Asked 24th Dec, 2014

Is anyone measuring phosphonatase activity in soil?

Can anybody suggest a reference papers on phosphonatase activity? Thank you so much.

Most recent answer

14th Jan, 2015
Yunuen Tapia-Torres
Universidad Nacional Autónoma de México
Thank you Dr. Sid James Nelson for your answer and suggestions. 

All Answers (5)

25th Dec, 2014
Rama Kant Dubey
National University of Singapore
Dear Yunuen Tapia,
Kindly find the paper entitled as " USE OF p-NITROPHENYL PHOSPHATE FOR ASSAY OF SOIL PHOSPHATASE ACTIVITY by  M. A. TABATABAI and J. M. BREMNER, if you have any problem then you can visit the page for article.
Good Luck.
28th Dec, 2014
Yunuen Tapia-Torres
Universidad Nacional Autónoma de México
Thank you very much!
31st Dec, 2014
Sid James Nelson
RAM Biochemicals, Inc.
Dr Abdel- Rhaheem points to a good paper that distinguishes the difference between phosphonate  ( P-C) and phosphate ( P-O) (bonding.). Note that the enzyme activity is monitored by the release of inorganic phosphate upon P-C cleavage but only after isolating the microbes on Phosphate free media, and measured in phosphate free buffer.  If one wants to measure the activity of a soil extract background phosphate would have to accounted for, and it might prove easier to measure appearance of the organic moeity from a substrate phosphonate. Otherwise an isotope technique might be used, to distinguish the P from substrate phosphonate.
1 Recommendation
14th Jan, 2015
Yunuen Tapia-Torres
Universidad Nacional Autónoma de México
Thank you Dr. Sid James Nelson for your answer and suggestions. 

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QPCR amplification curves show sudden linear course and low efficiency- what can this mean?
1 answer
  • Tina TuerlingsTina Tuerlings
In bee pathogen diagnostics, we often use different sets of primers to identify pathogens in bee guts.
One of the primer sets we have been using a while in PCR, we are now using to quantify pathogen loads via qPCR.
However, we discovered that a frequently used primer set, published years ago and used frequently in different papers, is showing a strange curve. It amplifies a fragment of the 18S rRNA gene, of Microsporidian pathogens belonging to the genus Nosema.
We tried different conditions and concentrations, but the curves stay the same.
In addition is a BioRad data file with the standard curves, and a picture for those who cannot open these types of files.
Efficiency is always low when using these primers, while R² is high, so we think it is due to the weird curves.
Does anyone know what could be the reason for the amplification curves?
Or the reason of the low efficiency?
More info:
Primer info:
Forward = Nosema universal, edited by Fernandez 2012, originally from Higes 2006 (TATGCCGACGATGTGATATG)
Reverse = Nosema universal, from Higes 2006 (CACAGCATCCATTGAAAACG)
qPCR Program:
2 min at 95°C, 39 times: 15 sec at 95°C, 30 sec at 56°C, 45 sec at 72°C
Fernández, J. M. et al. (2012) ‘Asymptomatic presence of Nosema spp. in Spanish commercial apiaries’, Journal of invertebrate pathology. J Invertebr Pathol, 111(2), pp. 106–110. doi: 10.1016/J.JIP.2012.06.008.
Higes, M., Martín, R. and Meana, A. (2006) ‘Nosema ceranae, a new microsporidian parasite in honeybees in Europe’, Journal of Invertebrate Pathology. Academic Press, 92(2), pp. 93–95. doi: 10.1016/J.JIP.2006.02.005.

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