Science of the Total Environment 649 (2019) 1467–1480

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Science of the Total Environment

journal homepage: www.elsevier.com/locate/scitotenv

The crucial factors of soil fertility and rapeseed yield - A five year field
trial with biochar addition in upland red soil, China☆
Zewen Jin a,1, Can Chen b,1, Xiaomin Chen a,⁎, Isaac Hopkins c, Xiaoling Zhang a, Zhaoqiang Han a,
Fei Jiang c, Grace Billy c
a
College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing 210095, the People's Republic of China
b
College of Applied Meteorology, Nanjing University of Information Science and Technology, Nanjing 210044, the People's Republic of China
c
Dept. of Ecosystem Science and Management, The Pennsylvania State Univ., University Park, PA, 16802, United States

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• The effects of 7 rates of biochar in an up-

land red soil were studied over 5 years.
• Soil pH and hydraulic property are
major influences on upland red soil fer-
tility.
• Biochar can improve rape yield by rais-
ing soil pH and water percolation and
retention.

a r t i c l e i n f o a b s t r a c t

Article history: Biochar has been used as an amendment to improve soil fertility and increase crop yield. However, the effects of
Received 5 July 2018 biochar on soil properties and rapeseed yield in upland red soil have not been thoroughly investigated, and the
Received in revised form 22 August 2018 factors crucial for rapeseed yield are not yet clear. A five-year field trial was conducted to investigate the effects
Accepted 29 August 2018
the of biochar (biochar application rates of 0, 2.5, 5, 10, 20, 30, and 40 t ha−1, respectively) on soil physicochem-
Available online 31 August 2018
ical and microbial properties as well as rapeseed yield in upland red soil in Jiangxi Province, China. Results
Editor: Baoliang Chen showed that biochar can significantly increase soil pH, available phosphorus, organic carbon, Ks, and water reten-
tion, however, the influences of biochar on these indexes declined over time. Soil total nitrogen increased signif-
Keywords: icantly when the dose of biochar exceeded 5 t ha−1, and the content of total nitrogen in the 40 t ha−1 biochar
Biochar treatment increased each year. While the application of biochar gradually increased the contents of NH+ 4 -N,
Soil nutrient contents NO− 3 -N and enhanced the soil microorganism and enzymatic activities during the first three years, they had
Soil hydraulic properties returned nearly to their starting values by the end of this study. Rapeseed yield and yield components were sig-
Soil microbial biomass and enzymatic activities nificantly improved relative to the control for all biochar amendments in the first year, but the rapeseed yield in
Rapeseed yield
all biochar treatments decreased steadily after 2012. According to the principal components analysis and path

☆ Project supported by the National Key R & D Program (2016 YFD 0200305), the Natural Science Foundation of Jiangsu Province, China (No. BK 20150909), State Key Laboratory of Soil
and Sustainable Agriculture (Institute of Soil Science, Chinese Academy of Sciences), grant number Y20160038, and China Postdoctoral Science Foundation (2016M591884).
⁎ Corresponding author.
E-mail address: xmchen@njau.edu.cn (X. Chen).
1
Zewen Jin and Can Chen have contributed equally.

https://doi.org/10.1016/j.scitotenv.2018.08.412
0048-9697/© 2018 Elsevier B.V. All rights reserved.
1468 Z. Jin et al. / Science of the Total Environment 649 (2019) 1467–1480
analysis, the most responsive parameters in the upland red soil were soil acidity and hydraulic properties, mean-
while, soil acidity and hydraulic properties had greater impacts on rapeseed yield than did other indexes. Taken
together, these results suggest that biochar can significantly improve soil fertility and rapeseed yield, but the im-
provements are not permanent. Soil acidity and hydraulic properties were the crucial factors that determined soil
fertility and rapeseed yield in upland red soil.
1. Introduction
Red soil (classified as Ultisols by the USDA) is widely distributed in
regions with abundant rainfall and high temperatures, such tropical
and sub-tropical South America, Africa, and Southeast Asia. It is the
most prominent and important soil resource in those areas (Li et al.,
2017). However, upland red soil has relatively low soil pH, poorly devel-
oped soil structure, a lack of available nutrients (especially soil available
P), and a single soil microbial community structure because of the
strong weathering, obvious aluminizing, torrential summer rains as
well as irresponsible cultivation (Xu et al., 2003; Ling et al., 2007;
Zhang et al., 2007; Six et al., 2004; Bronick and Lal, 2005). This combina-
tion results in poor productivity. Therefore, exploring effective ways of
improving soil fertility and developing sustainable agricultural systems
crop yield in upland red soil has important practical significance (Chen
et al., 2014).
Biochar is a carbon-rich material produced via pyrolysis of biomass
in an absence of oxygen at temperatures N 250 °C (Laird et al., 2009).
Soil fertility refers to the ability of a soil to sustain agricultural plant
growth (Kloss et al., 2014). Various studies have shown the beneficial
effects of biochar as an amendment to improve soil fertility with the
co-benefits of waste management, pollutant immobilization, crop pro-
ductivity increase and N2O emission reductions (Deenik et al., 2011;
Akhtar et al., 2014). Its strong alkalinity, well-developed pore structure
(Atkinson et al., 2010), large specific surface area and high physical-
biological stability allow biochar to ameliorate soil acidity (Yuan and
Xu, 2015), enhance soil porosity and tensile strength (Kinney et al.,
2012; Yanai et al., 2007), and increase soil saturated hydraulic conduc-
tivity (Ks) (Herath et al., 2013), soil water holding capacity, (Laird
et al., 2010a; Laird et al., 2010b), and soil available water content
(Atkinson et al., 2010).
Evidence shows that the application of biochar can play a significant
role in increasing soil organic carbon (SOC), total nitrogen, available
phosphorus, and ammonium (Hurni et al., 2015); controlling the rates
of nitrogen cycling and increasing the nutrient retention by reducing
the emission of N2O and the losses of nitrogen. Furthermore, the appli-
cation of biochar to soils might be a practical method to stimulate soil
microbes, increase microbial biomass, and enhance soil enzymatic ac-
tivities (Thies and Rillig, 2009) by absorbing relatively large amounts
of substrate, which promotes microbial growth and reproduction
(Lammirato et al., 2011).
Studies have repeated reported long-term positive changes in bio-
mass and crop yields resulting from biochar application. According to
a meta-analysis, the mean yield increases with biochar addition is 18%
(Hagemann et al., 2017). Biochar's beneficial role in increasing crop
yields is often attributed to (a) its porosity and sorption capacity (Gul
et al., 2015); (b) its liming effects in acidic soils (Peng et al., 2011);
(c) its improvement of soil water holding capacity, pore-size distribu-
tion, and soil structure stability (Chan et al., 2007); (d) its high rate of
nutrient transformations and utilization efficiency (Mizuta et al.,
2004); and (e) its stimulation of microbial and enzymatic activity
(Bhattacharjya et al., 2016). Most researchers believe that there is a sig-
nificant correlation between overall improvement of soil qualities and
yield increases (Agegnehu et al., 2015). However, pin-pointing the
exact mechanisms of soil fertility that influence rapeseed yields has
proven challenging. Haefele et al. (2011) hypothesized the 16–35% in-
crease in rice yield that they observed was a result of improvements
© 2018 Elsevier B.V. All rights reserved.
in water retention and available K and P after biochar was added in an
extensive four-season field trial in Thailand and Philippines.
Bhattacharjya et al. (2016) found that biochars derived from pine
needles and lantana increased the grain yield of wheat by 6.2–24.2%,
and that increase in yield was strongly associated with more efficient
N and P uptake by grains, and soil enzymatic activities. Relative to con-
trols, biochar increased rice yield when applied along with N-based fer-
tilizers due to its high agronomic N use efficiency (Huang et al., 2013).
While the immediate benefits of biochar have been well established,
the durations of biochar studies are often too short to determine the
long-term effects of biochar on crop yield, and laboratory tests con-
ducted over short time periods are of limited use in marking recom-
mendations to farmers who are looking for a reasonable plan for
biochar use (e.g. biochar dose rate, fertilizer/pesticide management,
etc.).
Thus, the objective of this study was to evaluate the effects of biochar
on soil fertility and rapeseed yield over a six-year period. Principal com-
ponents analysis (PCA) was used to investigate the most susceptible
variances; path analysis (PA) was used to determine the contributions
of those factors to rapeseed yield, and hierarchical cluster analysis
(HCA) was used to sort a set of data with similar characteristics into
groups (Dai et al., 2014). We used multivariate statistical techniques
to identify the most susceptible soil property parameters that were
most responsive to biochar application and to identify the factors
most critical to rapeseed yield.
2. Materials and methods
2.1. Study site
The field experiment was conducted from Sept. 2011 to Dec. 2016 at
the Institute of Red Soil (28°37′ N, 116°26′ E), in Jinxian County, Jiangxi
Province, China. The site is located in a flat area with an elevation of
26 m above sea level. The climate is typical of subtropical monsoon re-
gions, with distinct wet (March–June) and dry (July–September) sea-
sons. The average annual precipitation is 1549 mm, with 61–69%
falling between March and June. The average annual evaporation is
1100–1200 mm, of which 40–50% occurs during the dry season. The av-
erage annual temperature is 17.5 °C, with the lowest monthly average
temperature dropping to 5.1 °C in January and the highest monthly av-
erage temperature reaching 29.8 °C in July. The average annual sunshine
and the frost-free durations are 1900–2000 h and 282 d, respectively.
2.2. Biochar
Biochar was produced by pyrolysis of wheat straw at 450 °C in a ver-
tical kiln made of refractory bricks at Sanli New Energy Company in
Henan Province, China. About 35% of the wheat straw dry matter was
converted to biochar during the pyrolysis process. Detailed procedures
of the biochar production were reported by Pan et al. (2011). The bio-
char had an initial pH of 10.35, total N of 5.9 g kg−1, total P of
14.43 g kg−1, total K of 11.5 g kg−1, SOC of 467.2 g kg−1, and cation ex-
change capacity (CEC) of 21.7 cmol kg−1. To ensure a uniform mix with
the soil mass, the biochar was ground up to pass through a 2 mm sieve
before being incorporated into the soil.
 Z. Jin et al. / Science of the Total Environment 649 (2019) 1467–1480
2.3. Field experiment
Seven biochar application rates (0, 2.5, 5, 10, 20, 30, and 40 t ha−1)
were labeled as control (CK), C1, C2, C3, C4, C5 and C6, respectively.
Each trial plot was 20 m2 (4 m × 5 m), and all 21 plots (3 × 7) were ar-
ranged in a randomized complete block design with three replicates.
Biochar was spread uniformly on the surface of the red soil and then
mixed into the soil with spades to a depth of approximately 15 cm on
September 30th, 2011. The control plots were mixed in the same man-
ner as the other plots to maintain consistency, though without the addi-
tion of biochar. No further biochar was added for the duration of the
study. Two crops, rape and sweet potato, were grown in an annual rota-
tion; rape was planted in October and harvested in mid-May, and the
sweet potato was planted in late May and harvested in late September.
During the rape season, nitrogen fertilizer in the form of urea was
applied at a rate of 90 kg N ha−1, of which 40% was applied as a base fer-
tilizer prior to the seeding season and another 60% during the blossom
period. During the sweet potato season, the same dose of nitrogen fertil-
izer was broadcast before transplanting. Calcium superphosphate and
potassium chloride were also applied as basal fertilizers before
transplanting at a rate of 52.5 kg P2O5 ha−1 and 107 kg K2O ha−1 during
both rape and sweet potato seasons.
2.4. Soil sampling and measurement
Soil samples were collected each April during the rapeseed harvest
seasons from 2012 to 2016. Five replicated, undisturbed soil samples
were taken from each plot at depths of 0–15 cm using metal rings that
were 6.6 cm in inner diameter and 4.1 cm in height, after removing all
visible plant residues. The samples were mixed to form one representa-
tive sample for each plot. All soil samples were stored at 4 °C and
transported to the laboratory in aluminum containers within two
days. Basic soil nutrient contents were determined using the methods
suggested by Lu (2000). Soil pH (H2O) was determined with a glass
electrode in a solution of 5 g of soil in 25 ml of water. SOC was measured
by K2Cr2O7–H2SO4 oxidation (Lu, 2000). Total N was determined using a
TruSpec CN analyzer, and available P was measured by treatment with
0.5 mol l−1 NaHCO3 followed by molybdenum blue colorimetry (Lu,
2000). Bulk density was determined by extracting in 100 cm3 stainless
steel bulk density rings, drying at 105 °C for 24 h, correcting the weight
for any grains N 2 mm in diameter. The basic properties of the soil are
given in Table 1.
2.5. Soil microbial biomass carbon, nitrogen and enzymatic activities
The microbial biomass carbon (MBC) and microbial biomass nitro-
gen (MBN) were estimated by the chloroform fumigation-incubation
method (Jenkinson and Powlson, 1976). Urease activity was deter-
mined by spectrophotometry at 578 nm as the NH+ 4 –N released from
1.0 g of soil after a 24-h incubation at 37 °C with a 10% (w/v) urea solu-
tion in 20 ml of 1 M citrate buffer at pH 6.7 (Kandeler and Gerber, 1988).
Catalase activity in the soil was measured based on the back titration re-
sidual H2O2 with 0.1 M KMnO4. For invertase, 5 g of soil; 15 ml 8% of su-
crose solution, 5 ml phosphate buffer, and 0.25 ml toluene were
incubated in 50-ml Erlenmeyer flasks at 37 °C for 24 h. The mixture
was filtered through filter paper. After stopping the reaction with 3 ml
Table 1
Basic physical and chemical properties of the soil.
pH SOC TN
(g kg−1) (g kg−1)
Soil 4.24 ± 0.21 9.45 ± 0.86 1.06 ± 0.21
SOC: soil organic carbon.
TN: total nitrogen.
CEC: cation exchange capacity.
1469
of 3, 5-Dinitrosalicylic acid, the activity was measured at a wavelength
of 508 nm (Guan, 1986).
2.6. Saturated hydraulic conductivity and soil water characteristic curves
The constant-head method was employed to determine Ks (Wang
et al., 2008). Soil water characteristic curves were measured using the
Pressure-Plate Apparatus method (Richards and Fireman, 1943) with
sample dimensions (D = 5 cm, H = 2 cm). Key parameters, such as
field capacity (soil water content at 0.3 × 105 Pa), permanent wilting
point (soil water content at 1.5 × 106 Pa), and available water content
were recorded. The saturation moisture content (SMC) of the soil was
determined by saturating the undisturbed soil samples for 24 h (Sun
and Lu, 2014).
2.7. Harvest
Before harvest, five plants were collected from each plot in order to
determine rape yield components, including plant height, productive
tillers per plant, pod numbers per plant, biological yield, and rape
yield. To determine the total biological yield (t ha−1), the plants from
each plot were harvested, sundried (until constant weight) and
weighed.
2.8. Statistical analysis
An SPSS 20.0 statistical package was used to perform statistical anal-
ysis of the data. One-way ANOVA test was used to evaluate the effects of
different biochar application rates on the measured parameters, and the
differences between the treatments means were compared using least
significant difference testing (LSD) at p b 0.05.
3. Results
3.1. Soil chemical properties response to biochar addition
The responses of soil chemical properties including pH and nutrient
contents are presented in this study (Fig. 1). Overall, biochar had a sig-
nificant effect on soil pH in the first year when the addition of biochar is
N10 t ha−1. Relative to CK, the greatest increase in pH (0.53 units) was
observed for C6 treatment, followed by C5 (0.43 units) in 2012
(Fig. 1a). While, the influence of biochar on soil pH weakened over
time, with the pH value in the control and the C6 treatments decreasing
from 4.78 to 4.34 and from 5.31 to 4.66, respectively, over this five-year
period. The highest soil pH, 5.31, was found in the C6 treatment in 2012.
The addition of biochar to the soil resulted in significant increases in
soil nutrient content. Soil available P content was significantly (p b 0.05)
increased by all biochar treatments in the 2013 and 2014 sampling, but
not by the end of the trial period. The peak value of available P about
41.89 mg kg−1 was observed in the C6 treatment in 2012. However,
the influence of biochar on available P paralleled the pattern of soil pH
and gradually decreased with time in the biochar-amended plots
(Fig. 1b). With a few exceptions, the application of biochar significantly
increased (p b 0.05) the soil total nitrogen when the application of bio-
char exceeded 20 t ha−1 (Fig. 1c), and the difference between C5 and
the control gradually increased from 18.39% in 2012 to 27.92% in
CEC Bulk density Textural composition (%)
(cmol kg−1) (g cm−3)
Clay Silt Sand
15.2 ± 2.24 1.23 ± 0.13 31.6 ± 1.44 39.12 ± 1.78 29.28 ± 1.31
1470 Z. Jin et al. / Science of the Total Environment 649 (2019) 1467–1480

CK C1 C2 C3 C4 C5 C6 CK C1 C2 C3 C4 C5 C6
5.8 10 a

a d a a
a
a a
5.4 9

NH4+-N (mg kg-1)

ab a abab b a
b a a a bc b a a a
c b ab
ab ab ab
pH

5 c bc ab 8 b b b b b ab
c b
b
b b ab ab a a a a
c b b b b bc b
a b
c c a a c b
b b ab
4.6 b 7
c c c c

4.2 6
2012 2013 2014 2015 2016 2012 2013 2014 2015 2016

CK C1 C2 C3 C4 C5 C6
55
CK C1 C2 C3 a C4 C5 C6
b 15
Available P (mg kg-1)

a
45
e a
a a
abab 13 a
a a b
a a

NO3--N (mg kg-1)

ab a a
a a abab a
ab a
b ab ab ab b
35 b b c b
ab a 11 cd
bc b b a b bc
c c c
b a b b b
b b ab ab d d
b a a b
25 ab d c b b
c b b ab b ab 9

15 7
2012 2013 2014 2015 2016 2012 2013 2014 2015 2016

CK C1 C2 C3 C4 C5 C6 CK C1 C2 C3 C4 C5 C6
1.20 a 20
a a
c a a
a a 18 f
a a b
1.10 a ab ab
ab ab ab 16 a
TN (g kg-1)

ab
SOC (g kg-1)

a a a a
b b b a
b b b b 14 b
1.00 b b b c cc ab ab ab
b b
b c
b 12 b b
c c c c c b bc
c bc bc
c c c c c c c c c
0.90 10 c c c c

8
0.80 6

Fig. 1. Effect of biochar on soil pH (1a), available P (1b), total nitrogen (1c), ammonium (1d), nitrate (1e), and soil organic carbon (1f). Bars (standard deviation) followed by different
letters are significantly different at p b 0.05.

−
2016. NH+ 4 -N and NO3 -N are the primary forms of nitrogen taken up by after 2012, especially in the C5 and C6 treatments. The Ks in C6 de-
−
crops. Variation in biochar's effectiveness in boosting NH+4 -N and NO3 - creased N95% (from 8.44 to 0.40 × 10−4 cm s−1) within the five-year
N revealed an interesting pattern (Fig. 1d and e). In general, biochar period.
−
increased soil NH+4 -N and NO3 -N contents, and the effectiveness scaled The changes in saturated moisture content, field capacity, perma-
−
with the amount of biochar added, while, the NH+ 4 -N and NO3 -N nent wilting point, and available water content during the five-year pe-
contents in biochar-amended soil decreased after 2014. Changes in riod are shown in Table 2. Overall, biochar increased the soil water
SOC showed a threshold effect (Fig. 1f): SOC contents were significantly holding capacity. Averaged across samples, biochar application in-
(p b 0.05) higher at the high dosage of biochar (≥20 t ha−1) than those at creased the saturated moisture content and field capacity, respectively,
the low dosage of biochar (≤10 t ha−1). The highest SOC value, about from 0.4623 and 0.2219 cm3 cm−3 in the control to 0.5119 and
17.29 g kg−1, was obtained in C6 treatment in 2012. However, the influ- 0.2678 cm3 cm−3 in the treatment amended with 40 t biochar ha−1.
ence of biochar on SOC weakened over time, with the SOC in the C6 The available water content declined under each treatment from 2012
treatments decreasing from 17.29 g kg−1 to 12.69 g kg−1 over this to 2016, but remained significantly higher (p b 0.05) in the C6 than in
five-year period. the control. Given the influence of soil bulk density on these hydraulic
properties, it came as no surprise that the effects of biochar on soil sat-
3.2. Soil hydraulic property responses to biochar addition urated water content, field capacity, and available water content weak-
ened over time. Only the C5 and C6 treatments showed significantly
Biochar had a small but significant effect (p b 0.05) on soil bulk den- increased field capacity and available water content by 2016. Finally,
sity (Fig. 2a) with the greatest effect (a decrease in bulk density of there were no significant changes in the wilting coefficient among all
0.43 g cm−3) observed in 2012 in the C6 treatment. While, the influence treatments within the five-year period trial.
of biochar on soil bulk density weakened over time, only the C5 and C6
treatments still showed a significant lower bulk density in 2016 (p b 3.3. Soil microbial biomass and enzymatic activities response to biochar
0.05). In 2012, there were sharp increases in Ks in all biochar treat- addition
ments, and the C6 treatment significantly increased (p b 0.05) Ks by
8.08 × 10−4 cm s−1 (Fig. 2b). Although C1 and C2 had higher Ks than The average values of the samples showed that soil microbial bio-
the control, the differences were not significant in 2013 and 2014 (p N mass and enzymatic activities in biochar-amended treatments in-
0.05). In addition, Ks in biochar-amended treatments decreased sharply creased until 2014, and then decreased within the five-year period
 Z. Jin et al. / Science of the Total Environment 649 (2019) 1467–1480 1471

1.60
CK C1 C2 C3 C4 C5 C6
a
a a ab

Bluk density (g cm-3)

1.40 a
b a a a
a a ab a a
ab
bc
a a a a
b b b b b
1.20 b b b
bcbc b bc
c c c
1.00 d

0.80
2012 2013 2014 2015 2016

10 CK C1 C2 C3 C4 C5 C6

a
b
8

b
c
Ks (10-4 cm s-1)

6
a

4 b a
d

b
2
e c c c c a
ef
d d d b ab
f d d d b b b b a a a a a a a
0

Fig. 2. Effect of biochar on soil bulk density (2a) and saturated hydraulic conductivity (2b). Bars (standard deviation) followed by different letters are significantly different at p b 0.05.

trial. Soil microbial biomass nitrogen significantly increased (p b 0.05) 10 t ha−1 significantly increased (p b 0.05) rapeseed yield compared
when the addition of biochar was N10 t ha−1 within the five-year period to the control, while, there was no statistical difference in rapeseed
trial (Fig. 3a); the highest soil microbial biomass nitrogen concentration yield between the control and the C1 treatments in 2012 (p N 0.05).
of about 24.81 mg kg−1 was obtained in 2014. The soil microbial bio- Plant height, biological yield, and productive tillers per plant were sig-
mass carbon in C6 was markedly higher (p b 0.05) than the control, a nificantly higher (p b 0.05) in biochar plots than in the control plot,
difference of 15.91%, in 2014 (Fig. 3b). However, soil microbial biomass when the biochar application rates were higher than 10 t ha−1 in
carbon changed but was not sensitive to the biochar addition rate in any 2012. Furthermore, the number of pods per plant was significantly
given year. The C/N showed a significant decrease (p b 0.05) with bio- greater (p b 0.05) in biochar plots than in the control in 2012. However,
char addition, as C/N in C6 treatment decreased about 45.83% compared the rapeseed yield and yield components decreased year-by-year in
to the control in 2015. each treatment. In 2016, only the C5 and C6 treatments still significantly
Urease activity had the same time-series behavior as NH+ 4 -N and (p b 0.05) out-performed the control in terms of the rapeseed yield, the
NO− 3 -N (Fig. 4a), with the C3, C4, C5, and C6 treatments significantly in- pod numbers per plant, and the biological yield. The rapeseed yield in
creasing (p b 0.05) the urease activity by 33.19%, 39.2%, 44.58% and the C6 treatment was 1.11 t ha−1 in 2016, which was even lower than
65.20%, respectively, in 2012. While, the urease activity in biochar- the control yield in 2012.
amended soil decreased after 2014; the urease activity in C6 decreased
by 67.96% from 2014 to 2016. The activities of catalase and invertase in- 3.5. The key factors of soil fertility and rape yield
creased with biochar addition (C4, C5, and C6 treatments), but did not
show any regular with biochar addition rate in the same year (Fig. 4b The results of the PCA conducted for each soil treatment are shown
and c). Over time, catalase and invertase activity follow trends similar in Fig. 5, and the first three principal components (PCs) of each analysis
to urease activity. The highest activities of catalase and invertase, were used. These three PCs accounted for 82.19% of the total variance in
about 0.535 mg g−1 20 min−1 and 13.25 mg g−1 24 h−1, respectively, the original data for the upland red soil (Table 4), indicating that most
were obtained in the C5 treatments in 2014. information about soil fertility parameters was contained in the first
three PCs. PC1 explained 60.27% of the total variance, whereas PC2
3.4. Rapeseed yield and yield components response to biochar addition and PC3 explained 14.56 and 7.36%, respectively (Table 4).
Component score coefficients of the first three PCs are presented in
The effects of biochar amendment on rapeseed yield and yield com- Table 5. The component score coefficients explain the relative impor-
ponents are shown in Table 3. Overall, biochar had positive effects on tance of a parameter compared to other parameters in a single PC.
rape growth over the five-year period. Rapeseed yield varied among Thus, the component score coefficients whose absolute values are
the five experimental years, with the highest yield recorded in 2012 higher show the statistical importance of each PC. The results from
and the lowest yield recorded in 2016. Biochar application in excess of PC1 showed a preponderance of soil pH and hydraulic properties
1472 Z. Jin et al. / Science of the Total Environment 649 (2019) 1467–1480

Table 2
Saturated moisture content (cm3 cm−3), field capacity (cm3 cm−3), wilting coefficient (cm3 cm−3) and available water content (cm3 cm−3) with different biochar application rates from
2012 to 2016.

Year Treatments Saturated water content Field capacity Wilting coefficient Available water content

2012 CK 0.4653 ± 0.021d 0.2294 ± 0.011c 0.1260 ± 0.011a 0.1034 ± 0.014d

C1 0.4795 ± 0.024c 0.2373 ± 0.009c 0.1228 ± 0.009a 0.1145 ± 0.017c
C2 0.4762 ± 0.017c 0.239 ± 0.017c 0.1224 ± 0.010a 0.1166 ± 0.009c
C3 0.5259 ± 0.018b 0.2943 ± 0.019a 0.1288 ± 0.009a 0.1327 ± 0.006b
C4 0.5316 ± 0.032ab 0.2587 ± 0.025b 0.1271 ± 0.008a 0.1316 ± 0.012b
C5 0.5434 ± 0.036a 0.2579 ± 0.023b 0.1252 ± 0.013a 0.1655 ± 0.020a
C6 0.5424 ± 0.027a 0.2817 ± 0.026ab 0.1253 ± 0.012a 0.1564 ± 0.011ab
2013 CK 0.4637 ± 0.024c 0.2187 ± 0.017d 0.1268 ± 0.024a 0.0919 ± 0.007d
C1 0.4682 ± 0.019c 0.2264 ± 0.021 cd 0.1228 ± 0.021a 0.1036 ± 0.010d
C2 0.4681 ± 0.026c 0.2319 ± 0.023c 0.1234 ± 0.007a 0.1085 ± 0.008d
C3 0.5146 ± 0.027ab 0.2697 ± 0.031b 0.1266 ± 0.011a 0.1431 ± 0.011b
C4 0.5136 ± 0.031bc 0.2624 ± 0.014b 0.1257 ± 0.009a 0.1367 ± 0.011c
C5 0.5328 ± 0.033a 0.2516 ± 0.021bc 0.1252 ± 0.002a 0.1264 ± 0.012 cd
C6 0.5231 ± 0.018b 0.2932 ± 0.012a 0.1253 ± 0.014a 0.1679 ± 0.017a
2014 CK 0.4597 ± 0.022d 0.2172 ± 0.022d 0.1256 ± 0.012a 0.0916 ± 0.008d
C1 0.4669 ± 0.024c 0.2395 ± 0.028c 0.1252 ± 0.013a 0.1143 ± 0.014c
C2 0.4733 ± 0.025bc 0.2431 ± 0.021bc 0.1224 ± 0.008a 0.1207 ± 0.009bc
C3 0.5027 ± 0.029b 0.2265 ± 0.025 cd 0.1248 ± 0.012a 0.1017 ± 0.012d
C4 0.5026 ± 0.026b 0.2572 ± 0.011b 0.1283 ± 0.007a 0.1289 ± 0.012b
C5 0.4969 ± 0.018ab 0.2599 ± 0.019b 0.1278 ± 0.012a 0.1321 ± 0.016b
C6 0.5155 ± 0.011a 0.2781 ± 0.024a 0.1223 ± 0.015a 0.1558 ± 0.008a
2015 CK 0.4662 ± 0.023b 0.2212 ± 0.021b 0.1272 ± 0.006a 0.0940 ± 0.008c
C1 0.4613 ± 0.016b 0.2159 ± 0.025ab 0.1278 ± 0.013a 0.0881 ± 0.011c
C2 0.4795 ± 0.017b 0.2264 ± 0.018ab 0.1249 ± 0.008a 0.1015 ± 0.013b
C3 0.4821 ± 0.026a 0.2319 ± 0.023ab 0.1252 ± 0.013a 0.1067 ± 0.012b
C4 0.4716 ± 0.026ab 0.2307 ± 0.019ab 0.1221 ± 0.011a 0.1086 ± 0.009b
C5 0.4759 ± 0.021ab 0.2443 ± 0.011a 0.1238 ± 0.012a 0.1205 ± 0.016a
C6 0.4836 ± 0.018a 0.2467 ± 0.014a 0.1267 ± 0.017a 0.1200 ± 0.015a
2016 CK 0.4615 ± 0.019b 0.2232 ± 0.012b 0.1262 ± 0.007a 0.0970 ± 0.011b
C1 0.4612 ± 0.023b 0.2203 ± 0.017b 0.1248 ± 0.013a 0.0955 ± 0.014b
C2 0.4585 ± 0.016b 0.2309 ± 0.021ab 0.1273 ± 0.011a 0.1036 ± 0.012b
C3 0.4716 ± 0.017ab 0.2437 ± 0.022ab 0.1268 ± 0.014a 0.1169 ± 0.006a
C4 0.4853 ± 0.018a 0.2307 ± 0.018ab 0.1271 ± 0.009a 0.1036 ± 0.014ab
C5 0.4834 ± 0.022a 0.2393 ± 0.024a 0.1293 ± 0.017a 0.1100 ± 0.007a
C6 0.4822 ± 0.031a 0.2394 ± 0.019a 0.1214 ± 0.012a 0.1183 ± 0.011a

Different lowercase letters in the same column indicate significant differences (p b 0.05) among treatments.

(Table 5) (e.g., pH, Ks, saturation moisture content, field capacity and that all treatments could be separated into three distinct groups
available water content) over other parameters following biochar addi- (Fig. 7): Group 1 included control and C1 treatments; Group 2 included
tion. PC2 did the same for some soil nutrients, soil microbial biomass, C2, C3, and C4 treatments; and Group 3 included C5 and C6 treatments.
and enzymatic activities (e.g. total nitrogen, NO− +
3 -N, NH4 -N, pH, avail- The rape yield multiple linear regression model coefficients derived
able P, microbial biomass carbon and nitrogen, catalase activity, inver- from the results of PCA and PA are shown below, with their predictors in
tase activity and urease activity), and PC3 for SOC, wilting coefficient, order of importance based on their beta scores:
bulk density and C/N. Constant: B = −7.19;
Path analysis can reflect the direct and indirect effects of different Ks: B = 2069.23, β = 0.619;
factors and clarify the relationships that exist between characteristics. pH: B = 1.527, β = 0.433;
The results of the path analysis in Table 6 and Fig. 6 reveal that Ks, pH, SMC: B = 2.486, β = 0.382;
soil moisture content, field capacity, and available water content FC: B = 3.517, β = 0.379;
exerted the highest positive phenotypic direct influence on rapeseed AWC: B = 4.564, β = 0.322;
yield. Catalase activity, invertase activity, and total nitrogen had positive MBN: B = −0.045, β = −0.175;
but small direct effects on rapeseed yield. Conversely, soil organic mat- UA: B = 0.52, β = 0.154;
ter, bulk density, wilting coefficient and C/N exerted negative direct in- SOC: B = −0.083, β = −0.151;
fluence on rapeseed yield. Interestingly, soil nutrients, enzymatic MBC: B = 0.005, β = 0.141;
activities, and soil microbial biomass carbon and nitrogen had high pos- CA: B = 1.23, β = 0.087;
itive indirect effects through pH, Ks, field capacity, and available water The overall model fit was R2 = 0.937.
content; but negative indirect effects via soil organic matter, bulk den- Eq. (2) shows the derived regression equation:
sity, wilting coefficient and C/N. The decision coefficients are those
whose absolute values can be used to identify which variable is the prin- Yr ¼ −7:19 þ 2069:23 Ks þ 1:527 pH þ 2:486SMC þ 3:517FC
cipal decision variable and which variable is the restricted variable. Ac- þ 4:564AWC þ 0:005MBC−0:045MBN
cording to our results (Table 6), Ks has the greatest impact on rapeseed
yield (0.87), followed by pH, field capacity, saturated moisture content, þ0:52UA þ 1:23CA−0:083SOC ð2Þ
microbial biomass nitrogen, available water content and microbial bio-
mass nitrogen. Therefore, soil pH and hydraulic properties were more where SMC is saturated moisture content; AWC is available water con-
important indexes for predicting the rapeseed yield than other factors. tent; FC is field capacity; MBN is microbial biomass nitrogen; UA is urease
HCA was used to categorize all the biochar treatments into several activity; MBC is microbial biomass carbon; and CA is catalase activity.
groups with similar characteristics and to investigate the amelioration The model and the absolute beta scores show that the effects of soil
effects of the different application rates of biochar. The HCA revealed pH and hydraulic properties (e.g. Ks, SMC, AWC, and FC) on rape yield
 Z. Jin et al. / Science of the Total Environment 649 (2019) 1467–1480 1473

30 CK C1 C2 C3 C4 C5 C6

Soil microbial biomass nitrogen (mg kg-1)

a
25 a
ab
a
20 ab b a
b b c bc a
a
15 a b b a a a
c d cd b
ab a ab c b
ab
10 b b c c
c bc
c
5

0
2012 2013 2014 2015 2016

220
CK C1 C2 C3 C4 C5 C6
Soil microbial biomass carbon (mg kg-1)

a a a
200 b ab a
a
a ab ab ab
180 b b ab
b b abab b b b
b b
bc b bc b
160 c c
bc c c
d d
c
140 c

120

100
2012 2013 2014 2015 2016

30
CK C1 C2 C3 C4 C5 C6
a
c
25
ab
a a
b
20 a a
a a a
C/N

a c b
a a ab a a b
15 b cd b b b
b cd
bc ab d
b
c c b bc
10 c

Fig. 3. Effect of biochar on soil microbial biomass carbon (3a), microbial biomass nitrogen (3b), and C/N (3c). Bars (standard deviation) followed by different letters are significantly
different at p b 0.05.

were more significant than the influences of soil nutrients, soil microbial The model and the absolute beta scores show that the pod numbers
biomass, and enzymatic activities on rapeseed yield. per plant is the main controlling yield component of rape yield, followed
In addition, we attempted to find the multiple linear regression by biological yield.
model between rapeseed yield and yield components. The resulting These two models confirmed that high soil pH and good soil hydrau-
model coefficients are shown: lic properties can improve rape yield by increasing the pod number per
Constant: B = −0.269; plant.
PNPP: B = 0.0049, β = 0.508;
BY: B = 0.0053, β = 0.329; 4. Discussion
PTPP: B = 0.053, β = 0.108;
PH: B = 0.000223, β = 0.006.where PNPP is pod numbers per plant; 4.1. Biochar's effects on soil fertility and rapeseed yield
BY is biological yield; PTPP is productive tillers per plant; and PH is plant
height. The results of our research confirmed the effectiveness of biochar in
The overall model fit was R2 = 0.876. reducing soil acidity, enhancing soil hydraulic properties, and improv-
Eq. (3) shows the derived regression equation. ing soil nutrient contents, soil microbial biomass as well as enzymatic
activities. Several theories have been proposed to explain the mecha-
nism of biochar's role in improving soil pH and soil nutrient content.
Yr ¼ −0:269 þ 0:0049PNPP þ 0:0053BY þ 0:053PTPP The increase in the pH could be due to the alkaline materials such as
þ 0:000223PH ð3Þ wood ash to be used to produce biochar (Atkinson et al., 2010;
1474 Z. Jin et al. / Science of the Total Environment 649 (2019) 1467–1480

35 CK C1 C2 C3 C4 C5 C6
a a
a

Urease activity (mg NH4+-N g-1)

30
a a ab
a ab a
25 ab b b b
b b b abab
b
bc b bc a
20 c c c bc a
c c c ab a
15 b
bc
c
10

5
2012 2013 2014 2015 2016

0.6 CK C1 C2 C3 C4 C5 C6
a a
Catalase activity (mg g-1 20min-1)

b a a
a
0.5
ab
b
a b
a b a a
a a ab a a
0.4 ab
ab ab b c
bc b c c c b
b c c b
b
0.3 b

0.2
2012 2013 2014 2015 2016

CK C1 C2 C3 C4 C5 C6
14 a
a a ab a
Invertase activity (mg g-1 24h-1)

c
12 a
a ab b a
c c c
b
10
b ab
c a
b b
8 b ab
b b bc b
d b b b
bc
6 c c d
c

Fig. 4. Effect of biochar on urease activity (4a), catalase activity (4b), and invertase activity (4c). Bars (standard deviation) followed by different letters are significantly different at p b 0.05.

Clarholm, 1994; Mahmood et al., 2003). The basic cations from biochar A few investigations have reported on the effects of biochar on soil
and the negatively charged carboxyl, as well as phenolic functional total N under field conditions (Haefele et al., 2011; Laird et al., 2010a;
groups on the surface of biochar can also reduce the H+ concentration Mukherjee et al., 2014). In line with Borchard et al. (2014), the total N
in soil solutions. (Gaskin et al., 2010). content in biochar-amended soils was elevated relative to that in the
Biochar applications in soil have also been shown to have effects on control, which is known to promote N-immobilization by microorgan-
soil nutrient content. Soil pH is mainly responsible for controlling the isms (Atkinson et al., 2010; Sohi et al., 2010). One of the goals of biochar
fixation of P in biochar-amended soils, with the availability of P in acidic application is to stimulate the assimilation and transformation of N
soils strongly regulated by Fe-P (Han et al., 2016). Alkaline biochar can within the soil, instead of relying on exogenous nitrogen input (Darby
saturate Fe ions by increasing soil pH to enhance the effectiveness of P et al., 2016), and this effect was observed in our field trial. Our results
(Dume et al., 2017). Furthermore, biochar itself can be a potential P showed that biochar can increase the concentration of NH+ 4 -N and
source (Chintala et al., 2014). Zhao et al. (2013) found that the contents NO− 3 -N, demonstrating its ability to supply mineral N into soil and stim-
of P are high in crop biochar. Atkinson et al. (2010) also found that bio- ulate or change the intrinsic N cycling in the soil. Quilliam et al. (2012)
char contributed greatly toward soil P availability because of the high observed that re-addition of biochar can increase NH+ 4 -N concentra-
release rates of available P from biochar. tions by stimulating decomposition of organic matter. Laird et al.
SOC is a key metric of soil fertility (Andrews et al., 2004). As ex- (2010b) believe that the increase in NO− 3 -N content following the addi-
pected, the SOC content in biochar-amended soil was higher than that tion of biochar may be due to the fact that the negatively charged bio-
of the control. During the biomass pyrolysis process, the carbon in bio- char surface and high soil N mineralization were primarily responsible
mass becomes more aromatic in form, which increases the percentage for NO− 3 -N concentrations in soil.
of recalcitrant carbon in biochar (Liu et al., 2016). Depending on C stabil- Our experiment showed that biochar can decrease the soil bulk den-
ity, biochar can enhance SOC via either the release of biochar-derived sity, and increase Ks, saturated moisture content, field capacity, and soil
dissolved organic matter or by preserving the existing natural SOC available water content. These results are consistent with several previ-
(Al-Wabel et al., 2017; Aller et al., 2017). ous studies (Uzoma et al., 2011; Peake et al., 2014; Lim et al., 2016;
 Z. Jin et al. / Science of the Total Environment 649 (2019) 1467–1480 1475

Table 3
Biochar's effects on rape yield and yield components from 2012 to 2016.

Year Treatment Yield Plant height (cm) Productive tillers per plant Pod numbers per plant Biological yield (g plant−1)
(t ha−1)

2012 CK 1.4 ± 0.21d 136.33 ± 8.57c 7.66 ± 0.94c 101 ± 24.38e 83.33 ± 11.44d
C1 1.5 ± 0.16d 159.66 ± 12.81c 8.33 ± 1.24bc 144.66 ± 2.86d 122.66 ± 3.09d
C2 1.55 ± 0.17 cd 145.77 ± 3.75c 9.66 ± 0.94abc 167.33 ± 15.96d 104 ± 12.19d
C3 3.3 ± 0.18b 151.22 ± 17.54b 9.66 ± 1.24ab 241.66 ± 13.91c 166.66 ± 21.13c
C4 3.2 ± 0.19c 176 ± 7.78ab 10.33 ± 0.94ab 306 ± 15.29b 200 ± 17.68c
C5 3.95 ± 0.18a 188.66 ± 3.68a 11.66 ± 0.94a 371.33 ± 14.81a 241.66 ± 20.23b
C6 4.1 ± 0.18a 188.33 ± 12.11a 12 ± 0.81a 392 ± 7.34a 286.66 ± 30.34a
2013 CK 1.36 ± 0.22d 141.62 ± 19.15c 7.66 ± 0.47c 104 ± 13.88d 87.45 ± 4.32f
C1 1.25 ± 0.19d 139.33 ± 10.62c 8 ± 0.81bc 128.33 ± 9.97d 120.79 ± 4.8ef
C2 1.5 ± 0.14 cd 143.33 ± 25.1c 9.66 ± 0.47abc 155.66 ± 13.27 cd 111.61 ± 24.2de
C3 1.6 ± 0.18bc 149.33 ± 14.7bc 9.33 ± 0.47ab 202.66 ± 16.21c 148.53 ± 9.21 cd
C4 2.15 ± 0.16b 168.33 ± 9.74abc 9.66 ± 1.24ab 287.33 ± 32.96b 172.38 ± 18.93bc
C5 2.5 ± 0.15a 191 ± 8.64ab 10 ± 0.81a 289 ± 51.92b 200.96 ± 11.82ab
C6 2.35 ± 0.2ab 184.66 ± 18.19a 11 ± 0.81a 352.66 ± 6.59a 226.42 ± 14.55a
2014 CK 1.2 ± 0.17e 134 ± 14.51b 7.66 ± 0.94b 90 ± 1.41d 83.42 ± 4.15e
C1 1.6 ± 0.19 cd 134.66 ± 7.31b 6.33 ± 0.47ab 113.33 ± 7.4 cd 101.97 ± 9.41d
C2 1.6 ± 0.2 cd 133.33 ± 17.46b 7.66 ± 0.47ab 148.33 ± 31.2bcd 117.32 ± 7.76d
C3 1.55 ± 0.21d 156.66 ± 5.43ab 8 ± 1.41ab 179 ± 15.12bcd 134.86 ± 6.48c
C4 1.7 ± 0.18c 149 ± 3.26ab 8.33 ± 1.24ab 155.66 ± 29bc 153.48 ± 5.23b
C5 2.2 ± 0.19b 159.33 ± 6.59a 8.66 ± 1.69ab 206.33 ± 59.11b 168.11 ± 12.42b
C6 2.85 ± 0.17a 170.66 ± 10.53a 9 ± 0.81a 299 ± 59.75a 186.86 ± 5.5a
2015 CK 1.27 ± 0.19 cd 127.56 ± 5.5b 6 ± 0.81d 79 ± 9.09d 72.65 ± 8.53d
C1 1.11 ± 0.18d 131.44 ± 5.79b 6.33 ± 0.47 cd 91.33 ± 7.58d 84.32 ± 4.93 cd
C2 1.36 ± 0.21 cd 136.51 ± 11.22b 6.33 ± 0.47 cd 117.33 ± 12.91c 99.28 ± 9.33c
C3 1.34 ± 0.18 cd 149.78 ± 12.39ab 7 ± 0.81bcd 134.33 ± 13.47c 105.28 ± 9.5c
C4 1.43 ± 0.14c 148.36 ± 17.63ab 7.33 ± 0.47bcd 141 ± 7.25c 100.72 ± 3.19c
C5 1.87 ± 0.17b 153.78 ± 17.04a 8.33 ± 0.47ab 168.33 ± 11.89b 138.93 ± 14.34b
C6 2.12 ± 0.17a 165.71 ± 9.7a 8 ± 0a 218 ± 14.51a 167.12 ± 20.81a
2016 CK 0.61 ± 0.2c 109.84 ± 16.43d 5.66 ± 0.47c 78.33 ± 5.55c 72.81 ± 4.69b
C1 0.68 ± 0.18c 102.33 ± 12.18 cd 5.33 ± 0.94bc 78 ± 5.71c 77.88 ± 6.42b
C2 0.68 ± 0.11c 109.81 ± 6.23bcd 6 ± 0.81abc 84.66 ± 8.49c 83.53 ± 7.79b
C3 0.67 ± 0.2c 105.47 ± 8.7abcd 6.33 ± 0.47ab 85.33 ± 5.43c 84.12 ± 4.64b
C4 0.65 ± 0.17c 107.65 ± 11.6abc 6.66 ± 0.47ab 82 ± 8.04c 85.02 ± 6.44b
C5 0.88 ± 0.18b 116.61 ± 5.87ab 6 ± 0.81a 101.66 ± 8.95b 102.55 ± 7.05a
C6 1.11 ± 0.19a 113.81 ± 7.93a 6.33 ± 0.47a 110.33 ± 4.49a 113.97 ± 5.09a

Different lowercase letters in the same column indicate significant differences (p b 0.05) among treatments.

Atkinson et al., 2010; Mukherjee and Lal, 2013). Physical dilution effects
within the biochar-amended soils caused a reduction in bulk density,
which is in agreement with the findings of Busscher et al. (2011). In
line with Glaser et al. (2002) and Chen et al. (2010), biochar can change
the original conditions of soil pore structures and increase drainable po-
rosity because of its developed pore structure and large surface energy,
enabling it to absorb more water and improving soil water-holding ca-
pacity. Downie et al. (2009) observed that biochar contains extensive
macroporosity in the range of 1–10 μm in diameter, which can increase
water storage between −1,000,000 and −10,000 kPa. Our study

Table 4
Total variance explains.

Component Initial eigenvalues Extraction sums of squared

loadings

Total % of Cumulative Total % of Cumulative

variance % variance %

1 10.849 60.272 60.272 10.849 60.272 60.272

2 2.620 14.556 74.828 2.620 14.556 74.828
3 1.324 7.358 82.186 1.324 7.358 82.186
4 0.981 5.452 87.638
5 0.504 2.802 90.440
6 0.425 2.363 92.804
7 0.333 1.852 94.656
8 0.246 1.364 96.020
9 0.205 1.137 97.157
10 0.163 0.904 98.061
11 0.128 0.711 98.772
12 0.091 0.507 99.279
13 0.044 0.246 99.525
14 0.034 0.187 99.712
15 0.027 0.151 99.863
Fig. 5. Loading plot of PC1, PC2 and PC3. UA: Urease activity, SOC: Soil organic carbon, TN:
− − 16 0.015 0.083 99.946
Total nitrogen, NH+ +
4 : NH4 -N, NO3 :NO3 -N, BD: Bulk density, SMC: Saturation moisture
17 0.010 0.054 100.00
content, FC: Field capacity, PWP: Permanent wilting point, AP: Available P, MBC:
18 -4.2E-16 -2.3E-15 100.00
Microbial biomass carbon, MBN: Microbial biomass nitrogen, CA: Catalase activity, IA:
Invertase activity. Extraction method: principal component analysis.
1476 Z. Jin et al. / Science of the Total Environment 649 (2019) 1467–1480

4 -N, X7:NO3 -N, X8: Bulk density, X9: Saturation moisture content, X10: Field capacity, X11: Permanent wilting point, X12: Available water content, X13: Available
Table 5

coefficient

−0.20062
0.354367
0.303534
0.866848

0.044455
0.220108
0.089158
0.009867
0.264268
0.286069

0.178982
0.225374

0.035813

0.057229
−0.0209
Decision
Component score coefficients of the first three PCs.

0.26817

0.08185
0.2292
Original variables Component score coefficients

PC1 PC2 PC3

−0.007

−0.016
0.017
0.025
0.015
0.019
0.014
0.021
0.014

0.019
0.021
0.004
0.021
0.015

0.021

0.016
0.02
−0.060 −0.099

X18
pH 0.172
Ks 0.192 −0.134 0.103
−0.062

−0.042

−0.051
Saturated moisture content 0.155 0.036

0.029
0.056
0.009
0.035

0.046
0.061

0.031
0.036

0.025
0.017
0.054
0.062

−0.253 −0.035 0.044

Field capacity 0.134 −0.022 −0.008

0.05

0.01
X17
Available water content 0.136 −0.014 −0.059
Available phosphorus 0.107 −0.109 −0.097

−0.025
−0.047

−0.028
−0.038
−0.034
−0.041

−0.025
−0.032

−0.034
−0.016
−0.041

−0.267 −0.039
−0.01

−0.06
NH+

0.009
4 -N 0.029 0.086 0.038

0.02
X16
NO−3 -N −0.041 0.149 0.049
Microbial biomass carbon 0.312 0.813 0.136

−0.354
−0.306
−0.088
−0.403
−0.253
−0.353
−0.288

−0.371

−0.273
−0.303
−0.308
−0.102

−0.32
Microbial biomass nitrogen 0.226 0.931

0.171

0.026

0.338
X15
Catalase activity 0.095 0.801 0.141
Invertase activity 0.050 0.137 −0.075

−0.078
Urease activity 0.041 0.183 −0.114

−0.09
0.066
0.107
0.052
0.097
0.112
0.102
0.103

0.074
0.086

0.067
0.051

0.119

0.093
Total nitrogen −0.056 0.245 0.125

0.09
X14
SOC 0.089 −0.008 0.189

0
Bulk density 0.091 −0.092 −0.460

−0.049
−0.032
−0.045
−0.079

−0.022

−0.046
−0.044

−0.054

−0.018
−0.014

−0.026
−0.02
−0.04

−0.01
−0.011 −0.079

0.007

0.001

0.012
Wilting coefficient 0.516

X13
C/N 0.060 −0.206 0.243

0.2059
0.3645
0.2956
0.3099
0.3012
0.3262
0.3005
0.1692
0.2455
0.1849

0.2907
0.2075
0.2447
0.1946
0.3227
0.043

0.218
X12
demonstrated that application of biochar had no significant effect on the
permanent wilting point. Similar findings have been reported by Hardie

−0.083
−0.093
−0.005
−0.025
−0.028

−0.053

−0.004

−0.002

−0.069
−0.057
−0.057
−0.08
et al. (2014), who found that application of biochar at 47 Mg ha−1 had

0.034

0.086

0.007

0.034
X11
no significant effect on the permanent wilting point.

0
Biochar can increase the soil microbial biomass and enzymatic activ-

−0.1262

−0.1694
0.2888

0.2871
0.2159
0.2147
0.1749
0.1119

0.2224

0.2469
0.1969

0.1022

0.1423
0.2335
0.257

0.369

0.207
ities by creating microhabitats and introducting labile organic com-

X10
pounds for the growth of bacteria (Lehmann et al., 2011). A previous
study indicated that soil pH was the primary control over microbial ac-

−0.011

−0.011
0.026
0.021
0.026
0.025
0.016
0.025
0.016

0.028
0.005
0.028
0.027
0.016
0.014

0.011
0.019
tivity because pH controls microbial enzyme production, carbon and
X9

nutrient availabilities and the concentration of DOC (Tabatabai and −0.107

−0.011
−0.068
−0.036

−0.023
−0.055

−0.009

−0.005
−0.009
−0.004
−0.014
−0.012

−0.013
−0.006
Bremner, 1969). Our results confirmed these findings, with the opti-

−0.02

−0.06

0.008
mum value of pH for oxidoreductase being between 6.5 and 7.0

P, X14: Microbial biomass carbon, X15: Microbial biomass nitrogen, X16: C/N, X17: Catalase activity, X18: Invertase activity.
X8

(Frankenberger et al., 1983). The pH of biochar-amended soils was

−0.068

−0.007

−0.076
more suitable for the growth of microbes, especially for fungal hyphae,
0.064
0.082
0.045

0.096
0.085

0.067
0.075

0.076
0.053
0.088
0.094

0.086
0.057
0.08
X7

supporting the findings of Wuddivira et al. (2009). Other possible

mechanisms for the increase microbial biomass and enzymatic activities
−0.215
−0.111
−0.181
−0.126
−0.102

−0.107

−0.115
−0.122
−0.007
−0.122
−0.108
−0.098
−0.094

−0.074
−0.096
0.068

0.072
in biochar- amended soil could be the large surface area of biochar for
X6

soil microbial attachment, the porous structure of biochar to provide

−0.009

−0.006
protection for soil microorganisms, and the dissolved organic carbon

−
−0.233 0.005
−0.102 0.006
−0.219 0.005
0.009

−0.229 0.008
−0.293 0.009

−0.317 0.006
−0.221 0.007

0.007
−0.309 0.004
−0.096 0.009
−0.297 0.008

−0.058 0.006
−0.089 0.005
which can be used by microorganisms (Quilliam et al., 2012).
X5

−0.018 0

We observed that the effectiveness of biochar on soil pH, bulk den-

−0.114

sity, available P, SOC, Ks, saturated moisture content, field capacity,

−0.32
−0.139 −0.176 0.085
Indirectly pass through coefficient

X1: pH, X2: Urease activity, X3: Ks, X4: Soil organic carbon, X5: Total nitrogen, X6: NH+
−0.242 −0.093 0.06
and available water content started to fade after the first year. This tem-
X4

poral fading effect can probably be attributed to the decomposition of

0.207
0.311

0.308

0.285
0.123

0.129
0.184
−0.024 0.102
0.206

0.217
0.141

0.066
0.299
the biochar, itself, and leaching of the alkaline ashes in the humid sub-
0.25

0.13
X3

tropical climate with high rainfall (Glaser et al., 2002; Lehmann and
Rondon, 2006; Lian and Xing, 2017). Analogous to our study,
0.267

0.176
0.246
0.198
0.175

0.139
0.195

0.159
0.116
0.255
0.219

0.223
0.279

Cornelissen et al. (2018) tested two different biochars over five growing
0.23
X2

seasons in an Indonesian Ultisol, and found that biochar can increase

−0.083

−0.124

maize yield and soil pH, but that the effect faded after the third season.
0.254
0.174

0.142
0.172

0.185
0.223

0.175
0.209
0.149
0.134

0.182
0.16

0.17

0.11

Jones et al. (2012) found that after three years in the field, the alkalinity
X1

associated with the biochar had been fully neutralized, similar to our
pass by the
sum of the
coefficient

own observations, with the pH of the soil returning to 4.6.

−0.2127
Indirect

−0.133
0.1927
0.9085
0.3907
0.7398
0.8419
0.7161
0.3044

0.1549
0.1879

0.1949
0.3039
0.7287
0.6787

0.4269
0.9722
0.508

The physical and chemical degradation of biochar may trigger the

Path analysis of key factors of rape yield.

dissipation of the effects of biochar on soil hydraulic properties (Lian

and Xing, 2017). Biochar is likely to experience physical and chemical
coefficient
through

−0.151

−0.033

−0.021

−0.072

disintegration into smaller fragments including colloids and nanoparti-

Direct

0.433
0.154
0.619

0.026

0.122

0.382
0.379

0.322
0.217
0.141
0.175

0.087
0.029
coefficient pass

0.14

cles after multiple seasons (Wang et al., 2012). Doerr et al. (2000) found
that biochar will adsorb and fix inorganic ions and polar or nonpolar or-
ganic compounds over the course of several years, thereby forming or-
−0.266

−0.207
Factors Related

0.933**
0.679**
0.912**
0.806**

0.807**
0.448**

0.886**
0.834**

0.831**
0.918**
0.436**

0.568**
0.405*

0.077

0.297

0.274

ganic and inorganic composites and aggregates in the soil. This, may
decrease soil macropore space, resulting in the recovery of soil bulk
density and a decrease in soil water holding capacity. Prost et al.
Table 6

X10
X11
X12
X13
X14
X15
X16
X17
X18

(2013) found that the surface area and porosity of biochar decreased
X1
X2
X3
X4
X5
X6
X7
X8
X9
 Z. Jin et al. / Science of the Total Environment 649 (2019) 1467–1480 1477

Fig. 6. Network of path analysis. T1: pH, T3: Ks, T9: Saturation moisture content, T10: Field capacity, T12: Available water content, Y: Rapeseed yield. T indicates the direct path coefficient; P
indicates the indirect path coefficient.

over time, which undermines its effectiveness on soil saturated hydrau-
lic conductivity.
In our study, soil microbial biomass C, N, and enzymatic activities
showed an initial increase followed by a decrease for the high dose of
biochar treatment (C6) during the five-year study, and a similar trend
−
was identified in NH+ 4 -N and NO3 -N. Nielsen et al. (2018) found that
biochar can stimulate the internal processes of N assimilation and con-
−
version, as well as increasing the content of NH+4 -N and NO3 -N. How-
ever, the N stimulation by enhanced biochar may not be permanent:
nitrate levels in biochar treatments were no greater after 12 months
than treatments without any amendment. According to previous stud-
ies, the microbial decomposition of biochar in soils can be divided into
two stages: fast-degrading stage and slow-degrading stage (Ameloot
et al., 2013). The volatile fractions in biochar can be used as energy
sources for microbial communities and enhance the soil microbial bio-
mass, as well as enzymatic activities, in the fast-degrading stage
(Bhaduri et al., 2016). However, biochar may induce a negative priming
effect over a longer period of time, owing to the sorption of organic

Fig. 7. Hierarchical cluster analysis of upland red soils with different biochar application rates.
1478 Z. Jin et al. / Science of the Total Environment 649 (2019) 1467–1480
matter on biochar surfaces and/or into biochar-soil microaggregates
(Maestrini et al., 2015). Consistently, biochar has been observed induc-
ing a negative effect on soil microorganism and enzymatic activities
during later stages.
4.2. The key factors of rape yield
Biochar amendment can alter soil acidity and hydraulic properties in
ways that are favorable to agricultural productivity. The statistical tech-
nique of PCA was applied to explain the maximum information in all
variances by integrating the correlated original variables into a false var-
iable, the PA permits a critical examination of the specific factor that
produces a given correlation, they could be successfully employed in
formulating an effective selection strategy for biochar application. In
our research, the first principal component (PC1) accounted for
60.27% of the total variances and represented the great majority of the
soil properties in upland red soil. We concluded that the most suscepti-
ble parameters in the upland red soil were soil hydraulic and acidity
properties because of the high component score coefficients between
PC1 and soil pH, saturated moisture content, Ks, field capacity, as well
as available water content. The PA and linear regression equation of
soil properties and rapeseed yield showed that Ks, pH, soil moisture
content, field capacity, and available water content exerted the highest
positive phenotypic direct influence on rapeseed yield. The effects of pH
and soil hydraulic properties (e.g. Ks, saturated moisture content and
available water content) on crop yield were more significant than the
influences of soil nutrient content, soil microbial biomass, and enzy-
matic activity, which constitutes indirect evidence that the most suscep-
tible parameters in the upland red soil were soil hydraulic and acidity
properties following biochar addition. Low soil pH and aluminum toxic-
ity are two major obstacles that limit rape growth in upland red soil (Liu
et al., 2014). The exchangeable aluminum concentration in acidic soil
reaches its maximum values at pH ≈ 4.2 and declines as pH increases
(Gruba and Mulder, 2015). Aluminum inhibits the growth of crop
roots (Wang and Kao, 2007). A low soil pH also interferes with microbial
activity (Pietri and Brookes, 2008). As soil pH increases in response to
biochar application, microbial activity and the degree of nitrification
would increase and microbial factors would no longer be limited (Xu
et al., 2006). Biochar addition would then preferentially affect other pa-
rameters such as soil nutrients content. Thus, biochars could be utilized
to limit and reverse acidification in otherwise acidic soils (Yuan and Xu,
2015), allowing increased the rapeseed yield.
Soil hydraulic properties had a more significant impact on crop yield
according to the PCA and the linear regression equation compared with
the soil nutrient content, soil microbial biomass, and enzymatic activi-
ties. The nutrient content of the red soil is abundant, but the soil struc-
ture and hydraulic properties are poor. This “water problem” is mainly
reflected in its high bulk density, and in its low available water storable
capacity. The drought stress in July, August, and September is particu-
larly serious, the crop roots cannot easily penetrate through the soil to
absorb water because of the high bulk density. On the other hand, red
soil readily stores water in the deep soil, but such water is not available
for most plants, especially annual crops. Low Ks and available water
content limit the movement of moisture in the soil and the uptake of
water by crops (Xianliang, 1996). Lawes et al. (2009) found that there
was a significant positive correlation between available water content
and wheat yield. Thus, soil water infiltration and available water con-
tent become the crucial factors of soil fertility and rapeseed yield in re-
gions with red soil. Based on our results, biochar can increase soil water
holding capacity and prevent crops from dying by increasing field ca-
pacity and available water content.
Although the effects of soil nutrients and microbial and enzymatic
activities on rapeseed yield were not as good as those of soil hydraulic
properties, they also affected rape yield. It has been well-established
that biochar amendments can increase N and P availability to crops
and that high levels of SOC accumulation can enhance long-term N
efficiency and increase rice productivity in rice paddies (Pan et al.,
2011). According to the PA analysis, microbial biomass carbon and ni-
trogen also affect rape yield, but the influence is indirect through pH,
Ks, field capacity, and available water content. Soil microorganisms con-
tribute to the formation and stabilization of soil aggregates (Li et al.,
2017), which directly affects the soil's hydraulic properties. So, soil mi-
crobial biomass can affect rapeseed yield through soil hydraulic
properties.
In terms of yield components, the number of pods per plant is deci-
sive for rapeseed yield, followed by biological yield. The results of this
present study support the speculation by Diepenbrock (2000). During
the course of development, the pod number is ultimately determined
by the number of buds, branches, flowers, and young pods, all of
which are constrained by source capacity, the supply of nutrients and
water (Tayo and Morgan, 1979). External factors, such as the arrange-
ment of the stand, sowing date, and N fertilization also play a crucial
role in the control of pod number. We hypothesized that biochar
would continuously increase rapeseed yield because of its biological sta-
bility. Our results, however, showed that the rapeseed yield decreased
over time for each treatment. These results revealed directly that the re-
duction in pH, Ks, field capacity, and available water content were the
main causes of crop yield reduction in the biochar-amended soil.
5. Conclusions
The results presented here suggest that biochar can significantly en-
hance the soil pH, nutrient contents, hydraulics properties, soil micro-
bial biomass carbon, nitrogen and enzymatic activities. However, the
effects of biochar on these soil fertility indexes weakened over time.
Our results also warn that biochar can increase rapeseed yield, but it
loses potency with each year. The PCA, PA, and linear regression equa-
tion showed that soil pH and hydraulic properties have more significant
effects on soil fertility and rape yield than do soil nutrients, microbial
biomass, and enzymatic activities. The pod number per plant is the
main controlling yield component of rapeseed yield. We believe that
the soil hydraulic and acidity properties are crucial factors for soil fertil-
ity and rapeseed yield in upland red soil regions. Biochar can enhance
pod number per plant by enhancing soil pH and soil hydraulic proper-
ties, and then increasing rapeseed yield.
Acknowledgements
The authors thank Dr. Christopher Ogden and Dr. Melissa Miller for
their grammatical edits and comments on this paper. We also wish to
express our thanks to anonymous reviewers for providing useful com-
ments to improve the paper. This work was supported by National
Key R & D Program (2016 YFD 0200305), the Natural Science Founda-
tion of Jiangsu Province, China (No. BK 20150909), State Key Laboratory
of Soil and Sustainable Agriculture (Institute of Soil Science, Chinese
Academy of Sciences), grant number Y20160038, and China Postdoc-
toral Science Foundation(2016M591884).
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