Does anyone have advice for growing ammonia oxidizng archaea? I have twice purchased actively growing Nitrososphaera viennensis cultures from DSMZ, followed their culturing instructions careful, and get no growth. Alternately, does any one have a culture they could share? I need to isolate genomic DNA.Thanks!
Dear Dr.Rima,
You may try a modified Synthetic Freshwater Crenarchaeota Medium containing Synthetic Freshwater Crenarchaeota Medium.
Couple of papers are added for your reference.
1. Nitrososphaera viennensis, an ammonia oxidizing archaeon from soil Maria Tourna, Michaela Stieglmeier,Anja Spang,Martin Könneke, Arno Schintlmeister,Tim Urich,a Marion Engel, Michael Schloter, Michael Wagner,Andreas Richter,and Christa Schlepera,Proc Natl Acad Sci U S A. 2011 May 17; 108(20): 8420–8425.
2. Abundance of amoA genes of ammonia-oxidizing archaea and bacteria
in activated sludge of full-scale wastewater treatment plants,Tawan Limpiyakorn , Puntipar Sonthiphand,, Chaiwat Rongsayamanont, Chongrak Polprasert,2011, Bioresourse Technology.ELSEVIER.
Good luck!
How are you measuring "growth"? For the cultures in our lab, we measure growth by qPCR of 16S rRNA and/or amoA, or change in concentration of NOx and/or ammonia.
Hi Christian - We get essentially no change ammonium concentration and no DNA when we do an extraction. (Our goal is to get DNA for qPCR.) We've tried after relatively short incubations (~ 1 month) and longer ones (2 and 3 months).
With the caveat that we don't work with pure AOA cultures, the only other thing I can think of is that we incubate and transfer our enrichment cultures in the dark and w/o shaking (and at a lower temperature than the N. viennensis culture).
Eventually copper can be limiting when using low concentration of copper and high concentration of EDTA. Nitritoxidizing Archaea might have a higher demand on copper.
Usually I trust the recipies from DSMZ. However, you can try to calculate the amount of free copper for example with PhreeqC or compare it with the amounts in this paper:
Generally, they need time to multiply. DSMZ gives minimum of two weeks. It might take much longer.
S. A. Amin, J. W. Moffett, W. Martens-Habbena, J. E. Jacquot, Y. Han, A. Devol,
A. E. Ingalls, D. a. Stahl, and E. Virginia Armbrust. Copper requirements of the
ammonia-oxidizing archaeon Nitrosopumilus maritimus SCM1 and implications for
nitrification in the marine environment. Limnology and Oceanography, 58(6):2037–
2045, 2013.
It might help to contact Dr. Michaela Stieglmeier directly:
http://www.plantdevelopment.bio.lmu.de/personen/stieglmeier/index.html
Tourna, M.*, M. Stieglmeier*, A. Spang, M. Koenneke, A. Schintlmeister, T. Urich, M. Engel, M. Schloter, M. Wagner, A. Richter, and C. Schleper. 2011. Nitrososphaera viennensis, an ammonia oxidizing archaeon from soil. Proc Natl Acad Sci 108(20): 8420-5
Stieglmeier, M., M. Mooshammer, B. Kitzler, W. Wanek, S. Zechmeister-Boltenstern, A. Richter, and C. Schleper. 2014. Aerobic nitrous oxide production through N-nitrosating hybrid formation in ammonia-oxidizing archaea. ISME J. 8(5): 1135-46
Stieglmeier, M., A. Klingl , R. J. E. Alves, S. K.-M. R. Rittman, M. Melcher, N. Leisch, and C. Schleper. 2014. Nitrososphaera viennensis sp. nov., an aerobic and mesophilic ammonia-oxidizing archaeon from soil and member of the newly described archaeal phylum Thaumarchaeota phyl. nov. IJSEM. 64 (Pt8):2738-52
Taylor, A. E., K. Taylor, B. Tennigkeit, M. Palatinszky, M. Stieglmeier, D. D. Myrold, C. Schleper, M. Wagner, and P. J. Bottomley. 2015. Inhibitory effects of C2 - C10 1-alkynes on ammonia oxidation in two Nitrososphaera species. Appl Environ Microbiol. 81 (6): 1942-48
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology