Glomalin is a glycoprotein produced abundantly on hyphae and spores of arbuscular mycorrhizal (AM) fungi in soil and in roots.
The glomalin test measures the content of this protein produced by soil microorganisms recognized for their beneficial relationships with crops (arbuscular mycorrhizal fungi).
Glomalin-related soil protein (GRSP) extractions were based on the procedure described by Wright and Upadhyaya (1996). The two GRSPs extraction procedures were different in the extraction condition and autoclaving time.
The two procedures are:
1- Easily extracted glomalin-related soil protein (EEG)
2- Total glomalin-related soil protein (TG)
What is the best method? is there methods better?
I would appreciate your assistance.
Dear Amina,
The two most commonly used methods for measuring glomalin in soil are the easily extracted glomalin-related soil protein (EEG) method and the total glomalin-related soil protein (TG) method. Both methods are based on the procedure described by Wright and Upadhyaya (1996).
The EEG method involves extracting glomalin from soil using a solution of 0.1 M NaOH and 0.1% Tween 20, followed by centrifugation, precipitation with ethanol, and resuspension in water. This method is considered less labor-intensive and time-consuming than the TG method.
The TG method involves extracting glomalin from soil using a solution of 0.5 M NaOH and 0.1% Tween 20, followed by autoclaving, centrifugation, precipitation with ethanol, and resuspension in water. This method extracts total glomalin, including the tightly bound glomalin.
Both methods have been widely used and are reliable for quantifying soil glomalin. However, it is essential to note that the specific method used will depend on the research question and the studied soil characteristics. Also, it is necessary to consider that both approaches have limitations, such as being unable to distinguish between different forms of glomalin and differentiating between glomalin and other glycoproteins.
It is essential to consult with the lab where the samples will be analyzed to ensure that they can be processed using your chosen method. Also, you may consider using other methods like ELISA, western blot, etc. but the extraction methods are more commonly used.
I hope I was able to answer your question.
Yours sincerely,
Edgar M Cambaza
Thank you Edgar Manuel Cambaza for your answer, what I have found in some studies, that they used sodium Citrate buffer and Bradford reagent to determine the soil glomalin, I would appreciate your effort to explain more in your protocol that you have mentioned above
Could you please specify when we use each method EEG & TG?
Best Regards,
Dear Amina,
Thank you for your question. I'm glad I could help.
The sodium citrate buffer and Bradford reagent method you mentioned is a variation of the Total Glomalin-Related Soil Protein (TG) method. However, instead of NaOH, it uses sodium citrate buffer for extraction and Bradford reagent for protein quantification. The Bradford reagent is commonly used for determining the protein concentration of a sample by binding to the protein and producing a color change proportional to the protein concentration.
The choice between the Easily Extracted Glomalin-Related Soil Protein (EEG) method and the Total Glomalin-Related Soil Protein (TG) method will depend on the research question and the characteristics of the soil being studied.
The EEG method is generally used when a quick and straightforward estimation of glomalin in the soil is sufficient and can provide an overview of the glomalin levels in the ground. It is less labor-intensive and time-consuming compared to the TG method.
On the other hand, the TG method is used when a more in-depth and accurate estimation of glomalin in the soil is required. This method extracts all forms of glomalin, including the tightly bound glomalin, and it is more thorough, complex, and time-consuming than the EEG method.
It is essential to consult with the lab where the samples will be analyzed to ensure that they can be processed using the chosen method and to verify the availability of the necessary reagents, such as sodium citrate buffer and Bradford reagent.
I hope this additional information is helpful. If you have any further questions, please let me know.
Best regards,
Edgar M Cambaza
Dear Edgar Manuel Cambaza
Thank you so much for these instructive details.
For the reagents, I have collected all, but this technique I would try them for the first time, That's why I want to know more about this analysis
Best Regards,
Dear Amina,
I'm glad to hear that the information I provided was helpful to you. Since you mentioned that this is your first time using these methods, I recommend carefully reading through the protocols and following the instructions closely to ensure that you obtain accurate and reliable results.
If you have any questions or concerns while performing the analysis, reach out to the manufacturer of the reagents or other experts in the field who may be able to provide guidance.
Additionally, it is a good practice to include positive and negative controls in your analysis to verify the accuracy and sensitivity of the methods.
I wish you the best of luck with your analysis. Please let me know if you have any further questions or if there is anything else I can assist you with.
Yours sincerely,
Edgar M Cambaza
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