I cultured several flasks of one green microalgae species in TAP medium under irradiance of 90 umol.m-2.s-1,
I added 18g/L glucose in the medium, and after 9days of cutlure, I found the aglal culture in bad growth condition though the biomass accumulated was much higher than the control culture without glucose, the color of the algae showed a tendency of turning white!! I collected the cells and tested the pH value of the culture medium, and it was below pH4.5!! I think most of the algal cells must have died! The pH value in the control culture without glucose was 8.0. I checked the culture under microscope and no bacteria was found.
I think the whole thing is very confusing
The nitrogen source in TAP medium is NH4Cl, which should bring the pH down upon algal growth. So this could be a possible reason for the pH decrease in the above culture. But in another culture which also contained 18g/L glucose and under a much stronger irradiance of 800 umol.m-2.s-1. , the pH was 7.0 and with even more biomass accumulated!!! If NH4Cl is the criminal, then why the culture under stronger irradiance which should have taken in much more NH4Cl to produce biomass did not give a even lower pH??
Is it possible that the CO2 produced by respiration can cause a pH as acid as below 4.5?
The following is TAP recipe:
Tris - Acetate - Phosphate (TAP) Medium
(modified from Durnford Lab)
For One Liter of Medium:
2X Filnerís Beijernicks Solution 25ml
1M Potassium Phosphate 1ml
Trace mineral solution or 5ml
P IV solution 2X (see SVM) 3ml
Glacial Acetic Acid (17.4 mM acetate) 1ml
(pH should be 7.2)
2X Filnerís Beijernicks Solution (500 ml)
CaCl2 * 2H2O 1g
MgSO4 *7H2O 2g
Trace Mineral Solution (500 ml)
5 g disodium EDTA Ė dissolve in 400 ml water by heating and stirring
Neutralize to pH 6.5 with 5N NaoH
Add each of the following in order. Allow each to dissolve completely before adding the next.
0.5 g FeSO4*7H2O
2.2 g ZnSO4*7H2O
1.14 g H3BO3
0.51 g MnCl2*4H2O
0.016 g CuSO4*5H2O
0.073 g Na2MoO4*2H2O
0.016 g CoCl2*6H2O
1M Potassium Phosphate Stock (50 ml)
add: 20 ml 1M stock KH2PO4 (1M stock: 6.8g/50ml)
30 ml 1M stock K2HPO4 (1M stock: 8.7g/50ml)
pH 7.2 at RT
Edited by biobunny, 03 March 2009 - 11:44 PM.