iForest - Biogeosciences and Forestry


Is microbial biomass measurement by the chloroform fumigation extraction method biased by experimental addition of N and P?

Taiki Mori (1-2), Senhao Wang (1-3), Cong Wang (1-3), Jiangming Mo (1), Wei Zhang (1)   

iForest - Biogeosciences and Forestry, Volume 14, Issue 5, Pages 408-412 (2021)
doi: https://doi.org/10.3832/ifor3374-014
Published: Sep 04, 2021 - Copyright © 2021 SISEF

Short Communications

The chloroform fumigation extraction (CFE) method determines microbial biomass carbon (MBC) or nitrogen (MBN) by calculating the increase in extractable carbon (C) or nitrogen (N) due to microbial lysis during chloroform fumigation. In China, many studies have focused on the impacts of N and phosphorus (P) addition on soil MBC and MBN in forest ecosystems, where substantial atmospheric N deposition has strongly acidified soils. The addition of nutrients may alter the extraction process applied in the CFE method, potentially influencing the MBC and MBN determined by the CFE method independently of the actual microbial biomass. In this study, we tested whether the MBC and MBN determined by the CFE method were biased by the experimental addition of N and P in strongly acidified Chinese forest soils by adding N and P to the soils immediately before chloroform fumigation, which should not affect the actual microbial biomass. P addition significantly elevated the dissolved organic carbon (DOC) content, especially after fumigation, while N addition significantly reduced the dissolved nitrogen (DN) content. The added N was subtracted using blank samples without soil. However, the altered DOC and DN contents did not affect the MBC and MBN contents determined by the CFE method. In conclusion, our study suggests that the CFE is a relatively robust method to test the impacts of nutrient addition on microbial biomass in the strongly acidified soils of Chinese forests. We also suggested that: (i) even if a fertilization experiment results in an elevated DOC content following P addition, it does not necessarily indicate a stimulation of DOC production by microbes; and (ii) the soil adsorption capacity or the strength of microbial N uptake during the extraction procedure applied in the CFE method may affect the determination of MBN by influencing the DN extraction efficiency.


Chloroform Fumigation Extraction, Microbial Biomass, Nitrogen, Phosphorus, Soil, Tropical Forest

Authors’ address

Taiki Mori 0000-0002-1552-892X
Senhao Wang 0000-0001-6228-3999
Cong Wang
Jiangming Mo
Wei Zhang 0000-0002-6623-1341
Key Laboratory of Vegetation Restoration and Management of Degraded Ecosystems, South China Botanical Garden and Guangdong Provincial Key Laboratory of Applied Botany, Chinese Academy of Sciences, Guangzhou, 510650 (China)
Taiki Mori 0000-0002-1552-892X
Kyushu Research Center, Forestry and Forest Products Research Institute, FFPRI, Kurokami 4-11-16, Kumamoto, 860-0862 (Japan)
Senhao Wang 0000-0001-6228-3999
Cong Wang
University of Chinese Academy of Sciences, Beijing 100049 (China)

Corresponding author



Mori T, Wang S, Wang C, Mo J, Zhang W (2021). Is microbial biomass measurement by the chloroform fumigation extraction method biased by experimental addition of N and P?. iForest 14: 408-412. - doi: 10.3832/ifor3374-014

Academic Editor

Maurizio Ventura

Paper history

Received: Feb 13, 2020
Accepted: Jul 07, 2021

First online: Sep 04, 2021
Publication Date: Oct 31, 2021
Publication Time: 1.97 months

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