| CPC A01H 4/001 (2013.01) [A01G 17/005 (2013.01); A01G 24/15 (2018.02); A01G 24/22 (2018.02); A01H 4/008 (2013.01)] | 9 Claims |
|
1. A method of rapid propagation of blueberry stem segment tissue, comprising the following steps:
S1: inoculating the sterile blueberry stem segment with a bud point into the pre-culture medium to conduct pre-culture to obtain a pre-cultured blueberry stem segment, the pre-culture medium including
825 mg/L Ammonium Nitrate, 370 mg/L Magnesium Sulfate Heptahydrate, 950 mg/L Potassium Nitrate, 170 mg/L Potassium Dihydrogen, 220 mg/L Calcium chloride dihydrate, 6.2 mg/L Boric acid, 8.6 mg/L Zinc Sulfate Heptahydrate, 22.3 mg/L Manganese Sulfate Monohydrate, 0.25 mg/L Sodium Molybdate Dihydrate, 0.025 mg/L Cooper Sulfate Pentahydrate, 0.83 mg/L Potassium iodide, 0.025 mg/L Cobalt chloride hexahydrate, 40 mg/L ethylenediamine dihydroxyphenyl sodium iron acetate, 20 mg/L 2-(N-morpholine) ethanesulfonic acid, 100 mg/L Inositol, 2 mg/L Glycine, 0.1 mg/L Thiamine Hydrochloride, 0.5 mg/L Niacin, 0.5 mg/L Pyridoxine Hydrochloride, 0.1-0.5 mg/L IAA, 0.05-0.6 mg/L CPPU, 50-100 mg/L NaCl, 4-8 g/L agar and 15-30 g/L sucrose, with a pH in the range of 5.0-5.5;
S2: inoculating the pre-cultured blueberry stem segment into a rapid culture medium to conduct rapid culture to obtain rapid cultured blueberry seedlings, the rapid culture medium including
825 mg/L Ammonium Nitrate, 370 mg/L Magnesium Sulfate Heptahydrate, 950 mg/L Potassium Nitrate, 170 mg/L Potassium Dihydrogen, 220 mg/L Calcium chloride dihydrate, 6.2 mg/L Boric acid, 8.6 mg/L Zinc Sulfate Heptahydrate, 22.3 mg/L Manganese Sulfate Monohydrate, 0.25 mg/L Sodium Molybdate Dihydrate, 0.025 mg/L Cooper Sulfate Pentahydrate, 0.83 mg/L Potassium iodide, 0.025 mg/L Cobalt chloride hexahydrate, 40 mg/L ethylenediamine dihydroxyphenyl sodium iron acetate, 20 mg/L 2-(N-morpholine) ethanesulfonic acid, 100 mg/L Inositol, 2 mg/L Glycine, 0.1 mg/L Thiamine Hydrochloride, 0.5 mg/L Niacin, 0.5 mg/L Pyridoxine Hydrochloride, 0.2-0.5 mg/L ZT, 0.5-1.5 mg/L putrescine, 0.5-1.5 mg/L spermine, 0.5-1.5 mg/L spermidine, 4-8 g/L agar and 15-30 g/L sucrose, with a pH in the range of 5.0-5.5;
S3: inoculating the rapid cultured blueberry seedlings into a seedling growth medium to conduct continue culture to obtain further cultured blueberry seedlings, the seedling growth medium including
825 mg/L Ammonium Nitrate, 370 mg/L Magnesium Sulfate Heptahydrate, 950 mg/L Potassium Nitrate, 170 mg/L Potassium Dihydrogen, 220 mg/L Calcium chloride dihydrate, 6.2 mg/L Boric acid, 8.6 mg/L Zinc Sulfate Heptahydrate, 22.3 mg/L Manganese Sulfate Monohydrate, 0.25 mg/L Sodium Molybdate Dihydrate, 0.025 mg/L Cooper Sulfate Pentahydrate, 0.83 mg/L Potassium iodide, 0.025 mg/L Cobalt chloride hexahydrate, 40 mg/L ethylenediamine dihydroxyphenyl sodium iron acetate, 20 mg/L 2-(N-morpholine) ethanesulfonic acid, 100 mg/L Inositol, 2 mg/L Glycine, 0.1 mg/L Thiamine Hydrochloride, 0.5 mg/L Niacin, 0.5 mg/L Pyridoxine Hydrochloride, 0.5-1.5 mg/L 2-ip, 4-8mg/L agar and 15-30 g/L sucrose, with a pH in the range of 5.0-5.5; and
S4: inoculating the further cultured blueberry seedlings into a rooting medium to conduct continue culture to obtain blueberry tissue culture seedlings, the rooting medium including
950mg/L Potassium Nitrate, 166.1 mg/L Calcium Chloride, 825 mg/L Ammonium Nitrate, 90.35 mg/L Magnesium Sulfate, 85 mg/L Potassium Dihydrogen Phosphate, 0.025 mg/L Cooper Sulphate, 6.2 mg/L Boric Acid, 16.9 mg/L Manganese Sulfate, 0.25 mg/L Sodium Molybdate, 8.6 mg/L Zinc Sulfate, 0.025 mg/L Cobalt Chloride, 0.83 mg/L Potassium Iodide, 27.8 mg/L FeSO4·7H2O, 37.26 mg/L Na2-EDTA·2H2O, 2.0 mg/L Glycine, 100 mg/L Inositol, 0.50 mg/L Niacin, 0.1 mg/L Thiamine Hydrochloride, 0.5 mg/L Pyridoxine Hydrochloride, 1.0-3.0 mg/L NAA, 4-8 g/L agar and 15-30 g/L sucrose, with a pH in the range of 5.0-5.5.
|