Tissue Culture Of Bletilla Striata(Thunb.) Reichb.f And Plantlet Nutrition Regulation

The Chinese Bletillae Rhizoma of 2010 was contained Bletilla striata?Thunb.?Reichb.f.It is a traditional Chinese medicine,with the function nourishing lung,hemostasis,relieves rheumatism and chills,therapy of ventilating lung and relieving stuffy nose,etc.Because of the small seeds of B.striata must germinate wtih the specific symbiotic fungus,and low survival rate resulting in asexual propagation such as cuttage and division,the destruction of natural habitats,wild resources were damaged seriously.It is necessary that develop more perfect tissue culture and rapid propagation system of B.striata.To study seedling and transplanting technology,provide the theoretical basis for solving the above problems,this paper establish the tissue culture and rapid propagation with its seeds,isolate endophytic fungus in B.striata.from different habitats.Study on how to improve transplanting survival measures.And now there are some aspects:1.B.striata.seeds must germinate with light.1/2 MS is suitable seed germination medium.NAA and 6-BA is beneficial to germinate,especially when adding NAA 1.0 mg·L^-1 and 6-BA 1.6 mg·L^-1.PLBs monthly induction rare reach highly when add coconut juice instead of sucrose in the medium.It is noneffective when adding potato extracts.So 1/2 MS+1.0 mg·L^-1NAA+1.6 mg·L^-16-BA is the suitable germination medium to obtain strong seedlings,the germination rate can be up to 98%?50 d?.In order to obtain the PLBs,1/2 MS with 20%coconut juice is the optimum medium.2.MS as the basic culture medium can promote the growth of seedlings.Add 10 mg·L^-1 FA,or 10%coconut and 5%banana juice instead of the sucrose can promote B.striata.To grow healthily.The addition of 25 mg·L^-1 HA was benefit for rooting.Culture on various culture medium should not be too long.Effects of yeast and potato on growth of need further study to determine.3.Study on effects of NH₄⁺-N and NO₃^--N on seedling growth,founding that seedlings grew better when the ratio of ammonium to nitrate was 75:25.More ammonium nitrogen content than nitrate nitrogen would be better for seedlings growing.Total ammonium nitrogen or nitrate nitrogen were unfavorable.Leaves would have high physiological activity when atio of ammonium to nitrate was 50:50.4.B.striata seedlings grew well when adding 26.85 mmol·L^-1 K⁺.At the same time,the photosynthetic pigment content in leaves is highest.Enrichment of K⁺ in a certain range of concentration could enhance SOD and POD activities and soluble protein content in leaves and reduce the accumulation of MDA.In order to promote the growth of B.striata seedlings,increasing the content of K⁺ in culture medium was required,and the feat concentration was 26.85 mmol L.1.5.Organic additives played important roles in B.striata tissue culture,such as potato extract,coconut and banana juice.Different plant hormones regulated different function.NAA helped to promote B.striata seedlings to grow well and it could work better while adding 6-BA.2,4-D can successfully induce callus by B.striata seedlings.The most favorable concentration of KT to promote the growth of leaves was 1 mg·L^-1.6.In a certain range of concentration,the increasing of Ca²⁺ content in culture medium is conducive to the promotion of the growth of B.striata seedlings and produce higher physiological activity of leaves.When the concentration of Ca²⁺ was 4.10 mmol·L^-1,the seedlings grew well,with a high concentrations of photosynthetic pigments;and there POD activity of leaves reached the highest level.It increased the the content of soluble protein and activity of SOD in leaves and decreased MDA accumulation when Ca²⁺ concentration was 3.40 mmol L^-1.7.The B.striata seedlings stage had great influence on the survival rate.too high or too short were not benefit to survive.When the height is about 49.75mm,the transplanting survival rate was up to 64%after 2 months.The order that B.striata survival rate in a variety of basic matrix was:Northeast black soil>soil with fertilizer>vermiculitef>nutritive soil for fiowers.Adding sawdust could facilitate B.striata survive,especially with northeast black soil.8.We found two new B.striata endophytes T.Koningiopsis and H.Koningii using the method of ITS and morporlogy.