1. Media for freshwater, terrestrial, hot spring and salt water algae

AAF–6 Acid–CSi/5 AF–6
AF–6/2 AF–6/3 AFAC
Allen BBM BG–11
C C+10% Seawater C/6
C/6G C/10 CA
CAM Carefoot CB
CB–V CC CSi
CSi + Cu CSi/5 CT
CYT DH + Fe DY–V
HUT M–11 MA
MAF–6 M–Allen M–Allen(+ U)
M–ASP7 MBM M Chu No. 10
MDM MG MGM
Modified acetate medium (mAC) Modified BG–110 Modified Johnson's medium
Modified M–1 (mM–1) MW MW/5
N–Free O P 35
Pro SOT SW
TAP Tre URO
URO–H URO + Soil URO–T
VT VTAC VTYT
W WC 9:1 agar medium

2. Media for marine and brackish water microalgae

BESM BESM 2 ESM ESM + CB–V ESM2 f/2 f/2 + NH4Cl
IMK IMK + NaCl IMK + Soil Extract K K/2 K/2ET MF
mIMR ( modified IMR ) MKM MNK Modified SWM–3 (mSWM–3) PRO–99 WESM  

3. Bacteria-free check media for freshwater algae

B–I B–II B–III B–IV B–V YT

4. Bacteria-free check media for marine algae

Bf/2 MM 23 STP

5. Trace metals, vitamin mixtures and soil extracts

A2 Fe stock solution A2 trace element stock solution A5 solution Allen metals
C–Source Mix II D stock medium D trace mix DY trace metal solution
Fe ( as EDTA; 1:1 molar ) Fe solution f/2 metals Hutner ’s trace elements
K metals K/2 metals mIMR trace metals mM–1 Trace elements
mTYGM–9 P–1 metals P II metals P IV metals
PN metals PRO–99 trace metals Soil extract Trace metal mix A5 + Co
Vitamin mix S3 Waris EDTA Waris P-2 WC trace metals

6. Media for protozoa

AF–6 + Wheat ESM + mTYGM–9 + Rice ESM + Rice
f/2 + mTYGM–9 + Rice f/2 + Rice f/2 + Wheat
Helio LE PYG
SUY SUY 1/10 SUY 1/10 + mTYGM–9 + Rice
SUY 1/10 + Wheat URO + Wheat URO–H + Wheat
UYTS + Rice YE100ppm 1/2Waris H + Si

7. Media for freshwater red algae

Bold 3N

8. Media for Charales

mSWC–2 ( Modified SWC–2 ) SWC–1 SWCN–1 SWCN–2 SWCN–3 SWCN–4 1/3 Herbst ASW

1 Media for freshwater, terrestrial, hot spring and salt water algae

AAF–6

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Prepare as for AF–6 medium but adjust to pH 5.5–5.8.

1 Media for freshwater, terrestrial, hot spring and salt water algae

Acid–CSi/5

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Dilute CSi 1 ) medium with distilled water to one–fifth. Adjust to pH 3 with sulfuric acid.
1 ) See CSi

1 Media for freshwater, terrestrial, hot spring and salt water algae

AF–6

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NaNO3 14 mg
NH4NO3 2.2 mg
MgSO4 · 7H2O 3 mg
KH2PO4 1 mg
K2HPO4 0.5 mg
CaCl2 · 2H2O 1 mg
CaCO3 1 ) 1 mg
Fe–citrate 0.2 mg
Citric acid 0.2 mg
Biotin 0.2 μg
Thiamine HCl 1 μg
Vitamin B6 0.1 μg
Vitamin B12 0.1 μg
Trace metals 1 ) 0.5 mL
Distilled water 99.5 mL
pH 6.6 2 )

1 ) In the NIES–Collection, CaCO3 is removed and PIV metals are used instead of Trace metals.
2 ) In the NIES–Collection, 40 mg MES is added and pH is adjusted to 6.6.

Reference(s)
Kato, S. 1982 Laboratory culture and morphology of Colacium vesiculosum Ehrb. ( Euglenophyceae ). Jpn. J. Phycol., 30, 63-67 ( in Japanese with English summary ).

1 Media for freshwater, terrestrial, hot spring and salt water algae

AF–6/2

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AF–6 1 ) medium is diluted with distilled water to one-half.
1 ) See AF–6

1 Media for freshwater, terrestrial, hot spring and salt water algae

AF–6/3

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Mix 1 volume of AF–6 1 ) medium and 2 volumes of distilled water.
1 ) See AF–6

1 Media for freshwater, terrestrial, hot spring and salt water algae

AFAC

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To 100 mL AF–6 1 ) medium add 20 mg sodium acetate.
1 ) See AF–6

1 Media for freshwater, terrestrial, hot spring and salt water algae

Allen

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( NH4 )2SO4 132 mg
KH2PO4 27.2 mg
MgSO4 · 7H2O 24.6 mg
CaCl2 · 2H2O 7.4 mg
Allen metals 1 ) 0.01 mL
Distilled water 99.9 mL
pH 2.5 2 )

1 ) See Allen metals
2 ) pH is adjusted to 2.5 with 0.5 mol / L H2SO4.

Reference(s)
Allen, M. B. 1959 Studies with Cyanidium caldarium, an anomalously pigmented chlorophyte. Arch. Mikrobiol., 32, 270-277.
Starr, R. C., Zeikus, J. A. 1987 UTEX - The culture collection of algae at the University of Texas at Austin. J. Phycol., 23, Suppl. to Sept ., 1-106.

1 Media for freshwater, terrestrial, hot spring and salt water algae

BBM

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NaNO3 25 mg
KH2PO4 17.5 mg
K2HPO4 10 mg
MgSO4 · 7H2O 7.5 mg
CaCl2 · 2H2O 2.5 mg
NaCl 2.5 mg
KOH 3.1 mg
FeSO4 · 7H2O 0.498 mg
H3BO3 1.142 mg
ZnSO4 · 7H2O 0.882 mg
MnCl2 · 7H2O 0.144 mg
MoO3 0.071 mg
CuSO4 · 5H2O 0.157 mg
Co( NO3 )2 · 6H2O 0.049 mg
Na2EDTA 5 mg
Distilled water 100 mL

Add 1.2 g agar to 100 mL of medium to give a solid medium.

Reference(s)
Bischoff, H. W., Bold, H. C. 1963 Some soil algae from enchanted rock and related algal species. Phycological Studies IV.,Univ. No. 6318, Texas, p. 95.

1 Media for freshwater, terrestrial, hot spring and salt water algae

BG–11

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NaNO3 150 mg
K2HPO4 · 3H2O 4 mg
MgSO4 · 7H2O 7.5 mg
CaCl2 · 2H2O 3.6 mg
Citric acid 0.6 mg
Ferric ammonium citrate 0.6 mg
Na2EDTA – Mg 0.1 mg
Na2CO3 2 mg
Trace metal mix A5 + Co 1 ) 0.1 mL
Distilled water 99.9 mL
pH 7.4

Add 1.5 g agar to 100 mL of medium to give a solid medium.
1 ) See Trace metal mix A5 + Co

Reference(s)
Waterbury, J. B., Sranier, R. Y. 1981 Isolation and growth of cyanobacteria from marine and hypersaline environments. In The Prokaryotes, Springer Berlin Heidelberg, New York, pp. 221-223.

1 Media for freshwater, terrestrial, hot spring and salt water algae

C

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Ca( NO3 )2 · 4H2O 15 mg
KNO3 10 mg
β–Na2glycerophosphate · 5H2O 5 mg
MgSO4 · 7H2O 4 mg
Vitamin B12 0.01 μg
Biotin 0.01 μg
Thiamine HCl 1 μg
PIV metals 1 ) 0.3 mL
Tris ( hydroxymethyl ) aminomethane 50 mg
Distilled water 99.7 mL
pH 7.5

Add 1.5 g agar to 100 mL of medium to give a solid medium.
1 ) See PIV metals

Reference(s)
Ichimura, T. 1971 Sexual cell division and conjugation-papilla formation in sexual reproduction of Closterium strigosum. In Proceedings of the Seventh International Seaweed Symposium, University of Tokyo Press, Tokyo, p. 208-214.


1 Media for freshwater, terrestrial, hot spring and salt water algae

C+10% Seawater ( N. Tezuka, unpubl. )

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C 1 ) medium with 10% filtered seawater.
1 ) See C

1 Media for freshwater, terrestrial, hot spring and salt water algae

C/6

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Mix 1 volume of C 1 ) medium and 5 volumes of distilled water.
1 ) See C

1 Media for freshwater, terrestrial, hot spring and salt water algae

C/6G

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Mix 1 volume of C 1 ) medium and 5 volumes of Lake Nojiri water ( sterilized through GF/F filter, and store at 5°C ).
1 ) See C

1 Media for freshwater, terrestrial, hot spring and salt water algae

C/10

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Mix 1 volume of C 1 ) medium and 9 volumes of distilled water.
1 ) See C

1 Media for freshwater, terrestrial, hot spring and salt water algae

CA

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Ca( NO3 )2 · 4H2O 2 mg
KNO3 10 mg
NH4NO3 5 mg
β–Na2glycerophosphate · 5H2O 3 mg
MgSO4 · 7H2O 2 mg
Vitamin B12 0.01 μg
Biotin 0.01 μg
Thiamine HCl 1 μg
PIV metals 1 ) 0.1 mL
Fe ( as EDTA; 1:1 molar ) 2 ) 0.1 mL
HEPES 40 mg
Distilled water 99.9 mL
pH 7.2

1 ) See PIV metals
2 ) See Fe ( as EDTA; 1:1 molar )

Reference(s)
Ichimura, T., Watanabe, M. 1974 The Closterium calosporum complex from the Ryukyu Islands - Variation and taxonomical problems. Mem. Natn. Sci. Mus. Tokyo, 7, 89-102, pls. 13-14.

1 Media for freshwater, terrestrial, hot spring and salt water algae

CAM

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CA 1 ) medium with pH adjusted to 6.5 by buffering with MES instead of HEPES.
1 ) See CA

1 Media for freshwater, terrestrial, hot spring and salt water algae

Carefoot

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NaNO3 24.7 mg
CaCl2 · 2H2O 1.1 mg
MgSO4 · 7H2O 4.7 mg
K2HPO4 0.9 mg
KH2PO4 2.3 mg
NaCl 1.5 mg
PIV metals 1 ) 0.5 mL
Distilled water 99.5 mL
pH 7.5

In the NIES–Collection, 0.02 μg vitamin B12, 0.02 μg biotin and 2 μg thiamine HCl are added to this medium.
1 ) See PIV metals

Reference(s)
Carefoot, J. R. 1968 Culture and heterotrophy of the freshwater dinoflagellate, Peridinium cinctum fa. ovoplanum Lindeman. J. Phycol., 4, 129-131.

1 Media for freshwater, terrestrial, hot spring and salt water algae

CB

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To C 1 ) medium add Bicine instead of Tris ( hydroxymethyl ) aminomethane, and adjust pH to 9.0.
1 ) See C

1 Media for freshwater, terrestrial, hot spring and salt water algae

CB–V

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Make B–V 1 ) medium with C 2 ) medium.
1 ) See B–V
2 ) See C

1 Media for freshwater, terrestrial, hot spring and salt water algae

CC

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C 1 ) medium with pH adjusted to 3.0 by buffering with 1,2,3,4–cyclopentane tetracarboxylic acid instead of Tris ( hydroxymethyl ) aminomethane.
1 ) See C

Reference(s)
Ichimura, T., Itoh, T. 1977 17. Preservation methods of microalgae ( I ) [17. Bisaisôrui no hozonhô ( I )]. In Preservation methods of microorganisms [ Biseibutsu Hozonhô], Ed. by Nei, T., University of Tokyo Press, Tokyo, p. 355-373 ( in Japanese without English title ).


1 Media for freshwater, terrestrial, hot spring and salt water algae

CSi

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C 1 ) medium with pH adjusted to 7.0 by buffering with 50mg HEPES instead of Tris ( hydroxymethyl ) aminomethane. Thereafter, 10 mg Na2SiO3 · 9H2O is added.
1 ) See C

1 Media for freshwater, terrestrial, hot spring and salt water algae

CSi + Cu

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To 100 mL CSi 1 ) medium add 0.25 mg CuSO4 · 5H2O and 100 mg agar.
1 ) See CSi

1 Media for freshwater, terrestrial, hot spring and salt water algae

CSi/5

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Dilute CSi 1 ) medium with distilled water to 1/5.
1 ) See CSi

1 Media for freshwater, terrestrial, hot spring and salt water algae

CT

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C 1 ) medium with pH adjusted to 8.2 by buffering with 40mg TAPS instead of Tris ( hydroxymethyl ) aminomethane.
1 ) See C

Reference(s)
Watanabe, M. M., Ichimura, T. 1977 Fresh- and salt-water forms of Spirulina platensis in axenic cultures. Bull. Jpn. Soc. Phycol., 25, Suppl. ( Mem. Iss. Yamada ) , 371-377.

1 Media for freshwater, terrestrial, hot spring and salt water algae

CYT

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To 100 mL C 1 ) medium add 100 mg yeast extract and 200 mg tryptone.
1 ) See C

1 Media for freshwater, terrestrial, hot spring and salt water algae

DH + Fe ( I.I. Brown, unpubl. )

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D stock medium 1 ) 5 mL
HEPES 0.12 mg
FeCl3 · 6H2O 1.14 mg
Distilled water 95 mL
pH 8.24 – 8.26

After autoclaving, keep in room temperature overnight. Next day, adjust pH to 7.5–7.6 and add 1.5 g agar.
1 ) See D stock medium

1 Media for freshwater, terrestrial, hot spring and salt water algae

DY–V

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MES 20 mg
MgSO4 · 7H2O 5 mg
KCl 0.3 mg
NH4Cl 0.27 mg
NaNO3 2 mg
β–Na2glycerophosphate · 5H2O 0.22 mg
H3BO3 0.08 mg
Na2EDTA · 2H2O 0.8 mg
Na2SiO3 · 9H2O 1.4 mg
FeCl3 · 6H2O 0.1 mg
CaCl2 · 2H2O 7.5 mg
Vitamin B12 0.05 μg
Biotin 0.05 μg
Thiamine HCl 10 μg
DY trace metal solution 1 ) 0.1 mL
Distilled water 99.9 mL
pH 6.8

1 ) See DY trace metal solution

1 Media for freshwater, terrestrial, hot spring and salt water algae

HUT

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KH2PO4 2 mg
MgSO4 · 7H2O 2.5 mg
Sodium acetate 40 mg
Potassium citrate 4 mg
Polypeptone 60 mg
Yeast extract 40 mg
Vitamin B12 0.05 μg
Thiamine HCl 0.04 mg
Distilled water 100 mL
pH 6.4

Add 150 mg agar to 100 mL of medium to give a semi-solid medium.

Reference(s)
Ichimura, T. 1979 2. Isolation and culture methods of algae. 2.5.B. Freshwater algae [2. Sôrui no bunri to baiyôhô. 2.5.B. Tansui sôrui]. In Methods in Phycological Studies [Sôrui Kenkyûhô], Eds. by Nishizawa, K. & Chihara, M., Kyoritsu Shuppan, Tokyo, p. 301 (in Japanese without English title).

1 Media for freshwater, terrestrial, hot spring and salt water algae

M–11

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NaNO3 10 mg
K2HPO4 1 mg
MgSO4 · 7H2O 7.5 mg
CaCl2 · 2H2O 4 mg
Na2CO3 3 mg
FeSO4 · 7H2O 0.1 mg
Na2EDTA · 2H2O 0.1 mg
Distilled water 100 mL
pH 8.0

Reference(s)
Hagiwara, T., Yagi, O., Takamura, Y., Sudo, R. 1984 Isolation of bacteria-free Microcystis aeruginosa from Lake Kasumigaura. Jpn. J. Water Poll. Res., 7, 437-442 ( in Japanese with English summary ).
Yagi, O., Okada, M., Sudo, R. 1979 Cultivation of Microcystis and red-tide-organisms. Res. Rep. Natl. Inst. Environ. Stud., No. 6, 223-229 ( in Japanese with English summary ).

1 Media for freshwater, terrestrial, hot spring and salt water algae

MA

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Ca( NO3 )2 · 4H2O 5 mg
KNO3 10 mg
NaNO3 5 mg
Na2SO4 4 mg
MgCl2 · 6H2O 5 mg
β–Na2glycerophosphate · 5H2O 10 mg
Na2EDTA · 2H2O 0.5 mg
FeCl3 · 6H2O 0.05 mg
MnCl2 · 4H2O 0.5 mg
ZnCl2 0.05 mg
CoCl2 · 6H2O 0.5 mg
Na2MoO4 · 2H2O 0.08 mg
H3BO3 2 mg
Bicine 50 mg
Distilled water 100 mL
pH 8.6

Reference(s)
Ichimura, T. 1979 2. Isolation and culture methods of algae. 2.5.B. Freshwater algae [2. Sôrui no bunri to baiyôhô. 2.5.B. Tansui sôrui]. In Methods in Phycological Studies [Sôrui Kenkyûhô], Eds. by Nishizawa, K. & Chihara, M., Kyoritsu Shuppan, Tokyo, p. 294-305 ( in Japanese without English title ).

1 Media for freshwater, terrestrial, hot spring and salt water algae

MAF–6

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To 100 mL of AF–6 1 ) medium add 10 mg glucose and 10 mg sodium acetate.
1 ) See AF–6

1 Media for freshwater, terrestrial, hot spring and salt water algae

M–Allen

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( NH4 )2SO4 262 mg
KH2PO4 54 mg
MgSO4 · 7H2O 50 mg
CaCl2 · 2H2O 14 mg
A2 trace element stock solution 1 ) 0.2 mL
Distilled water 99.4 mL
pH 2.5 2 )

After autoclaving, add 0.4 mL of A2 Fe stock solution 3 ) ( filter-sterilized ).
1 ) See A2 trace element stock solution
2 ) pH is adjusted to 2.5 with 0.5 mol / L H2SO4.
3 ) See A2 Fe stock solution
Indicated as “MA” medium in reference.

1 Media for freshwater, terrestrial, hot spring and salt water algae

M–Allen( + U )

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To 100 mL M–Allen 1 ) medium add 50 mg uracil.
1 ) See M–Allen


1 Media for freshwater, terrestrial, hot spring and salt water algae

M–ASP7

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NTA 7 mg
NaCl 2.5 g
MgSO4 · 7H2O 900 mg
KCl 70 mg
CaCl2 · 2H2O 30 mg
NaNO3 5 mg
NaH2PO4 · 2H2O 2 mg
Vitamin B12 0.1 μg
Vitamin mix S3 1 ) 1 mL
Na2SiO3 · 9H2O 1 mg
PN metals 2 ) 3 mL
Tris ( hydroxymethyl ) aminomethane 100 mg
Distilled water 96 mL
pH 8.0

1 ) See Vitamin mix S3
2 ) See PN metals

Reference(s)
Watanabe, M. M., Satake, K. N. ( Eds. ) 1991 NIES-Collection. List of Strains, Third Edition, 1991, Microalgae and Protozoa. Microbial Culture Collection, National Institute for Environmental Studies, Tsukuba, 163 pp.k.pdf

1 Media for freshwater, terrestrial, hot spring and salt water algae

MBM

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KNO3 25 mg
MgSO4 · 7H2O 7.5 mg
K2HPO4 7.5 mg
KH2PO4 17.5 mg
NaCl 2.5 mg
CaCl2 · 2H2O 1 mg
Fe solution 1 ) 0.1 mL
A5 solution 2 ) 0.1 mL
Agar 1.5 g
Distilled water 99.8 mL
pH 6.0

1 ) See Fe solution.
2 ) See A5 solution .
Indicated as “Modified Bristol medium” in reference.

Reference(s)
Ichimura, T., Itoh, T. 1977 17. Preservation methods of microalgae ( I ) [17. Bisaisôrui no hozonhô ( I )]. In Preservation methods of microorganisms [Biseibutsu Hozonhô], Ed. by Nei, T., University of Tokyo Press, Tokyo, p. 355-373 ( in Japanese without English title ).

1 Media for freshwater, terrestrial, hot spring and salt water algae

M Chu No. 10

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Ca( NO3 )2 · 4H2O 2.0 mg
KH2PO4 0.62 mg
MgSO4 · 7H2O 2.5 mg
Na2CO3 2 mg
Na2SiO3 · 9H2O 2.5 mg
HCl ( 1 mol / L ) 1 ) 0.025 mL
Na2EDTA · 2H2O 0.2 mg
FeCl3 · 6H2O 0.1 mg
H3BO3 0.248 mg
MnCl2 · 4H2O 0.139 mg
( NH4 )6Mo7O24 · 4H2O 0.1 mg
Vitamin B12 1 μg
Thiamine HCl 0.1 μg
Biotin 0.1 μg
Distilled water 100 mL

1 ) In the NIES–Collection, pH is adjusted to 7.6 with 1 mol / L HCl.

Reference(s)
Chu, S. P. 1942 The influence of the mineral composition of the medium on the growth of planktonic algae. Part I. Methods and culture media. J. Ecol., 30, 284-325.

1 Media for freshwater, terrestrial, hot spring and salt water algae

MDM

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KNO3 100 mg
MgSO4 · 7H2O 25 mg
K2HPO4 25 mg
NaCl 10 mg
CaCl2 · 2H2O 1 mg
Fe solution 1 ) 0.1 mL
A5 solution 2 ) 0.1 mL
Agar 1.5 g
Distilled water 99.8 mL
pH 8.0

1 ) See Fe solution
2 ) See A5 solution

Reference(s)
Watanabe, A. 1960 List of algal strains in collection at the Institute of Applied Micro-biology, University of Tokyo. J. Gen. Appl. Microbiol., 6, 283-292.

1 Media for freshwater, terrestrial, hot spring and salt water algae

MG

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Ca ( NO3 )2·4H2O 2 mg
KNO3 10 mg
β–Na2 glycerophosphate·5H2O 3 mg
MgSO4 · 7H2O 2 mg
Vitamin B12 0.01 µg
Biotin 0.01 µg
Thiamine HCl 1 µg
PIV metals 1 ) 0.1  mL
Fe ( as EDTA; 1:1 molar ) 2 ) 0.1  mL
HEPES 40 mg
Distilled water 99.8  mL
pH 7.2

1 ) See PIV metals
2 ) See Fe ( as EDTA; 1:1 molar )

Reference(s)
Ichimura, T. 1973 The life cycle and its control in some species of Closterium, with special reference to the biological species problems. Thesis D. Sci., University of Tokyo, 69 pp., 11 tables, 40 figs.

1 Media for freshwater, terrestrial, hot spring and salt water algae

MGM

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MG 1 ) medium with pH adjusted to 6.5 by buffering with MES instead of HEPES.
1 ) See MG

1 Media for freshwater, terrestrial, hot spring and salt water algae

Modified acetate medium ( mAC )

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To 100 mL AF–6 1 ) medium, add 40mg glucose, yeast extract, tryptone, and sodium acetate.
1 ) See AF–6

Reference(s)
Nozaki, H., Wanababe, M. M. & Aizawa, K. 1995 Morphology and paedogamous sexual reproduction in Chlorogonium capillatum sp. nov. ( Volvocales, Chlorophyta ). J. Phycol., 31, 655-663.

1 Media for freshwater, terrestrial, hot spring and salt water algae

Modified BG–110

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1M HEPES–NaOH 1 ) 2 mL
K2HPO4 · 3H2O 4 mg
MgSO4 · 7H2O 7.5 mg
CaCl2 · 2H2O 3.6 mg
Citric acid 0.6 mg
Ferric ammonium citrate 0.6 mg
Na2EDTA – Mg 0.1 mg
Na2CO3 2 mg
Trace metal mix A5 + Co 2 ) 0.1 mL
Vitamin B12 0.1 µg
Biotin 0.1 µg
Thiamine HCl 200µg
Agar 1.2 g
Distilled water 97.9 mL

1 ) To 60 mL of Distilled water add 23.8 g HEPES, and pH is adjusted to 7.5. Bring to 100 mL with distilled water.v
2 ) See Trace metal mix A5 + Co

Reference(s)
Castenholz, R. W. 1988 Cultureing methods for cyanobacteria. Methods Enzymol., 167, 68-93.

1 Media for freshwater, terrestrial, hot spring and salt water algae

Modified Johnson's medium

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NaCl 3 mg
MgCl2 · 6H2O 150 mg
KNO3 100 mg
MgSO4 · 7H2O 50 mg
CaCl2 · 2H2O 1) 26.5 mg
KCl 20 mg
NaHCO3 4.3 mg
KH2PO4 3.5 mg
Tris ( hydroxymethyl ) aminomethane 245 mg
Distilled water 100 mL
pH 8.5

1 ) In the NIES-Collection, 20 mg CaCl2 is replaced by 26.5 mg CaCl2 · 2H2O.

Reference(s)
Johnson, M. K., Johnson, E. J., MacElroy, R. D., Speer, H. L., Bruff, B. S. 1968 Effects of salts on the halophilic alga Dunaliella viridis. J. Bacteriol., 95, 1461-1468.
Utsunomiya, A., Watanuki, T., Matsushita, K., Tomita, I. 1997 Toxic effects of linear alkylbenzene sulfonate, quaternaryalkylammonium chloride and their complexes on Dunaliellasp. and Chlorella pyrenoidosa. Environ. Toxicol. Chem., 16, 1247-1254.
Mihirogi, M., Kikuchi, M., Sawai, J. 2012 Development of screening algal growth inhibition test with Dunaliella sp. Jpn. J. Environ. Toxicol., 15, 11-16 (in Japanese with English summary).

1 Media for freshwater, terrestrial, hot spring and salt water algae

Modified M–1 ( mM–1 )

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CaCl2 · 2H2O 0.5 mg
NaNO3 2.5 mg
NH4Cl 0.5 mg
CaSO4 · 2H2O 0.4 mg
MgSO4 · 7H2O 0.5 mg
Na2SiO3 · 9H2O 0.2 mg
Fe ( as EDTA; 1:1 molar ) 1 ) 25 mL
mM–1 Trace elements 2 ) 0.1 mL
K2HPO4 6.96 mg
KH2PO4 266.5 mg
Distilled water 74.9 mL
pH 5.1 – 5.3

1 ) See Fe ( as EDTA; 1:1 molar )
2 ) See mM–1 Trace elements

Reference(s)
Hoham, R. W., Berman, J. D., Rogers, H. S., Felio, J. H., Ryba, J. B., Miller, P. R. 2006 Two new species of green snow algae from Upstate New York, Chloromonas chenangoensis sp. nov. and Chloromonas tughillensis sp. nov. ( Volvocales, Chlorophyceae ) and the effects of light on their life cycle development. Phycologia, 45, 319-330.

1 Media for freshwater, terrestrial, hot spring and salt water algae

MW

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Urea 0.85 mg
NaNO3 0.17 mg
NH4Cl 0.042 mg
Ca( NO3 )2 · 4H2O 10 mg
CaCO3 1 mg
CaCl2 · 2H2O 1.4 mg
KNO3 1 mg
KHCO3 0.9 mg
β–Na2glycerophosphate · 5H2O 2 mg
MgSO4 · 7H2O 1.5 mg
PIV metals 1 ) 0.05 mL
Vitamin B12 0.02 μg
Thiamine HCl 2 μg
Biotin 0.02 μg
Glycylglycine 10 mg
Distilled water 100 mL
pH 7.2

1 ) See PIV metals

Reference(s)
Sako, Y., Ishida, Y., Kadota, H., Hata, Y. 1984 Sexual reproduction and cyst formation in the freshwater dinoflagellate Peridinium cunningtonii. Bull. Jpn. Soc. Sci. Fish., 50, 743-750.

1 Media for freshwater, terrestrial, hot spring and salt water algae

MW/5

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MW 1 ) medium is diluted with distilled water to 1/5.
1 ) See MW

1 Media for freshwater, terrestrial, hot spring and salt water algae

N–Free

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NaCl 7 mg
MgSO4 · 7H2O 38 mg
CaCl2 · 2H2O 10.6 mg
K2HPO4 60 mg
Fe2( SO4 )3 · 6H2O 1 mg
Na2EDTA · 2H2O 2.7 mg
H3BO3 0.3 mg
MnSO4 · 4H2O 1 ) 0.2 mg
Na2MoO4 · 2H2O 0.8 mg
ZnSO4 · 7H2O 0.03 mg
CuSO4 · 5H2O 8 μg
CoCl2 · 6H2O 3.7 μg
Agar 1.5 g
Distilled water 100 mL
pH 7.5

1 ) In the NIES–Colletion, 0.2 mg MnSO4 · 4H2O is replaced by 0.22 mg MnSO4 · 5H2O.


1 Media for freshwater, terrestrial, hot spring and salt water algae

O

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Glucose 100 mg
Tryptone 100 mg
Yeast extract 100 mg
Beef extract 1 ) 50 mg
Agar 150 mg
Distilled water 100 ml

1 ) In the NIES-Collection, beef extract is removed.
Indicated as “Ochomonas medium” in reference.

Reference(s)
Ichimura, T., Itoh, T. 1977 17. Preservation methods of microalgae ( I ) [17. Bisaisôrui no hozonhô ( I )]. In Preservation methods of microorganisms [Biseibutsu Hozonhô], Ed. by Nei, T., University of Tokyo Press, Tokyo, p. 355-373 ( in Japanese without English title ).
Starr, R. C. 1964 The culture collection of algae at Indiana University. Amer. J. Bot., 51, 1013-1044.

1 Media for freshwater, terrestrial, hot spring and salt water algae

P 35

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NH4NO3 10 mg
MgSO4 · 7H2O 4 mg
KCl 5 mg
CaCl2 · 2H2O 7.4 mg
β–Na2glycerophosphate · 5H2O 5 mg
Sodium acetate 100 mg
Vitamin B12 0.01 μg
Biotin 0.01 μg
Thiamine HCl 1 μg
PIV metals 1 ) 0.3 mL
Tris ( hydroxymethyl ) aminomethane 50 mg
Distilled water 99.7 mL
pH 8.0

1 ) See PIV metals

Reference(s)
Ichimura, T. 1979 2. Isolation and culture methods of algae. 2.5.B. Freshwater algae [2. Sôrui no bunri to baiyôhô. 2.5.B. Tansui sôrui]. In Methods in Phycological Studies [Sôrui Kenkyûhô ], Eds. by Nishizawa, K. & Chihara, M., Kyoritsu Shuppan, Tokyo, p. 294-305 ( in Japanese without English title ).

1 Media for freshwater, terrestrial, hot spring and salt water algae

Pro

Print

To 100 mL MBM 1 ) medium add 100 mg proteose peptone.
1 ) See MBM
Indicated as “Proteose medium” in reference.

Reference(s)
Ichimura, T., Itoh, T. 1977 17. Preservation methods of microalgae ( I ) [17. Bisaisôrui no hozonhô ( I )]. In Preservation methods of microorganisms [Biseibutsu Hozonhô], Ed. by Nei, T., University of Tokyo Press, Tokyo, p. 355-373 ( in Japanese without English title ).
Starr, R. C. 1964 The culture collection of algae at Indiana University. Amer. J. Bot., 51, 1013-1044.

1 Media for freshwater, terrestrial, hot spring and salt water algae

SOT

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NaHCO3 1.68 g
K2HPO4 50 mg
NaNO3 250 mg
K2SO4 100 mg
NaCl 100 mg
MgSO4 · 7H2O 20 mg
CaCl2 · 2H2O 4 mg
FeSO4 · 7H2O 1 mg
Na2EDTA · 2H2O 8 mg
A5 solution 1 ) 0.1 mL
Distilled water 99.9 mL

1 ) See A5 solution

Reference(s)
Ogawa, T., Terui, G. 1970 Studies on the growth of Spirulina platensis. ( I ) On the pure culture of Spirulina platensis. J. Ferment. Technol., 48, 361-367.

1 Media for freshwater, terrestrial, hot spring and salt water algae

SW

Print

Put a small amount of dried soil into a test tube and add 20mL distilled water.

Reference(s)
Pringsheim, E. G. 1946 The biphasic or soil-water culture method for growing algae and flagellata. J. Ecol., 33, 193-204.

1 Media for freshwater, terrestrial, hot spring and salt water algae

TAP

Print
NH4Cl 40 mg
CaCl2 · 2H2O 5.1 mg
MgSO4 · 7H2O 10 mg
K2HPO4 11.9 mg
KH2PO4 6.03 mg
Hutner ’s trace elements 1 ) 0.1 mL
Acetic acid 0.1 mL
Tris ( hydroxymethyl ) aminomethane 242 mg
Agar 1.5 g
Distilled water 99.8 mL

1 ) See Hutner ’s trace elements

1 Media for freshwater, terrestrial, hot spring and salt water algae

Tre

Print

To 100 mL MBM 1 ) medium add 1 g proteose peptone and 2 g glucose.
1 ) See MBM
Indicated as “Trebouxia medium” in reference.

Reference(s)
Ichimura, T., Itoh, T. 1977 17. Preservation methods of microalgae ( I ) [17. Bisaisôrui no hozonhô ( I )]. In Preservation methods of microorganisms [Biseibutsu Hozonhô], Ed. by Nei, T., University of Tokyo Press, Tokyo, p. 355-373 ( in Japanese without English title ).
Starr, R. C. 1964 The culture collection of algae at Indiana University. Amer. J. Bot., 51, 1013-1044.

1 Media for freshwater, terrestrial, hot spring and salt water algae

URO

Print
NH4NO3 0.5 mg
β–Na2glycerophosphate · 5H2O 0.4 mg
MgSO4 · 7H2O 1 mg
CaCl2 · 2H2O 1 mg
KCl 0.1 mg
Thiamine HCl 1 μg
Vitamin B12 0.01 μg
Biotin 0.01 μg
Fe-EDTA 0.05 mg
PIV metals 1 ) 0.1 mL
Distilled water 99.9 mL
pH 7.5 2 )

1 ) See PIV metals
2 ) pH is adjusted to 7.5 with 0.1 mol / L HCl.

Reference(s)
Kimura, B., Ishida, Y. 1985 Photophagotrophy in Uroglena americana, Chrysophyceae. Jpn. J. Limnol., 46, 315-318.
Nakahara, H., Sako, Y. 1987 2. Life history of freshwater phytoplankton [2. Tansui syokubutsu purankuton no seikatsushi]. In Freshwater red tide [Tansui Akashio ], Ed. by Kadota, H., Kôseisya-Kôseikaku, Tokyo, p. 21-77 ( in Japanese without English title ).

1 Media for freshwater, terrestrial, hot spring and salt water algae

URO–H

Print

To 100 mL URO 1 ) medium add 40 mg HEPES.
1 ) See URO

1 Media for freshwater, terrestrial, hot spring and salt water algae

URO + Soil

Print

Prepare as for 100 mL URO 1 ) medium with 89.9 mL instead of 99.9 mL distilled water, add 10 mL Soil extract 2 ) and adjust pH 7.5.
1 ) See URO
2 ) See Soil extract

1 Media for freshwater, terrestrial, hot spring and salt water algae

URO–T

Print

To 100 mL URO 1 ) medium add 50 mg Tris ( hydroxymethyl ) aminomethane.
1 ) See URO

1 Media for freshwater, terrestrial, hot spring and salt water algae

VT

Print
Ca( NO3 )2 · 4H2O 11.78 mg
β-Na2glycerophosphate · 5H2O 5 mg
MgSO4 · 7H2O 4 mg
KCl 5 mg
Vitamin B12 0.01 μg
Biotin 0.01 μg
Thiamine HCl 1 μg
PIV metals 1 ) 0.3 mL
Glycylglycine 50 mg
Distilled water 99.7 mL
pH 7.5

1 ) See PIV metals

Reference(s)
Provasoli, L., Pintner, I. J. 1959 Artificial media for fresh-water algae: problems and suggestions. In The Ecology of Algae. Spec. Pub. No. 2,, Eds. by Tryon, C. A., Jr. & Hartmann, R. T., Pymatuning Laboratory of Field Biology, University of Pittsburgh, Pittsburgh, p. 84-96.
Starr, R. C. 1973 Special methods-dry soil samples. In Handbook of Phycological Methods. Culture Methods and Growth Measurements, Ed. by Stein, J. R., Cambridge University Press, Cambridge, p. 159-167.

1 Media for freshwater, terrestrial, hot spring and salt water algae

VTAC

Print

To 100 mL VT 1 ) medium add 20 mg sodium acetate.
1 ) See VT

Reference(s)
Nozaki, H., Kuroiwa, H., Mita, T., Kuroiwa, T. 1989 Pleodorina japonica sp. nov. ( Volvocales, Chlorophyta ) with bacteria-like endosymbionts. Phycologia, 28, 252-267.

1 Media for freshwater, terrestrial, hot spring and salt water algae

VTYT

Print

To 100 mL VT 1 ) medium add 10 mg yeast extract and 20 mg tryptone.
1 ) See VT

Reference(s)
Ichimura, T., Itoh, T. 1977 17. Preservation methods of microalgae ( I ) [17. Bisaisôrui no hozonhô ( I ) ]. In Preservation methods of microorganisms [Biseibutsu Hozonhô], Ed. by Nei, T., University of Tokyo Press, Tokyo, p. 355-373 ( in Japanese without English title ).

1 Media for freshwater, terrestrial, hot spring and salt water algae

W

Print
Ca( NO3 )2 · 4H2O 10 mg
KNO3 1 mg
MgSO4 · 7H2O 1.5 mg
β–Na2glycerophosphate · 5H2O 2 mg
Urea 1.7 mg
Thiamine HCl 0.2 μg
Vitamin B12 0.002 μg
Biotin 0.002 μg
PIV metals 1 ) 0.05 mL
Glycylglycine 10 mg
Distilled water 99.95 mL
pH 7.5

1 ) See PIV metals

Reference(s)
Watanabe, M. M. 1983 Growth characteristics of freshwater red tide alga, Peridinium based on axenic culture. Establishment of synthetic culture medium [Junsuibaiyôhô niyoru tansuiakashio Peridinium no zôsyokutokusei no kaiseki-gôseibaichi no kakuritsu ]. Res. Data Natl. Inst. Environ. Stud., No. 24, 111-121 ( in Japanese without English title ).

1 Media for freshwater, terrestrial, hot spring and salt water algae

WC

print
NaNO3 8.5 mg
CaCl2 · 2H2O 3.67 mg
MgSO4 · 7H2O 3.69 mg
NaHCO3 1.26 mg
Na2SiO3 · 9H2O 2.84 mg
K2HPO4 0.87 mg
WC trace metals 1 ) 0.1 mL
Vitamin B12 0.05 μg
Biotin 0.05 μg
Thiamine HCl 10 μg
Distilled water 99.9 mL
pH 7.5

1 ) See WC trace metals

Reference(s)
Guillard, R. R. L., Lorenzen, C. J. 1972 Yellow-green algae with chlorohyllide C. J. Phycol., 8, 10-14.
Andersen, R. A. E, Berges, J. A., Harrison, P. J., Watanabe, M. M. 2005 Appendix A―Recipes for freshwater and seawater media. In Algal culturing techniques, Ed. by Andersen, R. A. E, Elsevier Academic Press, p. 474.

1 Media for freshwater, terrestrial, hot spring and salt water algae

9:1 agar medium

Print
NaNO3 25 mg
KH2PO4 17.5 mg
K2HPO4 10 mg
MgSO4 · 7H2O 7.5 mg
CaCl2 · 2H2O 2.5 mg
NaCl 2.5 mg
KOH 3.1 mg
Na2EDTA· 2H2O 5 mg
FeSO4 · 7H2O 0.498 mg
H3BO3 1.142 mg
ZnSO4 · 7H2O 0.882 mg
MnCl2 · 7H2O 0.144 mg
MoO3 0.071 mg
CuSO4 · 5H2O 0.157 mg
Co( NO3 )2 · 6H2O 0.049 mg
Vitamin B12 0.01 μg
Biotin 0.01 μg
Thiamine HCl 1 μg
Soil extract 1 ) 10 mL
Distilled water 100 mL

Add 1.5 g agar to 100 mL of medium to give a solid medium.
1 ) See Soil extract

Reference(s)
Bischoff, H. W., Bold, H. C. 1963 Some soil algae from enchanted rock and related algal species. Phycological Studies IV.,Univ. No. 6318, Texas, p. 95.
Watanabe, S., Fučíková, K., Lewis, L. A., Lewis P.O. 2016 Hiding in plain sight: Koshicola spirodelophila gen. et sp. nov. (Chaetopeltidales, Chlorophyceae), a novel green alga associated with the aquatic angiosperm Spirodela polyrhiza. Am. J. Bot., 103, 865-875.

2 Media for marine and brackish water microalgae

BESM

Print

Make diluted seawater by mixing 27.5 mL seawater and 70 mL distilled water. Make ESM 1 ) medium by using this diluted seawater instead of original seawater.
1 ) See ESM

2 Media for marine and brackish water microalgae

BESM 2

Print

Make diluted seawater by mixing 47.5 mL seawater and 50 mL distilled water. Make ESM 1 ) medium by using this diluted seawater instead of original seawater.
1 ) See ESM


2 Media for marine and brackish water microalgae

ESM*

Print
NaNO3 12 mg
K2HPO4 0.5 mg
Vitamin B12 0.1 μg
Biotin 0.1 μg
Thiamine HCl 10 μg
Fe – EDTA 25.9 μg
Mn – EDTA 33.2 μg
Tris ( hydroxymethyl ) aminomethane 100 mg
Soil extract 1 ) 2.5 mL
Seawater 97.5 mL
pH 8.0

Add 1.5 g agar to 100 mL of medium to give a solid medium.
1 ) The amount of Soil extract depends on the quality of the soil. In the NIES–Collection, Soil extract was reduced from 5 mL to 2.5 mL after 2002. See Soil extract.

Reference(s)
Okaichi, T., Nishio, S., Imatomi, Y. 1982 Collection and mass culture [Shiryô no saisyu to baiyô]. In Toxic phytoplankton - Occurrence, mode of action, and toxins [Yûdoku Purankuton -Hassei, Sayôkikô, Dokuseibun ], Ed. by Jpn. Fish. Soc., Kôseisya-Kôseikaku, Tokyo, p. 22-34 ( in Japanese without English title ).

2 Media for marine and brackish water microalgae

ESM + CB–V

Print

Mix 100 mL of ESM 1 ) medium and 0.25 mL of CB–V 2 ) medium.

1 ) See ESM
2 ) See CB–V

2 Media for marine and brackish water microalgae

ESM2

Print

Prepare as for 100 mL ESM 1 ) medium with 95.5mL instead of 97.5 mL seawater and with 5 mL instead of 2.5 mL Soil extract 2 ).
1 ) See ESM
2 ) See Soil extract

2 Media for marine and brackish water microalgae

f/2

Print
NaNO3 7.5 mg
NaH2PO4 · 2H2O 0.6 mg
Vitamin B12 0.05 μg
Biotin 0.05 μg
Thiamine HCl 10 μg
Na2SiO3 · 9H2O 1 mg
f/2 metals 1 ) 0.1 mL
Seawater 99.9 mL

1 ) See f/2 metals

Reference(s)
Guillard, R. R. L., Ryther, J. H. 1962 Studies of marine planktonic diatoms. I. Cyclotella nana Hustedt, and Detonula confervacea ( Cleve ) Gran. Can. J. Microbiol., 8, 229-239.

2 Media for marine and brackish water microalgae

f/2 + NH4Cl

Print

To 100 mL f/2 1 ) medium add 2.67 mg NH4Cl.
1 ) See f/2

2 Media for marine and brackish water microalgae

IMK

Print

Into 100 mL seawater dissolve 25.2 mg powder medium of Daigo IMK ( Nihon Pharmaceutical Co., Ltd. ). In the NIES–Collection, IMK medium is used after autoclaving ( 121°C, 20 min ).

2 Media for marine and brackish water microalgae

IMK + NaCl

Print

Into 100 mL seawater dissolve 25.2 mg powder medium of Daigo IMK ( Nihon Pharmaceutical Co., Ltd. ). Adjust to salinity 10 percent with NaCl. In the NIES-Collection, IMK + NaCl medium is used after autoclaving ( 121°C, 20 min ).

2 Media for marine and brackish water microalgae

IMK + Soil Extract

Print

Into 100 mL seawater dissolve 25.2 mg powder medium of Daigo IMK ( Nihon Pharmaceutical Co., Ltd. ). Thereafter, 2 mL Soil extract 1 ) is added. In the NIES–Collection, IMK + Soil Extract medium is used after autoclaving ( 121°C, 20 min ).
1 ) See Soil extract

2 Media for marine and brackish water microalgae

K

Print
Tris-base (  pH 7.2) 1 ) 0.1 mL
NaNO3 7.5 mg
NH4Cl 0.267 mg
β–Na2glycerophosphate · 5H2O 0.216 mg
H2SeO3 0.129 μg
Vitamin B12 0.05 μg
Biotin 0.05 μg
Thiamine HCl 0.01 mg
Na2SiO3 · 9H2O2 ) 1.535 mg
K metals 3 ) 0.1 mL
Seawater 99.8 mL

1 ) To 100 mL of Distilled water add 12.11 g Tris-base, and pH is adjusted to 7.2.
2 ) In the NIES-Collection, Na2SiO3 · 9H2O is removed.
3 ) See K metals

Reference(s)
Keller, M. D., Guillard, R.R.L. 1985 Factors significant to marine diatom culture. pp. 113-6. In Anderson, D.M., White, A.W., Baden, D.G. (eds.) Toxic Dinoflagellates. Elsevier, New York.
Keller, M.D., Selvin, R.C., Claus, W., Guillard, R.R.L. 1987. Media for the culture of oceanic ultraphytoplankton. J. Phycol., 23: 633-638.

2 Media for marine and brackish water microalgae

K/2 (K/2-modified by Ian Probert)*

Print
NaNO3 2.44 mg
NH4Cl 0.0267 mg
KH2PO4 0.244 mg
Fe-EDTA 0.215 mg
Vitamin B12 0.05 μg
Biotin 0.0005 μg
Thiamine HCl 0.01 mg
K/2 metals 1 ) 0.05 mL
Seawater 99.95 mL
pH 8.2

K medium is diluted half strength and modified by Ian. This medium should not be autoclaved but filter-sterilized.
1 ) See K/2 metals

Reference(s)
Keller, M. D., Guillard, R.R.L. 1985 Factors significant to marine diatom culture. pp. 113-6. In Anderson, D.M., White, A.W., Baden, D.G. (eds.) Toxic Dinoflagellates. Elsevier, New York.
Keller, M.D., Selvin, R.C., Claus, W., Guillard, R.R.L. 1987. Media for the culture of oceanic ultraphytoplankton. J. Phycol., 23: 633-638.

2 Media for marine and brackish water microalgae

K/2ET

Print
NaNO3 2.44 mg
NH4Cl 0.0267 mg
KH2PO4 0.244 mg
Fe-EDTA 0.215 mg
Vitamin B12 0.05 μg
Biotin 0.0005 μg
Thiamine HCl 0.01 mg
K/2 metals 1 ) 0.05 mL
Soil extract 2 ) 1.5 mL
Seawater 98.45 mL
pH 8.2

K medium is diluted half strength and modified by Ian. This medium should not be autoclaved but filter-sterilized.
1 ) See K/2 metals
2 ) See Soil extract

Reference(s)
Keller, M. D., Guillard, R.R.L. 1985 Factors significant to marine diatom culture. pp. 113-6. In Anderson, D.M., White, A.W., Baden, D.G. (eds.) Toxic Dinoflagellates. Elsevier, New York.
Keller, M.D., Selvin, R.C., Claus, W., Guillard, R.R.L. 1987. Media for the culture of oceanic ultraphytoplankton. J. Phycol., 23: 633-638.

2 Media for marine and brackish water microalgae

MF

Print

f/2 1 ) medium with Na2SiO3 · 9H2O replaced by 1 mL Soil extract 2 ) and adjusted to pH 8.0 by buffering with 100 mg Tris ( hydroxymethyl ) aminomethane.
1 ) See f/2
2 ) See Soil extract

2 Media for marine and brackish water microalgae

mIMR ( modified IMR )

Print
KNO3 1.26 mg
K2HPO4 0.22 mg
Na2SiO3 1 ) 0.61 mg
mIMR trace metals 2 ) 0.1 mL
Thiamine 20 μg
Vitamin B12 1 μg
Biotin 0.1 μg
Seawater 100 mL

1 ) In the NIES–Collection, Na2SiO3 is replaced by Na2SiO3·9H2O.
2 ) See mIMR trace metals

Reference(s)
Paasche, E., Bruback, S., Skattebol, S., Young, J. R., Green, J. C 1996 Growth and calcification in the coccolithophorid Emiliania huxleyi ( Haptophyceae ) at low salinities. Phycologia, 35, 394-403.

2 Media for marine and brackish water microalgae

MKM

Print
KNO3 75 mg
KH2PO4 2.5 mg
MgSO4 · 7H2O 2 mg
Fe–citrate 250 μg
Agar 1.5 g
Seawater 50 mL
Distilled water 50 mL

Reference(s)
Watanabe, A. 1960 List of algal strains in collection at the Institute of Applied Micro-biology, University of Tokyo. J. Gen. Appl. Microbiol., 6, 283-292.

2 Media for marine and brackish water microalgae

MNK

Print
NaNO3 2 mg
K2HPO4 0.1 mg
Na2HPO4 · 12H2O 0.028 mg
Vitamin B12 0.015 μg
Biotin 0.015 μg
Thiamine HCl 2 μg
CoSO4 · 7H2O 0.12 μg
ZnSO4 · 7H2O 0.24 μg
MnCl2 · 4H2O 0.9 μg
CuSO4 · 5H2O 0.006 μg
Na2SeO3 0.003 μg
Na2MoO4 · 2H2O 0.07 μg
Na2EDTA · 2H2O 0.37 μg
Fe–EDTA 2.6 μg
Mn–EDTA 3.3 μg
Seawater 100 mL

Vitamins should be added at the end of the preparation. This medium should not be autoclaved but filter–sterilized.

Reference(s)
Noël, M.-H., Kawachi, M., Inouye, I. 2004 Induced dimorphic life cycle of a coccolithophorid, Calyptrosphaera sphaeroidea ( Prymnesiophyceae, Haptophyta ). J. Phycol., 40, 112-129.

2 Media for marine and brackish water microalgae

Modified SWM–3 ( mSWM–3 )

Print
NaNO3 17 mg
NaH2PO4· 2H2O 1.56 mg
Na2SiO3· 9H2O 5.68 mg
Na2EDTA· 2H2O 1.12 mg
Fe-EDTA 0.084 mg
Na2SeO3 0.0346 mg
P–1 metals 1 mL
Vitamin B12 0.2 μg
Vitamin mix S3 0.2 mL
Tris ( hydroxymethyl ) aminomethane 50 mg
Seawater 98.8 mL
pH 7.7 – 7.8

1 ) See P–1 metals
2 ) See Vitamin mix S3

Reference(s)
Imai, I. 2012 High growth rates of the red tide flagellate Chattonella marina (Raphidophyceae) observed in culture. Hokudai Suisan Ihou, 62, 71-74 ( in Japanese with English summary ).

2 Media for marine and brackish water microalgae

PRO–99

Print
NaH2PO4 · 2H2O 0.779 mg
NH4Cl 4.28 mg
PRO–99 trace metals 1 ) 0.01 mL
Seawater 100 mL

1 ) See PRO–99 trace metals

2 Media for marine and brackish water microalgae

WESM

Print

Make diluted seawater by mixing 87.5 mL seawater and 10mL distilled water. Make ESM 1 ) medium by using this diluted seawater instead of seawater.
1 ) See ESM

3 Bacteria-free check media for freshwater algae

B–I

Print
Appropriate medium 100 mL
Proteose peptone 100 mg

Reference(s)
Ichimura, T., Watanabe, M. M. 1977 An axenic clone of Microcystis aeruginosa Kutz. emend. Elenkin from Lake Kasumigaura. Bull. Jpn. Soc. Phycol., 25, 177-181.

3 Bacteria-free check media for freshwater algae

B–II

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Appropriate medium 100 mL
Yeast extract 500 mg

Reference(s)
Ichimura, T., Watanabe, M. M. 1977 An axenic clone of Microcystis aeruginosa Kutz. emend. Elenkin from Lake Kasumigaura. Bull. Jpn. Soc. Phycol., 25, 177-181.

3 Bacteria-free check media for freshwater algae

B–III

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Appropriate medium 100 mL
Peptone 500 mg
Beef extract 300 mg

Reference(s)
Ichimura, T., Watanabe, M. M. 1977 An axenic clone of Microcystis aeruginosa Kutz. emend. Elenkin from Lake Kasumigaura. Bull. Jpn. Soc. Phycol., 25, 177-181.

3 Bacteria-free check media for freshwater algae

B–IV

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Appropriate medium 100 mL
Glucose 100 mg
Peptone 100 mg

Reference(s)
Ichimura, T., Watanabe, M. M. 1977 An axenic clone of Microcystis aeruginosa Kutz. emend. Elenkin from Lake Kasumigaura. Bull. Jpn. Soc. Phycol., 25, 177-181.

3 Bacteria-free check media for freshwater algae

B–V

Print
Appropriate medium 100 mL
Sodium acetate 50 mg
Glucose 50 mg
Tryptone 50 mg
Yeast extract 30 mg

Reference(s)
Ichimura, T., Watanabe, M. M. 1977 An axenic clone of Microcystis aeruginosa Kutz. emend. Elenkin from Lake Kasumigaura. Bull. Jpn. Soc. Phycol., 25, 177-181.

3 Bacteria-free check media for freshwater algae

YT

Print
Appropriate medium 100 mL
Yeast extract 100 mg
Tryptone 200 mg

Reference(s)
Ichimura, T., Itoh, T. 1977 17. Preservation methods of microalgae ( I ) [17. Bisaisôrui no hozonhô ( I ) ]. In Preservation methods of microorganisms [Biseibutsu Hozonhô], Ed. by Nei, T., University of Tokyo Press, Tokyo, p. 355-373 ( in Japanese without English title ).

4 Bacteria-free check media for marine algae

Bf/2

Print
ASP 7 1 ) 100 mL
Trypticase 50 mg
Yeast extract 5 mg

1 ) In the NIES-Collection, ASP 7 is replaced by f/2 medium. See f/2.

Reference(s)
Yoshida, Y., Kawaguchi, K. 1983 Buoyancy and phototaxis of Chattonella antiqua ( Hada ) Ono. Bull. Plankton Soc. Jpn., 30, 11-19 ( in Japanese with English summary ).

4 Bacteria-free check media for marine algae

MM 23 ( M. Tatewaki, pers. comm. )

Print
NaCl 1.8 g
MgSO4 · 7H2O 500 mg
KCl 60 mg
NaNO3 100 mg
CaCl2 · 2H2O 36.7 mg
K2HPO4 6 mg
Sucrose 400 mg
PII metals 1 ) 2 mL
FeCl3 · 6H2O 48 μg
Thiamine HCl 10 μg
Biotin 0.1 μg
Vitamin B12 0.2 μg
C–Source Mix II 2 ) 1 mL
Tris ( hydroxymethyl ) aminomethane 100 mg
Distilled water 97 mL
pH 8.0

1 ) See PII metals
2 ) See C–Source Mix II

4 Bacteria-free check media for marine algae

STP

Print
NaNO3 20 mg
K2HPO4 1 mg
Sodium glutamate 50 mg
Glucose 20 mg
Glycine 10 mg
D,L–Alanine 10 mg
Vitamin mix 8 1 ) 0.1 mL
Trypticase 20 mg
Yeast autolysate 2 ) 20 mg
Sucrose 100 mg
Soil extract 3 ) 5 mL
Seawater 80 mL
Distilled water 15 mL
pH 7.5

1 ) In the NIES–Collection, Vitamin mix 8 is replaced by

Vitamin mix S3. See Vitamin mix S3 .
2 ) In the NIES–Collection, yeast autolysate is replaced by yeast extract.
3 ) See Soil extract

Reference(s)
Provasoli, L., McLaughlin, J. J. A., Droop, M. R. 1957 The development of artificial media for marine algae. Arch. Mikrobiol., 25, 392-428.

5 Trace metals, vitamin mixtures and soil extracts

A2 Fe stock solution

Print
EDTA · 2Na 700 mg
FeCl3 · 6H2O 400 mg
Distilled water 100 mL

Sterilize by passing through a Millipore filter ( 0.22 μm ).


5 Trace metals, vitamin mixtures and soil extracts

A2 trace element stock solution

Print

Mix 1 volume of C 1 ) medium and 9 volumes of distilled water.
1 ) See C

5 Trace metals, vitamin mixtures and soil extracts

A5 solution

Print
H3BO3 286 mg
MnSO4 · 7H2O 1 ) 250 mg
ZnSO4 · 7H2O 22.2 mg
CuSO4 · 5H2O 7.9 mg
Na2MoO4 · 2H2O 2.1 mg
Distilled water 100 mL

1 ) In the NIES–Collection, 250 mg MnSO4 · 7H2O is replaced by 217 mg MnSO4 · 5H2O.

Reference(s)
Holm-Hansen, O., Gerloff, G. C., Skoog, F. 1954 Cobalt as an essential element for blue-green algae. Physiol. Planta., 7, 665-675.

5 Trace metals, vitamin mixtures and soil extracts

Allen metals

Print
Fe–EDTA 30.16 g
MnCl2 · 4H2O 1.79 g
H3BO3 2.86 g
ZnSO4 · 7H2O 220 mg
CuSO4 · 5H2O 79 mg
( NH4 )6Mo7O24 · 4H2O 130 mg
NH4VO3 23 mg
Distilled water 100 mL

In the NIES–Collection, Allen metals are used after dilution with distilled water to 1/1,000.

Reference(s)
Allen, M. B. 1959 Studies with Cyanidium caldarium, an anomalously pigmented chlorophyte. Arch. Mikrobiol., 32, 270-277.

5 Trace metals, vitamin mixtures and soil extracts

C–Source Mix II ( M. Tatewaki, pers. comm. )

Print
Glycine 100 mg
D,L-Alanine 100 mg
L-Asparagine 100 mg
Sodium acetate · 3H2O 1 ) 200 mg
Glucose 200 mg
L-Glutamic acid 200 mg
Distilled water 100 mL

1 ) In the NIES–Collection, 200 mg sodium acetate · 3H2O is replaced by 120 mg sodium acetate, anhydrous.


5 Trace metals, vitamin mixtures and soil extracts

D stock medium

Print
NTA 0.2 g
D trace mix 1 ) 1 mL
FeCl3 · 6H2O 0.58 mg
CaSO4 · 2H2O 120 mg
MgSO4 · 7H2O 200 mg
NaCl 16 mg
KNO3 200 mg
NaNO3 1.4 g
Na2HPO4 2 ) 220 mg
Distilled water 99 mL

1 ) See D trace mix
2 ) In the NIES-Collection, 220 mg Na2HPO4 is replaced by 550 mg Na2HPO4· 12H2O.

Reference(s)
Castenholz, R. W. 1969 Thermophilic blue-green algae and the thermal environment. Bacteriol. Rev., 33, 476-504.

5 Trace metals, vitamin mixtures and soil extracts

D trace mix

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conc · H2SO4 0.05 mL
MnSO4 · H2O 1 ) 228 mg
ZnSO4 · 7H2O 50 mg
H3BO3 50 mg
CuSO4 · 5H2O 2.5 mg
Na2MoO4 · 2H2O 2.5 mg
CoCl2 · 6H2O 4.5 mg
Distilled water 100 mL

1 ) In the NIES–Collection, 228 mg MnSO4 · H2O is replaced by 349 mg MnSO4 · 5H2O.

Reference(s)
Castenholz, R. W. 1969 Thermophilic blue-green algae and the thermal environment. Bacteriol. Rev., 33, 476-504.

5 Trace metals, vitamin mixtures and soil extracts

DY trace metal solution

Print
MnCl2 · 4H2O 20 mg
ZnSO4 · 7H2O 4 mg
CoCl2 · 6H2O 0.8 mg
Na2MoO4 · 6H2O 1 ) 2 mg
Na3VO4 · 10H2O 2 ) 0.2 mg
H2SeO3 0.2 mg
Distilled water 100 mL

1 ) In the NIES–Collection, Na2MoO4 · 6H2O is replaced by Na2MoO4 · 2H2O.
2 ) In the NIES-Collection, 0.2 mg Na3VO4 · 10H2O is replaced by 0.1 mg Na3VO4.

5 Trace metals, vitamin mixtures and soil extracts

Fe ( as EDTA; 1:1 molar )

Print
Fe( NH4 )2( SO4 )2 · 6H2O 70.2 mg
Na2EDTA · 2H2O 66 mg
Distilled water 100 mL

1 mL of   this solution contains 0.1 mg Fe.

Reference(s)
Provasoli, L. 1966 Media and prospects for the cultivation of marine algae. In Cultures and Collections of Algae, Eds. by Watanabe, A. & Hattori, A., Proc. U.S.-Japan Conf., Hakone, Sept. 1966., Jpn. Soc. Plant Physiol., p. 63-75.

5 Trace metals, vitamin mixtures and soil extracts

Fe solution

Print
FeSO4 · 7H2O 200 mg
Distilled water 100 mL
Conc · H2SO4 0.026 mL 1 )

1 ) 2 drops/500 mL

Reference(s)
Ichimura, T., Itoh, T. 1977 17. Preservation methods of microalgae ( I ) [17. Bisaisôrui no hozonhô ( I )]. In Preservation methods of microorganisms [Biseibutsu Hozonhô], Ed. by Nei, T., University of Tokyo Press, Tokyo, p. 355-373 ( in Japanese without English title ).

5 Trace metals, vitamin mixtures and soil extracts

f/2 metals

Print
Na2EDTA · 2H2O 440 mg
FeCl3 · 6H2O 316 mg
CoSO4 · 7H2O 1.2 mg
ZnSO4 · 7H2O 2.1 mg
MnCl2 · 4H2O 18 mg
CuSO4 · 5H2O 0.7 mg
Na2MoO4 · 2H2O 0.7 mg
Distilled water 100 mL

Reference(s)
Guillard, R. R. L., Ryther, J. H. 1962 Studies of marine planktonic diatoms. I. Cyclotella nana Hustedt, and Detonula confervacea ( Cleve ) Gran. Can. J. Microbiol., 8, 229-239.

5 Trace metals, vitamin mixtures and soil extracts

Hutner ’s trace elements

Print
Na2EDTA · 2H2O 5 g
ZnSO4 · 7H2O 2.2 g
H3BO3 1.14 g
MnCl2 · 4H2O 506 mg
FeSO4 · 7H2O 499 mg
CoCl2 · 6H2O 161 mg
CuSO4· 5H2O 157 mg
( NH4 )6Mo7O24 · 4H2O 110 mg
Distilled water 100 mL

Adjust the pH to 6.5-6.8 with KOH ( ~1.6 g ). Store the solution in a refrigerator ( 5°C ). The solution should have turned to violet color before use. This process takes a while and is necessary.

5 Trace metals, vitamin mixtures and soil extracts

K metals

Print
Na2EDTA · 2H2O 3.722 g
Fe-EDTA 0.493 g
FeCl3 · 6H2O 0.315 g
MnCl2 · 4H2O 17.8 mg
ZnSO4 · 7H2O 2.3 mg
CoSO4 · 7H2O 1.405 mg
Na2MoO4 · 2H2O 0.726 mg
CuSO4 · 5H2O 0.25 mg
Distilled water 100 mL

Reference(s)
Keller, M. D., Guillard, R.R.L. 1985 Factors significant to marine diatom culture. pp. 113-6. In Anderson, D.M., White, A.W., Baden, D.G. (eds.) Toxic Dinoflagellates. Elsevier, New York.
Keller, M.D., Selvin, R.C., Claus, W., Guillard, R.R.L. 1987. Media for the culture of oceanic ultraphytoplankton. J. Phycol., 23: 633-638.

5 Trace metals, vitamin mixtures and soil extracts

K/2 metals

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1 ) In the NIES–Collection, 0.149 mg NiCl2 · 6H2O is replaced by 0.812 mg NiCl2.

Reference(s)
Guillard, R. R. L., Ryther, J. H. 1962 Studies of marine planktonic diatoms. I. Cyclotella nana Hustedt, and Detonula confervacea ( Cleve ) Gran. Can. J. Microbiol., 8, 229-239.

5 Trace metals, vitamin mixtures and soil extracts

mIMR trace metals

Print
FeCl3 · 6H2O 100 mg
MnSO4 · 4H2O 85.5 mg
ZnSO4 · 7H2O 25 mg
Na2MoO4 · 2H2O 13 mg
CoCl2 · 6H2O 0.4 mg
CuSO4 · 5H2O 0.4 mg
Na2SeO3 0.173 mg
Na2EDTA 1 ) 600 mg
Distilled water 100 mL

1 ) In the NIES–Collection, Na2EDTA is replaced by Na2EDTA · 2H2O.


5 Trace metals, vitamin mixtures and soil extracts

mM–1 Trace elements

Print
CuSO4 · 5H2O 10 mg
MnCl2 · 4H2O 10 mg
Br ( 1mol / L solution ) 0.01 mL
ZnSO4 · 7H2O 10 mg
CoCl2 · 6H2O 5 mg
BaCl2 · 2H2O 1 mg
H3BO3 10 mg
FeCl3 · 6H2O 10 mg
Na2MoO4 · 2H2O 5 mg
Distilled water 100 mL

Indicated as “Trace elements” in reference.

Reference(s)
Hoham, R. W., Berman, J. D., Rogers, H. S., Felio, J. H., Ryba, J. B., Miller, P. R. 2006 Two new species of green snow algae from Upstate New York, Chloromonas chenangoensis sp. nov. and Chloromonas tughillensis sp. nov. ( Volvocales, Chlorophyceae ) and the effects of light on their life cycle development. Phycologia, 45, 319-330.

5 Trace metals, vitamin mixtures and soil extracts

mTYGM–9

Print

Pre–solution

K2HPO4 0.28 g
KH2PO4 40 mg
Casein Digest 0.2 g
Yeast extract 0.1 g
NaCl 0.75 g
Mucin, gastric 0.2 g
Distilled water 97 mL

Sterilize Pre–solution by autoclaving ( 121°C, 15 min ) , add aseptically 3 mL horse serum and 50 μL 10% Tween 80 dissolved in absolute ethanol ( both filter–sterilized ). Keep in a cool place.


5 Trace metals, vitamin mixtures and soil extracts

P II metals

Print
Na2EDTA · 2H2O 100 mg
H3BO3 114 mg
FeCl3 · 6H2O 4.9 mg
MnSO4 · 4H2O 16.4 mg
ZnSO4 · 7H2O 2.2 mg
CoSO4 · 7H2O 480 μg
Distilled water 100 mL

Reference(s)
Provasoli, L. 1963 Growing marine seaweeds. In Proceedings of the Fourth International Seaweed Symposium, University of Tokyo Press, Tokyo, p. 9-17.

5 Trace metals, vitamin mixtures and soil extracts

P–1 metals

Print
H3BO3 618.3 mg
MnCl2 · 4H2O 69.25 mg
ZnCl2 5.45 mg
CoCl2 · 6H2O 238 μg
CuCl2 · 2H2O 0.1704 μg
Distilled water 100 mL

1 ) 1) In the NIES-Collection, CuCl2 · 2H2O is removed.

Reference(s)
Imai, I. 2012 High growth rates of the red tide flagellate Chattonella marina (Raphidophyceae) observed in culture. Hokudai Suisan Ihou, 62, 71-74 (in Japanese with English summary).

5 Trace metals, vitamin mixtures and soil extracts

P IV metals

Print
Na2EDTA · 2H2O 100 mg
FeCl3 · 6H2O 19.6 mg
MnCl2 · 4H2O 3.6 mg
ZnCl2 1 ) 1.04 mg
CoCl2 · 6H2O 0.4 mg
Na2MoO4 · 2H2O 0.25 mg
Distilled water 100 mL

1 ) In the NIES–Collection, 1.04mg ZnCl2 is replaced by 2.2mg ZnSO4 · 7H2O.

Reference(s)
Provasoli, L., Pintner, I. J. 1959 Artificial media for fresh-water algae: problems and suggestions. In The Ecology of Algae. Spec. Pub. No. 2,, Eds. by Tryon, C. A., Jr. & Hartmann, R. T., Pymatuning Laboratory of Field Biology, University of Pittsburgh, Pittsburgh, p. 84-96.

5 Trace metals, vitamin mixtures and soil extracts

PN metals

Print
Na2EDTA · 2H2O 100 mg
H3BO3 113 mg
FeCl3 · 6H2O 6.3 mg
CoSO4 · 7H2O 0.093 mg
ZnSO4 · 7H2O 4.66 mg
MnCl2 · 4H2O 3.2 mg
Distilled water 100 mL

Reference(s)
Watanabe, M. M., Satake, K. N. ( Eds. ) 1991 NIES-Collection. List of Strains, Third Edition, 1991, Microalgae and Protozoa. Microbial Culture Collection, National Institute for Environmental Studies, Tsukuba, 163 pp.

5 Trace metals, vitamin mixtures and soil extracts

PRO–99 trace metals

Print
Na2EDTA 1 ) 0.58 g
FeCl3 · 6H2O 0.422 g
ZnSO4 · 7H2O 2.933 mg
CoCl2 · 6H2O 1.332 mg
MnCl2 · 4H2O 24 mg
Na2MoO4 · 2H2O 0.839 g
Na2SeO3 2.306 mg
NiCl2 · 6H2O 0.37 mg
Distilled water 100 mL

1 ) In the NIES–Collection, 0.58 g Na2EDTA is replaced by 0.637 g Na2EDTA · 2H2O.


5 Trace metals, vitamin mixtures and soil extracts

Soil extract

Print

To 1000 mL distilled water add 200 mL of soil ( soil from undisturbed deciduous woodland is best ) and heat by autoclaving for 1 h at 105°C. When cool, heat by autoclaving for 1 h at 105°C again. Pass the supernatant through a GF/C filter and Celite, and then pass the filtrate through a GF/F filter. Adjust to 1000 mL by adding distilled water. Dispense 10 mL of the final filtrate into each test tube and sterilize by autoclaving for 20min at 121°C. Keep in a cool place.

Reference(s)
Provasoli, L., McLaughlin, J. J. A., Droop, M. R. 1957 The development of artificial media for marine algae. Arch. Mikrobiol., 25, 392-428.

5 Trace metals, vitamin mixtures and soil extracts

Trace metal mix A5 + Co

Print
H3BO3 286 mg
MnCl2 · 4H2O 181 mg
ZnSO4 · 7H2O 22.2 mg
Na2MoO4 · 2H2O 39 mg
CuSO4 · 5H2O 7.9 mg
Co( NO3 )2·6H2O 4.9 mg
Distilled water 100 mL

5 Trace metals, vitamin mixtures and soil extracts

Vitamin mix S3

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Thiamine HCl 5 mg
Nicotinic acid 1 mg
Calcium pantothenate 1 mg
p–Aminobenzoic acid 0.1 mg
Biotin 0.01 mg
Inositol 50 mg
Folic acid 0.02 mg
Thymine 30 mg
Distilled water 100 mL

Reference(s)
Provasoli, L. 1963 Growing marine seaweeds. In Proceedings of the Fourth International Seaweed Symposium, University of Tokyo Press, Tokyo, p. 9-17.

5 Trace metals, vitamin mixtures and soil extracts

Waris EDTA

Print
EDTA 0.522 g
FeSO4 · 7H2O 0.498 g
KOH (1 mol/L solution) 5.4 mL
Distilled water 94.6 mL

EDTA and FeSO4 · 7H2O is heated for 30 min (100°C ); KOH is added to the cooled mixture.

Reference(s)
McFadden, G. I., Melkonian, M. 1986 Use of Hepes buffer for microalgael culture media and fixation for eledtron microscopy. Phycology, 25, 551-557.

5 Trace metals, vitamin mixtures and soil extracts

Waris P-2

Print
Na2EDTA· 2H2O 0.3 g
H3BO3 0.114 g
MnCl2· 4H2O 14.4 mg
ZnSO4·7H2O 2.1 mg
CoCl2·6H2O 0.4 mg
Distilled water 100 mL

Reference(s)
McFadden, G. I., Melkonian, M. 1986 Use of Hepes buffer for microalgael culture media and fixation for eledtron microscopy. Phycology, 25, 551-557.

5 Trace metals, vitamin mixtures and soil extracts

WC trace metals

print
Na2EDTA· 2H2O 0.43 g
FeCl3 · 6H2O 0.31 mg
CuSO4 · 5H2O 1 mg
ZnSO4 · 7H2O 2.2 mg
CoCl2 · 6H2O 1 mg
MnCl2 · 4H2O 1 mg
Na2MoO4 · 2H2O 18 mg
H3BO3 0.6 mg
Distilled water 100 mL

Reference(s)
Guillard, R. R. L., Lorenzen, C. J. 1972 Yellow-green algae with chlorohyllide C. J. Phycol., 8, 10-14.
Andersen, R. A. E, Berges, J. A., Harrison, P. J., Watanabe, M. M. 2005 Appendix A — Recipes for freshwater and seawater media. In Algal culturing techniques, Ed. by Andersen, R. A. E, Elsevier Academic Press, p. 474.

6 Media for protozoa

AF–6 + Wheat

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Beforehand, sterilize wheat grains by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add a grain of sterile wheat to 10 mL AF–6 1 ) medium.
1 ) See AF–6


6 Media for protozoa

ESM + mTYGM–9 + Rice

Print

Beforehand, sterilize polished rice by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add 500 μL mTYGM–9 1 ) and a grain of sterile rice to 10 mL ESM 2 ) medium.
1 ) See mTYGM–9
2 ) See ESM

6 Media for protozoa

ESM + Rice

Print

Beforehand, sterilize polished rice by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add a grain of sterile rice to 10 mL ESM 1 ) medium.
1 ) See ESM


6 Media for protozoa

f/2 + mTYGM–9 + Rice

Print

Beforehand, sterilize polished rice by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add 500 μL mTYGM–9 1 ) and a grain of sterile rice to 10 mL f/2 2 ) medium.
1 ) See mTYGM–9
2 ) See f/2


6 Media for protozoa

f/2 + Rice

Print

Beforehand, sterilize polished rice by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add a grain of sterile wheat to 10 mL f/2 1 ) medium.
1 ) See f/2

6 Media for protozoa

f/2 + Wheat

Print

Beforehand, sterilize wheat grains by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add a grain of sterile wheat to 10 mL f/2 1 ) medium.
1 ) See f/2

6 Media for protozoa

Helio

Print
Sodium acetate 10 mg
Polypeptone 10 mg
Tryptone 20 mg
Yeast extract 20 mg
CaCl2 · 2H2O 0.1 mg
Sea water 10 mL
Distilled water 90 mL

6 Media for protozoa

LE

Print

L solution: White part of lettuce is dried at 90ºC for 16–18 h without scorching; 300 mg of the dried lettuce is added to 100 mL boiling water ( 9:1 distilled water to tap water ) and boiled for 30 min, while stirring. The supernatant is passed through cotton wool.
E solution: 300 mg of crushed yolk of hardboiled egg is added to 100 mL water ( 9:1 distilled water to tap water ) and boiled for 30 min, while stirring. The supernatant is passed through cotton wool.
Equal quantities of L and E solutions are mixed. The pH is adjusted to 6.8–7.0 with 1 mol / L NaOH, and 100 mL of the solution is dispensed into each 200–mL Erlenmeyer flask and sterilized by autoclaving ( 121ºC, 15 min ).

6 Media for protozoa

PYG

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Proteose Peptone 0.15 g
Yeast extract 0.1 g
Glucose 0.5 g
Distilled water 100 mL
pH 7.0

PPYG medium is diluted half strength and reduced proteose peptone.

Reference(s)
Mori, K., Yomo, T., Kashiwagi, A. 2011 Single-cell isolation and cloning of tetrahymena thermophila cells with a fluorescence-activated cell sorter. J. Eukaryotic Microbiol., 58(1), 37-42.

6 Media for protozoa

SUY

Print

Prepare as for 100 mL URO 1 ) medium with seawater instead of distilled water. Add 10 mg yeast extract and 20 mg tryptone. Indicated as " URO–YT " in reference.
1 ) See URO

Reference(s)
Moriya, M., Nakayama, T., Inouye, I. 2000 Ultrastructure and 18S rDNA sequence analysis of Wobblia lunata gen. et sp. nov., a new heterotrophic flagellate ( Stramenopiles, incertae sedis ). Protist, 151, 41-55.

6 Media for protozoa

SUY 1/10

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Prepare as for 100 mL URO 1 ) medium with seawater instead of distilled water. Add 1 mg yeast extract and 2 mg tryptone. Indicated as " URO–1/10 YT " in reference.
1 ) See URO

Reference(s)
Moriya, M., Nakayama, T., Inouye, I. 2002 A new class of the Stramenopiles, Placididea Classis nova: description of Placidia cafeteriopsis gen. et sp. nov. Protist, 153, 143-156.

6 Media for protozoa

SUY 1/10 + mTYGM–9 + Rice

Print

Beforehand, sterilize polished rice by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add 500 μL mTYGM–9 1 ) and a grain of sterile rice to 10 mL SUY 1/10 2 ) medium.
1 ) See mTYGM–9
2 ) See SUY 1/10


6 Media for protozoa

SUY 1/10 + Wheat

Print

Beforehand, sterilize wheat grains by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add a grain of sterile wheat to 10 mL SUY 1/10 1 ) medium.
1 ) See SUY 1/10


6 Media for protozoa

URO + Wheat

Print

Beforehand, sterilize wheat grains by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add a grain of sterile wheat to 10 mL URO 1 ) medium.
1 ) See URO


6 Media for protozoa

URO–H + Wheat

Print

Beforehand, sterilize wheat grains by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add a grain of sterile wheat to 10 mL URO–H 1 ) medium.
1 ) See URO–H


6 Media for protozoa

UYTS + Rice

Print

Prepare as for 100 mL URO 1 ) medium with 99.2 mL instead of 99.9 mL distilled water. Adjust to pH 7.5 with 0.1 mol / L HCl, and add 10 mg yeast extract, 20 mg tryptone and 0.3 ml horse serum ( UYTS medium ). Beforehand, sterilize polished rice by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add a grain of sterile rice to 10 mL UYTS medium.
1 ) See URO


6 Media for protozoa

YE100ppm

Print

Into 100 mL distilled water 9 mg powder medium of Daigo's Artificial SeawaterSP
( Nihon Pharmaceutical Co., Ltd. ) and 10 mg yeast extract.


6 Media for protozoa

1/2Waris H+Si

Print
HEPES 11.915 mg
KNO3 5 mg
MgSO4 · 7H2O 1 mg
(NH4)2HPO4 1 mg
Ca(NO3)2 · 4H2O 5 mg
Waris P-2 0.05 mL
Waris EDTA 0.05 mL
Vitamin B12 1 μg
Biotin 0.05 μg
Thiamine HCl 5 μg
Niacinamide 0.005 μg
Soil estract 3 ) 0.5 mL
Na2SiO3 · 9H2O 2.842 mg
Distilled water 99.4 mL
pH 7.0

Waris H is diluted half strength and added Na2SiO3 · 9H2O.
1 ) See Waris P-2
2 ) See Waris EDTA
3 ) See Soil extract

Reference(s)
McFadden, G. I., Melkonian, M. 1986 Use of Hepes buffer for microalgael culture media and fixation for eledtron microscopy. Phycology, 25, 551-557.

7 Media for freshwater red algae

Bold 3N

Print
NaNO3 75 mg
CaCl2 · 2H2O 2.5 mg
MgSO4 · 7H2O 7.5 mg
K2HPO4 7.5 mg
KH2PO4 1 ) 17.5 mg
NaCl 2.5 mg
Vitamin B12 2 ) 0. 015 μg
PIV metals 3 ) 0.6 mL
Soil extract 4 ) 4 mL
Distilled water 96.4 mL

1 ) In the NIES–Collection, the amount of KH2PO4 is reduced from 17.5 mg to 10.5 mg.
2 ) In the NIES–Collection, the amount of Vitamin B12 is increased from 0.015 μg to 0.02 μg.
3 ) See PIV metals
4 ) See Soil extract


8 Media for Charales

mSWC–2 ( Modified SWC–2 )

Print

Put leaf mould into a glass vessel to make a thin bottom layer, and add river sand onto the bottom layer up to one–quarter to one–fifth from the bottom. Add a pinch of garden lime to the river sand before use. Dampen the soil with deionized water ( or distilled water ). Cover the glass vessel with a plastic cap or aluminum foil, and autoclave it twice with overnight rest in between ( 121ºC, 20 min ). After cooling the mixture to room temperature, pour sterilized deionized water ( or sterilized distilled water ) into it carefully ( so as not to disturb the soil ). In the case of brackish water strains, deionized water is replaced by about one–third–diluted Herbst artificial seawater ( 1/3 Herbst ASW ).

Reference(s)
Sakayama, H., Hara Y., Nozaki, H. 2004 Taxonomic re-examination of six species of Nitella ( Charales, Charophyceae ) from Asia, and phylogenetic relationships within the genus based on rbcL and atpB gene sequences. Phycologia, 43, 91-104.

8 Media for Charales

SWC–1

Print

Put leaf mould into a glass vessel to make a thin bottom layer, and add black soil onto the bottom layer up to one-quarter to one–fifth from the bottom. Dampen the soil with deionized water ( or distilled water ). Cover the glass vessel with a plastic cap or aluminum foil, and autoclave it twice with overnight rest in between ( 121ºC, 20 min ). After cooling the mixture to room temperature, pour sterilized deionized water ( or sterilized distilled water ) into it carefully ( so as not to disturb the soil ).

Reference(s)
Sakayama, H., Hara Y., Nozaki, H. 2004 Taxonomic re-examination of six species of Nitella ( Charales, Charophyceae ) from Asia, and phylogenetic relationships within the genus based on rbcL and atpB gene sequences. Phycologia, 43, 91-104.

8 Media for Charales

SWCN–1

Print

Put bottom mud from a paddy field into a glass vessel up to one–quarter to one–fifth from the bottom. Dampen the mud with deionized water ( or distilled water ). Cover the glass vessel or jar with a plastic cap or aluminum foil, and autoclave it twice with overnight rest in between ( 121ºC, 20 min ). After cooling the mixture to room temperature, pour sterilized deionized water ( or sterilized distilled water ) into it carefully ( so as not to disturb the soil ).

8 Media for Charales

SWCN–2

Print

Put leaf mould into a glass vessel to make a thin bottom layer, and add a mixture of black soil and river sand onto the bottom layer up to one–quarter to one–fifth from the bottom. Dampen the soil with deionized water ( or distilled water ). Cover the glass vessel or jar with a plastic cap or aluminum foil, and autoclave it twice with overnight rest in between ( 121ºC, 20 min ). After cooling the mixture to room temperature, pour sterilized deionized water ( or sterilized distilled water ) into it carefully ( so as not to disturb the soil ). In the case of brackish water strains, deionized water is replaced by about one–third–diluted Herbst artificial seawater ( 1/3 Herbst ASW ).

8 Media for Charales

SWCN–3

Print

Put a mixture of black soil and bottom mud from a paddy field into a glass vessel up to one–quarter to one–fifth from the bottom. Dampen the soil with deionized water ( or distilled water ). Cover the glass vessel or jar with a plastic cap or aluminum foil, and autoclave it twice with overnight rest in between ( 121ºC, 20 min ). After cooling the mixture to room temperature, pour sterilized deionized water ( or sterilized distilled water ) into it carefully ( so as not to disturb the soil ).

8 Media for Charales

SWCN–4

Print

Put a mixture of river sand and bottom mud from a paddy field into a glass vessel up to one–quarter to one–fifth from the bottom. Dampen the soil with deionized water ( or distilled water ). Cover the glass vessel or jar with a plastic cap or aluminum foil, and autoclave it twice with overnight rest in between ( 121ºC, 20 min ). After cooling the mixture to room temperature, pour sterilized deionized water ( or sterilized distilled water ) into it carefully ( so as not to disturb the soil ).

8 Media for Charales

1/3 Herbst ASW

Print
NaCl 3.0 mg
KCl 1 ) 81.4 mg
CaCl2 1 ) 132 mg
MgSO4 1 ) 660 mg
NaHCO3 1 ) 504 mg
Distilled water 100 mL

1 ) In the NIES–Collection, the amount of KCl is reduced from 81.4 mg to 80.0 mg, 132 mg CaCl2 is replaced by 172 mg CaCl2 · 2H2O, 660 mg MgSO4 is replaced by 1.35 g MgSO4 · 7H2O, and the amount of NaHCO3 is reduced from 504 mg to 49.5 mg.

Reference(s)
Okazaki, Y., Shimmen, T., Tazawa, M. 1984 Turgor regulation in a brackish charophyte, Lamprothamnium succinctum I. Artificial modification of intracellular osmotic pressure. Plant Cell Physiol., 25, 565-571.

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