World J Microbiol Biotechnol (2014) 30:1085–1092
DOI 10.1007/s11274-013-1528-5
ORIGINAL PAPER
Bacterial and fungal taxon changes in soil microbial community
composition induced by short-term biochar amendment in red
oxidized loam soil
Liao Hu • Lixiang Cao • Renduo Zhang
Received: 13 August 2013 / Accepted: 14 October 2013 / Published online: 18 October 2013
Ó Springer Science+Business Media Dordrecht 2013
Abstract To take full advantage of biochar as a soil
amendment, the objective of this study was to investigate
the effects of biochar addition on soil bacterial and fungal
diversity and community composition. Incubation experi-
ments with a forest soil (a red oxidized loam soil) with and
without biochar amendment were conducted for 96 days.
The culture-independent molecular method was utilized to
analyze soil bacterial and fungal species after the incuba-
tion experiments. Results showed that bacteria and fungi
responded differently to the biochar addition during the
short-term soil incubation. Twenty four and 18 bacterial
genara were observed in the biochar amended and una-
mended soils, respectively, whereas 11 and 8 fungal genera
were observed in the biochar amended and unamended
soils, respectively. Microbial taxa analysis indicated that
the biochar amendment resulted in significant shifts in both
bacterial and fungal taxa during the incubation period. The
shift for bacteria occurred at the genus and phylum levels,
while for fungi only at the genus level. Specific taxa, such
as Actinobacteria of bacteria and Trichoderma and Pae-
cilomyces of fungi, were enriched in the biochar amended
soil. The results reveal a pronounced impact of biochar on
soil microbial community composition and an enrichment
of key bacterial and fungal taxa in the soil during the short
time period.
L. Hu  R. Zhang (&)
Guangdong Provincial Key Laboratory of Environmental
Pollution Control and Remediation Technology, School of
Environmental Science and Engineering, Sun Yat-sen
University, Guangzhou 510275, China
e-mail: zhangrd@mail.sysu.edu.cn
L. Cao
School of Life Science, Sun Yat-sen University,
Guangzhou 510275, China
Keywords Biochar  Bacterial community  Fungal
community  Diversity
Introduction
Biochar is a product of incomplete combustion of biomass
in the pyrolysis process. Biochar amendment to soils has
been shown as a possible strategy of carbon sequestration
to mitigate the climate change (Lehmann 2007a; Lehmann
et al. 2009). Biochar amendment can also improve soil
quality (Chan et al. 2007; Laird 2008; Novak et al. 2009;
Sohi et al. 2010). The enhancement of soil fertility by
biochar is attributable to soil pH increase (van Zwieten
et al. 2010) and cation adsorption of nutrients (Liang et al.
2006). The effects may also be related to changes in the
microbial community composition.
Research on ancient soil management practices, which
are known as pyrogenic carbon-enriched soils in tropical
forests, has shown considerable community composition
differences. In the Terra preta, more diverse bacterial
groups are observed compared to adjacent non-anthropic
soils (Kim et al. 2007; O’Neill et al. 2009; Grossman et al.
2010). Compared to unamended soils, biochar addition
results in lower bacterial diversity in tropical forest soils
(Khodadad et al. 2011) and lower fungal diversity in
temperate region soils (Jin 2010). However, little infor-
mation is available on the soil microbial taxon changes
induced by biochar in subtropical forest soils. Moreover,
short-term and long-term biochar additions may lead to
different bacterial communities (Khodadad et al. 2011).
Greater microbial diversities are found in long-term bio-
char-enriched soils (Kim et al. 2007; O’Neill et al. 2009;
Grossman et al. 2010), whereas lower bacterial diversities
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are observed in short-term biochar-amended soils (Jin
2010; Khodadad et al. 2011). The labile substances in
biochar are usually mineralized within a short period of
time (Cheng et al. 2006), which should influence microbial
community greatly in short-term amendment (Steiner et al.
2008). Nevertheless, the effect of short-term biochar
additions on the microbial taxa is poorly understood. Red
oxidized loam soil is a typical type in the subtropical forest
soil. It should be interesting to study how biochar enhances
the fertility of this soil type (Glaser 2007; Novak et al.
2009).
The aim of this study was to investigate the effects of
biochar amendment on bacterial and fungal community
structures in a red oxidized loam soil in a subtropical forest
during a short-term incubation. It was hypothesized that
biochar addition affected bacteria and fungi differently in
terms of both diversity and community composition during
the short-term incubation.
Materials and methods
Soil sampling and biochar production
Plant materials for biochar production and bulk soil sam-
ples (10 kg) were collected from the surface and the sur-
face layer without organic litter (0–10 cm), respectively.
The sampling process was conducted in a mixed forest in
the Dinghushan Nature Reserve (23o090 2100 –23o110 3000 N,
112o300 3900 –112o330 4100 E, 100–700 mH), in Guangdong
Province of South China. Fresh soil samples were put in
sterilized sealing plastic bags and then stored at -80 °C in
the lab for following DNA extraction and experimental
treatments. Determined using the pipette method (Day
1965), the soil texture was loam with 41 % sand, 37 % silt,
and 22 % clay. A mixture of 2 g dried soil (dried at 105 °C
for 24 h) and 5 mL distilled water was shaken for 5 min
and set for 1 h, then was used to measure soil pH with a pH
electrode. The measured soil pH was 3.7.
Biochar was produced using the forest litter collected
from the soil surface. The litter was dried at 40 °C, ground
with a mortar and pestle, and sieved with a 2 mm sieve.
The plant materials were pyrolyzed under N2 in a pipe
furnace with a heating rate of 5 °C min-1 and the final
temperature at 400 °C for 1 h. The biochar particles were
then ground and passed a 250 lm sieve.
Experimental design
Two treatments were set up, including the soil with biochar
addition (denoted by BC) and without biochar addition
(used as the control and denoted by CK). For each treat-
ment, coarse materials were picked out from the soil, and
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World J Microbiol Biotechnol (2014) 30:1085–1092
20 g of soil were added into a column covered with gas
permeable plastic film. For the BC treatment, 1 g of bio-
char (i.e., a moderate application rate of 5 % w/w) was
added and mixed with the soil. Sterilized deionized water
was added to each column to bring the soil to 60 % of the
water holding capacity and the soil moisture was adjusted
every 6–7 days. Triple replicates were set up for each
treatment. The soil columns were incubated at 25 °C in
dark for 96 days. At the end of incubation, samples of the
BC and CK treatments were collected for the following
analyses. To assess the parent microbial phylotypes in the
soil, three original soil samples without incubation (deno-
ted by OS) were used for the following analyses.
DNA extraction
Each soil sample (0.5 g) of the OS, CK, and BC treatments
was frozen in liquid nitrogen and ground into fine powder
in a sterilized and precooled mortar. The powder was then
gently placed into a sterilized 15 mL centrifuge tube. The
total DNA was extracted from the soil powder using a
modified E.Z.N.A.TM Soil DNA kit (Omega Bio-tek., Inc.,
USA) according to the manufacturer’s instructions. Finally,
the DNA extracts were stored in 100 lL of elution buffer at
-20 °C for further studies.
PCR amplification of bacterial and fungal populations
For each soil DNA product, the 16S rRNA V3 genes of
bacteria (hereafter denoted by OSB, CKB, and BCB for the
OS, CK, and BC treatments, respectively) were amplified
using primer 27F and the reverse primer 1492R (Lane
1991). The ITS1-5.8S-ITS2 regions of fungi (hereafter
denoted by OSF, CKF, and BCF for the OS, CK, and BC
treatments, respectively) were amplified with the fungal
specific ITS1F and ITS4 primers (Gardes and Bruns 1993).
The PCR reaction mixture (50 lL) contained 1 9 PCR
buffer (Takara, Dalian, China), 2.5 U Taq DNA polymer-
ase (Takara, Dalian, China), 200 lM dNTP, 0.2 lM of
each primer, 3 mM MgCl2. The reaction conditions
included pre-denaturation at 94 °C for 2 min followed by
30 thermal cycles of denaturation, each at 94 °C for 30 s,
annealing at 56 °C for 30 s, and elongation at 72 °C for
1.5 min. The last step was extended at 72 °C for 5 min.
To confirm repeatability and minimize the PCR bias,
PCR products of the triple replicates for each treatment
were mixed together. The mixed PCR products were
checked by 1 % w/v agarose gel electrophoresis stained
with SYBR-Green I (Sigma, St. Louis, USA), based on the
band sizes of bacteria and fungi about 1.5 kb and 700 bp,
respectively. The bands were excised and purified using the
E.Z.N.A. gel extraction Kit according to the manufac-
turer’s instructions (Omega Bio-tek., Inc., USA).
World J Microbiol Biotechnol (2014) 30:1085–1092 1087

Cloning and sequencing Table 1 The proportions (%) of bacterial 16S sequences showing the
highest similarity (B 93 to 99 %) to GenBank database in clone
libraries from the original soil (OSB), unamended soil (CKB), and
The purified DNA products of each subset were ligated into
biochar amended soil (BCB)
the pMD 18-T Vector (Takara, Dalian, China) and the pMD
20-T Vector (Takara, Dalian, China) for fungal and bacterial Library 99 % 98 % 97 % 96 % 95 % 94 % B93 %
fragments, respectively, and transformed into the competent OSB 4 0 9 17 4 17 50
cells of Escherichia coli DH5a. The recombinants of colo- CKB 6 4 2 9 24 17 38
nies were identified through the blue-white color selection BCB 7 8 8 4 26 19 27
on Luria–Bertani (LB) agar plates supplemented with
ampicillin (100 lg mL-1; Sigma, St. Louis, USA), X-gal
(100 lg mL-1; Takara, Dalian, China), and IPTG (0.5 mM; Table 2 The proportions (%) of fungal 16S sequences, showing the
Takara, Dalian, China). The OSB, OSF, CKB, CKF, BCB, highest similarity (B 94 to 100 %) to GenBank database in clone
libraries from the original soil (OSF), unamended soil (CKF), and
and BCF clone libraries were separately constructed by
biochar amended soil (BCF)
picking 250 white clones randomly in each clone library.
Plasmid DNA was isolated from these selected clones Library 100 % 99 % 98 % 97 % 96 % 95 % B94 %
from each library and was reamplified by the vector primer OSF 5 66 4 9 2 2 12
pair RVM and M13-47 (Zhang et al. 2011). The PCR CKF 4 77 2 7 4 4 2
reaction mixture (50 lL) and conditions were used as BCF 0 80 6 0 0 3 11
above. PCR products of clones containing inserts of the
expected sizes were further analyzed by restriction frag-
ment length polymorphism (RFLP) analysis. PCR aliquots
(each of 10 lL) were characterized by digestion with the Table 3 Diversity indexes of bacterial and fungal species from the
unamended soil (CKB and CKF, respectively) and the biochar
restriction endonucleases HinfI and MspI for fungal frag- amended soil (BCB and BCF, respectively)
ments and EcoreI and HindI for bacterial fragments. The
digested products were classified based on the agarose gel Index CKB BCB CKF BCF
electrophoresis. Clones with different restriction patterns Shannon-wiener index 2.96 3.36 2.66 2.22
were randomly chosen for sequencing. Sequencing was Simpsons index 24.8 35.5 12.4 7.33
performed by Major Biotechnology, Ltd. (Shanghai, Chao I index 29.9 47.4 23.5 18.2
China). The sequences were examined for possible chi-
meras by the program DECIPHER’s Find Chimeras web
tool (http://decipher.cee.wisc.edu/FindChimeras.html) to (Good 1953). To quantify bacterial and fungal diversities,
remove chimeric sequences. the commonly used diversity indexes, including Shannon-
The sequences data in present study have been deposited wiener index, Simpsons index, and Chao I index, were
in the GenBank database. Clones from bacteria and fungi calculated using the program EstimateS.
were assigned accession numbers of KF22593–KF226104
and KF225791–KF225935, respectively.
Results
Sequences and phylogenetic analyses
Characteristics of libraries
To infer their approximately phylogenetic affiliations, the
bacterial sequences were compared initially with those in Total DNAs were extracted from the original soil (without
GenBank using EzBioCloud analysis (http://eztaxon-e. incubation), and unamended soil and amended soil with
ezbiocloud.net/), and the fungal sequences were com- biochar after incubation for 96 days. By using the bacte-
pared with those in GenBank using BLAST algorithm rium-specific PCR primer pair 27F-1492R and fungus-
(http://blast.ncbi.nlm.nih.gov/Blast.cgi) of the National specific primer pair ITS1F-ITS4, six libraries named as
Center for Biotechnology Information (Altschul et al. OSB, CKB, BCB, OSF, CKF, and BCF were constructed.
1997). Analyses of the diversity calculation and rarefaction Totally 169 clones for bacteria (53, 45, 71 in OSB, CKB,
curves to observed operational taxonomic units (OTUs) and BCB, respectively) and 145 clones for fungi (57, 52, 36
were carried out using the program EstimateS v.8.0 (http:// in OSF, CKF and BCF, respectively) were sequenced.
viceroy.eeb.uconn.edu/estimates/). The library coverage Library coverage values of OSB, CKB, and BCB were
values were calculated by [1-(n/N)], where n is the 0.81, 0.76, and 0.76, respectively, while the coverage value
number of OTUs representing a single clone and N is the of OSF, CKF, and BCF were 0.77, 0.85, and 0.81,
number of total OTUs representing the clones in the library respectively. The coverage value of each library was

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Table 4 Taxa of bacterial clone sequences from libraries of the original soil (OSB), unamended soil (CKB), and biochar amended soil (BCB)
Phylum OSB library CKB library BCB library
a b a b
Species N Species N Speciesa Nb

Proteobacteria Stella vacuolata 1 Pseudolabrys taiwanensis 6 Pseudolabrys taiwanensis 2

Rhodoplanes elegans
Methylosinus trichosporium
Bradyrhizobium lablabi
Shigella flexneri
Proteobacteria sp.
Steroidobacter denitrificans
Stella vacuolata
Rhodopila globiformis
Acidobacteria Solibacter usitatus
Koribacter versatilis
Acidobacteria sp. 1
Acidobacteria sp. 2
Actinobacteria Aciditerrimonas ferrireducens
Actinoallomurus purpureus
Conexibacter woesei
Conexibacter arvalis
Solirubrobacter ginsenosidimutans
Planctomycetes Gemmata sp.
Singulisphaera rosea
Verrucomicrobia Pedosphaera parvula
Firmicutes
Bacteroidetes
In the OSB, CKB, and BCB libraries, 53, 45 and 71 clones were randomly selected for sequenceing, respectively
The OTUs were defined by [ 97 % sequence similarity
a
The species in Genbank database are most closely related to the sequenced clones belong to the same OTU
b
The number of clones matched
c
Sequences showed the same most closely related species in Genbank database but belong to the different OTUs
7 Rhodoplanes elegans 4 Rhodoplanes elegans 5
1 Rhodoplanes piscinae 2 Methylocystis echinoides 2
1 Bradyrhizobium pachyrhizi 1 Methylosinus trichosporium 1
1 Bradyrhizobium denitrificans 2 Bradyrhizobium japonicum 2
4 Bradyrhizobium iriomotense 1
3 Gamma proteobacterium sp. 2 Bradyrhizobium rifense 1
1 Steroidobacter denitrificans 2 Bradyrhizobium pachyrhizi 2
1 Burkholderia graminis 1
Burkholderia tuberum 1
Burkholderia acidipaludis 1
Massilia namucuoensis 4
Nitrobacter vulgaris 1
2 Koribacter versatilis 4 Solibacter usitatus 2
5 Bryocella elongata 1 Acidobacterium capsulatum 2
5 Edaphobacter aggregans 2 Edaphobacter modestus 1c 5
1 Edaphobacter modestus 1c 1 Edaphobacter modestus 2c 2
c
Edaphobacter modestus 2 2 Edaphobacter aggregans 3
Granulicella arctica 1 Bryocella elongata 1
Telmatobacter bradus 1
Acidipila rosea 3
6 Aciditerrimonas ferrireducens 1c 1 Aciditerrimonas ferrireducens 5
2 Aciditerrimonas ferrireducens 2c 3 Actinoallomurus purpureus 3
4 Conexibacter arvalis 1 Actinoallomurus amamiensis 2
3 Mycobacterium cookii 1 Actinoallomurus spadix 1
1 Actinokineospora soli 1 Conexibacter arvalis 1
Mycobacterium kyorinense 1
Streptomyces misakiensis 1
Actinocorallia aurantiaca 1
1 Planctomycetales sp. 3 Singulisphaera rosea 6
2 Gemmata sp. 2 Aquisphaera giovannonii 2
Singulisphaera rosea 1
Pirellula sp. 1
1 Pedosphaera parvula 1 Pedosphaera parvula 2
Bacillus fumarioli 1
Chitinophaga ginsengisegetis 1
Niastella populi 1

approximately 0.8, suggesting that the libraries represented criteria are presented in Tables 1 and 2 for bacterial and
the major bacterial and fungal phyla in the soils. In fungal sequences, respectively. In general, fungal sequen-
the study, the OTUs were defined by [ 97 % sequence ces matched higher similarity to the sequences in Genbank
similarity. The proportions of phylotypes under matched database than bacterial sequences.

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Table 5 Taxa of fungal clone sequences from libraries of the original soil (OSF), unamended soil (CKF), and biochar amended soil (BCF)
Plylum OSF library CKF library BCF library
a b a b
Species N Species N Speciesa Nb

Chytridiomycota Kochiomyces dichotomus 1

Ascomycota Penicillium sp.
Aspergillus cervinus
Paecilomyces carneus
Hypocrea koningii 1c
Hypocrea koningii 2c
Chaunopycnis alba
Trichoderma gamsii
Pseudallescheria fimeti
Preussia sp.
Helotiales 1 RB-2011
Gloeotinia temulenta
Basidiomycota Cryptococcus podzolicus
Thelephoraceae sp.
Pseudozyma sp.
Leucocoprinus birnbaumii
Mortierellomycotina Mortierella humilis
Mortierella kuhlmanii
Mortierella sp.
Mortierella hyalina
Others Fungi sp.
In the OSF, CKF, and BCF libraries, 57, 52, and 36 clones were randomly selected for sequenceing, respectively
The OTUs were defined by [ 97 % sequence similarity
a
The species in Genbank database are most closely related to the sequenced clones belong to the same OTU
b
The number of clones matched
c
Sequences showed the same most closely related species in Genbank database but belong to the different OTUs
1 Penicillium sp. 1c 2 Penicillium herquei 1
1 Penicillium sp. 2c 2 Penicillium sp. 1c 1
c c
1 Penicillium pinophilum1 2 Penicillium sp. 2 1
1 Penicillium pinophilum2c 6 Penicillium pinophilum 2
1 Penicillium adametzii 1 Paecilomyces carneus 4
1 Penicillium olsonii 1 Trichoderma sp. 2
7 Paecilomyces carneus 1 Trichoderma koningiopsis 1
1 Aspergillus penicillioides1c 2 Trichoderma gamsii 5
1 Aspergillus penicillioides2c 2 Trichoderma viride 1
1 Aspergillus sp. 1c 1 Hypocrea sp. 1
4 Aspergillus sp. 2c 1 Cladosporium sp. 2
Chaunopycnis alba 1
Trichoderma gamsii 3
Trichoderma sp. 1
Cladosporium sp. 1c 2
Cladosporium sp. 2c 4
Sordariomycetes sp. 1
16 Cryptococcus podzolicus 12 Cryptococcus podzolicus 12
2 Wallemia sp. 5 Cryptococcus sp. 2
3 Sterigmatomyces sp. 2 Amanita spissacea 1
1
2
8
2
1
1

Bacterial and fungal diversities
For bacteria, the Shannon-wiener index values were dif-
ferent between the biochar-amended and unamended soils
(2.96 and 3.36, respectively). The values of Simpsons
Index and Chao I between the two treatments were also
different (Table 3). For fungi, the Shannon-wiener index
values of CKF and BCF libraries were 2.66 and 2.22,
respectively (Table 3). The index differences between the
two treatments for bacteria were larger than those for fungi.
The CKB library comprised 23 OTUs belonging to 18
genera, while the BCB comprised 35 OTUs belonging to
24 genera (Table 4). The CKF library included 20 OTUs
belonging to 11 genera and the BCF included 14 OTUs
belonging to 8 genera (Table 5). These results suggested
that the diversity of bacteria and fungi in the soil was
affected by the biochar addition differently. According to
rarefaction curves (Fig. 1), the separation of OTU richness
between different libraries was also obvious as the clone
number in each library was greater than about 20. Bacteria
in the biochar-amended soil had greater overall diversity
than in the unamended soil. However, fungi in the biochar-
amended soil had lower diversity than in the control.
Shifts in community composition of bacteria
Besides inducing changes in soil microbial diversity, the
biochar addition also affected the community composition.

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Fig. 1 Rarefaction analyses of (a) bacterial sequences from the
unamend (CKB) and biochar amended soils (BCB), (b) fungal
sequences from the unamend (CKF) and biochar amended soils (BCF)
For bacteria, both CKB and BCB comprised of Proteo-
bacteria, Acidobacteria, Actinobacteria, Planctomycetes,
and Verrucomicrobia. These phyla were all detected in
OSB, indicating that the major taxa in the treatments were
mainly related to the parent bacterial community compo-
sition in the forest soil. Bacteroidetes and Firmicutes were
only detected in BCB (Table 4). The proportion of Ac-
tinobacteria in BCB was 5.6 % higher than that in CKB,
whereas Proteobacteria and Planctomycetes were detected
with lower percentages (Fig. 2a). Although the proportion
of Proteobacteria decreased in the soil amended with
biochar, more taxa of Proteobacteria was detected in the
BCB library.
Shifts in community composition of fungi
Differences between fungal communities were found
mainly at the levels of genera and species (Fig. 2b). The
OSF library comprised of five phyla, but only Ascomycota
and Basidiomycota of these phyla were detected in CKF
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World J Microbiol Biotechnol (2014) 30:1085–1092
Fig. 2 Proportions of (a) bacterial phyla in the unamended (CKB)
and biochar amended soils (BCB), (b) fungal phyla in the unamended
(CKF) and biochar amended soils (BCF)
and BCF, which suggested that fungal growth of other
phylotypes might not adapt to the incubation conditions.
Compared to the control, the proportion of Basidiomycota
in the biochar-amended soil increased slightly. At the
genus level, Cryptococcus, Talaromyces, Cladosporium,
Trichoderma, Penicillium, and Paecilomyces were detected
in the CKF and BCF libraries. The proportion of Tricho-
derma in BCF was 14.5 % higher than that in CKF, and the
proportion of Paecilomyces in BCF was 9.2 % higher than
that in CKF.
Discussion
The results supported our hypothesis that biochar addition
affected bacteria and fungi differently in terms of diversity
and community composition during the short-term incu-
bation. After 3 months of biochar application, the overall
diversity of bacteria prominently increased. The results
were consistent with previous long-term studies in biochar-
enriched Terra preta soils (Kim et al. 2007; O’Neill et al.
2009). However, the diversity of fungi declined in the
biochar amended soil. The different diversity results sug-
gested that bacteria were more adaptive than fungi to the
World J Microbiol Biotechnol (2014) 30:1085–1092
change of soil environment induced by biochar (Lehamann
et al. 2011). Another possible reason for the different
results was that bacteria might be more readily to utilize
the nutrients and mineral elements by sorbing to biochar
surface or colonizing in biochar pores (Liang et al. 2008;
Thies and Rillig 2009). Both bacterial and fungal com-
munity structures altered markedly due to the biochar
amendment, with fungi mainly at the genus level and
bacteria at both genus and phylum levels. This may indi-
cate a higher sensitivity of bacteria to biochar in this soil
type. In our study, the bacterial diversity was different from
the previous study of forest soils amended with oak or
grass biochar (Khodadad et al. 2011). Possible reasons for
the inconsistent results might be related to the different
treatments of soils and the types of biochar (Lehmann
2007b), or to the diverse parent bacterial communities
between the two studies (Lehamann et al. 2011).
Most of bacterial sequences in both biochar-amended
and unamended soils matched those in the databases with
\ 97 % similarity, indicating that these soils contained
taxonomically novel bacterial phylotypes. The proportion
of bacterial sequences with [ 97 % similarity was higher
in BCB than that of CKB, and the proportion of sequences
with \ 93 % similarity was lower in BCB. These results
showed that the biochar treatment might not serve a good
habitat to novel bacterial population in the soil but sup-
ported the growth of original bacterial groups. On the
contrary, it was found that most of sequences in OSF, CKF,
and BCF were with [ 98 % similarity, and the proportions
of sequences with\ 94 % similarity in OSF and BCF were
12 and 11 %, respectively, higher than that in CKF. The
result indicated that the biochar addition had some
advantageous effects to ensure the growth of novel fungal
phylotypes in the soil.
Taxa analysis of bacteria showed that both relative
abundance and diversity of Actinobacteria increased with
the biochar treatment, which was consistent with the
studies on char layers of forest soils (Bääth et al. 1995),
Terra preta (O’Neill et al. 2009), and soils amended with
pyrogenic carbon (Khodadad et al. 2011). These results
demonstrated that Actinobacteria was the representative
species in recalcitrant carbon-rich soils. This is the first
report that the phylum of Bacteroidetes, which is com-
monly found in animal centeric canals and able to degrade
polysaccharose and cellulose, was detected in the soil
amended with biochar. Fungal taxa were altered mainly at
the genus level. In contrast to the findings of Jin (2010),
Zygomycota was not detected in this study. The difference
might be attributable to the different parent populations in
the two soil types or the different incubation time periods
(long vs. short). Both Trichoderma and Paecilomyces
showed high proportions in the biochar amended soil.
These two taxa of fungi have been widely studied and
1091
applied in biological control to promote plant growth and
enhance soil quality. Therefore, the biochar-induced
enhancement of such fungal groups may show its potential
effects in biological control. As very little is known about
changes of specific soil microbes to biochar addition
(Graber et al. 2010), to take full advantage of biochar as a
soil amendment, further research is necessary to focus on
biochar influences on targeted microbial groups with spe-
cific soil functions.
Acknowledgments This work was partly supported by grants from
the Chinese National Natural Science Foundation (Nos. 51039007
and 51179212).
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