medium no. 5: Jouanneau and Péaud-Lenoël medium, 1 µM 1-naphthaleneacetic acid

Chemicals and stock solutions

(All stock solutions are stored at 4°C)
  • JPL_A’

    Chemical Concentration (mg/mL)
    KNO3 65.5
    CaCl2·2H2O 4.4
    MgSO4·7H2O 3.7
    KH2PO4 1.7
  • JPL_B

    Chemical Concentration (mg/mL)
    H3BO3 6.2
    MnSO4·4H2 O 22.3
    ZnSO4·7H2 O 10.6
    KI 0.83
    Na2MoO4·2H2O 0.25
    CuSO4·5H2O 0.025
    CoCl2·6H2O 0.025
  • JPL_C

    Chemical Concentration (mg/mL)
    FeSO4·7H2O 2.78
    Na2-EDTA 3.73

    Heat at 80°C for 3–4 hours for chelating Fe

  • JPL_D

    Chemical Concentration (mg/mL)
    Glycine 0.2
    myo-Inositol 10
  • JPL_VT

    Chemical Concentration (mg/mL)
    Nicotinic acid 0.5
    Pyridoxine·HCl 0.5
    Thiamine·HCl 0.4
  • JPL_P

    Stock solution Volume (mL)
    200 mM KH2PO4 19.5
    200 mM Na2HPO4 30.5
    H2O 50
  • NAA (1 mM)

    Chemical Concentration (mg/mL)
    NAA·K 0.224

    Potassium 1-naphthylacetate, Wako Pure Chemical Industries (#161-04021)

  • Sucrose

  • Casein hydrolysate, vitamin free

    Casamino acids vitamin assay, Difco (#228820)

  • KOH (1 N)

  • HCl (1 N)

Medium preparation

  1. Prepare three solutions as follows.
  • JPL mineral solution (1 L)

    Stock solution Volume (mL)
    JPL_A’ 37.5
    JPL_B 0.375
    JPL_C 2.5

    Adjust the pH to 5.7 with KOH (1 N)

  • JPL organic solution (100 mL)

    Chemical/Stock solution Amount
    JPL_D 10 mL
    JPL_VT 1 mL
    Casein hydrolysate 0.1 g

    Adjust the pH to 5.7 with HCl (1 N)

  • JPL sucrose solution (100 mL)

    Chemical/Stock solution Amount
    JPL_P 1 mL
    NAA (1 mM) 1 mL
    Sucrose 15 g
  1. Autoclave the solutions at 121°C for 20 min.
  2. Add 800 mL of JPL mineral solution, 100 mL of JPL organic solution, and 100 mL of JPL sucrose solution asceptically.

Formulation

Chemical Concentration (mg/L)
KNO3 1965
CaCl2·2H2O 132
MgSO4·7H2O 111
KH2PO4 56.308
Na2HPO4 8.66
H3BO3 1.86
MnSO4·4H2O 6.69
ZnSO4·7H2O 3.18
KI 0.249
Na2MoO4·2H2O 0.075
CuSO4·5H2O 0.0075
CoCl2·6H2O 0.0075
FeSO4·7H2O 5.56
Na2-EDTA 7.46
Nicotinic acid 0.5
Pyridoxine·HCl 0.5
Thiamine·HCl 0.4
Glycine 2
myo-Inositol 100
Casein hydrolysate 100
Sucrose 15000
NAA·K 0.224