L zeatin, pH 5.8

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
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
2,4-D (0.2 mg/mL)

Chemical

Concentration (mg/mL)

2,4-D sodium monohydrate

0.236

(2,4-Dichlorophenoxy)acetic acid sodium salt monohydrate, Sigma-Aldrich (D6679)
NAA (1 mg/mL)

Chemical

Concentration (mg/mL)

NAA·K

1.2

Potassium 1-naphthylacetate, FUJIFILM Wako Pure Chemical Corporation (#161-04021)
Zeatin (1 mg/mL)

Chemical

Concentration (mg/mL)

trans-Zeatin

1

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

Medium preparation

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

Chemical

Amount

MS salt mix

1 bag (1 L)

Sucrose

30 g
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

2,4-D (0.2 mg/mL)

1

NAA (1 mg/mL)

2

Zeatin (1 mg/mL)

0.2
Adjust the pH of the solution to 5.8 with KOH (1 N), and fill up to 1 L with distilled water.
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

0.5

Pyridoxine·HCl

0.5

Thiamine·HCl

0.1

Glycine

2

myo-Inositol

100

Sucrose

30000

2,4-D sodium monohydrate

0.236

NAA·K

2.4

trans-Zeatin

0.2

Chemical	Amount
MS salt mix	1 bag (1 L)
Sucrose	30 g

Stock solution	Volume (mL)
MS_VT	1
MS_inositol	2.5
2,4-D (0.2 mg/mL)	1
NAA (1 mg/mL)	2
Zeatin (1 mg/mL)	0.2