Callus cells typically multiply under relatively low temperatures and light
intensity, but differentiation into sporophytes under relatively high temperatures and
light intensity was found in culturing results using leaf body tissue from kelp. This
was later found to be related to rising or falling melanosis within the cells (Fig. 6.3)
(Borowitzka et al. 2009).
Table 6.3 Preparation of sterile detection media for major seaweeds
STP
ST 3
ESS B1
Seawater
750 ml
700 ml
900 ml
Distilled water
200 ml
250 ml
70 ml
Undistilled ESS solution
a
–
–
10 ml
Soil extract
50 ml
50 ml
25 ml
Sodium nitrate (NaNO 3 )
200 mg
50 mg
–
Dipotassium phosphate (K 2 HPO 4 )
1 0m g
–
–
Na 2 -glycerol phosphate
–
10 mg
–
Bacto peptone
–
–
5 mg
Hy-case (Scheffielde Chemical)
200 mg
20 mg
Casein lysates
–
–
5 mg
Yeast rate (Difco)
200 mg
–
–
Yeast extract
–
10 mg
5 mg
Malt lysates
–
–
5 mg
Liver (Oxo L25, Oxo Ltd)
–
20 mg
–
Beef extract
–
–
5 mg
Vitamin B 12
–
0.1 µg
–
Vitamin mixture 8A
b
1 ml
1 ml
–
Sucrose
1 g
–
–
Glucose
2 g
–
–
Carbon source mixture II
c
–
20 ml
5 ml
Na H-glutamic acid
0.5 g
–
–
D, L-alanine
100 mg
–
–
Glycine
100 mg
–
–
Glycylglycine
–
400 mg
–
Agar
(4 g)
(4 g)
(10 g)
pH
7.5–7.6
7.9
7.8
a See Table 6.1
b
Preparation for vitamin mixture 8A (in 1 ml): 0.2 mg thiamine hydrochloride, 0.1 mg nicotinic
acid, 0.04 mg putresine-2HCl, 0.1 mg calcium pantothenate, 5 µg riboflavin, 0.04 mg
pyridoxine-2HCl, 0.02 mg pyridoxamine-2HCl, 0.01 mg p-aminobenzoic acid, 0.5 lg biotin,
0.5 mg choline citrate, 1 mg inositol, 0.8 mg thymine, 0.26 mg orotic acid, 0.05 lg vitamin B 12 ,
0.2 lg folinic acid, 2.5 lg folate
c 1 ml includes 1 mg glycine, 1 mg D, L-alanine, 1 mg L-asparagine, 2 mg sodium acetate, 2 mg
glucose, and 2 mg L-glutamic acid
6.3 Tissue Culture
153
intensity, but differentiation into sporophytes under relatively high temperatures and
light intensity was found in culturing results using leaf body tissue from kelp. This
was later found to be related to rising or falling melanosis within the cells (Fig. 6.3)
(Borowitzka et al. 2009).
Table 6.3 Preparation of sterile detection media for major seaweeds
STP
ST 3
ESS B1
Seawater
750 ml
700 ml
900 ml
Distilled water
200 ml
250 ml
70 ml
Undistilled ESS solution
a
–
–
10 ml
Soil extract
50 ml
50 ml
25 ml
Sodium nitrate (NaNO 3 )
200 mg
50 mg
–
Dipotassium phosphate (K 2 HPO 4 )
1 0m g
–
–
Na 2 -glycerol phosphate
–
10 mg
–
Bacto peptone
–
–
5 mg
Hy-case (Scheffielde Chemical)
200 mg
20 mg
Casein lysates
–
–
5 mg
Yeast rate (Difco)
200 mg
–
–
Yeast extract
–
10 mg
5 mg
Malt lysates
–
–
5 mg
Liver (Oxo L25, Oxo Ltd)
–
20 mg
–
Beef extract
–
–
5 mg
Vitamin B 12
–
0.1 µg
–
Vitamin mixture 8A
b
1 ml
1 ml
–
Sucrose
1 g
–
–
Glucose
2 g
–
–
Carbon source mixture II
c
–
20 ml
5 ml
Na H-glutamic acid
0.5 g
–
–
D, L-alanine
100 mg
–
–
Glycine
100 mg
–
–
Glycylglycine
–
400 mg
–
Agar
(4 g)
(4 g)
(10 g)
pH
7.5–7.6
7.9
7.8
a See Table 6.1
b
Preparation for vitamin mixture 8A (in 1 ml): 0.2 mg thiamine hydrochloride, 0.1 mg nicotinic
acid, 0.04 mg putresine-2HCl, 0.1 mg calcium pantothenate, 5 µg riboflavin, 0.04 mg
pyridoxine-2HCl, 0.02 mg pyridoxamine-2HCl, 0.01 mg p-aminobenzoic acid, 0.5 lg biotin,
0.5 mg choline citrate, 1 mg inositol, 0.8 mg thymine, 0.26 mg orotic acid, 0.05 lg vitamin B 12 ,
0.2 lg folinic acid, 2.5 lg folate
c 1 ml includes 1 mg glycine, 1 mg D, L-alanine, 1 mg L-asparagine, 2 mg sodium acetate, 2 mg
glucose, and 2 mg L-glutamic acid
6.3 Tissue Culture
153