510 APPENDIX 



Chloropseiidomonas ethylicum strain 2K. 



Recipe for synthetic liquid medium used by J. M, Olson, based on 

 the medium of Shaposhnikov, Kondrat'eva, Krasil'nikova and Ramen- 

 skaya {Doklady - Biol. Sci. Sect. (English Transl.), 129, 1047 (1959)), 

 which, in turn, is a modification of Larsen's inorganic medium [J. 

 Bacteriol., 64, 187 (1952)) for green sulfur bacteria. 



Stock cultures: stabs (1.5% agar) in the medium specified below, 

 except that the final concentration of Na2S • 9 H2O is 0.1% (w/v). 



Stock Solutions 



1. Versenol iron solution: see medium B for Chromatium. 



2. Trace elements solution: 

 FeCl3 



Na2B407 ' 10 H2O 

 ZnS04 • 7 H2O 

 C0SO4 • 7 H2O 

 CuCl2 • 2 H2O 

 MnS04 • H2O 

 Versenol iron solution 

 Distilled water to 1 liter, 



3. Double strength basal salts: 

 KH2PO4 

 NH4CI 



MgCl2 • 6 H2O 

 NaCl 

 CaCl2 

 Trace elements solution (add last) 



Final Medium 

 I, Double strength basal salts 500 ml 



Distilled water 450 ml 



II. 10% (w/v) Na2S • 9 H2O 2 ml 



ni. 10% (w/v) NaHC03 40 ml 



IV. 0.05% (w/v) FeS04 • 7 H2O in 0.3 N HCl 5 ml 



V. 70% ethanol (sterilized by filtration) 3 ml 



Autoclave solutions I to IV separately. Cool and mix. Add 

 solution V, and adjust pH to 7.3 with cone. HCl. 

 Incubation temperature: 30°C. 

 Light intensity should be less than 100 foot-candles. 



