medium no. 53: modified Gamborgʼs B5 medium, 0.25% (w/v) gellan gum, pH 5.7

Plant cell lines

• rpc00067: Fragaria × ananassa SB489 callus culture

Chemicals and stock solutions

(All stock solutions are stored at 4°C)

• B5 salt mix

  Gamborgʼs B5 Medium Salt Mixture, FUJIFILM Wako Pure Chemical Corporation (#399-00621)

• Sucrose

• MS_VT

  Chemical	Concentration (mg/mL)
  Nicotinic acid	0.5
  Pyridoxine·HCl	0.5
  Thiamine·HCl	0.1
  Glycine	2

• MS_inositol

  Chemical	Concentration (mg/mL)
  myo-Inositol	40

• Gellan gum

• KOH (1 N)

Medium preparation

1. Dissolve the following chemicals in approximately 800 mL of distilled water.

   Chemical	Amount
   B5 salt mix	1 bag (1 L)
   Sucrose	20 g

2. Add following stock solutions, and fill up to approximately 950 mL with distilled water.

   Stock solution	Volume (mL)
   MS_VT	1
   MS_inositol	2.5

3. Adjust the pH of the solution to 5.7 with KOH (1 N), and fill up to 1 L with distilled water.

4. Add 2.5 g of gellan gum to the medium.

5. Autoclave the medium at 121°C for 20 min.

Formulation

Chemical	Concentration (mg/L)
KNO3	2500
(NH4)2SO4	134
CaCl2·2H2O	150
MgSO4·7H2O	250
NaH2PO4·H2O	150
H3BO3	3
MnSO4·H2O	10
ZnSO4·7H2O	2
KI	0.75
Na2MoO4·2H2O	0.25
CuSO4·5H2O	0.025
CoCl2·6H2O	0.025
FeSO4·7H2O	27.8
Na2-EDTA	37.3
Nicotinic acid	0.5
Pyridoxine·HCl	0.5
Thiamine·HCl	0.1
Glycine	2
myo-Inositol	100
Sucrose	20000
Gellan gum	2500