulum, a fractional disinfection procedure was developed. Based on the principle that vegetative microbial cells are more easily destroyed than spores, fractional disinfection induces spore germination before the treatment of explants with a sodium hypochlorite solution. This study compares the effectiveness of fractional disinfection to standard disinfection methods for club moss explants. When standard methods were used, 100% of H. lucidulum apical tips disinfected for 10, 20, or 30 minutes with 1% sodium hypochlorite/0.15% Tween 20 either quickly showed evidence of microbial contamination or turned brown. In contrast, when one 24 h cycle of fractional disinfection was used to surface sterilize H. lucidulum explants, 22% remained contamination free for over 5 weeks. Even better, when club moss explants were treated with two consecutive 24 h cycles of fractional disinfection, 50% remained contamination free for over five weeks. Because the fractional disinfection procedure was more effective than standard disinfection methods, it was applied to the original club moss explants, 81% of which had remained contaminated even after multiple standard disinfections. Applying the fractional disinfection method to these contaminated cultures produced an additional 15.6% contamination free club moss apical tips. After successful surface sterilization, H. lucidulum apical tips and gemmae established in vitro grew normally compared to conventionally propagated club moss cuttings and gemmae in nature.

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