Study On Rapid Propagation Tissue Culture Process Of Dendrobium Officinale Homologous Tetraploid

Based on dendrobium homologous tetraploid Flower-2 which from the two strains by tip induction in laboratory,depending on stems break propagation and protocorm induced proliferation and differentiation methods,establishing ploidy strain bacteria free high frequency fast propagation system,providing dendrobium homologous tetraploid strain large scale breeding,laying excellent ploidy strain's production and promotion foundation.Using stomata,Chloroplast,Chromosome count and flow cytometry 4 methods to judge from the Flower-2 homozygous,comparing two technical solutions's proliferation efficiency of stems break propagation and protocorm induced proliferation and differentiation methods,and re-identify protocorm differentiation to get test tube seedlings ploidy stability.Confirming to get Flower-2 strains is homologous tetraploid by tip induction in laboratory.Stem segments spreading optimal proliferation medium is:MS + 2.0 mg·L-16-BA + 0.5 mg·L-1NAA+15%banana juice,but this way has low proliferation coefficient,so it doesn't satisfy factory production needs.Young stem segments can induced production numerous to protocorm,and stem segments induced production the protocorm's optimal medium is MS+0.5 mg·L-16-BA+0.5 mg·L-1NAA+0.2 mg·L-1 KT,induction rate is 76.66%;Dendrobium the protocorm's proliferation optimal medium is MS+1.0 mg·L-16-BA+0.5 mg·L-1NAA+15%potato extract;Dendrobium protocorm's differentiation optimal medium is 1/2MS+0.5 mg·L-1NAA+20%potato extract;Differentiation rate is 85.70%;Dendrobium protocorm's rooting optimal medium is 1/2 MS + 0.5 mg·L-1 NAA + 15%banana juice,rooting rate is 92.77%;Bottle of seedlings transplanting's optimal refining period is 5 days.Test establishing dendrobium homologous tetraploid strain high frequency and fast propagation system,laying tetraploid test tube seedlings' factory production and production promotion technical foundation.