Scientia Horticulturae 176 (2014) 70–78

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Scientia Horticulturae
journal homepage: www.elsevier.com/locate/scihorti

Biochar and humic acid amendments improve the quality of

composted green waste as a growth medium for the ornamental plant
Calathea insignis
Lu Zhang a , Xiang-yang Sun a,∗ , Yun Tian b , Xiao-qiang Gong a
a
College of Forestry, Beijing Forestry University, Beijing 100083, PR China
b
Institute of Desertification Studies, Chinese Academy of Forestry, Beijing 100091, PR China

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

Article history: The effects of different percentages of biochar (BC) (at 0, 20, and 35%) and/or humic acid (HA) (at 0,
Received 30 October 2013 0.5, and 0.7%) on the physical and chemical characteristics of composted green waste (CGW) and on the
Received in revised form 9 May 2014 growth and nutrition of the ornamental plant Calathea insignis were investigated. The highest quality
Accepted 18 June 2014
growth medium and the best C. insignis growth were achieved when the CGW was amended with the
Available online 17 July 2014
combination of 20% BC and 0.7% HA; the lowest quality medium and the least growth were obtained
with nonamended CGW. The optimum combination not only improved the particle-size distribution and
Keywords:
adjusted the bulk density (BD), porosity, and water-holding capacity (WHC) into ideal ranges but also
Coarseness index
Microbial biomass
decreased pH and electrical conductivity (EC) and increased macro- and micro-nutrient contents and
Particle-size distribution microbial biomass C and N of the growth media. In comparison with the 100% CGW, shoot fresh weight,
Photosynthetic pigment shoot dry weight, root fresh weight, root dry weight, plant height, crown breadth, number of leaves, and
total root length obtained with the optimal media were increased by 57.3, 79.7, 64.5, 82.0, 45.2, 89.1,
31.1, and 94.1%, respectively, and total nitrogen, total phosphorus, total potassium, and total chlorophyll
contents of plant leaves were increased by 66.4, 55.8, 72.1, and 47.8%, respectively.
© 2014 Elsevier B.V. All rights reserved.

1. Introduction Biochar (BC) is a solid, charred, and carbon-rich residue derived

Most potted, ornamental plants are grown in soilless media in
which peat has traditionally been the main component (Tullio et al.,
2012). In recent years, environmental and economic concerns have
caused growers to seek alternatives to peat, and compost has been
frequently considered as a substitute for peat in the media used
to produce potted, ornamental plants (Cai et al., 2010; Jayasinghe,
2011). However, many compost products on the market have an
inappropriate pH and soluble salt content, a low nutrient content,
and a low cation exchange capacity (CEC), resulting in poor plant
performance (Medina et al., 2009; Zhang et al., 2013a). For these
reasons, compost may require organic amendments to improve
their ability to support ornamental plant production (Jayasinghe,
2012a; Molineux et al., 2009).
∗ Corresponding author. Tel.: +86 13366950662/+86 01062338103;
fax: +86 01062338103.
E-mail addresses: zhanglu1211@gmail.com (L. Zhang), sunxy@bjfu.edu.cn
(X.-y. Sun), bjxtty@gmail.com (Y. Tian), styybl@163.com (X.-q. Gong).
from the pyrolysis at a relatively low temperature (<700 ◦ C) of
organic materials like wood, grass, manure, or agricultural residue
(Lehmann and Rhodon, 2006). BC has attracted widespread atten-
tion as a growth medium amendment that enhances CEC, nutrient
retention, and water retention, and that neutralizes acidity (Karami
et al., 2011; Lehmann and Rhodon, 2006; Sun et al., 2012). Further-
more, the addition of BC to growth media or soil represents a form of
long-term carbon sequestration (Dias et al., 2010; Dumroese et al.,
2011; Hartley et al., 2009).
The supply of available nutrients for potted plants mainly
depends on the nutrient content of the growth medium itself. If
the nutrients in the medium are insufficient, they can be increased
by adding fertilizer or other amendments (Hartwigsen and Evans,
2000). One method of adding nutrients is to add humic acid (HA).
The positive effects of HA on plant growth have long been recog-
nized (Pal and Biswas, 2005). HA has been extensively studied as
an amendment to plant growth media including soil and compost
(Senn and Kingman, 1973). HA is a major component of organic
matter and has both direct and indirect positive effects on plant
growth and health. The direct effects result from the uptake of HA
by the plant. The indirect effects result from the improvement of

http://dx.doi.org/10.1016/j.scienta.2014.06.021
0304-4238/© 2014 Elsevier B.V. All rights reserved.
 L. Zhang et al. / Scientia Horticulturae 176 (2014) 70–78
growth media properties, such as aggregation, aeration, perme-
ability, and nutrient transport and availability (Mohamed, 2012;
Salman et al., 2005). Positive effects of HA addition on plant growth
have been reported for oriental lily (Chang et al., 2012), maize
(Hanafi and Salwa, 1998), and watermelon (Salman et al., 2005). The
positive effects of HA on root initiation and growth, shoot devel-
opment, and the uptake of macro- and micro-elements by plants
have been summarized by Chen and Aviad (1990) and Varanini and
Pinton (1995).
Although research has indicated that BC or HA improve growth
media characteristics and plant growth, the combined effects of
BC and HA addition on the growth of the potted ornamental plant
Calathea insignis in composted green waste (CGW) have not been
reported. C. insignis, which belongs to the Marantaceae or arrow-
root family, is a perennial evergreen herb that is native to Brazil and
other countries of tropical America. The plant generally requires
acidic growth media or humic soil. It also requires a growth medium
that has high porosity, that is well-drained but retains sufficient
water, and that is rich in nutrients (Zhang et al., 2013b). C. insig-
nis is a very popular ornamental in China, where it is used indoors
and is highly valued for its beautiful foliage. The plant is also an
important source of arrowroot starch. With the rapid growth of the
population in China, the research on and development and use of
starch plants have increased. C. insignis is known for the high qual-
ity of starch in its tubers and roots and its low-cost of production
(Tran et al., 2013). Moreover, the aerial part of C. insignis and the
residues after starch extraction can be used as the animal feed or
as raw material by the paper industry.
The aim of the current research was to investigate the influences
of BC and/or HA addition, in various proportions, on the physi-
cal, chemical, and microbial characteristics of the CGW and on the
growth, nutrient content, and photosynthetic pigment content of
the ornamental plant C. insignis.
2. Materials and methods
2.1. Material preparation
The compost (CGW) used in this experiment was made from
green waste by a two-stage composting method (with primary and
secondary fermentations) and with addition of fermentation addi-
tives (brown sugar and calcium superphosphate) as described by
Zhang et al. (2013a). The green waste mostly consisted of fallen
leaves and branch cuttings collected during greening maintenance
in Beijing, China in the spring of 2012. Before composting, the green
waste was reduced to 1-cm particle size, and then the moisture
content was adjusted to 60% and the C/N ratio was adjusted to
between 25 and 30. After that, the raw material was subjected
to primary fermentation (PF) followed by secondary fermentation
(SF). The PF occurred in a digester cell. The materials in the digester
cell were turned every day, and the moisture content was checked
once every day; water was added to maintain the water content
at 60%. During the PF, the fermentation temperature increased to
a maximum of 57 ◦ C on day 3. When the temperature dropped to
45–55 ◦ C on day 6, the PF was considered complete. Then, materi-
als were removed and arranged in windrows (2 m long, 1.5 m wide,
and 1 m high) for the SF. Windrows were turned manually every 3
days for aeration, and water was added during the turning process
as needed to maintain the moisture content (60%). The SF included
the combined addition of 0.5% brown sugar and 6% calcium super-
phosphate every 6 days beginning on day 6. The second highest
temperature during the SF was 60 ◦ C on day 15, which was higher
than the maximum in the PF. The composting process was consid-
ered complete when the fermentation temperature was similar to
that of the surrounding atmosphere. In the two-stage composting,
71
the thermophilic periods (50–60 ◦ C) last for 4 days in the PF and for
12 days in the SF. The production of mature compost requires only
30 days.
In the present study, CGW was mixed with different quantities of
BC and/or HA. BC was made from coir (coconut husk fiber) and was
produced at approximately 450 ◦ C in a low-temperature pyrolysis
facility. Before BC was used in the study, it was passed through a
2.0-mm soil sieve. The HA consisted of black, regular, and water-
soluble granules, and the pH and EC values of HA were 5.04 and
0.21 mS cm−1 , respectively. HA was dissolved in water before it was
added to the CGW. The HA used in this study was purchased from
the Beijing Kaiyin Organic Fertilizer Production Co., Beijing, China.
2.2. Greenhouse experiment
An experiment was carried out to investigate the effects of
adding different proportions of BC and/or HA to CGW on the proper-
ties of the potting medium and on the growth and nutrient content
of C. insignis. BC and/or HA were added to the CGW as indicated
by the nine treatments described in Table 1. The experiment was
conducted in a controlled-temperature greenhouse at the Shunyi
District Nursery of the Beijing Green Garden Group (Beijing, China)
from 31 March 2012 to 31 October 2012. The greenhouse temper-
ature was 20–25 ◦ C in the day and 18–20 ◦ C at night, with 65–75%
relative humidity and 7000 lx light intensity.
First, the different quantities of BC and CGW needed for each
treatment were mechanically blended. Second, the different quan-
tities of HA required for each treatment were mixed thoroughly
with 2 L of water, and the diluted HA was then evenly sprayed on the
corresponding treatments according to Table 1. Then, the mixtures
were mixed again and air dried. The experiment used 2000 cm3
plastic pots with holes in the bottom. As described by Jayasinghe
(2011), the air-dried growth media (T1–T9) were added to each
pot (leaving a distance of 1 cm from the top of the pot) without
compaction. Each of the nine growth media was represented by
30 replicate pots. Altogether, there were 270 pots in the experi-
ment, which were arranged in a completely randomized design on
the benches in the greenhouse. The pots were saturated with opti-
mized tap water (water hardness ≤10 d, pH between 6.8 and 7.1,
and NaCl content <2 mmol L−1 ) and then kept for 48 h to attain their
respective field capacities. C. insignis seedlings were produced by
tissue culture. When the plants were about 15 cm tall and had a
fresh shoot weight of about 42 g and a fresh root weight of about
15 g, they were transplanted into the pots (one plant per pot). The
irrigation schedule and the kind and quantity of irrigation water
added were identical for all treatments. No additional fertilization
was added.
The experiment was terminated 7 months after planting (from
31 March 2012 to 31 October 2012), when the transplants had
grown to commercial size. All of the plants were harvested, and
their growth parameters and nutrient content were determined as
described in Section 2.4.
Table 1
Orthogonal design L9 (34 ) of the experiment.
Medium Amendment (% in each medium based on dry weight)
Biochar Humic acid
T1 0 0
T2 0 0.5
T3 0 0.7
T4 20 0
T5 20 0.5
T6 20 0.7
T7 35 0
T8 35 0.5
T9 35 0.7
72 L. Zhang et al. / Scientia Horticulturae 176 (2014) 70–78
2.3. Growth media analysis
Before planting, the physical, chemical, and microbial character-
istics of the nine growth media were determined for three replicate
samples per medium. Each sample was divided into three parts:
the first part was air-dried (3–5% moisture content) and used for
determination of physical characteristics, pH, electrical conductiv-
ity (EC), CEC, total organic carbon (TOC), total Kjeldahl nitrogen
(TN), and total phosphorus (TP). The second part was oven-dried
at 65 ◦ C and used for determination of contents of total potassium
(TK), mineral N (NH4 + N and NO3 − N), Ca, Mg, Na, Fe, and Mn. The
third part was stored at 4 ◦ C for analysis of microbial biomass C and
N. The physical and chemical properties of BC were determined by
the same methods used to determine the properties of the growth
media.
2.3.1. Physical characteristics
Bulk density (BD), water-holding capacity (WHC), total porosity
(TPS), aeration porosity (AP), and water-holding porosity (WHP)
were determined according to the ring knife method as described
by Zhang et al. (2013a).
Particle-size distribution of the growth media was determined
according to Jayasinghe et al. (2010b) and Jayasinghe et al. (2009).
Samples of air-dried media were passed through a series of sieves
(0.10, 0.25, 0.50, 1.00, 2.00, and 12.00 mm; 3 min of shaking on each
sieve size). The material retained on each sieve was weighed. The
coarseness index (CI), expressed as a percentage (based on air-dried
weight) of particles >1 mm was determined (Jayasinghe, 2012b).
2.3.2. Chemical characteristics
pH and EC values were measured in a 1:10 water soluble extract
(w/v) using an MP521 pH/EC meter (Shanghai, China) (Zhang et al.,
2013a). CEC was determined with 1 M ammonium acetate at pH 7
(Belyaeva and Haynes, 2012). Mineral N was extracted with 2 M KCl
(1:100 ratio for 1 h) followed by colorimetric analysis of NH4 + N
and NO3 − N using a Seal automated discrete analyzer (Belyaeva
and Haynes, 2012). TOC was determined with a ‘Liqui TOC total
organic carbon analyzer (Zhang et al., 2013b). TN was measured by
the modified micro-Kjeldahl procedure with an automatic Kjeldahl
apparatus (KDY-9830, China) (Dede et al., 2010). TP was mea-
sured by the Anti-Mo–Sb spectrophotometry method (Jayasinghe,
2012a). TK was determined by flame photometry (Jayasinghe et al.,
2007). Ca, Mg, Na, Fe, and Mn were digested with sulfuric acid, and
the digested liquid was analyzed by inductively coupled plasma
mass spectrometry (Prodigy, America) (Jayasinghe et al., 2010b).
2.3.3. Microbial biomass C and N
According to the methods of Brookes et al. (1985) and Vance
et al. (1987), microbial biomass C and N were estimated based on
the difference between total organic C and N extracted with 0.5 M
K2 SO4 from chloroform-fumigated vs. unfumigated samples using
a KC factor of 0.38 and a KN factor of 0.54.
2.4. Calathea insignis growth parameters and nutrient content
analysis
Growth parameters and nutrient content of C. insignis were
determined for 30 plants per treatment. Plant height was mea-
sured with a measuring tape from the growth medium surface
to the tops of the three highest leaves for each plant of each
treatment, and then the average was calculated. Crown width
was measured twice with a measuring tape for each plant (the
second measurement was at a right angle to the first), and the
results were averaged. Leaf number was also recorded. After that,
shoots and roots were washed thoroughly with deionized water to
remove media residues, and fresh weights were determined. Then,
following the method of Ahmed and El-Araby (2012), the photo-
synthetic pigments (chlorophylls a, b, and carotenoids) of fresh
leaves were quantified. Anti-Mo-Sb spectrophotometry was used
for extinction measurements of the alcohol (95%) extracts of the
different photosynthetic pigments. Measurements were performed
on four randomly chosen leaves per replicate, each measurement
was performed twice, and the mean was used for statistical anal-
ysis; the sample weight was constant for all determinations of
pigment content. The relative value of total chlorophyll content
(SPAD value) was also measured with an SPAD-YLS-1 chlorophyll
meter (Shandong, China). After that, roots were scanned to analyze
root morphology (in terms of total root length, total root surface
area, average root diameter, total root volume, and total number
of root tips) using the WinRHIZO image analysis software (Regent
Instruments, WinRHIZO-EC, Canada). The shoots and roots of the
same plant were then oven-dried at 90 ◦ C for 30 min and then at
65 ◦ C. When constant weights were obtained, the dry weights were
recorded. Finally, the oven-dried shoots were crushed in a small
grinder and passed through a 1.0-mm soil sieve for analysis of nutri-
ent content. Contents of TN, TP, TK, the macro-nutrients Ca and
Mg, and the micro-nutrients Fe and Mn in shoots were determined
using the same methods described for analysis of the growth media
(Medina et al., 2009). Contents of the micro-nutrients Cu, Zn, and B
in shoots were determined by the same method used to determine
the contents of Ca, Mg, Na, Fe, and Mn in the growth media.
2.5. Statistical analysis
One-way analyses of variance (ANOVAs) were used to deter-
mine how treatments affected the characteristics of the growth
media and the growth parameters and nutrient content of C. insignis
plants. When ANOVAs were significant, means were separated with
an LSD test. All statistical analyses were performed with SPSS16.0.
3. Results
3.1. Physical characteristics of the growth media
3.1.1. Bulk density, water-holding capacity, and porosity
BD value was significantly (p < 0.05) reduced by addition of BC
and HA alone and in combination (Table 2). BD values of media
T2, T3, T4, T5, T6, T7, T8, and T9 were decreased by 7.5, 11.1, 12.6,
22.4, 29.2, 22.6, 34.1, and 46.1%, respectively, compared to the CGW
without amendment (medium T1). The BD values in media T2 to T9
were within the optimal range (<0.4000 g cm−1 ). In contrast, addi-
tion of BC or HA, especially the combined addition of BC and HA,
greatly (p < 0.05) increased WHC, TPS, AP, and WHP. In comparison
with medium T1, WHC values in media T2, T3, T4, T5, T6, T7, T8, and
T9 were increased by 3.3, 7.6, 12.9, 17.8, 22.3, 25.3, 28.4, and 31.1%,
respectively, and TPS values in those media were increased by 8.9,
16.7, 27.4, 35.1, 40.1, 41.5, 48.9, and 58.8%, respectively. WHC, TPS,
and WHP values for media T4 to T8 were within the acceptable
ranges, and AP values for media T3 to T8 were within the optimal
range of 20 to 30%.
3.1.2. Particle-size distribution and coarseness index
BC and/or HA addition significantly (p < 0.05) increased the per-
centage of particles between 0.25 and 2.00 mm. This was especially
true for the combination of BC and HA: in media T2, T3, T4, T5, T6, T7,
T8, and T9, the percentage of particles between 0.25 and 2.00 mm
was increased by 15.5, 17.6, 15.2, 45.7, 79.7, 30.9, 66.0, and 71.8%,
respectively, compared to the 100% CGW medium (medium T1)
(Table 3). Medium T6 had the highest percentage between 0.25 and
2.00 mm while medium T1 had the lowest percentage. Similarly,
the CI of the growth media was increased (p < 0.05) with addition
of BC and/or HA. In comparison with the medium T1, the CI in media
 L. Zhang et al. / Scientia Horticulturae 176 (2014) 70–78 73

Table 2
Physical properties of the growth media.

Medium BD (g cm−3 ) WHC (%) TPS (%) AP (%) WHP (%)

BC 0.1352(0.0400) 89.36(0.44) 96.88(0.41) 33.20(0.17) 63.68(0.29)

T1 0.4027(0.0200)a 64.91(0.35)i 58.05(0.26)h 16.41(0.25)g 41.64(0.36)i
T2 0.3725(0.0500)b 67.02(0.41)h 63.20(0.32)g 18.09(0.23)f 45.11(0.12)h
T3 0.3582(0.0200)c 69.81(0.29)g 67.73(0.43)f 20.34(0.12)e 47.39(0.37)g
T4 0.3518(0.0100)c 73.29(0.11)f 73.95(0.37)e 22.92(0.31)d 51.03(0.45)f
T5 0.3126(0.0300)d 76.45(0.24)e 78.42(0.48)d 24.72(0.24)c 53.70(0.28)e
T6 0.2851(0.0600)e 79.39(0.43)d 81.30(0.16)c 26.38(0.37)b 54.92(0.19)d
T7 0.3117(0.0800)d 81.31(0.28)c 82.16(0.27)c 25.14(0.26)c 57.02(0.42)c
T8 0.2654(0.0200)f 83.34(0.32)b 86.44(0.34)b 27.01(0.19)b 59.43(0.21)b
T9 0.2172(0.0900)g 85.09(0.18)a 92.19(0.40)a 30.59(0.20)a 61.60(0.16)a
IMa <0.4000 70.00–85.00 70.00–90.00 20.00–30.00 50.00–60.00

Values are means (SD), n = 3. Means in a column followed by the same letter are not significantly different at p ≤ 0.05 according to LSD.
BD = bulk density; WHC = water-holding capacity; TPS = total porosity; AP = aeration porosity; WHP = water-holding porosity.
a
IM = ideal medium, according to de Boodt and Verdonck (1972), Hicklenton et al. (2001), and Jayasinghe et al. (2010a).

Table 3
Particle-size distribution and coarseness index of the growth media. The values indicate the percentage of each particle size (in mm) for each treatment.

Medium >12.00 (mm) 12.00–2.00 1.00–2.00 0.50–1.00 0.50–0.25 0.25–0.10 <0.10 0.25–2.00 CI (>1.00)

T1 0 10.01(1.03) 14.02(0.65) 7.16(0.43) 12.42(1.09) 37.90(0.50) 18.49(1.97) 33.60(2.90)g 24.03(2.09)i

T2 0 18.19(0.97) 11.07(0.98) 10.63(0.65) 17.09(1.54) 24.56(0.89) 18.46(1.76) 38.79(2.52)f 29.26(3.11)h
T3 0 19.82(1.22) 12.63(0.63) 11.14(0.89) 15.73(1.87) 25.61(0.31) 15.07(2.03) 39.50(2.86)f 32.45(3.98)f
T4 0 17.22(1.09) 13.34(0.65) 5.97(0.32) 19.38(1.43) 27.47(0.65) 16.62(1.89) 38.69(2.65)f 30.56(3.65)g
T5 0 19.28(0.76) 18.36(0.51) 16.73(0.54) 13.87(1.05) 15.35(0.73) 16.41(1.65) 48.96(2.08)d 37.64(3.09)d
T6 0 29.11(0.44) 14.76(0.76) 18.32(0.98) 27.29(0.88) 7.54(0.32) 2.98(1.54) 60.37(2.54)a 43.87(2.91)a
T7 0 19.17(1.18) 15.45(0.93) 12.85(0.60) 15.68(1.46) 23.72(0.76) 13.13(1.06) 43.98(2.27)e 34.62(3.53)e
T8 0 24.56(1.54) 14.91(1.04) 13.06(0.21) 27.80(1.50) 11.32(0.95) 8.35(2.11) 55.77(2.62)c 39.47(3.67)c
T9 0 22.33(1.76) 19.42(1.16) 20.55(0.87) 17.75(1.77) 14.11(0.74) 5.84(1.00) 57.72(1.76)b 41.75(3.98)b
IMa – – – – – – – – 30.00–45.00

Values are means (SD), n = 3. Means in a column followed by the same letter are not significantly different at p ≤ 0.05 according to LSD.
CI = coarseness index.
a
IM = ideal medium, according to Jayasinghe (2012a).

T2, T3, T4, T5, T6, T7, T8, and T9 was increased by 21.7, 35.0, 27.2,
56.6, 82.6, 44.1, 64.3, and 73.7%, respectively. Except for media T1
and T2, the CIs of all other media were within the established ideal
range (30.00–45.00).
3.2. Chemical characteristics of the growth media
The pH and EC values of the 100% CGW medium were 7.90 and
0.69, respectively (Table 4). BC applied alone significantly (p < 0.05)
increased pH and EC values relative to the control, and pH and EC
values were highest in medium T7. In contrast, pH and EC values
were reduced by application of HA. The pH and EC values were opti-
mal in media T2, T3, and T6, but were much higher than optimum
in the other media. CEC (at pH 7.0) values were greatly (p < 0.05)
increased by addition of BC and HA alone or in combination and
were highest in medium T9 and lowest in medium T1.
Addition of BC and HA alone or in combination significantly
(p < 0.05) increased concentrations of mineral N (NH4 + N and
NO3 − N), TOC, TK, Mg, Na, and Fe in the media, and the concen-
trations were highest in medium T9 and lowest in medium T1.
Although BC contained large amounts of mineral N, TOC, TK, Mg,
Na, and Fe, it contained little TN, TP, Ca, and Mn (Table 4). Thus,
compared with the control (medium T1), media amended with BC
alone contained higher levels of NH4 + N and NO3 − N, TOC, TK, Mg,
Na, and Fe but lower levels of TN, TP, Ca, and Mn. In contrast, the
concentrations of all nutrients listed in Table 4 increased with HA
addition. The levels of TN, TP, Ca, and Mn were highest in medium
T3 and lowest in medium T7.
3.3. Microbial biomass C and N in the growth media
Microbial biomass C and N in the growth media were signifi-
cantly (p < 0.05) increased by addition of BC or HA and especially
by the combined addition of BC and HA (Fig. 1). Microbial biomass C
of media T1, T2, T3, T4, T5, T6, T7, T8, and T9 were 16, 25, 36, 43, 54,
76, 58, 73, and 90 mg C kg−1 , respectively, and microbial biomass
N of those media were 3, 8, 13, 20, 32, 46, 29, 42, and 53 mg kg−1 ,
respectively, indicating that microbial biomass C and N were high-
est in medium T9, followed by media T6 and T8, all three of which
had been amended with both BC and HA. The values were lowest
in medium T1.
3.4. Calathea insignis growth parameters
C. insignis biomass was significantly (p < 0.05) increased by addi-
tion of BC or HA alone and especially by the combined addition of
BC and HA (Fig. 2). Shoot fresh weights of media T1, T2, T3, T4,
T5, T6, T7, T8, and T9 were 239.1, 266.4, 289.6, 280.0, 339.2, 376.1,
265.8, 278.2, and 299.2 g, respectively. Shoot dry weights of those
media were 39.0, 42.0, 59.5, 51.3, 66.2, 70.0, 51.4, 60.5, and 63.5 g,
respectively. Root fresh weights of those media were 212.4, 248.9,
264.1, 252.4, 303.6, 349.2, 279.6, 289.2, and 313.1 g, respectively.
Root dry weights of those media were 33.0, 37.5, 45.6, 48.3, 57.0,
60.0, 47.2, 52.1, and 56.4 g, respectively. Fresh and dry weights of
shoots and roots were highest with medium T6 and lowest with
medium T1. Similar results were obtained for plant height, length
of the longest root, crown breadth, and leaf number (Table 5) and
also for root morphology parameters (Table 6). Compared with the
CGW without amendment (medium T1), shoot fresh weight, shoot
dry weight, root fresh weight, root dry weight, plant height, longest
root length, crown breadth, number of leaves, total root length, total
root surface area, average root diameter, total root volume, and
total number of root tips of C. insignis obtained from the medium
T6 were increased by 57.3, 79.7, 64.5, 82.0, 45.2, 63.8, 89.1, 31.1,
94.1, 48.6, 91.0, 91.5, and 58.7%, respectively.
74 L. Zhang et al. / Scientia Horticulturae 176 (2014) 70–78

Table 4
Chemical properties of the growth media.

Medium pHa EC CEC NH4 + N NO3 − N TOC (%)a TN (%)a

(mS cm−1 )a (cmol kg−1 )a (mg kg−1 )b (mg kg−1 )b

BC 8.21(0.20) 1.04(0.11) 152.8(3.8) – – 91.42(1.35) 1.28(0.10)

T1 7.90(0.17)b 0.69(0.13)b 77.3(2.5)g 201.3(8.7)h 27.9(1.3)h 28.59(1.24)h 3.95(0.09)c
T2 6.40(0.11)e 0.41(0.04)f 88.2(2.7)f 223.9(6.2)g 33.8(0.9)g 32.66(1.35)g 4.64(0.03)b
T3 6.07(0.12)f 0.29(0.10)g 97.9(3.1)e 256.5(7.3)e 38.3(2.0)e 35.74(2.09)f 5.36(0.11)a
T4 7.95(0.23)ab 0.78(0.02)a 89.7(1.9)f 240.1(10.1)f 36.3(1.6)f 39.02(1.87)e 2.93(0.14)f
T5 6.83(0.08)d 0.59(0.08)cd 111.8(1.1)d 263.8(8.9)d 40.5(2.3)d 46.21(1.54)d 3.05(0.16)e
T6 6.39(0.36)e 0.48(0.09)e 136.2(2.5)b 354.6(7.6)b 46.2(1.7)b 53.92(1.09)b 3.42(0.06)d
T7 8.00(0.21)a 0.82(0.10)a 97.4(2.9)e 259.7(6.3)e 39.0(0.5)e 46.90(1.65)d 2.68(0.18)g
T8 7.15(0.29)c 0.63(0.03)c 123.2(2.1)c 323.4(9.2)c 43.9(2.3)c 51.02(1.30)c 3.91(0.19)c
T9 7.08(0.15)c 0.55(0.06)d 149.6(1.3)a 362.2(10.4)a 48.1(1.4)a 56.11(1.32)a 3.12(0.07)e
IMc 5.2–6.5 ≤0.5 – – – – –
TP (%)a TK (%)b Ca (%)b Mg (%)b Na (%)b Fe (%)b Mn (%)b
BC 0.17(0.04) 1.89(0.09) 0.33(0.09) 0.73(0.04) 4.83(0.98) 0.83(0.07) 0.14(0.03)
T1 0.29(0.01)c 0.84(0.01)f 1.46(0.04)c 0.26(0.03)f 1.34(1.02)g 0.20(0.02)g 0.31(0.06)c
T2 0.41(0.03)b 0.95(0.02)e 1.58(0.03)b 0.35(0.01)d 2.67(0.24)f 0.34(0.01)e 0.37(0.09)b
T3 0.50(0.05)a 1.08(0.07)d 1.63(0.08)a 0.42(0.04)c 2.93(0.56)e 0.45(0.04)c 0.46(0.02)a
T4 0.15(0.09)g 0.96(0.04)e 1.05(0.09)f 0.31(0.05)e 2.88(0.68)e 0.30(0.06)f 0.18(0.01)fg
T5 0.20(0.10)ef 1.22(0.03)c 1.19(0.01)e 0.42(0.06)c 3.65(0.13)b 0.41(0.03)d 0.24(0.04)d
T6 0.26(0.03)cd 1.34(0.04)a 1.31(0.05)d 0.50(0.03)ab 3.82(1.00)a 0.52(0.07)b 0.29(0.05)c
T7 0.13(0.04)g 1.12(0.10)d 1.01(0.02)f 0.36(0.02)d 3.21(0.58)d 0.35(0.09)e 0.15(0.04)g
T8 0.17(0.06)fg 1.28(0.08)b 1.16(0.07)e 0.48(0.09)b 3.49(0.34)c 0.48(0.02)c 0.20(0.07)ef
T9 0.22(0.01)de 1.37(0.05)a 1.28(0.04)d 0.53(0.08)a 3.91(0.21)a 0.56(0.05)a 0.23(0.08)de
IMc – – – – – – –

Values are means (SD), n = 3. Means in a column followed by the same letter are not significantly different at p ≤ 0.05 according to LSD.
EC = electrical conductivity (at 25 ◦ C); CEC = cation-exchange capacity (pH = 7.0); TOC = total organic carbon; TN = total Kjeldahl nitrogen; TP = total phosphorus; TK = total
potassium.
a
Percentages are based on air-dried weight.
b
Percentages are based on oven-dried weight.
c
IM = ideal medium, according to Jayasinghe (2011) and Jayasinghe et al. (2010a).

Fig. 1. Microbial biomass C and N in the growth media. Values are means ± SD (n = 3). Bars within each panel with different letters are significantly different according to LSD
at p ≤ 0.05.

Table 5 3.5. Nutrient content of Calathea insignis leaves

Plant height, the length of the longest root, crown breadth, and leaf number of
Calathea insignis as affected by growth medium.
The effects of addition of BC and/or HA on the nutrient con-
Medium Plant height Longest root Crown Leaf number tent of C. insignis shoots were similar to the effects on the growth
(cm) length (cm) breadth (cm)
parameters, i.e., the contents of macro-nutrients (TN, TP, TK, Ca, and
T1 49.8(1.3)g 33.4(1.1)g 24.8(1.7)g 45(3)g Mg) and micro-nutrients (Fe, Cu, Mn, Zn, and B) were remarkably
T2 54.7(0.9)e 36.6(1.2)f 26.2(0.8)f 47(2)efg (p < 0.05) increased by addition of BC and HA alone and especially by
T3 58.2(1.2)c 40.3(0.7)e 30.4(0.6)e 52(3)bc
BC and HA in combination; contents were highest for plants grown
T4 55.9(0.7)d 42.3(0.9)d 33.3(1.3)d 49(1)de
T5 63.6(1.5)b 46.8(1.5)c 38.1(1.4)c 53(2)b in medium T6 and lowest in medium T1 (Table 7). Compared with
T6 72.3(0.4)a 54.7(0.8)a 46.9(1.3)a 59(4)a medium T1, medium T6 increased the contents of TN, TP, TK, Ca,
T7 53.6(1.0)f 41.2(0.3)e 34.0(1.5)d 46(2)fg Mg, Fe, Cu, Mn, Zn, and B in C. insignis leaves by 66.4, 55.8, 72.1,
T8 56.4(0.6)d 46.3(1.4)c 38.7(0.9)c 48(1)def
122.5, 109.9, 49.7, 62.4, 24.1, 45.8, and 27.0%, respectively.
T9 59.1(1.8)c 50.0(1.1)b 41.2(1.2)b 50(1)cd

Values are means (SD), n = 30. Means in a column followed by the same letter are
not significantly different at p ≤ 0.05 according to LSD.
3.6. Photosynthetic pigment content in Calathea insignis leaves

As was the case for most of the other parameters, the contents of
photosynthetic pigments (chlorophyll-a, chlorophyll-b, carotenoid,
 L. Zhang et al. / Scientia Horticulturae 176 (2014) 70–78 75

Fig. 2. Biomasses (fresh and dry) of shoots and roots of Calathea insignis as affected by growth medium. Values are means ± SD (n = 30). For each parameter, bars within each
panel with different letters are significantly different according to LSD at p ≤ 0.05.

Table 6
Characteristics of Calathea insignis root systems as affected by growth medium.

Medium Total root Total root surface Average root Total root Total number of
length (cm) area (cm2 ) diameter (mm) volume (cm3 ) root tips

T1 919(26)g 449.41(10.06)g 1.56(0.07)g 17.50(0.51)g 5067(69)h

T2 1178(13)f 490.76(11.20)f 1.82(0.11)f 20.32(0.62)f 5872(45)g
T3 1372(10)d 527.34(9.33)e 2.21(0.32)d 24.21(0.99)d 6721(67)d
T4 1235(21)e 565.55(10.09)c 1.94(0.19)e 22.80(0.24)e 6033(43)f
T5 1502(15)c 603.20(13.15)b 2.35(0.12)c 27.33(0.46)c 7130(54)b
T6 1784(27)a 667.87(9.24)a 2.98(0.06)a 33.52(0.23)a 8043(34)a
T7 1267(17)e 553.21(8.32)d 2.02(0.43)e 23.59(0.67)de 5934(61)fg
T8 1346(19)d 570.22(12.00)c 2.33(0.18)c 27.81(1.00)c 6480(35)e
T9 1582(11)b 607.03(9.33)b 2.55(0.22)b 30.10(0.43)b 6992(45)c

Values are means (SD), n = 30. Means in a column followed by the same letter are not significantly different at p ≤ 0.05 according to LSD.

and total chlorophyll content) in C. insignis leaves were significantly
(p < 0.05) increased by the addition of BC or HA and especially by the
combined addition of BC and HA; contents were highest for plants
grown in medium T6 and lowest in medium T1 (Table 8). The same
results were obtained for SPAD values. Compared with the control
(medium T1), medium T6 increased chlorophyll-a, chlorophyll-b,
carotenoid, total chlorophyll content, and SPAD values by 43.5, 69.2,
75.1, 47.8, and 55.8%, respectively.
4. Discussion
4.1. Effects of organic amendments on the physical characteristics
of the growth media
4.1.1. Bulk density
The BD values in all media except the control (100% CGW) were
lower than 0.4000 g cm−3 , indicating that addition of BC and/or HA
optimized media compaction (Abad et al., 2001; Jayasinghe et al.,
2010a). This was consistent with a previous report that BC addi-
tion reduces BD (Ogawa and Okimori, 2010). BD is of course closely
related to media porosity, and inappropriate AP or WHP values may
limit air exchange or water retention of growth media and thereby
inhibit plant growth (Hicklenton et al., 2001).
4.1.2. Water-holding capacity and porosity
The amendments not only increased water retention but also
optimized porosity. Addition of BC or HA and especially addition
of BC and HA together shifted values for WHC, TPS, AP, and WHP
into the optimal ranges and increased water retention and poros-
ity. Similar results were obtained by Belyaeva and Haynes (2012)
and Dumroese et al. (2011), who reported that BC or HA improved
the particle structure of the growth media. WHC increased with
addition of BC and/or HA because of the capillary pores of the BC
and the hydrophilic groups of the HA (Montoneri et al., 2007; Senn
76 L. Zhang et al. / Scientia Horticulturae 176 (2014) 70–78

Table 7
Contents of macro-nutrients (TN, TP, TK, Ca, and Mg) and micro-nutrients (Fe, Cu, Mn, Zn, and B) in Calathea insignis leaves as affected by growth medium.

Medium Content of macro-nutrient

TN (%)a TP (%)a TK (%)b Ca (%)b Mg (%)b

T1 1.780(0.020)h 0.120(0.013)h 2.021(0.013)h 0.382(0.010)h 0.192(0.012)g

T2 1.934(0.034)g 0.133(0.022)g 2.377(0.020)g 0.492(0.008)g 0.243(0.009)f
T3 2.213(0.098)e 0.150(0.014)d 2.602(0.025)e 0.536(0.009)f 0.278(0.008)d
T4 2.092(0.102)f 0.143(0.015)d 2.410(0.011)f 0.634(0.012)e 0.256(0.013)e
T5 2.458(0.026)c 0.162(0.011)c 2.763(0.017)c 0.732(0.011)c 0.329(0.011)b
T6 2.962(0.088)a 0.187(0.020)a 3.479(0.020)a 0.850(0.015)a 0.403(0.015)a
T7 2.103(0.109)f 0.139(0.019)f 2.407(0.018)f 0.540(0.007)f 0.249(0.007)ef
T8 2.335(0.074)d 0.148(0.017)de 2.668(0.015)d 0.677(0.014)d 0.270(0.014)d
T9 2.587(0.077)b 0.169(0.018)b 2.782(0.014)b 0.759(0.016)b 0.304(0.009)c
Content of micro-nutrient
Fe (×10−3 %)b Cu (×10−3 %)b Mn (×10−3 %)b Zn (×10−3 %)b B (×10−3 %)b
T1 10.024(0.770)i 0.575(0.012)i 12.573(0.018)i 1.278(0.009)h 2.199(0.190)g
T2 10.986(0.278)h 0.659(0.022)h 13.102(0.011)h 1.390(0.036)g 2.346(0.202)f
T3 11.653(0.109)g 0.743(0.057)f 13.789(0.024)f 1.482(0.024)d 2.459(0.187)d
T4 12.575(0.662)e 0.772(0.093)e 13.976(0.036)e 1.459(0.049)e 2.480(0.211)c
T5 13.682(0.374)c 0.840(0.062)b 14.820(0.045)c 1.634(0.023)b 2.603(0.166)b
T6 15.007(0.280)a 0.934(0.101)a 15.607(0.088)a 1.863(0.011)a 2.792(0.159)a
T7 11.906(0.331)f 0.701(0.046)g 13.622(0.051)g 1.401(0.027)f 2.200(0.172)g
T8 13.445(0.290)d 0.785(0.039)d 14.209(0.063)d 1.468(0.053)e 2.431(0.213)e
T9 14.062(0.443)b 0.800(0.073)c 14.833(0.039)b 1.536(0.028)c 2.602(0.120)b

Values are means (SD), n = 30. Means in a column followed by the same letter are not significantly different at p ≤ 0.05 according to LSD.
a
Percentages are based on air-dry weight.
b
Percentages are based on oven-dry weight.

Table 8
Photosynthetic pigment contents (based on fresh weight) in Calathea insignis leaves as affected by growth medium.

Medium Chlorophyll-a (mg g−1 ) Chlorophyll-b (mg g−1 ) Total chlorophyll (mg g−1 ) SPAD Carotenoid (mg g−1 )

T1 7.689(1.345)i 1.524(0. 234)i 9.213(1.098)i 46.6(1.2)i 2.505(0.047)i

T2 8.536(0.962)g 1.726(0.146)h 10.262(1.022)h 54.2(1.4)f 2.903(0.064)g
T3 9.005(1.420)e 1.859(0.302)f 10.864(0.987)e 61.3(1.7)c 3.105(0.099)e
T4 8.765(1.525)f 1.883(0.298)e 10.648(0.833)f 55.9(0.8)e 2.937(0.101)f
T5 9.972(0.856)b 2.136(0.112)b 12.108(0.609)b 63.2(0.9)b 3.434(0.058)b
T6 11.035(0.724)a 2.579(0.098)a 13.614(0.911)a 72.6(1.3)a 4.385(0.072)a
T7 8.450(1.223)h 1.832(0.291)g 10.282(0.103)g 47.3(1.5)h 2.610(0.055)h
T8 9.231(0.689)d 1.940(0.116)d 11.171(0.875)d 52.9(1.7)g 3.118(0.043)d
T9 9.922(1.007)c 2.010(0.300)c 11.932(0.981)c 59.7(0.6)d 3.409(0.070)c

Values are means (SD), n = 30. Means in a column followed by the same letter are not significantly different at p ≤ 0.05 according to LSD.

and Kingman, 1973). Enhanced WHC of the growth media will also
increase nutrient retention because of reduced leaching (Glaser,
2007).
4.1.3. Particle-size distribution and coarseness index
The particle-size distribution of a growth medium is important
because it determines pore space, gas exchange, and water-holding
capacities (Abad et al., 2001). Jayasinghe et al. (2010b) reported
that an excess of larger particles may lead to excessive aeration
and inadequate water retention and that an excess of fine parti-
cles may clog pores and decrease air-filled porosity. Growth media
with a high percentage of particles between 0.25 and 2.00 mm are
optimal for potted plants (Jayasinghe, 2011). The results indicate
that the combined addition of BC and HA to CGW increased the
percentage of particles in the optimal range and that the highest
percentage of particles in the optimal size range occurred when
CGW was amended with 20% BC and 0.7% HA. CI, which is defined
as the percentage of particles >1.0 mm in diameter, provides a value
for particle-size distribution that can be correlated with physi-
cal properties, and the ideal CI range is considered to be 30–45%
(Jayasinghe, 2012a). In the current study, CI was optimized by BC
or HA addition and especially by the combined addition of BC and
HA to CGW. Abad et al. (2005) reported that TPS increased with
increasing values of CI, which is consistent with the finding in the
present study that the combined addition of BC and HA to CGW
increased both CI and TPS.
4.2. Effects of organic amendments on the chemical
characteristics of the growth media
Although the addition of BC alone increased the pH and EC values
of the growth media to levels that could be unfavorable for plant
growth, these undesirable effects were countered by addition of HA.
The HA has acid groups, which would decrease pH and EC values
of the growth media (Montoneri et al., 2007). Also, by increasing
medium aggregation and water movement, HA could increase the
leaching of excessive soluble salts (Varanini and Pinton, 1995). The
increases in CEC in media amended with BC and/or HA, especially
the combination of BC and HA, could be explained by the high CEC
values of both amendments. BC has a high CEC because it is porous
and has a high surface area (Thies and Rillig, 2009). Increases in
CEC by BC or HA amendment to soil have been previously reported
(Hanafi and Salwa, 1998; Nigussie et al., 2012).
4.3. Effects of organic amendments on the nutrient content of the
growth media
Because CGW generally does not provide all of the nutrients
required for optimal plant growth, addition of BC and/or HA could
help overcome its nutrient limitations (Medina et al., 2009).
4.3.1. NH4 + N and NO3 − N
Mineral N (NH4 + N and NO3 − N) retention was optimized by
BC or HA addition, especially the combination of BC and HA. The
 L. Zhang et al. / Scientia Horticulturae 176 (2014) 70–78
retention of NH4 + and NO3 − was increased by BC because of its
positively and the negatively charged sites. An increase in NH4 +
and NO3 − storage in soil amended with BC was previously reported
(Clough and Condron, 2010). The increases in NH4 + and NO3 − in
media amended with HA could be explained by the high contents of
NH4 + and NO3 − in HA. The increased NH4 + and NO3 − contents could
be also explained in part by the increased CEC of growth media
caused by BC and/or HA addition (Nigussie et al., 2012).
4.3.2. Microbial biomass C and N
Microbial biomass C and N were significantly increased by addi-
tion of BC and/or HA and were more increased by the combined
addition of BC and HA. Thies and Rillig (2009) suggested that
because of its porous structure, its high internal surface area, and
its ability to adsorb soluble organic matter, gases, and inorganic
nutrients, BC was a highly suitable habitat for microbial growth.
In a previous study, an increase in microbial biomass and activity
after HA addition to soil was attributed to the effects of Fe in the HA
(Senn and Kingman, 1973). Other studies reported a rapid increase
in microbial biomass and activity and also changes in the bacte-
rial community after humus application (Pant et al., 2012; Siddiqui
et al., 2008; Sorrenti et al., 2012).
4.3.3. Other important nutrients
BC increased the contents of TOC, TK, Mg, Na, and Fe in the
growth media but did not provide sufficient levels of TN, TP, Ca,
or Mn. However, addition of HA to the growth media compensated
for the nutrients that were inadequately supplied by BC and CGW.
4.4. Effects of organic amendments on Calathea insignis growth
parameters
Changes in allocation patterns among roots and shoots largely
determine the ability of plants to capture resources, and plants may
change their allocation patterns in response to the environment
and especially in response to the availability of nutrients in the
growth medium (Zaller, 2007). C. insignis growth (fresh and dry
weights of shoots and roots, plant height, longest root length, crown
breadth, and leaf number) was best in medium T6 (CGW with 20%
BC and 0.7% HA). The improved growth with the combined addition
of BC and HA could be explained by an improved balance of water
availability and aeration and by an increased supply of macro- and
micro-nutrients. These observations agree with those of Glaser et al.
(2002) and Williams (1987), who reported that addition of BC or HA
to soil increased plant production.
4.5. Effects of organic amendments on Calathea insignis root
growth
As mentioned before, root development was greatly improved
in media containing BC and HA and was best with the combination
of 20% BC and 0.7% HA. The positive effects of BC and HA on root
development may be due to the nutrients supplied by these amend-
ments or to the effects of the amendments on nutrient and water
retention, pH, aeration, and other physical and chemical character-
istics of the media. The addition of BC or HA has been previously
reported to enhance root growth and development (Ogawa and
Okimori, 2010; Senn and Kingman, 1973).
4.6. Effects of organic amendments on the nutrients in Calathea
insignis leaves
The contents of nutrients in C. insignis leaves were significantly
increased when the plants were grown in media containing BC and
HA, and especially when they were grown in the medium with 20%
BC and 0.7% HA. The results indicated that application of BC and HA
77
increased nutrient availability and therefore increased the nutri-
ent content of C. insignis leaves. BC has been previously shown to
result in high concentrations of bioavailable nutrients that support
increases in the contents of nutrients in plant tissues (Lehmann and
Rhodon, 2006). HA addition also increased the contents of nutrients
in maize in a previous study (Turan et al., 2011). The increased nutri-
ent content caused by BC and HA addition could also be explained
in part by the increased CEC of the growth media, which would
have reduced the leaching of nutrients (Nigussie et al., 2012).
4.7. Effects of organic amendments on photosynthetic pigments
in Calathea insignis leaves
Adding an appropriate combination of BC and HA to the CGW,
and especially 20% BC and 0.7% HA, increased the contents of
photosynthetic pigments probably by providing optimal media
conditions for plant growth and photosynthesis and by increas-
ing chloroplast production and improving chloroplast function. BC
and HA evidently provided the nutrients required to support pho-
tosynthesis. For example, N is required for chlorophyll formation
and is essential for photosynthesis. Also, Mg and Fe are required
for pigment biosynthesis and are thought to be involved in chloro-
plast formation via protein synthesis (Lallawmsanga et al., 2012). In
addition to increasing the availability of these nutrients, BC and HA
could have increased photosynthetic pigment contents by improv-
ing the physical and chemical characteristics of the growth media
(Elhindi, 2012).
5. Conclusion
This paper is the first to report on the combined effects of BC
and HA on the physical, chemical, and nutritional characteristics of
CGW and on the ability of CGW to support the growth of C. insig-
nis. The paper also provides new information concerning the use of
growth media without peat for the cultivation of ornamental plants.
CGW properties were significantly better and C. insignis biomass
was significantly greater when the CGW was amended with a com-
bination of BC and HA than with BC or HA alone. The best results
were obtained with a combination of 20% BC and 0.7% HA. The find-
ings indicate that, when amended with a combination of BC and
HA, CGW is a viable substitute for peat in the soilless culture of
ornamental plants in pots.
Acknowledgments
This study was supported by the Fundamental Research
Funds for the Central Universities (no. BLYJ201308); the Major
Project of the Science and Technology of Beijing, PR China
(D09040903780801) “the Study and Demonstration of the Key
Technology of Recycling Green Waste to Product Flowers and Trees
Growth Media of Beijing, PR China”; and the National Level Project
Promotion of forestry scientific and technological achievements of
the National Forestry Bureau (no. 39, 2012) “the Promotion and
Demonstration of the Agricultural and Forestry Wastes Production
Flowers and Trees Growth Media Technology”. We also thank Prof.
Bruce Jaffee for his linguistic modification of this paper.
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