Patent ID: 6905876

Claim:
A method of in vitro cultivation of Polygonatum comprising the steps of: contacting a Polygonatum seed with a first medium comprising a MS basal culture medium and gibberellic acid (GA 3 ) present in an amount from about 10 mg/L to about 100 mg/L; upon emergence of a hypocotyl of a seedling, contacting said seedling with a second medium comprising a MS basal culture medium, 6-benzyl-aminopurine present in an amount from about 3 mg/L to about 6 mg/L, and naphthalene acetic acid (NAA) present in an amount from 0.5 mg/L to about 1.0 mg/L; and upon emergence of a first foliage leaf of said seedling, contacting said seedling with a third medium comprising a MS basal culture medium, 6-benzyl-aminopurine present in an amount of about 2.0 mg/L, naphthalene acetic acid present in an amount of about 1.0 mg/L, and GA 3 present in an amount from about 5 mg/L to about 20 mg/L, wherein the MS basal culture medium is selected from the group consisting of MS basal culture medium I and MS basal culture medium II, wherein MS basal culture medium I comprise 2.2 g/L NH 4 NO 3 , 2.0 g/L KNO 3 , 0.44 g/L CaCl 2 .2H 2 O, 0.37 g/L MgSO 4 .7H 2 O, 0.17 g/L KH 2 PO 4 , 37.25 mg/L Na 2 EDTA, 27.8 mg/L FeSO 4 .7H 2 O, 0.83 mg/L KI, 6.2 mg/L H 3 BO 3 , 22.3 mg/L MnSO 4 .4H 2 O, 8.6 mg/L ZnSO 4 .7H 2 O, 0.25 mg/L Na 2 MoO 4 .2H 2 O, 0.025 mg/L CuSO 4 .5H 2 O, 0.025 mg/L CoCl 2 .6H 2 O, 250 mg/L myo-inositol, 0.5 mg/L nicotinic acid, 0.5 mg/L pyridoxine HCl, 3 mg/L thiamine HCl, 2 mg/L glycine, and 30 g/L sucrose and MS basal culture medium II comprises 1.65 g/L NH 4 NO 3 , 1.9 g/L KNO 3 , 0.44 g/l CaCl 2 .2H 2 O, 0.37 g/L MgSO 4 .7H 2 O, 0.17 g/L KH 2 PO 4 , 37.25 mg/L Na 2 EDTA, 27.8 mg/L FeSO 4 .7H 2 O, 0.83 mg/L KI, 6.2 mg/L H 3 BO 3 , 22.3 mg/L MnSO 4 .4H 2 O, 8.6 mg/L ZnSO 4 .7H 2 O, 0.25 mg/L Na 2 MoO 4 .2H 2 O, 0.025 mg/L CuSO 4 .5H 2 O, 0.025 mg/L CoCl 2 .6H 2 O, 100 mg/L myo-inositol, 0.5 mg/L nicotinic acid, 0.5 mg/L pyridoxine HCl, 0.1 mg/L thiamine HCl, 2 mg/L glycine, and 30 g/L sucrose.