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Article history: Green waste compost (GWC) is a value-added product generated by the composting of
Received 7 April 2021 urban green waste. Although GWC could potentially be used as a growth medium for
Received in revised form 24 August 2021 soilless culture of plants, GWC alone has high salinity, high pH, and other undesirable
Accepted 13 September 2021
physicochemical properties. In the current study, we therefore assessed how addition of
Available online 16 September 2021
vermiculite (VMT: 0, 3.5, and 7%) and humic acid (HA: 0, 0.5, and 1 g/100 g) affected
Keywords: the physicochemical properties of GWC and its use as a growth medium for cornflower
Green waste compost (Centaurea cyanus L.). GWC modified by the combination of 3.5% VMT and 0.5 g/100 g HA
Vermiculite had the best physicochemical properties (bulk density, porosity, water holding capacity,
Humic Acid electrical conductivity, pH, and nutrient content) and resulted in the best growth of
Growth medium cornflower in terms of plant height, maximum root length, flower number, and biomass.
The results therefore indicate that GWC modified by 3.5% VMT and 0.5 g/100 g HA
powder is an excellent soilless growth medium.
© 2021 Elsevier B.V. All rights reserved.
1. Introduction
With the rapid development of the flower production industry, the demand for growth media is increasing. Ornamental
plants have traditionally been grown in peat or soil. Peat, however, is a slowly renewable natural resource, and its
exploitation requires carbon emission, which may affect the climate (Boldrin et al., 2010). Soil as a growth medium
requires the use of pesticides and fertilizers. As a result, researchers have searched for alternatives to peat or soil, and
one promising alternative is compost (Bustamante et al., 2021).
Compost products are produced by the recycling of organic wastes, which are humus-like and harmless substances that
are rich in organic matter and nutrients, and that can therefore usually be used as growth media or organic fertilizers
(Zhang et al., 2013). In addition to supporting or improving plant growth (Grigatti et al., 2007), compost products can
inhibit the spread of soil-borne pathogens (Kazamias et al., 2016). Although compost products can improve the chemical
characteristics and physical characteristics of growth media (Cacini et al., 2020), compost products alone often have high
salinity (Sáez et al., 2016), high pH (Bustamante et al., 2021), and other undesirable physicochemical properties (Santos
et al., 2016). For example, the growth of carob (Ceratonia siliqua L.), geranium (Pelargonium zonale L.), and calendula
(Calendula officinalis L.) was generally poor in 100% green waste compost (GWC) (Bakry et al., 2013; Gong et al., 2018).
Researchers have therefore suggested that compost should be acidified and that its salt content should be reduced via
leaching before it is used as a growth medium (Gong et al., 2018; Bustamante et al., 2021). Unfortunately, acidification
and leaching are time consuming and water consuming. An alternative approach is to improve compost properties by
∗ Correspondence to: College of Forestry, Beijing Forestry University, P.O. Box 111, Beijing 100083, PR China.
E-mail addresses: fxq13269896811@163.com (X. Feng), zhanglu1211@bjfu.edu.cn (L. Zhang).
https://doi.org/10.1016/j.eti.2021.101945
2352-1864/© 2021 Elsevier B.V. All rights reserved.
X. Feng and L. Zhang Environmental Technology & Innovation 24 (2021) 101945
mixing it with other organic or inorganic wastes. In the current research, we determined whether addition of vermiculite
and humic acid improved the quality of GWC as a growth medium for cornflower (Centaurea cyanus L.).
Vermiculite (VMT), a 2:1 clay mineral with permanent, diffuse negative charges generated as a consequence of the
isomorphic substitution of Si (IV) by Al (III) (Fernando Henrique and Masini, 2014), has high adsorption and ion exchange
properties (Zhu et al., 2011). Its multilayer structure, which includes voids between the layers, is not destroyed by contact
with water (Shinzato et al., 2020). Because of this unique characteristic, VMT has superior ventilation, water permeability,
and buffering against environmental change. The high adsorption capacity of VMT causes the nutrients in the growth
medium to be slowly released, which reduces leaching and facilitates nutrient utilization by plants (Shinzato et al., 2020).
These properties suggest that VMT could be a useful additive to compost.
Humic acid (HA), which is composed of aromatic and various functional groups of organic polymers, has high adsorp-
tion and exchange capacities and a high capacity for forming complexes (Zhang et al., 2012). Researchers (e.g., Chang
et al., 2012) have reported that addition of HA to a growth medium can improve plant growth because of its positive
effects on the cation exchange capacity, buffering capacity, and pH of the medium. HA can also reduce the salt content
of growth media (Liu et al., 2020). Bustamante et al. (2021) considered that high salt concentration often limits the use
of compost as a plant growth medium, because most plants and especially young plants have a low tolerance of salinity
(Fornes et al., 2010). In addition, HA was found to promote the early flowering and to increase the leaf area of a number
of plants by improving nutrient absorption and hormone secretion (Luciano et al., 2015).
Because cornflower (Centaurea cyanus L.) is a good nectar source and has medicinal uses, it has considerable economic
value. As an ornamental plant, cornflower is commonly used in parterre and green belts in Beijing, China. For cornflower
cultivation in China, compost products are often used as growth media or as additives to growth media (personal
observation).
Previous studies have assessed the effects of VMT (Nhung Ngoc et al., 2019) or of HA on compost quality or plant
growth. However, the combined effects of VMT and HA addition on the physicochemical and nutritional characteristics
of GWC and on the performance of GWC as a growth medium for cornflower have not been considered. The objectives of
this study were to: (i) evaluate how adding VMT or/and HA affects the nature of GWC, (ii) the effects of improved GWC
as a growth medium on performance of cornflower.
The green waste (GW) used in this experiment was collected from the Beijing Botanical Garden. Before composting,
the GW was crushed into 1 to 2 cm particles. Urea was added to adjust the C/N ratio to 25–30. Each 1 kg of GW was
mixed with 5 mL EM microbial agent (a complex flora comprising photosynthetic flora, lactic acid flora, yeasts, gram-
positive actinomycetes, and filamentous flora of fermentations, which was purchased from the Nanhai Chantai Animal
pharmaceutical Co., LTD., Foshan, Guangdong) uniformly. At the beginning of the composting, feedstock mass was stacked
into a windrow, with a basal area of 1 m2 and a height of 0.75 m. The material was turned over every 3 days, and water
was added as needed to maintain the moisture content at about 60%. After 45 days, the compost was well decomposed
based on a germination test and on its physical appearance. The initial properties of GWC (T1) are described in Tables 2–3.
At that time, WMT and HA were added to the GWC as described in Section 2.2.
The VMT used in this study was a massive magnesium-hydro-aluminosilicate secondary metamorphic mineral, 3–
6 mm in diameter; it was obtained from the Rainbow Horticulture Co., Lingshou County, Shijiazhuang, Hebei. The main
properties are as follows: pH = 7.03, EC = 0.23, TPS = 74.42%, TN = 0.53%, TP = 0.58%. The HA used in this study was
purchased from Songben Foreign Trade Organic Raw Material Wholesale Co. Before it was added to the compost, the HA
was made into an aqueous solution. The main properties are as follows: PH = 6.87, EC = 1.69. Cornflower seeds were
obtained from the Beijing Forestry Technology Seed Co., LTD. Seedlings of uniform size were selected for the experiment.
The experiment included 9 treatments, including a control without VMT and HA (Table 1). Before the experiment
began, the GWC was air-dried to 10% moisture content; the VMT and HA were also dried to enable accurate weighing.
As indicated in Table 1, the materials were weighed and mixed together before they were added to about 2000 cm 3
pots (14.5 cm in diameter and 12.5 cm in height). A small hole in the bottom of each pot facilitated drainage. Cornflower
seedlings (about 15 cm tall; germinated and initially grown in seedling dish) were transplanted into the pots (one seedling
per pot). Each treatment was represented by 5 replicate pots.
The experiment was conducted in a greenhouse of the Beijing Forestry Technology Co., Ltd. The temperature in the
greenhouse ranged from 18 ◦ C to 25 ◦ C during the day and from 10 ◦ C to 20 ◦ C at night; the relative humidity ranged
from 60% to 75%; and the daily illumination period (at 7000 lx) ranged from 10 to 12 h (Zhang et al., 2014). The pots with
cornflower plants were watered as needed and were not fertilized. Three months after the seedlings were transplanted,
the experiment was ended. Media and plant properties were assessed as described in the following sections.
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X. Feng and L. Zhang Environmental Technology & Innovation 24 (2021) 101945
Table 1
Orthogonal design L9 (34 ) of the experiment
Treatment Additive
VMT (% in each medium based on dry weight) HA (g/100 g)
T1 0 0
T2 0 0.5
T3 0 1
T4 3.5 0
T5 3.5 0.5
T6 3.5 1
T7 7 0
T8 7 0.5
T9 7 1
Table 2
Physical characteristics of the growth media.
Treatment TPS (%) APS (%) WHP (%) BD (g cm−3 ) WHC (%)
T1 61.29(0.24)f 8.25(0.20)d 53.04(0.72)g 0.46(0.01)a 111.26(3.25)e
T2 63.50(0.24)e 8.38(0.16)cd 55.12(0.09)e 0.43(0.01)cd 119.41(4.34)d
T3 63.55(0.20)e 8.09(0.26)d 54.17(0.08)f 0.43(0.01)cd 119.03(3.74)d
T4 65.19(0.40)d 8.73(0.09)c 56.46(0.31)d 0.45(0.01)ab 118.03(1.73)d
T5 69.58(0.39)a 10.56(0.22)a 59.02(0.27)a 0.40(0.01)fg 137.79(1.72)a
T6 65.59(0.18)d 9.56(0.34)b 56.03(0.51)d 0.39(0.02)g 135.15(3.08)ab
T7 67.03(0.36)c 10.43(0.17)a 56.60(0.41)cd 0.44(0.01)bc 115.52(3.78)de
T8 67.45(0.48)c 10.44(0.35)a 57.00(0.26)bcd 0.42(0.01)de 126.96(3.07)c
T9 68.31(0.15)b 10.80(0.39)a 57.52(0.26)bc 0.41(0.01)ef 131.58(3.62)bc
IMa 50%–95% 10%–38% 30%–76% 0.4 >120
Values are means (SD), n = 3. Means in a column followed by the same letter are not significantly
different at p ⩽ 0.05 by LSD. a IR = ideal range according to Sun et al. (2016) and Garcia-Gomez et al.
(2002).
At the end of the experiment, the cornflower plants were carefully removed from the pots for determination of plant
height (HP), the length of the longest root (RL), flower number (FN), total fresh weight (TFW), and total dry weight (TDW).
HP and RL were measured with a measuring tape. For determination of TFW, the plants were washed with deionized water,
blotted dry, and weighed. For determination of TDW, the same plants were dried at 105 ◦ C for 30 min and then dried to
constant weight at 70 ◦ C (Zhang et al., 2014).
SPSS 22.0 was used to conduct one-way analyses of variance (ANOVAs) to assess the effect of treatment on the
growth media properties and the cornflower growth parameters. When ANOVAs were significant (p ≤ 0.05), means were
compared with an LSD test (p ≤ 0.05). A redundancy analysis with Canoco 4.5 was used to assess the correlations between
plant parameters and media properties. Figures were drawn with Microsoft Excel 2019 (Microsoft Corporation, USA).
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X. Feng and L. Zhang Environmental Technology & Innovation 24 (2021) 101945
Table 3
Chemical characteristics of the growth media.
Treatment pH EC (mS cm−1 ) AP (mg kg−1 ) AK (mg kg−1 )
T1 7.56(0.05)a 2.94(0.01)a 209.24(14.03)c 3589.15(352.13)f
T2 7.46(0.02)c 2.58(0.13)d 215.26(5.22)c 5086.39(44.49)c
T3 7.41(0.02)d 2.74(0.06)bc 215.86(14.45)c 4615.57(18.29)d
T4 7.40(0.03)d 2.81(0.01)b 214.05(16.26)c 4511.58(33.26)e
T5 7.20(0.02)f 2.19(0.05)e 271.38(4.70)a 6294.69(90.99)a
T6 7.24(0.01)f 2.75(0.13)b 270.65(6.34)a 5716.74(318.60)b
T7 7.50(0.01)b 2.79(0.07)b 214.66(2.76)c 4454.13(45.50)e
T8 7.38(0.03)d 2.54(0.09)d 223.09(15.75)c 4922.25(179.11)c
T9 7.33(0.01)e 2.62(0.02)cd 244.16(10.27)b 4928.79(78.80)c
IM 5.0–8.5 0.1–2.0 – –
TN (%) TP (%) TK (%) OM (g kg−1 )
T1 1.42(0.02)e 0.23(0.01)b 0.92(0.01)b 433.13(3.82)e
T2 1.68(0.04)ab 0.24(0.01)ab 0.93(0.03)b 437.00(6.41)de
T3 1.64(0.01)b 0.25(0.02)ab 0.97(0.01)a 438.70(7.70)cde
T4 1.55(0.07)c 0.25(0.01)ab 0.92(0.02)b 413.88(4.21)f
T5 1.74(0.01)a 0.26(0.00)a 0.99(0.01)a 485.26(1.49)a
T6 1.66(0.04)b 0.26(0.01)a 0.98(0.01)a 460.89(10.09)b
T7 1.43(0.05)de 0.25(0.02)ab 0.90(0.01)b 395.46(6.06)g
T8 1.50(0.04)c 0.25(0.01)ab 0.90(0.01)b 438.54(1.46)cde
T9 1.49(0.02)cd 0.26(0.01)a 0.93(0.02)b 445.01(8.43)cd
IM – – – –
Values are means (SD), n = 3. Means in a column followed by the same letter are not significantly
different at p ⩽ 0.05 by LSD. All percentages are based on air-dry weight. a IR = ideal range according
to Sun et al. (2016).
Table 4
RDA analysis of cornflower growth parameters and physicochemical properties of the growth media.
Statistic Axis 1 Axis 2 Axis 3 Axis 4
EV 0.762 0.140 0.061 0.029
SEC 1.000 1.000 1.000 1.000
CVSP (%) 76.2 90.2 96.3 99.2
CVSER (%) 76.2 90.2 96.3 99.2
SEV 1.000
Sum 1.000
3. Results
Fig. 1. Growth parameters of cornflower plants. HP = plant height; RL = the longest root length; FN = flower number; TFW = total fresh weight;
TDW = total dry weight. Values are means ± SE. Means with the same letters are not significantly difference at P ≤ 0.05 according to LSD.
Treatments are described in Table 1.
were significantly (p ≤ 0.05) better than those of other treatments. The optimal value of EC appeared at T5, which was
significantly (p ≤ 0.05) different from other treatments.
A redundancy analysis (RDA) was conducted to investigate the relationships between the physical and chemical
characteristics of the media and the cornflower growth parameters (Fig. 2,Table 4). The first two axes of the RDA plot
accounted for 90.2% of the variation in cornflower growth parameters. TDW, TFW, FN, RL, and HP were negatively
correlated with pH, EC, and BD, and were positively correlated with APS, TPS, WHP, TP, AP, AK, WHC, TK, TN, and OM.
The positive correlations between growth parameters and physicochemical properties were strongest between TFW and
APS, HP and TN. The correlation was weak between TDW and OM but was strong between TFW and porosity (TPS, APS,
and WHP).
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X. Feng and L. Zhang Environmental Technology & Innovation 24 (2021) 101945
Fig. 2. Bi-dimensional ordering chart of the RDA of relationships between cornflower parameters and physical and chemical characteristics of media.
The blue arrows point to cornflower growth parameters, and the red arrows point to the physical and chemical properties of the growth media.
The angle between the arrows (alpha) indicates the correlation between the growth parameters and media characteristics: an acute angle indicates
a positive correlation, and an obtuse angle indicates a negative correlation. (For interpretation of the references to color in this figure legend, the
reader is referred to the web version of this article.)
4. Discussion
4.1. Effects of VMT and HA on the physical properties of the growth media
The purpose of this study was to determine whether addition of VMT and HA improves the quality of GWC as a growth
medium for cornflower. We selected VMT and HA because many studies have shown that VMT and HA have high specific
surface areas, porosities, cation exchange capacities, available nutrient contents, and biological activities (Shinzato et al.,
2020; Zhang et al., 2012; Zhu et al., 2011).
4.2. Effects of VMT and HA on the chemical properties of the growth media
4.2.2. Contents of nutrient elements and organic matter in the growth media
The contents of nutrients in the growth media increased with the increased addition of VMT and HA but tended to
peak with the quantities of additives in T5; with higher quantities, the nutrient contents declined or at least increased at
a lower rate. These effects could be attributed to the physicochemical properties of HA and VMT. VMT has high adsorption
and high ion exchange capacities (Zhu et al., 2011), and the immobilized nutrients can be released when needed by the
plant (Shinzato et al., 2020). VMT also contains a variety of trace elements that may function as nutrients. HA contains
carboxyl and phenolic hydroxyl groups, which can function in ion exchange (Zhang et al., 2012) and metal complexation
or chelation (Asgharzade and Babaeian, 2012). HA can therefore combine with many mineral elements and can release
these elements when they are needed by the plant (Russo and Berlyn, 1990). As a macromolecule and a humic substance
(Canellas et al., 2017), HA promotes the formation of aggregates and thereby improves the environment of the growth
media for microbial activity (Zhang and Sun, 2017).
The combined addition of VMT and HA was more effective in improving the growth media than the single addition
of VMT or HA (Table 3), means synergistic effect of the two additives was confirmed. This may be because HA forms
aggregates with clay minerals (such as VMT) through a variety of mechanisms including ion exchange, hydrogen bonding,
water and/or metal bridging, ligand exchange, and van der Walls interactions (Zhang et al., 2012). The adsorption of HA
on VMT could increase the subsequent adsorption of metal ions because HA has abundant complexing/adsorption sites
(Fernando Henrique and Masini, 2014).
As noted earlier, the largest dose of additives did not provide the best results in the current study. The best treatment
was 3.5% VMT and 0.5 g/100 g HA powder (Table 3). When the dosage exceeded these amounts, the physicochemical
properties of the growth media deteriorated. For example, the lowest OM content was obtained with 7% VMT (T7).
The addition of a large amount of VMT may have greatly increased the aeration capacity of the growth medium, which
promoted the degradation of OM. In addition, AP, TP, and TK values did not significantly differ between T5 and T6 (which
differed only in the quantity of HA added) suggests that VMT may be more useful than HA for improving these media
properties.
The growth of plants is closely related to the growth media, because the physical and chemical properties of the
growth media (porosity, pH, EC, and nutrient content, etc.) will determine the extent of root development and proliferation
(Aghhavani Shajari et al., 2020). The VMT and HA modified media (T2–T9) provided a more suitable physical and chemical
environment for cornflower growth, resulting in higher values for HP, RL, FN, and biomass (Fig. 1). HA can improve
the quality of the growth medium, because HA contains acidic functional groups, which increase the bioavailability and
solubility of nutrients (Zhang et al., 2012). This also explains the higher values of AP, TP, and TK when a large amount of
HA was added (Table 3), and FN was positively correlated with the amount of HA added in the absence of VMT addition
(Fig. 1). Because HA can also promote the absorption and utilization of mineral nutrients by plant roots (Canellas et al.,
2015), adding an appropriate amount of HA to the growth medium can stimulate seed germination, increase plant growth,
and increase plant resistance to environmental stress (Hatami et al., 2018). Giulio et al. (2020) suggested that HA may
increase the stability of plant membranes; this would help roots to absorb water and inorganic ions under osmotic stress
and to synthesize proteins (Canellas et al., 2015). The negative effects of the high initial EC of GWC may be reduced by
HA induction of enzymes that increase plant defense against salt stress.
Addition of VMT to growth media can reduce pests and pathogens in the media (Leite et al., 2018). VMT is also
conducive to flower growth, which may help explain why FN was higher in T4–T9 than in the other treatments. Humus
(the main component of GWC) can adsorb NH4 + –N and thereby reduce NH4 + –N mobility and its uptake by roots. This
problem can be solved by addition of VMT because NH4 + –N adsorbed by VMT can be transformed into NO3 − –N through
nitrification (Lv et al., 2013). NO3 − –N is negatively charged and can be evenly distributed in the growth medium. The
uniform distribution of nitrogen in the growth media is beneficial to root growth. This could explain the significant
difference in RL, HP, FN, TFW, and TDW between 100% GWC (T1) and the treatments with VMT (T4–T9). The decomposition
of adsorbed NH4 + -N is slower under normal humidity conditions than under high humidity conditions (Lv et al., 2013).
This suggests that the high WHC values obtained in T5 can further promote NH4 + -N utilization. As shown in Fig. 1, T5
had the highest values of WHC, RL, HP, FN, TFW, and TDW.
Cornflower growth was better with the combined addition of VMT and HA than with addition of VMT and HA alone
(Fig. 1). This further confirms the synergistic effect between the two different additives. Considering each factor (such
as pH, EC, porosity, nutrient content, etc.) as a point, many points form a complex multidimensional cube, and the
optimization of each point (which can be thought of as an extension of the distance between two points) results in an
exponential increase in the volume of the entire multidimensional cube (which can be understood as a comprehensive
optimization of the growth media).
4.4. Correlation between the growth parameters of cornflower and the physicochemical properties of the growth media
Fig. 2 suggests that EC, pH, and BD were the main factors limiting cornflower growth in the current study. This is
reasonable because a high EC value of the medium makes it difficult for the plant roots to absorb water and nutrients.
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X. Feng and L. Zhang Environmental Technology & Innovation 24 (2021) 101945
Solis-Mejia et al. (2012) believed that BD was negatively correlated with plant growth, because a high BD indicates physical
resistance to root growth and reduced aeration. As shown in Tables 2–3 and Fig. 1, higher BD and EC values (T1, T4, and
T7) correspond to a smaller HP. This is consistent with Aghhavani Shajari et al. (2020), who reported that the increased
energy required for the emergence of flowers and leaves emergence in heavy soils leads to lower yields. In addition, high
pH can affect the activity of certain microorganisms and enzymes in the growth medium, thus inhibiting plant growth and
development, because the optimal pH of enzymes is usually close to neutral. As shown in Fig. 2, RL is positively correlated
with phosphorus content (AP and TP), which is consistent with Ma et al. (2021). This correlation may be due to the fact
that energy conversion during root development requires phosphorus for the synthesis of mitochondria. By comparing
TN and TK, we infer that high TN and TK correspond to high HP; T7 had the second lowest TN and the lowest HP (Table 3
and Fig. 2). The may be due to the fact that N can increase leaf photosynthesis (Zhou et al., 2020), and K can regulate
osmotic pressure and improve plant water-use efficiency, both of which are conducive to plant growth.
In this study, nutrients (other than AK) contributed less than pH, EC, BD, and porosity to plant growth. This is
consistent with Gouin (1998), who found that plant growth in compost was more affected by compost physicochemical
properties than by compost nutrient content. In addition, the raw material of the growth medium used in the current study
experiment was compost, which already contained substantial quantities of nutrient elements and OM, such that nutrient
elements and OM were no longer the main factors limiting plant growth. Although nutrient levels did not greatly affect
the results of this study, they are clearly important but can be more readily modified (via fertilization) than other media
properties. AK may have been found to be strongly related to plant growth because of its low original content in GWC, but
the addition of VMT increases AK. VMT is a potassium-rich mineral that can be used as a source of potassium (slow-release
fertilizer, etc.) for agriculture (Silva et al., 2014) and is not easy to be lost, resulting in nutrient loss (Rezaeinejad et al.,
2021). At the same time, HA increases potassium availability.
4.5. Prospects
The current study suggests how GWC can be improved as a soilless growth medium. The popularization of composting
with additives can not only promote the development of the flower cultivation industry, but can also facilitate the
recycling of waste resources. Future research should focus on finding more economically accessible substances, preferably
waste, to be used as growth media or as additives to growth media. Future research should also conduct longer-term
experiments to determine how additives can improve GWC and other composts as growth media and the reusability of
the improved growth media.
5. Conclusions
The best growth of cornflower plants was obtained when GWC was amended with 3.5% vermiculite and 0.5 g/100 g
humic acid. This may be related to the excellent physical structure, high adsorption performance, and biological activity
of the additives. Better results were obtained with the combined addition of vermiculite and humic acid than with
addition of either vermiculite or humic acid. Redundancy analysis showed that plant growth parameters (biomass, flower
number, plant height, and root length) were negatively correlated with pH, EC, and BD; that the nutrient content of the
media was positively correlated with plant growth. The effects of pH, EC, and BD were stronger than nutrients. Thus,
in the development of soilless growth media, the nutrient contents of the growth media are less important than other
environmental factors because low nutrient levels can be corrected via fertilization. Pore structure, pH, and salinity, in
contrast, are difficult to correct.
The authors declare that they have no known competing financial interests or personal relationships that could have
appeared to influence the work reported in this paper.
Acknowledgments
This study was supported by the National Natural Science Foundation of China (No. 32171751; No. 31700537).
All authors approved the version of the manuscript to be published.
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