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Seed CultureEdit

Knudson C formulaEdit

  • Calcium Nitrate Ca(NO3)2•4H2O = 1.0 gram
  • Magnesium Sulfate MgSO4•7H2O = 0.25 gram
  • Ammonium Sulfate (NH4)2•SO4 = 0.50 gram
  • Ferric Sulfate FeSO4•7H2O = 0.50 gram
  • Sucrose C12H22O11 = 20 gram
  • Agar = 12 to 15 gram
  • Potassium Phosphate Buffer 0.1M, pH 5.3 to 5 =18 milliliter
  • Distilled Water DH2O = 1 liter

Meristem CultureEdit

Murashige and Skoog mediumEdit

Murashige and Skoog medium or (MSO or MS0 (MS-zero)) is a plant growth medium used in the laboratories for cultivation of plant cell culture. MSO was invented by plant scientists Toshio Murashige and Folke K. Skoog during Murashige's search for a new plant growth regulator. It is the most commonly used medium in plant tissue culture experiments.

As Skoog's doctoral student, Murashige originally set out to find an as-yet undiscovered growth hormone present in tobacco juice. No such component was discovered; instead, analysis of juiced tobacco and ashed tobacco revealed higher concentrations of specific minerals in plant tissues than were previously known. A series of experiments demonstrated that varying the levels of these nutrients enhanced growth substantially over existing formulations. It was determined that nitrogen in particular enhanced growth of tobacco in tissue culture.

Macronutrients

  • Ammonium nitrate (NH4NO3) 1,650 mg/l= 20.6 ml/L
  • Boric acid (H3BO3) 6.2 mg/l
  • Calcium chloride (CaCl2 · 2H2O) 440 mg/l
  • Cobalt chloride (CoCl2 · 6H2O) 0.025 mg/l
  • Magnesium sulfate (MgSO4 · 7H2O) 370 mg/l
  • Cupric sulfate (CuSO4 · 5H2O) 0.025 mg/l
  • Potassium phosphate (KH2PO4) 170 mg/l
  • Ferrous sulfate (FeSO4 · 7H2O) 27.8 mg/l
  • Potassium nitrate (KNO3) 1,900 mg/l
  • Manganese sulfate (MnSO4 · 4H2O) 22.3 mg/l
  • Potassium iodide (KI) 0.83 mg/l
  • Sodium molybdate (Na2MoO4 · 2H2O) 0.25 mg/l
  • Zinc sulfate (ZnSO4 · 7H2O) 8.6 mg/l
  • Na2EDTA · 2H2Oa 37.2 mg/lb

Common organic additives

  • i-Inositol 100 mg/l
  • Niacin 0.5 mg/l
  • Pyridoxine · HCl 0.5 mg/l
  • Thiamine · HCl 0.1 mg/l
  • Indole-3-acetic acid 1–30 mg/l
  • Kinetin 0.04–10 mg/l
  • Glycine (recrystallized) 2.0 g/l
  • Edamine1.0 g/l
  • Sucrose 20 g/l
  • Agar 10 g/l

ReferencesEdit

  • Charles R., Harrison, Joseph Arditti. "Growing Orchids from Seed." Orchid Digest Vol. 34-7 (September 1970): 199 -204.

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