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

Plant cell lines

• rpc00008: Arabidopsis thaliana T87 cell suspension culture

Chemicals and stock solutions

(All stock solutions are stored at 4°C)

• JPL_Aʼ

  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
  MnSO4·5H2O	24.1
  ZnSO4·7H2O	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, FUJIFILM Wako Pure Chemical Corporation (# 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_Aʼ	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
MnSO4·5H2O	7.23
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, vitamin free	100
Sucrose	15000
NAA·K	0.224