medium no. 70: BSS medium, 0.54 µM 1-naphthaleneacetic acid, 0.2 mg/L kinetin, 0.3% (w/v) gellan gum, pH 5.7

Plant cell lines

Chemicals and stock solutions

(All stock solutions are stored at 4°C)

  • MS salt mix

    Murashige and Skoog Plant Salt Mixture, FUJIFILM Wako Pure Chemical Corporation (#392-00591)

  • Sucrose

  • BSS_VT

    Chemical

    Concentration (mg/mL)

    Nicotinic acid

    5

    Pyridoxine·HCl

    5

    Thiamine·HCl

    0.1

    Glycine

    2

  • MS_inositol

    Chemical

    Concentration (mg/mL)

    myo-Inositol

    40

  • NAA (1 mM)

    Chemical

    Concentration (mg/mL)

    NAA·K

    0.224

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

  • Kinetin (0.2 mg/mL)

    Chemical

    Concentration (mg/mL)

    Kinetin

    0.2

    Dissolve kinetin in small volume of KOH (1 N), and fill up with distilled water

  • Gellan gum

    Gellan gum, FUJIFILM Wako Pure Chemical Corporation (#073-03071)

  • KOH (1 N)

Medium preparation

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

    Chemical

    Amount

    MS 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)

    BSS_VT

    1

    MS_inositol

    2.5

    NAA (1 mM)

    0.54

    Kinetin (0.2 mg/mL)

    1

  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 3 g of gellan gum to the medium.

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

Formulation

Chemical

Concentration (mg/L)

KNO3

1900

NH4NO3

1650

CaCl2·2H2O

440

MgSO4·7H2O

370

KH2PO4

170

H3BO3

6.2

MnSO4·4H2O

22.3

ZnSO4·7H2O

8.6

KI

0.83

Na2MoO4·2H2O

0.25

CuSO4·5H2O

0.025

CoCl2·6H2O

0.025

FeSO4·7H2O

27.8

Na2-EDTA

37.3

Nicotinic acid

5

Pyridoxine·HCl

5

Thiamine·HCl

0.1

Glycine

2

myo-Inositol

100

Sucrose

20000

NAA·K

0.121

Kinetin

0.2

Gellan gum

3000