CCALA Culture Collection
of Autotrophic Organisms

Galdieria medium - modified

Authors: Gross& Schnarrenberger, 1995

Dissolve chemicals in 500 ml of distilled water. Add 2,07 ml of Fe-EDTA solution and 2,00 ml trace metal solution. Make up to 1000 ml. Autoclave.

Adjust medium to final pH of 1,8 or as desired with 0.5 M H₂SO₄ 

Fe-EDTA solution 

To 50 ml od dH₂O add following component and fill in with dH₂O do 100 ml.

Trace metals solution 

Literautre: 

• Gross, W., & Schnarrenberger, C. (1995). Heterotrophic growth of two strains of the acido-thermophilic red alga Galdieria sulphuraria. Plant and cell physiology, 36(4), 633-638.

S medium

Authors: Gerasimenko et al., 1996; Samylina et al., 2021

Dissolve the chemicals in 500 mL of distilled water. Add 1,00 ml of trace metal solution and make up to 1000 ml with distilled water. Autoclave.

Trace metals solution 

Literature:

• Gerasimenko, L. M., Dubinin, A. V., & Zavarzin, G. A. (1996). Alkaliphilic cyanobacteria from soda lakes of Tuva and their ecophysiology. Microbiology (Mikrobiologiya), 65(6), 736-740.
• Samylina, O. S., Sinetova, M. A., Kupriyanova, E. V., Starikov, A. Y., Sukhacheva, M. V., Dziuba, M. V., & Tourova, T. P. (2021). Ecology and biogeography of the ‘marine Geitlerinema’cluster and a description of Sodalinema orleanskyi sp. nov., Sodalinema gerasimenkoae sp. nov., Sodalinema stali sp. nov. and Baaleninema simplex gen. et sp. nov.(Oscillatoriales, Cyanobacteria). FEMS Microbiology Ecology, 97(8), fiab104.

Z medium

Authors: Staub, 1961

To 900 mL of dH₂O, add the appropriate volumes of all stock solutions. Bring the final volume to 1,000 ml with dH₂O. Autoclave. The final pH should be 6.6.

Fe-EDTA solution 

Trace metals solution 

Literature:

• Staub, R. (1961). Ernährungsphysiologisch-autökologische Untersuchungen an der planktischen Blaualge Oscillatoria rubescens DC. Schweizerische Zeitschrift für Hydrologie, 23, 82–198. DOI: 10.3929/ethz-a-00009-2335.

Dunaliella medium

Author: Schlösser, 1982

To 900 mL of dH₂O, add the appropriate volumes of all stock solutions and soil extract. Bring the final volume to 1,000 mL with dH₂O. Autoclave.

Soil extract

Fill about one third of a flask with air-dried garden or leaf soil. Avoid soil with high humus or clay content, and do not use soil contaminated with fertilizers, herbicides, or fungicides. Add deionized water until the water level is about 5 cm above the soil. Sterilize the mixture for 1 hour, then repeat the sterilization after 24 hours. Let the soil settle and separate the clear extract by decanting, filtration, or centrifugation. Dispense the extract into small containers, autoclave for 20 minutes, and store in a refrigerator.

Double-strength artificial seawater

Literature:

• Schlösser, U. G. (1982). Sammlung von algenkulturen. Berichte der Deutschen Botanischen Gesellschaft, 95(1), 181-276.

Spirulina medium

Authors: Aiba & Ogawa, 1977; Schlösser, 1994

Prepare Solutions I and II separately and bring each solution to a final volume of 500 mL with dH₂O. Autoclave separately and allow to cool. Combine the solutions aseptically, then add 1 mL of cyanocobalamin (vitamin B₁₂) solution aseptically.

Trace metals solution 

Vitamin Solution

Dissolve the cyanocobalamin in 1 liter dH₂O. Filter-sterilize and store frozen.
 

Literauture: 

• Aiba, S., & Ogawa, T. (1977). Assessment of growth yield of a blue—green alga, Spirulina platensis, in axenic and continuous culture. Microbiology, 102(1), 179-182.
• Schlösser, U. G. (1994). SAG‐sammlung von algenkulturen at the University of Göttingen catalogue of strains 1994. Botanica acta, 107(3), 113-186.

Jaworski’s Medium

Author: Schlösser, 1982

For freshwater algae.

To approximately 900 mL of dH₂O, add the appropriate volumes of all stock solutions. Bring the final volume to 1,000 mL with dH₂O.

EDTA-solution

Trace metals solution

Vitamins solution

Literature:

• Schlösser, U. G. (1982). Sammlung von algenkulturen. Berichte der Deutschen Botanischen Gesellschaft, 95(1), 181-276.

Dystrophic soil-peat medium 

Author: Schlösser 1982 (= Desmidiacean medium) 

Into 930 ml of dH₂O add the stock solutions, soil extract, peat extract and micronutrient solution. Autoclave. Add vitamin B₁₂ as sterile solution after cooling.
 

Soil extract

Fill about one third of a flask with air-dried garden or leaf soil. Avoid soil with high humus or clay content, and do not use soil contaminated with fertilizers, herbicides, or fungicides. Add deionized water until the water level is about 5 cm above the soil. Sterilize the mixture for 1 hour, then repeat the sterilization after 24 hours. Let the soil settle and separate the clear extract by decanting, filtration, or centrifugation. Dispense the extract into small containers, autoclave for 20 minutes, and store in a refrigerator.

Peat Exctract

Prepare peat extract in the same way as soil extract, using peat or dried Sphagnum moss instead of soil. After extraction and cooling, filter the extract and sterilize it in small portions by autoclaving.

Micronutrient solution

Autoclave the components separately in two solutions and combine them after cooling:

• Solution I: 881 ml of dH₂O + stock solutions of salts 
• Solution II: 100 ml of dH₂O + FeSO₄ + EDTA

Literature:

• Schlösser, U. G. (1982). Sammlung von algenkulturen. Berichte der Deutschen Botanischen Gesellschaft, 95(1), 181-276.

WC medium

Authors: Guillard & Lorenzen, 1972

For planktonic algae and cyanobacteria.

To approximately 900 mL of dH₂O, add the appropriate volumes of all stock solutions and TRIS buffer, except for the vitamin solution. Bring the final volume to 1,000 mL with dH₂O and autoclave. After cooling, add the sterile-filtered vitamin solution aseptically.

Trace metals solution

Vitamins solution

Sterile by filtration

Literature:

• Guillard, R. R., & Lorenzen, C. J. (1972). Yellow-green algae with chlorophyllide c. Journal of Phycology, 8(1).

Porphyridium medium

Authors: Brody & Emerson 1959, modif. Pekárková (pers. comm.)

Prepare Solution 1 and bring the final volume to 1,000 mL with dH₂O. Add 0.5 mL of Solution 2 and 0.5 mL of Solution 3. Autoclave.

Solution 1.

Solution 2.

Solution 3.

Literature:

• Brody, M., & Emerson, R. (1959). The effect of wavelength and intensity of light on the proportion of pigments in Porphyridium cruentum. American Journal of Botany, 46(6), 433-440.

Euglena medium

Author: Schlösser, 1982

To approximately 950 mL of dH₂O, add the appropriate amounts of all components while stirring continuously. Bring the final volume to 1,000 mL with dH₂O.

Literature:

• Schlösser, U. G. (1982). Sammlung von algenkulturen. Berichte der Deutschen Botanischen Gesellschaft, 95(1), 181-276.

Brackish medium

Author: Schlösser, 1982

Combine dH₂O and artificial seawater, then add the appropriate volumes of all stock solutions, soil extract, and trace metal solution. Bring the final volume to 1,000 mL with dH₂O.

Soil extract

Fill about one third of a flask with air-dried garden or leaf soil. Avoid soil with high humus or clay content, and do not use soil contaminated with fertilizers, herbicides, or fungicides. Add deionized water until the water level is about 5 cm above the soil. Sterilize the mixture for 1 hour, then repeat the sterilization after 24 hours. Let the soil settle and separate the clear extract by decanting, filtration, or centrifugation. Dispense the extract into small containers, autoclave for 20 minutes, and store in a refrigerator.

Trace metals solution 

Artificial seawater 

Literature:

• Schlösser, U. G. (1982). Sammlung von algenkulturen. Berichte der Deutschen Botanischen Gesellschaft, 95(1), 181-276.

Polytoma medium

Author: Schlösser, 1982

To 950 mL of dH₂O, add the appropriate volumes of all stock solutions and soil extract. Autoclave.

Soil extract

Fill about one third of a flask with air-dried garden or leaf soil. Avoid soil with high humus or clay content, and do not use soil contaminated with fertilizers, herbicides, or fungicides. Add deionized water until the water level is about 5 cm above the soil. Sterilize the mixture for 1 hour, then repeat the sterilization after 24 hours. Let the soil settle and separate the clear extract by decanting, filtration, or centrifugation. Dispense the extract into small containers, autoclave for 20 minutes, and store in a refrigerator.

Literature:

• Schlösser, U. G. (1982). Sammlung von algenkulturen. Berichte der Deutschen Botanischen Gesellschaft, 95(1), 181-276.

HS medium for Chlamydomonas

Author: Sueoka, 1960

To 500 mL of dH₂O, add the appropriate volumes of stock solutions 1–4 . Bring the volume to 990 mL with dH₂O and autoclave. After cooling, add the appropriate volume of stock solution 5 aseptically.

Trace metals solution 

Literature:

• Sueoka, N. (1960). Mitotic replication of deoxyribonucleic acid in Chlamydomonas reinhardi. Proceedings of the National Academy of Sciences, 46(1), 83-91.

Medium for non-seed plants

Authors: Baslerová & Dvořáková, 1962

Mosses, liverworts, and ferns are maintained in 250 mL Erlenmeyer flasks on agar medium in an air-conditioned room at 12–15 °C.

To 900 mL of dH₂O, add the appropriate volumes of all stock solutions for the selected medium variant (M or JKL) as indicated in the table. Bring the final volume to 1,000 mL with dH₂O and autoclave.

Literature:

• Baslerová, M., & Dvořáková, J. (1962). Algarum, Hepaticarum, Muscorumque in culturis collectio. Nakl. Československé akademie věd.

Gorham’s solution

Authors: Koeman & van den Hoek, 1981

Dissolve the appropriate amounts of all components in dH₂O  add the soil extract. Bring the final volume to 1,000 mL with dH₂O.

Soil extract

1/3 of 6 litre flask fill with garden or leaf soil (not too great humus or clay content), and deionized water until it stands 5cm above the soil. Sterilize by heating in a steamer one hour, twice in a 24h interval. Separate the decanted extract by centrifugation. Fill into small containers, autoclave 20 minutes/1 atmosphere; store in refrigerator.

Literature: 

• Koeman, R. P. T., & van den Hoek, C. (1981). The taxonomy of Ulva (Chlorophyceae) in the Netherlands. British Phycological Journal, 16(1), 9-53.

BBM medium

Authors: Bold 1949; Bischoff and Bold 1963

To 900 mL of dH₂O, add the appropriate volumes of all stock solutions. Bring the final volume to 1,000 mL with dH₂O and autoclave. The final pH should be 6.6.

EDTA solution 

Acidified Iron Solution

Trace metals solution 

Medium modification:

• 3N-BBM modification: For this modification, add 30 ml of stock solution No. 1 when preparing the medium.

Literature:

• Bischoff, H. W. (1963). Phycological studies IV. Some soil algae from Enchanted Rock and related algal species. University of Texas Publication, 6318, 1.
• Bold, H. C. (1949). The morphology of Chlamydomonas chlamydogama, sp. nov. Bulletin of the Torrey Botanical Club, 101-108.

Low-nutrient medium

Author: Schlösser, 1982 (= Basal medium)

To 905 mL of dH₂O, add the appropriate volumes of all stock solutions, soil extract, and micronutrient solution. Autoclave.

Soil extract

Fill about one third of a flask with air-dried garden or leaf soil. Avoid soil with high humus or clay content, and do not use soil contaminated with fertilizers, herbicides, or fungicides. Add deionized water until the water level is about 5 cm above the soil. Sterilize the mixture for 1 hour, then repeat the sterilization after 24 hours. Let the soil settle and separate the clear extract by decanting, filtration, or centrifugation. Dispense the extract into small containers, autoclave for 20 minutes, and store in a refrigerator.'
 

Micronutrient solution

Autoclave the components separately in two solutions and combine them after cooling:

• Solution I: 881 ml of dH₂O + stock solutions of salts 
• Solution II: 100 ml of dH₂O + FeSO₄ + EDTA

Literature:

• Schlösser, U. G. (1982). Sammlung von algenkulturen. Berichte der Deutschen Botanischen Gesellschaft, 95(1), 181-276.

BG11 medium

Authors: Allen, 1959, 1968; Rippka et al., 1979

To 800 mL of dH₂O, add the appropriate volumes of all stock solutions and the trace metals solution. Autoclave. The final pH should be 7.4 after cooling. 

Trace metals solution 

Medium modifications:

• Oxynema medium modification: For Oxynema (CCALA 960), add 30 g of NaCl to 1 litre of BG₁₁ medium.
• BG₁₁-0/BG₁₁-N modification: For nitrogen-fixing cyanobacteria, prepare standard BG₁₁ medium without Stock 1.

Literature:

• Allen, M. B. (1959). Studies with Cyanidium caldarium, an anomalously pigmented chlorophyte. Archiv für Mikrobiologie, 32(3), 270-277.
• Allen, M. M. (1968). Simple conditions for growth of unicellular blue‐green algae on plates 1, 2. Journal of phycology, 4(1), 1-4.
• Rippka, R., Deruelles, J., Waterbury, J. B., Herdman, M., & Stanier, R. Y. (1979). Generic assignments, strain histories and properties of pure cultures of cyanobacteria. Microbiology, 111(1), 1-61.

1/2 SŠ medium

Authors: Zachleder & Šetlík, 1982 

To 970 mL of dH₂O, add the appropriate volumes of all stock solutions and autoclave. After cooling, add 1.5 mL of 1 N NaOH aseptically

Trace metals solution 

Medium modification:

• Euhalothece medium modification: For Euhalothece (CCALA 947), add 100 g of Na₂CO₃ to 1 litre of ½ SŠ medium.

Literature: 

• Zachleder, V., & Šetlík, I. (1982). Effect of irradiance on the course of RNA synthesis in the cell cycle of Scenedesmus quadricauda. Biologia Plantarum, 24(5), 341-353.