Tissue Culture Regenerated Plant And Genetic Stability Analysis Of Dendrobium Candidium Wall. Ex Lindl

Dendrobium candidium Wall. Ex Lindl refers to a types of orchid perennial epiphytes, is one of the well known traditional Chinese medicines. Most of Dendrobium candidium Wall. Ex Lindl distribute in the following five major areas of China:ZheJiang,YunNan,HuNan and GuiZhou.According to the current researches,it contains dendrobium polysaccharide, alkaloids, amino acids, trace elements and some novel compounds,with higher medicinal and economic value.In this paper, a systematic and comprehensive study has been did for the whole regeneration system of Dendrobium candidium Wall though tissue culture,the analysis of RAPD polymorphism between seed-derived seedling and regenerated plant. These researches provide a significant scientific basis for the development and utilization of the medicinal value of Dendrobium candidium Wall. Ex Lindl.The main results obtained from the experiment as follows:1. The system for regeneration of Dendrobium candidium Wall. Ex Lindl(1)It was disinfected with70%ethanol to30s, brushing with sterile water2times, then treated with0.2%HgCl2for7min,brushing with sterile water5times,the ratio of survival was89%for Root hills;It was disinfected with70%ethanol to20s, brushing with sterile water2times, then treated with0.2%HgCl2for3min,brushing with sterile water5times,the ratio of survival was89%for Stem.(2)The best callus induction medium:MS+6-BA0.5mg/L+NAA0.1mg/L+2,4-D1.0mg/L+Sucrose30g/L+Agar7.5g/L, root hills induction rate reached93.33%; Stem induction rate reached95.56%.(3)The best proliferation medium:MS+6-BA0.5mg/L+IBA1.0mg/L+KT1.0mg/L+Agar7.5g/L, the callus grew fast, the proliferation coefficient reached4.3after30d cultivation.(4)The best differentiation medium:1/2MS+NAA0.5mg/L+6-BA1.0mg/L+banana juice50g/L+Agar7.5g/L+Sucrose30g/L, the differentiation rate reached97.78%.(5) The best rooting induction medium:1/2MS+NAA0.5mg/L+Agar7.5g/L+Sucrose30g/L+10%banana juice, the rooting rate reached99%. (6)The better basal material was humus soil:moss:peanut shel1(1:1:1) and the rate of survival was96.9%.2. The results of RAPD analysisRAPD molecular markers was used to analy the genetic stability between regenerated plant and wild seedling.The results show that the regenerated plant and wild seedling,compared to the DNA level on acertain extent the percentage variation for3.33%,At the same time that also exists between the regenerated plants,the percent variation for2.08%.