Study On The Cultivation Of Callus Tissue Rapid Propagation Technique And Secondary Metabolite Of Glycyrrhiza Uralensis

As a large amount medicinal materials and economic crop, liquorice has relevantrelationship with people’ live. But with the increase of market requirement and excessiveexcavation of wild liquorice,licorice germplasm resources is seriously damaged. If usingartificial cultivation, seed growth cycle is long and breeding slow, which will restrict thedevelopment of glycyrrhiza industry. Therefore, rapid propagation of tissue culturetechnology of radix glycyrrhizae is good for the improvement of the liquorice breedingspeed, direct extraction of effective ingredients, and the preservation of the germplasmresources and breeding of improved varieties. By orthogonal experimental design,we studiedthe different explant callus induction rate and growth condition has effected by differenthormone combinations. The effective component content in the different explant callus ofinfluence and impact on cutting rapid propagation of licorice stem section by differenthormone combinations, in order to determine the best combination of hormones, for theapplication of licorice rapid reproduction technology to provide theoretical and technicalbasis. The follow are experimental results:1. This paper researched effects of the inducement on callus with different hormoneconcentration. By orthogonal experiment, in the condition of different hormoneconcentration ratio, comparing the effects of the inducement and growth situation ofhypocotyledonary axis, cotyledon, radical and stem callus. The suitable medium for licoricehypocotyl callus induction is6-BA1.5mg.L^-1+2,4-D0.5mg.L^-1+KT0.5mg.L^-1+NAA0.2mg.L^-1ï¼› the suitable medium for licorice cotyledon callus induction is6-BA0.5mg.L^-1+2,4-D0.5mg.L^-1+KT0.5mg.L^-1+NAA0.4mg.L^-1ï¼›the suitable medium for licoriceradical callus induction is6-BA1.5mg.L^-1+2,4-D0.5mg.L^-1+KT1.0mg.L^-1+NAA0.2mg.L^-1;the suitable medium for licorice stem callus induction is6-BA0.5mg.L^-1+2,4-D0.5mg.L^-1+KT1.5mg.L^-1+NAA0.4mg.L^-1. These explained the key factor of the effects ofliquorice different explants callus induction is different hormone concentration.2. This paper researched the effects of the active ingredients in liquorice differentexplants callus with different hormone concentration. In the condition of different hormoneconcentration ratio, comparing the effects of their secondary metabolites ofhypocotyledonary axis, cotyledon, radical and stem callus. The medium with glycyrrhizinicacid and total flavones for licorice with hypocotyl explants callus induction is6-BA1.5 mg.L^-1+2,4-D0.5mg.L^-1+KT1.5mg.L^-1+NAA0.4mg.L^-1，the average yield ofglycyrrhizinic acid is27.6638mg/g, the average yield of flavones is2.2586mg/g; the mediumwith glycyrrhizinic acid for licorice with cotyledon explants callus induction is6-BA0.5mg.L^-1+2,4-D1.0mg.L^-1+KT1.0mg.L^-1+NAA0.4mg.L^-1, the average yield ofglycyrrhizinic acid is24.6521mg/g, the medium with licoflavone for licorice with cotyledonexplants callus induction is6-BA1.5mg.L^-1+2,4-D0.5mg.L^-1+KT1.5mg.L^-1+NAA0.4mg.L^-1，the average yield of licoflavone is2.1277mg/g; the medium with glycyrrhizinic acidfor licorice with radical explants callus induction is6-BA1.5mg.L^-1+2,4-D1.5mg.L^-1+KT1.0mg.L^-1+NAA0.2mg.L^-1，the average yield of glycyrrhizinic acid is21.0844mg/g; themedium with licoflavone for licorice with radical explants callus induction is6-BA1.0mg.L^-1+2,4-D1.0mg.L^-1+KT1.5mg.L^-1+NAA0.2mg.L^-1，the average yield of licoflavoneis0.8791mg/g; the medium with glycyrrhizinic acid for licorice with stem explants callusinduction is6-BA1.5mg.L^-1+2,4-D0.5mg.L^-1+KT1.5mg.L^-1+NAA0.4mg.L^-1, theaverage yield of glycyrrhizinic acid is16.5345mg/g; the medium with licoflavone forlicorice with stem explants callus induction is6-BA1.5mg.L^-1+2,4-D1.5mg.L^-1+KT1.0mg.L^-1+NAA0.2mg.L^-1, the average yield of licoflavone is1.1126mg/g. So the key factor ofthe effects of liquorice different explants secondary metabolite is different hormoneconcentration.3. By orthogonal experiment, we researched the effects of growth situation of test-tubeplantlet with different hormone concentration and providing fundamental basis for licoricepropagation technology. The medium for test-tube plantlet taking root is NAA0.2mg.L^-1+IBA0.2mg.L^-1+KH₂PO₄13.5mg.L^-1and IAA0.5mg.L^-1+IBA0.5mg.L^-1+KH₂PO₄13.5mg.L^-1ï¼›the medium for test-tube plantlet growing is NAA0.2mg.L^-1+IBA0.4mg.L^-1+KH₂PO₄13.5mg.L^-1and IAA1.0mg.L^-1+IBA1.0mg.L^-1+KH₂PO₄13.5mg.L^-1. So thetest-tube growing fast when KH₂PO₄is13.5mg.L.