Seedling And Tissue Culture Of Callus And Adventitious Root Of Epimedium

Epimedium,as a traditional medicinal plant,has been used for more than 2000 years in China,modern researchers have found a variety of chemical components with strong physiological activities,such as total flavonoids,polysaccharides,alkaloids,etc.It is widely used in traditional medicine and modern medicine in present.Now,the market demand of epimedium is about 5000 tons / year,the way of digging wild plants has been caused great damage to the natural environment,and adverse to product development and further scientific research.In order to meet the market demand,purify the quality of medicinal materials,protect the wild germplasm resources of epimedium and its surrounding environment,the experiment of epimedium biological characteristics was carried out to study the dormancy-released,seedling and tissue culture,the research methods,main works and related conclusions of the experiment are as follows:1.Release of seed dormancyBy orthogonal experiment,selected the best medicine combination which can relieve the dormancy of epimedium seeds effectively,The results show that,the combination of sodium tungstate1320mg/L+fluridone2.5mg/L+gibberellin300mg/L+sodium nitro-prusside298mg/L+ethephon144.5mg/l had the best effect on seed dormancy release,which can released seed dormancy within 60 days,and the total germination rate is 80%.The results of significance analysis and range R showed that the effects of sodium tungstate,fluridone,gibberellin,sodium nitroprusside and ethephon were ex-tremely significant(P <0.01)on the germination rate of epimedium seeds.and it can be seen that the order of influence was: fluridone> sodium tungstate >ethephon> gibbere-llin> sodium nitroprusside.2.Cultivation of seedlingBy orthogonal experiment,selected the seedbed substrate for epimedium seedlings,the results show that,The survival rate of seedlings,which were seeded on the seedbed with two parts of peat,three parts of vermiculite and one part of perlite,is the highest,the results of significance analysis and range R showed that the effect of peat on the survival rate of seedlings reached a significant level(P<0.05),and it can be seen that the influence among three factors,from large to small,was: peat >vermiculite >perlite.3.Tissue culture of callus and adventitious root(1)Callus induction: use epimedium stem as explants and design orthogonal experiment,screen out the optimal combination of hormone and exogenous additive for callus induction,2,4-D1.5mg/L+6-BA3.0mg/L+KT1.5mg/L+NAA0.5mg/L+ coconut juice70ml/L+casein0.5 g/L,under this combination,the callus formation rate of explants treated can reach 77.40%.The results of significance analysis and range R showed that the use of KT reached a significant level(P<0.05),while the use of 2,4-Dreached extre-mely significant(P <0.01),and the influence among each factor on the results was: 2,4-D >KT > casein > 6-BA > NAA> coconut juice.(2)Root induction: use epimedium callus as explants and design orthogonal experiment,screen out the best combination of hormone for root induction,From the experimental results,it can be seen that the hormone NAA0.3mg/L+IBA0.5mg/L+6-BA0.3mg/L+IAA0.6mg/L,under this combination,it was the best for the root formation of callus,the rooting rate can reach 90.71%.According to the results of significance analysis and range R,the use of IAA reached extremely significant(P < 0.01),and the order of influence of several hormones was: IAA > NAA > 6-BA > IBA.(3)Induction of total flavonoids synthesis: use epimedium callus and root as explants and design orthogonal experiment,screen out the best elicitor combination of callus and root.The experimental results show that,salicylic acid 0.9mg/l+yeast extract1.0mg/ml+methyl jasmonate5.0mg/l,it could promote the synthesis of total flavonoids in callus,and the content could reach 5.73%;salicylic acid 0.7mg/l+yeast extract1.0mg/ml+methyl jasmonate4.0mg/l,it could promote the synthesis of total flavonoids in root,and the content could reach 5.76%.According to the results of significance analysis and range R,salicylic acid to callus and root on the synthesis of total flavonoids,the level was significant(P < 0.05),in order to improve synthesis of total flavonoids in callus,the influence of several inducers was in the order of large to small,and the influence on roots was: salicylic acid > yeast extract > methyl jasmonate.