Biomass and Bioenergy 144 (2021) 105928

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Biomass and Bioenergy

journal homepage: http://www.elsevier.com/locate/biombioe

Effect of digestate and digestate supplemented with biochar on switchgrass

growth and chemical composition
Moon-Sub Lee a, Meltem Urgun-Demirtas b, Yanwen Shen b, 1, Colleen Zumpf a,
Eric K. Anderson c, A. Lane Rayburn a, D.K. Lee a, *
a
Department of Crop Sciences, University of Illinois at Urbana-Champaign, 1102 S. Goodwin Ave, Urbana, IL, 61801, USA
b
Applied Materials Division, Argonne National Laboratory, 9700 S Cass Ave, Lemont, IL, 60439, USA
c
Michigan State University Extension, 612 E. Main St., Centreville, MI, 49032, USA

A R T I C L E I N F O A B S T R A C T

Keywords: Digestate, a by-product of anaerobic digestion, can be an excellent source of nutrients for bioenergy crop pro­
Digestate duction, while biochar, resulting from pyrolyzing biomass, can valorize the nutrients of digestate. The objective
Biochar of this study was to evaluate the nutritional value of digestate and digestate + biochar for switchgrass pro­
N fertilization
duction. The greenhouse experiment was conducted to determine the effects of a nitrogen (N) source including 0
Switchgrass
N (control), urea (nutrient control), sewage sludge digestate, and sewage sludge digestate + corn stover biochar
Bioenergy crop production
on switchgrass growth and feedstock composition. Nitrogen sources were found to significantly increase plant
weight at the 32-week harvest, with weight ranging from 94.2 to 125.2 g plant− 1, depending on the N source,
compared to 70.7 g plant− 1 from the negative control. Moreover, switchgrass treated with digestate + biochar
showed higher plant weight than samples treated only with digestate. Nitrogen application of urea, digestate,
and digestate + biochar had positive effects on plant height and total leaf area in the 32-week harvests and
chlorophyll content in the 8-week harvests. Feedstock quality traits including hemicellulose, cellulose, and lignin
content were not affected by N sources in either the 8 or 32-week harvests with the exception of ash concen­
tration in the 32-week harvests. Our results indicate that digestate and digestate + biochar can be a potential
substitute for chemical fertilizer, resulting in increased biomass yield.

1. Introduction
Switchgrass (Panicum virgatum L.) is a C4 warm-season grass native to
North American prairies and is a model perennial grass for bioenergy as
well as the most advanced herbaceous perennial bioenergy feedstock
due to its high yield potential and low agronomic inputs [1–5]. During
the last several decades, switchgrass biomass productivity has been
greatly improved through high yielding cultivar development and
implementation of best agronomic management practices including ni­
trogen (N) fertility and harvest [4,5]. Switchgrass biomass yields can be
significantly impacted by soil nutrient availability. Nitrogen is particu­
larly important for switchgrass biomass production as N is the primary
limiting factor for biomass yield, and exogenous application of N fer­
tilizer contributes significantly to the cost of production [6,7].
The recommended N rate for switchgrass production depends on site,
cultivar, and management practices [6]. Optimal N rates for switchgrass
grown as pasture or hay ranged from 50 to 120 kg N ha− 1 in the central
Great Plains and Midwest [8], while ‘Alamo’ switchgrass grown for
biomass production required 168 kg N ha− 1 in Texas [9]. Conversely,
Mulkey et al. [10] suggested 56 kg N ha− 1 was adequate for switchgrass
production on Conservation Reserve Program (CRP) land in South
Dakota. Anderson et al. [11] indicated site-specific N fertility manage­
ment based on soil types and available soil N need to be considered to
improve N use efficiency for switchgrass biomass production. In addi­
tion, the feedstock composition, which is affected by management
practices including N fertilization, determines the conversion efficiency
of biomass to liquid fuels or heat energy based on conversion technol­
ogies [12–14]. Consequently, it is important to understand how fertil­
ization rates impact nutrient dynamics in harvested biomass, which
ultimately affects the sustainability of biomass energy production sys­
tems [15–17].
Anaerobic digestion (AD) is one of the most commonly adopted

* Corresponding author. University of Illinois at Urbana-Champaign, 1102 S. Goodwin Ave., Urbana, IL, 61801, USA.
E-mail address: leedk@illinois.edu (D.K. Lee).
1
Present address: School of Environmental Science and Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China.

https://doi.org/10.1016/j.biombioe.2020.105928
Received 18 September 2019; Received in revised form 4 December 2020; Accepted 6 December 2020
Available online 21 December 2020
0961-9534/© 2020 Elsevier Ltd. All rights reserved.
M.-S. Lee et al.
technologies in waste treatment practices and energy recovery because
of its simplicity and applicability for treatment of a wide range of
organic waste streams, such as sewage sludge, municipal solid waste,
and agricultural residues. The main driving force for the deployment of
AD technologies is waste stabilization through degradation of organic
compounds in waste streams and energy recovery in the form of biogas.
Biogas can be upgraded to biomethane (i.e. renewable natural gas) to
produce power and heat or can be directly used as a fuel in the trans­
portation sector [18]. Digestate is a post-AD byproduct consisting of
microbial cell mass and not readily biodegradable matter. Digestate is
also rich in nutrients (e.g. N, phosphorus, and potassium) and its valo­
rization may significantly enhance the overall economic benefits of the
AD industry [19]. Digestate can potentially be used as a fertilizer for
crop production instead of being deposited into landfills [20].
Recently a novel waste-to-energy system was developed by Shen
et al. [21]. This Integrated Waste to Energy and Nutrient Production
System (IWENPS) aims to produce the pipeline-quality renewable
methane (biomethane) and fertilizer-grade digestate (i.e. leftover from
anaerobic digestion of wastes) for soil application and reduction of
greenhouse gas (GHG) emissions and waste volumes. The system con­
sists of a sequential biomass pyrolysis for energy production and biochar
production and in situ AD of organic wastes with biochar for renewable
methane and digestate production, which could be used as a fertilizer.
Additional studies further demonstrated that biochar serves as an
effective additive to enhance AD performance in terms of methane
productivity and process stability [21–23]. The IWENPS is an attractive
process that permits the conversion of a large range of organic wastes
into renewable fuel. However, the digester effluent, digestate, needs to
be fully valorized to create a closed-loop system. Digestate valorization
and its integration in a sustainable biorefinery concept are crucial to
demonstrating a self-sustaining IWENPS utilizing digestate for the
cultivation of energy crops.
Upon addition of biochar into the digester, functional groups on the
biochar surface can interact with organic and inorganic compounds
during AD via various mechanisms, such as adsorption, electrostatic
interaction, ion exchange, and precipitation [24]. Such dynamics may
affect the bioavailability of the nutrients in the biochar-digestate matrix
for crop growth and nutrient uptake. The compositional and structural
analyses of digestate from biochar-supplemented digesters suggest that
it can be a suitable nutrient supply for the cultivation of energy crops
[21]. The utilization of digestate amended with biochar for the culti­
vation of energy crops can create an economic opportunity and addi­
tional income for the AD industry while preventing the flow of nutrient
pollution into the natural water bodies.
Although biochar has been extensively reported for its application as
a soil amendment and has shown benefits for plant growth and soil
quality improvement [25], biochar, which was added to the AD process,
has not been tested for its value for crop production. The agronomic
value of digestate + biochar from IWENPS must be quantified if it is to
be used as low-cost fertilizer for bioenergy feedstock production.
Therefore, the objective of this study was to compare whether there is
the effect of digestate with and without biochar on switchgrass yield and
composition as well as to determine an optimum application rate for
sustainable and cost-effective switchgrass biomass production.
2. Materials and methods
2.1. Digestate material and production
The digestate sources originated from sewage sludge produced by
the Woodridge Greene Valley Wastewater Facility (Woodridge, IL). The
semi-continuous two-stage anaerobic digestion (AD) experiments were
performed through a two-stage fermenter system (BioFlo 110, Eppen­
dorf, New Brunswick) which separates the acid and methane formation
stages. The system consisted of two identical 14-L stirred tank reactors
with individual control of pH, temperature, and agitation. The two-stage
2
Biomass and Bioenergy 144 (2021) 105928
AD systems were set up and run based on the operating conditions
specified by Shen et al. [23]. Each reactor had a working volume of 9.4
L. Sludge samples containing 1.86% (w/v) total solids and 1.47% (w/v)
volatile solids were taken from a local digester facility in Illinois to
inoculate the digesters. A digestate sample from a methanogenic
digester was taken daily to determine the pH, solid content, and nutrient
content. The chemical properties of the digestate are shown in Table 1.
The biochar used for the experiments was produced by the National
Renewable Energy Laboratory (NREL, Golden, CO). It was generated
from the pyrolysis of corn stover using NREL’s pilot-scale Thermo­
chemical Process Development Unit. Pyrolysis was conducted at
600–875 ◦ C. Specific information on the procedure of biochar produc­
tion was described in Carpenter et al. [26]. The reference of biochar
henceforth will refer to corn stover biochar. Chemical properties of the
biochar and digestate are shown in Table 2, with additional physical and
chemical properties described in Shen et al. [21].
2.2. Plant material
Switchgrass cultivar ‘Kanlow’ (USDA Natural Resources Conserva­
tion Services, Manhattan Plant Materials Center, KS, USA) was used for
the experiment. Each seed was individually planted at a depth of 3 mm
in a propagation tray liner (Nursery Supplies Inc, PA, USA) using Sun­
shine Metro-Mix1950 (Sun Gro Horticulture Distribution Inc., MA, USA)
as the growing medium. After planting, seedlings were maintained in a
greenhouse. Six-week-old plants with a fully expanded first leaf
(approximately 20 cm plant height) were selected for transplanting.
2.3. Plant growth condition
Tree-pots (8.69 L, 25.7 cm diameter × 23.2 cm deep, classic 1000,
Nursery Supplies Inc., PA, USA) were lined with a coffee filter to prevent
the potting mix from leaching out of the drainage holes. Each pot was
filled with a uniform volume of 8 L consisting of either soil or soil mixed
with fertilizer. The treatments included controls (no fertilizer), nutrient
controls (granular urea, 46-0-0), digestate, and digestate + biochar.
Application rates of digestate, digestate + biochar, and urea corre­
sponded to 0, 50, 100, and 150 kg N ha− 1. Digestate and digestate +
biochar application rates were calculated based on total N concentration
of digestate samples (Table 1). The required amount of digestate,
digestate + biochar, or urea for each application rate was mixed with
soil, which was prepared by mixing torpedo sand (0.0–0.6 cm size) and
Flanagan silt loam (Fine, smectitic, mesic Aquic Argiudolls) at a ratio of
2:1 (wt/wt). The base soil was designed to determine the sole effect of
digestate and digestate + biochar and minimize any nutrient derived
from soils. Basic chemical properties of the soil medium are presented in
Table 3, whereafter the mixing in the digestate and digestate + biochar,
the soil pH ranged from 7.9 to 8.0. After gently removing the potting mix
around roots, switchgrass seedlings were transplanted into each pot
filled with the premixed soil. Each pot was placed on a plastic plant
saucer (25.4 cm × 3 cm deep, Curtis Wagner plastics, TX, USA) and
periodically watered from below and misted from above as needed to
maintain moist soil conditions. The greenhouse temperature was
maintained between 27 ◦ C (day) and 16 ◦ C (night). A14 photoperiod was
kept with high-pressure sodium lamps providing 400 μmol m− 2 s− 1
photon flux at canopy level when necessary. The greenhouse experiment
was completely randomized with 50 switchgrass pots (5 replications per
treatment) that were established and harvested after 8 weeks (with two
replications) and after 32 weeks (three replications) (Suppl. Fig. 2). The
pots were randomly positioned on benches and rotated every week. The
entire experiment was repeated twice over time.
2.4. Determination of agronomic traits
The number of tillers and leaves along with the height of the tallest
tiller (from soil level to the tip of the tallest leaf) were recorded weekly
M.-S. Lee et al. Biomass and Bioenergy 144 (2021) 105928

Table 1
Chemical characteristics of digestate and digestate supplemented with biochar used in this experiment.
Total N NH4–N NO3–N P2O5 K2O S B Zn Mn Fe
− 1
————————————————————————————————————————————— mg kg
——————————————————————————————————————————————
Digestate 4133 632.3 88.7 496.9 104.6 85.5 0.7 7.6 6.4 87.2
D. Biochara 4200 989.3 51.3 899.1 1676.0 166.7 1.2 17.4 14.0 225.1
————————————————————————————————————————————— mg kg− 1
——————————————————————————————————————————————
Cu Ca Mg Na Al Cd Cr Pb Co Ni
Digestate 5.8 275.4 69.6 35.0 43.5 <0.03 1.0 3.3 <0.05 0.5
D. Biochar 11.9 629.8 177.0 469.0 127.9 <0.03 1.5 1.4 0.075 0.7
a
D.Biochar: digestate + biochar.

Table 2
Chemical characteristics of corn-stover biochar used in this experiment (adapted from Shen et al. [21]).
Biochar chemical characteristics

————————————————————————————————————————————— mg kg-1
—————————————————————————————————————————————
Total N C H O S
5000 527800 3300 2500 5200000
Biochar ash chemical characteristics
————————————————————————————————————————————— mg kg-1
——————————————————————————————————————————————
SiO2 Al2O3 TiO2 Fe2O3 CaO MgO Na2O K2O P2O5 SO3 Cl CO2
605800 56500 2700 19300 38700 42300 7400 141700 21900 2200 10100 11700

2.5. Analysis of feedstock chemical composition

Table 3
Chemical characteristics of soil used in this greenhouse experiment.
Organic matter NO3–N NH4–N P2O5 K2O
g kg− 1
————————————————— mg kg− 1
—————————————————
Soil 10.7 7 8 23 62
during the experimental period. After each harvest, additional agro­
nomic traits from each pot were measured including total plant fresh
weight (g plant− 1), wet and oven-dry weights of above- and below-
ground biomass, plant height (cm), tiller weight (g tiller− 1), and num­
ber of tillers (tiller plant− 1). Root samples were processed by using a
volume displacement method described by Harrington et al. (1994). Dry
matter content (g plant− 1) was determined by drying at 60 ◦ C for up to
72 h until the dry weight was constant, and subsamples from each pot
were used for analyzing feedstock chemical composition. Leaf area was
calculated with the method of O’Neal et al. [27]. 32-week-old leaves
were collected and then placed on a white background board to take
digital images with a l × 1 cm2 grid through a camera (ELPH 300HS
Cannon Inc., Tokyo, Japan). The digital images of leaves were calculated
using Image J v1.51p for Windows (https://imagej.nih.gov/ij/down
load.html). Chlorophyll a and b content were measured following the
method described by Mustafiz et al. [28]. Eight-week-old leaves were
collected, weighed, and ground with 80% acetone. The extract was
transferred into a 2 ml micro-centrifuge tube and maintained on ice
under dark conditions. After centrifuging at 10,000 rpm for 15 min, the
supernatant was transferred into a new 2 ml micro-centrifuge tube and
kept on ice. The absorbance at 645 and 663 nm was measured with a
spectrometer (G10S UV–Vis, Thermo Fisher Scientific, Madison, WI,
USA). The equation was used to calculate chlorophyll a and b:Where
A663 and A645 are the absorbances of chlorophyll extract at 663 nm and
645 nm respectively. V is the final volume of solution (mL) and w is the
fresh weight of leaf tissue in grams [28]. Chlorophyll absorbance was
estimated using a SPAD chlorophyll meter (MINOLTA SPAD-502, Min­
olta Camera Co., Ltd., Osaka, Japan).
Above-ground biomass samples were ground to pass through a 1 mm
screen using a cutting mill (Retsch SM2000, Haan, Germany). Ash
concentration in biomass samples was estimated using a muffle furnace
incineration at 600 ◦ C for 4 h. Fiber analyses of biomass samples were
performed sequentially using an Ankom 200 Fiber Analyzer
(Ankom21Technology, Macedon, NY, USA) for neutral detergent fiber
(NDF) and acid detergent fiber (ADF). Acid detergent lignin (ADL) was
determined by using an Ankom Daisy II incubator. The fiber analysis was
conducted according to the manufacturer’s protocol (Ankom Technol­
ogy, Macedon, NY, USA) [29–31]. The NDF, ADF, and ADL data were
utilized to estimate lignocellulosic concentrations in biomass. Cellulose
was estimated by calculating the difference between ADF and ADL,
while hemicellulose was estimated by calculating the difference be­
tween NDF and ADF. To measure total macro and micronutrients, 0.2 g
of the ground samples were treated with 5.0 ml of hydrochloric acid and
2.5 ml of nitric acid and set in microwave for 30 min. After adding 50 ml
of distilled water, the solutions were filtered and analyzed by induc­
tively coupled plasma mass spectrometry (iCAPTM 6500 Duo ICP,
Thermo Fisher Scientific Inc., Waltham, MA, USA) [32]. Nutrient
removal and mineral removal by plant biomass were determined by
multiplying biomass yield by the corresponding concentration of each
element.
2.6. Statistical analysis
Normality of the residuals and equality of the variances was tested
using a box plot in the UNIVARIATE procedure in SAS software (SAS
System for Windows, Version 9.4, SAS Institute Inc., Cary, NC, USA).
The data were analyzed using the MIXED procedure of SAS, with α =
0.05. The PDMIX800 macro in SAS was used to generate letter groupings
of similar means [33]. There was no significant difference between the
trials and the data were pooled across experiments. N source and rate
were considered as fixed variables. Contrast statements were used to test
if responses to fertilization were different among N sources using the
MIXED procedure. Correlations were examined between feedstock
chemical composition and N rate using the CORR procedure in SAS. The
quadratic + plateau model [34] was applied to calculate the optimal N

3
M.-S. Lee et al. Biomass and Bioenergy 144 (2021) 105928

fertilization rate using the NLIN procedure in SAS.

3. Results

All nutrients in the digestate + biochar treatment were higher than

digestate without biochar except NO3–N (Table 1). Phosphorus as P2O5
was almost two times higher and potassium (K) as K2O was 15 times
higher with digestate + biochar than digestate alone. With regard to the
effect of N source, plant weight was significant improved for the 32-
week harvest (P < .0001), with plant weight varying from 70.7 to
125.3 g plant− 1 depending on N source (Table 3). For the 32-week
harvest, all N treatments produced higher plant weight than the con­
trol (P < .0001), and switchgrass treated with digestate + biochar
showed higher plant weight than switchgrass treated solely with
digestate (Table 4). Plant weight was not significantly affected by N
source in the 8-week harvest, but plant weight, averaged across nutrient
control, digestate, and digestate + biochar, was higher than in the
controls (Table 4). Plant tissue N removal for each N source was highly
correlated with N application rate, and the rate of increase was greatest
with urea and least with digestate (Fig. 1). The biomass yield of the
nutrient control increased with increasing N rates until a plateau was
reached. Based on plant weight at the 32-week harvest, the optimal N Fig. 1. Relationships between N application rate and switchgrass N removal
fertilization rate of the urea control was 90 kg N ha− 1, according to the (A) and switchgrass biomass yield (B). ( ) - Urea, ( ) - Digestate, and ( ) -
quadratics + plateau model. Below ground biomass measurements Digestate + biochar.
†Nitrogen removal was calculated by multiplying biomass yield by the corre­
including root weight, root volume, and R:S ratio did not differ across N
sponding concentration of each element biomass.
sources in the 8- or 32-week harvests (Table 4).
With regard to agronomic traits, plant height was significantly
affected by N rates in the 32-week harvest (P = 0.0047). No other traits for digestate applied at a rate of 50 kg N ha− 1, and total leaf area of
varied significantly across N sources in the 8- or 32-week harvest digestate + biochar samples was not different from digestate samples.
(Table 5). All pots treated with N had higher plant height, tiller weight, Chlorophyll a and b contents did not respond to N fertilization, but
and number of leaves than the control, but there were no differences chlorophyll b content was higher in samples from the nutrient control
among the nutrient control, digestate, and digestate + biochar treat­ than in samples from the digestate treatment when averaged across the
ments in the 32-week harvest (Table 5). rates (Table 6).
In terms of leaf characteristics, there were significant effects of N Structural carbohydrates including hemicellulose, cellulose, and
treatment on chlorophyll absorbance (P = 0.0390) (Table 6). Chloro­ lignin were not affected by N fertilization in the 8- or 32-week harvest
phyll absorbance was not found to differ between nutrient controls and with the exception of ash concentration (P = 0.0004) in the 32-week
digestate + biochar treatments. Total leaf area was impacted by N rates harvest (Table 7). Ash concentration in the 32-week harvest varied be­
(P = 0.0198), and total leaf area varied between 2,356 cm2 and 4,787 tween 36.1 and 44.1 g kg− 1. Switchgrass without N fertilization had
cm2. higher ash concentration than plants grown with N except for digestate
Total leaf area was greater in samples from the nutrient control, at the rate of 50 kg N ha− 1. In addition, switchgrass treated with either
digestate, and digestate + biochar treatments than in the control except digestate or digestate + biochar had higher ash concentration than

Table 4
Above- and below-ground biomass yield of switchgrass affected by application of urea, digestate, and digestate + biochar.
Treatment N ratea Plant weight Root weight Root volume R:Sb

8wk 32wk 8wk 32wk 8wk 32wk 8wk 32wk

− 1 − 1 − 1 3 − 1
kg N ha g plant g plant cm plant

Control (C) 0 18.9 70.7e 75.0 205.1 64.6 227.5 6.2 2.8
Nutrient control (N) 50 31.0 113.2ab 144.2 362.8 137.9 341.3 4.8 3.3
100 34.4 122.4a 125.0 292.0 128.6 288.8 4.2 2.4
150 28.1 125.3a 106.7 261.3 143.2 341.3 5.2 2.1
Digestate (D)a 50 27.0 82.6de 188.3 321.7 174.1 291.7 6.9 4.0
100 23.8 94.1cd 100.8 296.3 88.9 291.7 9.2 3.0
bc
150 32.7 102.2 168.3 270.6 152.6 274.2 6.8 2.7
cd
Digestate + biochar (DB) 50 27.2 94.2 150.0 348.4 134.0 344.2 5.5 3.7
bc
100 31.1 101.7 142.5 395.2 114.4 297.5 6.9 2.7
150 35.4 110.3ab 170.8 274.1 84.0 280.2 6.2 3.6
N fertility contrast ———————————————————————————————————————————————— P > F
————————————————————————————————————————————————
C vs. (N + D + DB) 0.0117 <.0001 0.1713 0.0425 0.1342 0.0553 0.9974 0.6358
N vs. D 0.3049 <.0001 0.4286 0.8146 0.4263 0.4814 0.0043 0.1463
N vs. DB 0.9961 0.0028 0.4614 0.4206 0.3952 0.5740 0.1325 0.0830
D vs. DB 0.3027 0.0380 0.9540 0.2939 0.9539 0.2051 0.1330 0.7490

Mean with different letters are significantly different within a column at a = 0.05.
a
N rate of digestate and digestate + biochar treatment was calculated based on total N of digestate and digestate + biochar; Granular urea (46-0-0) was used for the
source of nutrient control at rates of 50, 100, and 150 kg N ha− 1
b
R:S, Root and shoot ratio.

4
M.-S. Lee et al. Biomass and Bioenergy 144 (2021) 105928

Table 5
Effects of N fertilization with digestate and digestate + biochar on switchgrass agronomic traits.
Treatment N rateb Plant height Total tiller number Tiller weight Leaf number

8wk 32wk 8wk 32wk 8wk 32wk 8wk 32wk

1 1 1 1
kg N ha− cm tillers plant− g plant− leaves plant−

Control (C) 0 165.0 168.5ba 10.0 15.7 2.0 5.5 5.0 6.5
Nutrient control (N) 50 171.0 252.7a 12.5 15.2 2.5 7.7 5.5 7.7
100 189.0 251.7a 11.5 16.2 3.1 8.0 5.5 8.5
150 168.8 252.5a 14.5 16.8 1.9 7.3 5.0 7.0
Digestate (D)b 50 175.0 239.8a
11.5 11.7 2.5 7.7 5.5 7.5
a
100 157.0 245.8 10.5 15.2 2.4 7.2 5.5 7.7
a
150 167.0 241.8 11.5 16.8 2.9 6.2 5.5 7.5
Digestate + biochar (DB) 50 174.5 238.7a 11.25 14.7 2.5 7.3 5.0 7.3
100 193.8 253.8a 12.75 15.2 2.4 8.2 6.0 8.5
150 174.5 233.4a 11.75 14.3 2.9 8.8 5.5 7.7
N fertility contrast ———————————————————————————————————————————————— P > F
————————————————————————————————————————————————
C vs. (N + D + DB) 0.3059 <.0001 0.2188 0.7633 0.1614 0.0154 0.1925 0.0113
N vs. D 0.1707 0.3829 0.1807 0.2804 0.8829 0.3016 0.5235 0.4452
N vs. DB 0.5139 0.3701 0.4569 0.2492 0.6508 0.9012 0.5235 0.6962
D vs. DB 0.0477 0.9693 0.5421 0.9260 0.7596 0.3635 1.0000 0.2482
a
Mean with different letters are significantly different within in a column at a = 0.05.
b
N rate of digestate and digestate + biochar treatment was calculated based on total N of digestate and digestate + biochar; Granular urea (46-0-0) was used for the
source of nutrient control at rates of 50, 100, and 150 kg N ha− 1

traits in the present study, and digestate and digestate + biochar were
Table 6
relatively comparable to the chemical fertilizer, urea (Table 4). Like­
Effects of N fertilization with urea, digestate, and digestate + biochar on chlo­
wise, plant weight responded positively to digestate + biochar in our
rophyll a and b, and SPAD of switchgrass harvested eight weeks after
study and was equal to or was enhanced in digestate + biochar treat­
transplanting.
ments compared to treatments with only digestate. Optimal N fertilizer
N ratea Chlorophyll Chlorophyll Chlorophyll
rates varied for urea, digestate, and digestate + biochar in our experi­
a b absorbance
ment. Even though the recommendations for switchgrass differ
1 1
kg N mg g FW− mg g FW− depending on location and harvest timing, generally 50–110 kg N ha− 1
ha− 1
Control (C) 0 0.105 0.055 30.1c
is required to maximize biomass [6,11,36] which is in agreement with
Nutrient control 50 0.088 0.085 34.7bc our result from the inorganic fertilizer treatment. Even though appli­
(N) 100 0.076 0.109 37.1ab cation rates of digestate and digestate + biochar did not reach the
150 0.102 0.151 40.2a plateau, we assumed optimal application rates of digestate and diges­
Digestate (D)a 50 0.116 0.071 32.7bc
tate + biochar might be higher than the rate estimated for urea appli­
100 0.095 0.097 31.6bc
150 0.078 0.061 32.8bc cation. It should be noted that total N in digestate samples consisted of
Digestate + 50 0.095 0.093 34.4bc 17–25% NH4–N and NO3–N, and 75–83% organic nitrogen. Organic N is
biochar (DB) 100 0.101 0.088 35.8ab not easily taken up by plants compared to urea. Eghball and Power [37]
150 0.087 0.074 33.2bc reported that the annual N mineralization rate of field applied organic
N fertility contrast ————————————————————— P >
F ————————————————————
waste was approximately 40% of total N applied during the first year.
C vs. (N + D + DB) 0.3993 0.1184 0.0272 However, in this experiment, even though the initial concentration of
N vs. D 0.4906 0.0416 0.0032 total N, NH4–N, and NO3–N was not different between digestate and
N vs. DB 0.5912 0.1138 0.0757 digestate + biochar, switchgrass yield tended to be higher in digestate +
D vs. DB 0.8778 0.6207 0.1830
biochar than digestate. It assumed that digestate + biochar appeared to
Mean with different letters are significantly different within a column at a = have more accessible N content than digestate alone. The reason for this
0.05. observation is that biochar can help to increase soil microbial biomass
Plant tissue samples harvested after 8 weeks were analyzed for the experiment. and activity associated with N cycling. Warnock et al. [38] addressed
a
N rate of digestate and digestate + biochar treatment was calculated based how biochar could affect the abundance or activity of mycorrhizal fungi
on total N of digestate and digestate + biochar; Granular urea (46-0-0) was used
in soils and plant roots. One explanation is that the application of bio­
for the source of nutrient control at rates of 50, 100, and 150 kg N ha−
char can affect soil physical and chemical properties such as pH, soil
cation exchange capacity, water holding capacity, and bulk density
switchgrass treated with urea. As the N application rate increased across [39–41]. Consequently, the modifications can have influences on soil
all treatments, ash concentration in switchgrass decreased (Suppl. nutrient availabilities including N, P, and metal ions [39,42–44], and the
Fig. 1A). Feedstock mineral concentrations responded differently to alterations of nutrient availability would be a primary driver of
urea, digestate, and digestate + biochar (Table 8). Additionally, ash mycorrhizal fungal abundance. Moreover, biochar is well-known as a
concentration was positively correlated with K concentration (R2 = soil amendment and biochar is a highly porous structure which provides
0.3818, P < .0001) across N sources (Suppl. Fig. 1B). micro-organisms with habitats [24]. For instance, one of the most crit­
ical nutrient movements is the N cycling process including N fixation,
4. Discussion mineralization, nitrification, and denitrification. Ducey et al. [45]
investigated how switchgrass-biochar affects microbial N cycling gene
Nitrogen and water are critical for the achievement of full biomass abundances and found soil microbial N cycling communities were
yield potential in warm-season perennial grasses, including switchgrass increased by applying biochar. In particular, the application of biochar
[6], even though switchgrass has high N and water use efficiency [35]. to switchgrass had positive impacts on relative gene abundance nifH and
Nitrogen fertilization increased switchgrass biomass along with growth nirS which encode the important enzymes in N fixation and

5
M.-S. Lee et al. Biomass and Bioenergy 144 (2021) 105928

Table 7
Switchgrass biomass feedstock composition affected by application of urea, digestate, and digestate + biochar.
Treatment N rateb Hemicellulose Cellulose Lignin Ash

8wk 32wk 8wk 32wk 8wk 32wk 8wk 32wk

1
kg N ha− ———————————————————————————————————————————————— g
kg− 1————————————————————————————————————————————————
Control (C) 0 342.2 306.4 355.3 396.9 54.2 64.8 72.3 44.1aa
Nutrient control (N) 50 333.6 290.1 355.3 411.8 57.9 70.3 73.6 36.1c
100 341.6 316.6 305.4 401.5 39.5 68.4 81.9 30.2d
150 350.5 316.6 354.4 405.6 45.8 77.4 78.8 29.4d
Digestate (D)b 50 342.4 309.7 350.9 426.4 38.3 69.9 78.5 42.5ab
100 351.2 313.8 359.4 394.9 52.6 67.3 77.7 36.3c
150 334.5 306.8 330.2 398.7 59.6 66.6 72.1 36.1c
Digestate + biochar (DB) 50 352.1 313.4 293.5 417.2 42.0 69.2 79.2 37.2bc
100 340.8 313.7 354.9 408.8 45.0 71.2 77.4 37.6bc
150 339.8 306.5 349.1 387.7 44.7 66.6 77.6 35.4cd
N fertility contrast ———————————————————————————————————————————————— P > F
————————————————————————————————————————————————
C vs. (N + D + DB) 0.9182 0.6883 0.4637 0.3884 0.8562 0.1599 0.2518 0.0004
N vs. D 0.8819 0.7711 0.6156 0.9658 0.4399 0.1185 0.5689 0.0005
N vs. DB 0.6685 0.5958 0.7368 0.7967 0.8744 0.1532 0.9928 0.0086
D vs. DB 0.7763 0.8036 0.4114 0.7607 0.5513 0.9090 0.5853 0.3592
a
Mean with different letters are significantly different within in a column at a = 0.05.
b
N rate of digestate and digestate + biochar treatment was calculated based on total N of digestate and digestate + biochar; Granular urea (46-0-0) was used for the
source of nutrient control at rates of 50, 100, and 150 kg N ha− 1

Table 8
Effects of N fertilization with urea, digestate, and digestate + biochar on feedstock mineral concentrations of switchgrass harvested 32 weeks after transplanting.
Treatment N rateb N K P S Na Ca Mg

– kg ha− 1– ————————————————————————————————————— mg kg− 1

——————————————————————————————————————

Control (C) 0 2233b-da 7550a 217 467 383 3183 1500c

Nutrient control (N) 50 1950d 5467c-e 100 400 367 3083 1817bc
100 2750a-c 5150de 167 467 467 3133 2100ab
150 3267a 4643e 105 474 627 3204 2200a
Digestate (D)b 50 2767a-c 6867ab 183 450 433 2867 1517c
100 2850ab 6783ab 222 433 427 2750 1667c
150 2980b-d 5317e 105 450 400 2933 1800bc
Digestate + biochar (DB) 50 2600b-d 6367a-c 133 417 350 2817 1633c
100 2117cd 6583a-c 133 383 300 2683 1617c
150 2333b-d 6163b-d 137 374 327 2784 1560c
N fertility contrast —————————————————————————————————————— P > F
——————————————————————————————————————
C vs. (N + D + DB) 0.1148 0.0003 0.0059 0.2686 0.5997 0.1829 0.0527
N vs. D 0.4859 0.0009 0.0221 0.7749 0.1224 0.0730 0.0016
N vs. DB 0.1035 0.0003 0.7337 0.0701 0.0004 0.0174 0.0002
D vs. DB 0.0025 0.7586 0.0484 0.1246 0.2835 0.5320 0.5333
Treatment N rate‡ Fe Cu Mn B Zn Al
– kg ha− 1– ———————————————————————————————————— mg kg− 1
————————————————————————————————————
Control (C) 0 44ab 11 175ab 12 19 152
Nutrient control (N) 50 35cd 10 108bc 14 21 153
100 40a-d 9 87c 13 17 140
150 48a 10 79c 13 22 147
Digestate (D)‡ 50 43a-c 9 210a 10 17 143
a a
100 46 8 217 11 22 144
ab a
150 46 10 189 13 21 169
a-d c
Digestate + biochar (DB) 50 40 9 85 12 20 148
100 34d 10 72c 11 20 184
150 37b-d 10 87c 12 22 155
N fertility contrast ———————————————————————————————————— P > F
————————————————————————————————————
C vs. (N + D + DB) 0.3448 0.0190 0.1092 0.9079 0.3253 0.9302
N vs. D 0.1147 0.2553 <.0001 0.0719 0.6914 0.7407
N vs. DB 0.0967 0.8111 0.6919 0.0873 0.6000 0.2237
D vs. DB 0.0019 0.3666 <.0001 0.9224 0.8988 0.3734
a
Mean with different letters are significantly different within in a column at a = 0.05.
b
N rate of digestate and digestate + biochar treatment was calculated based on total N of digestate and digestate + biochar; Granular urea (46-0-0) was used for the
source of nutrient control at rates of 50, 100, and 150 kg N ha− 1

6
M.-S. Lee et al.
denitrification [45]. In addition, Ker et al. [46] demonstrated that
switchgrass plots inoculated with plant growth-promoting rhizobacteria
including Paenibacillus polymyxa, Pseudomonas species, Rahnella species
and Serratia species, had increased biomass yields than uninoculated
switchgrass plots.
Our results showed that digestate and digestate + biochar contained
macronutrients including N, P, K, Ca, Mg, and S and micronutrients such
as B, Cu, Fe, Mn, Ni, and Zn. Although the nutrient concentration of
digestate and digestate + biochar did not generally deviate from the
range reported by other authors [47–49], a difference was observed
between digestate and digestate + biochar. This observation can be
explained by the following; the application of biochar for anaerobic
digestate can adsorb and concentrate the nutrients during the biogas
process, thereby containing higher concentrations of macro- and
micro-nutrients in digestate + biochar than digestate alone [21,50].
Moreover, the in situ biogas upgrading process with biochar amendment
uses corn stover biochar to remove contaminants including carbon di­
oxide and hydrogen sulfide from raw biogas [21]. Corn stover biochar
can absorb contaminants in this system, which consequently influence
the concentrations of macro- and micro-nutrients in digestate + biochar.
Particularly, Ca and K concentration was higher in digestate + biochar
than digestate (Table 1). As discussed above, N is considered as the most
limiting nutrient for optimal plant biomass and growth, but P, K, and
other nutrients are also important for switchgrass growth and yield [51].
Even though the effect of P and K on switchgrass is little or no response
to applying P and K, McMuphy et al. [52] addressed the greatest
switchgrass yields achieved with the highest rate of N plus P, indicating
P can be a limiting factor for yield. Similarly, Taylor and Allinson [53]
suggested that switchgrass obtaining either N and P or N, P, and K fer­
tilizer had increased biomass yield than switchgrass obtaining only N
fertilizer. However, in this experiment, overall feedstock mineral com­
positions of switchgrass were not found to be different between treat­
ments. One of the explanations for the observation is linked with
nutrient remobilization because aboveground biomass was harvested at
the end of growing season in this experiment. Perennial grass species,
such as switchgrass, adopted the strategy to translocate some nutrients
from shoots to roots prior to or during senescence, thereby reducing
nutrient concentrations and removal rates from harvesting [17,54–56].
Biomass feedstocks containing high percentages of cellulose and
hemicellulose and low levels of N and minerals are favorable for con­
version processes if switchgrass is grown for bioenergy production [11,
57]. Lemus et al. [58] found that concentrations of cellulose and lignin
increased as N fertilization rate increased, whereas hemicellulose and
ash decreased. These changes are ideal, considering the greater energy
content in cellulose and lignin compared to hemicellulose and the
potentially negative effects of ash for energy conversion processing [58,
59]. In this study, neither hemicellulose nor cellulose responded to N
sources. However, as N fertility increased, ash concentration decreased,
a pattern that is comparable to the results of other studies [11,58–60].
The decline in ash concentration is desirable for biomass quality. The
ash is mainly composed of Si, K, Ca, S, and Cl [61], and ash interferes
with thermochemical processing and the power plant internal system
due to slagging, deposit formation, and corrosion [58]. In addition, the
change of ash concentration resulting from N fertilization might be
related to the plant biomass fractions including leaves, stems, and in­
florescences. Lanning and Eleuterius [62] reported silica concentration
in switchgrass was higher in leaves and inflorescences than in stems.
Moreover, N fertilization led to an increase in tiller weight and stem
development for switchgrass [8]. Consequently, it can be assumed that
the tiller and stem proportion is increased compared to leaves as N
fertility increases [63]. The change in proportion could impact the
quality of switchgrass.
In addition, in our study, a positive correlation was observed be­
tween ash and K concentration (Supp. Fig. 1). Adler et al. [64] showed
that the change of plant nutrient elements including Cl, K, P, and Mg
accounted for the decrease in ash concentration. The concentration of K
7
Biomass and Bioenergy 144 (2021) 105928
and P in biomass was different depending on N sources in the present
study. Potassium concentrations in biomass were higher in the negative
control than in the fertilized treatments. The finding is similar to Wood
et al. [65] who reported that the decrease in K concentration was
observed in bermudagrass with increasing rates of ammonium-nitrate
application.
5. Conclusion
Perennial grasses such as switchgrass can be an excellent feedstock
for bioenergy production and are better for soil carbon sequestration
than annual crops like corn. The primary idea underlying this experi­
ment was to maximize carbon storage and mitigate greenhouse gases by
combining bioenergy crop and waste management. Even though
switchgrass has high N use efficiency, N fertilization is necessary for
sustainable biomass production. When bioenergy feedstocks are
continuously harvested, N is removed from the system and should be
replenished by an external source of N. Therefore, N fertility manage­
ment, including the choice of N source and the application rate, is
important for developing switchgrass as a bioenergy feedstock not only
economically, but also for the environment. Biochar derived from bio­
energy conversion processes and digestates produced from anaerobic
digestion processes could be used as N sources while improving soil
quality and C sequestration. Our results can be considered in future
research developing new agronomic strategies for sustainable bioenergy
production and assessing the recycling of nutrients from digestate and
digestate + biochar. However, the readers are cautioned that the
observation in this study should be viewed as preliminary and that
additional experimentation is required to confirm these results in field-
scale plots. For example, in this experiment, the rate of digestate and
digestate plus biochar did not reach a plateau yield. In order to find the
plateau, it will probably be necessary to expand the range of digestate
and digestate plus biochar concentrations. Also, further testing is needed
to determine if there are any negative effects on switchgrass produc­
tivity when digestate and digestate plus biochar concentrations are
increased.
Acknowledgement
The authors gratefully acknowledge the US Department of Energy’s
Bioenergy Technologies Office (BETO) for funding this work. Argonne
National Laboratory, a US Department of Energy Office of Science
Laboratory, is operated under contract no. DE-AC02-06CH11357. The
US Government retains for itself, and others acting on its behalf, a paid-
up nonexclusive, irrevocable worldwide license in said article to
reproduce, prepare derivative works, distribute copies to the public, and
perform publicly and display publicly, by or on behalf of the govern­
ment. The funding source for the work reported here did not have a role
in study design, data collection, analysis, data interpretation, writing, or
in the decision to publish.
Appendix A. Supplementary data
Supplementary data related to this article can be found at https://
doi.org/10.1016/j.biombioe.2020.105928.
References
[1] S.B. McLaughlin, M.E. Walsh, Evaluating environmental consequences of
producing herbaceous crops for bioenergy, Biomass Bioenergy 14 (1998) 317–324.
[2] D.K. Lee, J.J. Doolittle, V.N. Owens, Soil carbon dioxide fluxes in established
switchgrass land managed for biomass production, Soil Biol. Biochem. 39 (2007)
178–186.
[3] D.K. Lee, V.N. Owens, J.J. Doolittle, Switchgrass and soil carbon sequestration
response to ammonium nitrate, manure, and harvest frequency on conservation
reserve program land, Agron. J. 99 (2007) 462–468.
[4] K.P. Vogel, R.B. Mitchell, M.D. Casler, G. Sarath, Registration of
‘liberty’switchgrass, J. Plant Registrations 8 (2014) 242–247.
M.-S. Lee et al.
[5] R.B. Mitchell, M.R. Schmer, W.F. Anderson, V. Jin, K.S. Balkcom, J. Kiniry,
A. Coffin, P. White, Dedicated energy crops and crop residues for bioenergy
feedstocks in the Central and Eastern USA, Bioenergy Res 9 (2016) 384–398.
[6] K.P. Vogel, J.J. Brejda, D.T. Walters, D.R. Buxton, Switchgrass biomass production
in the Midwest USA, Agron. J. 94 (2002) 413–420.
[7] J.M. Albaugh, E.B. Sucre, Z.H. Leggett, J.C. Domes, J.S. King, Evaluation of
intercropped switchgrass establishment under a range of experimental site
preparation treatments in a forested setting on the Lower Coastal Plain of North
Caroline, USA, Biomass Bioenergy 46 (2012) 673–682.
[8] J.J. Brejda, J.R. Brown, G.W. Wyman, W.K. Schumacher, Management of
switchgrass for forage and seed production, J. Range Manag. 47 (1994) 22–27.
[9] J.P. Muir, M.A. Sanderson, W.R. Ocumpaugh, R.M. Jones, R.L. Reed, Biomass
production of ‘Alamo’switchgrass in response to nitrogen, phosphorus, and row
spacing, Agron. J. 93 (2001) 896–901.
[10] V.R. Mulkey, V.N. Owens, D.K. Lee, Management of switchgrass-dominated
conservation reserve program lands for biomass production in South Dakota, Crop
Sci. 46 (2006) 712–720.
[11] E.K. Anderson, A.S. Parrish, T.B. Voigt, V.N. Owens, C.H. Hong, D.K. Lee, Nitrogen
fertility and harvest management of switchgrass for sustainable bioenergy
feedstock production in Illinois, Ind. Crop. Prod. 48 (2013) 19–27.
[12] I. Lewandowski, A. Kicherer, Combustion quality of biomass: practical relevance
and experiments to modify the biomass quality of Miscanthus × giganteus, Eur. J.
Agron. 6 (1997) 163–177.
[13] P. McKendry, Energy production from biomass (part 1): overview of biomass,
Bioresour. Technol. 83 (2002) 37–46.
[14] M.A. Sanderson, N.P. Martin, P. Adler, Biomass, energy, and industrial uses of
forages, in: K.J. Moore, R.F. Barnes, C.J. Nelson, M. Collins (Eds.), Forages: the
Science of Grassland Agriculture, Blackwell, Ames, 2007, pp. 635–647.
[15] J.H. Reynolds, C.L. Walker, M.J. Kirchner, Nitrogen removal in switchgrass
biomass under two harvest systems, Biomass Bioenergy 19 (2000) 281–286.
[16] J.L. Propheter, S. Staggenborg, Performance of annual and perennial biofuel crops:
nutrient removal during the first two years, Agron. J. 102 (2010) 798–805.
[17] J.A. Guretzky, J.T. Biermacher, B.J. Cook, M.K. Kering, J. Mosali, Switchgrass for
forage and bioenergy: harvest and nitrogen rate effects on biomass yields and
nutrient composition, Plant Soil 339 (2011) 69–81.
[18] Y. Shen, L. Jarboe, R. Brown, Z. Wen Z, A thermochemical–biochemical hybrid
processing of lignocellulosic biomass for producing fuels and chemicals,
Biotechnol. Adv. 33 (2015) 1799–1813.
[19] W. Fuchs, B. Drosg, Assessment of the state of the art of technologies for the
processing of digestate residue from anaerobic digesters, Water Sci. Technol. 67
(2013) 1984–1993.
[20] T. Al Seadi, B. Drosg, W. Fuchs, D. Rutz, R. Janssen, Biogas digestate quality and
utilization, in: A. Wellinger, J.D. Murphy, D. Baxter (Eds.), The Biogas
Handbook–Science, Production and Applications, Woodhead Publishing Limited,
Oxford, 2013, pp. 267–301.
[21] Y. Shen, J.L. Linville, M. Urgun-Demirtas, R.P. Schoene, S.W. Snyder, Producing
pipeline-quality biomethane via anaerobic digestion of sludge amended with corn
stover biochar with in-situ CO2 removal, Appl. Energy 158 (2015) 300–309.
[22] Y. Shen, J.L. Linville, P.A.A. Ignacio-de Leon, R.P. Schoene, M. Urgun-Demirtas,
Towards a sustainable paradigm of waste-to-energy process: enhanced anaerobic
digestion of sludge with woody biochar, J. Clean. Prod. 135 (2016) 1054–1064.
[23] Y. Shen, S. Forrester, J. Koval, M. Urgun-Demirtas, Yearlong semi-continuous
operation of thermophilic two-stage anaerobic digesters amended with biochar for
enhanced biomethane production, J. Clean. Prod. 167 (2017) 863–874.
[24] M. Ahmad, A.U. Rajapaksha, J.E. Lim, M. Zhang, N. Bolan, D. Mohan,
M. Vithanage, S.S. Lee, Y.S. Ok, Biochar as a sorbent for contaminant management
in soil and water: a review, Chemosphere 99 (2014) 19–33.
[25] D.A. Laird, P. Fleming, D.D. Davis, R. Horton, B. Wang, D.L. Karlen, Impact of
biochar amendments on the quality of a typical Midwestern agricultural soil,
Geoderma 158 (2010) 443–449.
[26] D.L. Carpenter, R.L. Bain, R.E. Davis, A. Dutta, C.J. Feik, K.R. Gaston, W. Jablonski,
S.D. Phillips, M.R. Nimlos, Pilot-scale gasification of corn stover, switchgrass,
wheat straw, and wood: 1. Parametric study and comparison with literature, Ind.
Eng. Chem. Res. 49 (2010) 1859–1871.
[27] M.E. O’ Neal, D.A. Landis, R. Isaacs, An inexpensive, accurate method for
measuring leaf area and defoliation through digital image analysis, J. Econ.
Entomol. 95 (2002) 1190–1194.
[28] A. Mustafiz, K.K. Sahoo, S.L. Singla-Pareek, S.K. Sopory, Metabolic engineering of
glyoxalase pathway for enhancing stress tolerance in plants, in: R. SunKar (Ed.),
Plant Stress Tolerance: Methods and Protocols, Humana press, New York, 2010,
pp. 95–118.
[29] Ankom Technology, Method for Determining Acid Detergent Lignin in the Daisy II
Incubator, Ankom Technology Corporation, Fairport, NY, 2002.
[30] Ankom Technology, Method for Determining Neutral Detergent Fiber, Ankom
Technology Corp., Fairport, NY, 2003.
[31] Ankom Technology, Method for Determining Acid Detergent Fiber, Ankom
Technology Corp., Fairport, NY, 2003.
[32] R. Gavlak, D. Horneck, R.O. Miller, J. Kotuby-Amacher, Soil, Plant and Water
Reference Methods for the Western Region. WCC-103 Publication, WREP-125.
Wetland Reserve Enhancement Program, US Dept, Agriculture, Corvallis, OR,
2003, pp. 17–36.
[33] A.M. Saxton, A macro for converting mean separation output to letter groupings in
Proc Mixed. Proc. 23rd SAS Users Group Intl., Nashville, TN, March 22–25, SAS
Institute, Cary, NC, 1998, pp. 1243–1246.
8
Biomass and Bioenergy 144 (2021) 105928
[34] D.G. Bullock, D.S. Bullock, Quadratic and quadratic-plus-plateau models for
predicting optimal nitrogen rate of corn: a comparison, Agron. J. 86 (1994)
191–195.
[35] S.B. McLaughlin, L.A. Kszos, Development of switchgrass (Panicum virgatum) as a
bioenergy feedstock in the United States, Biomass Bioenergy 28 (2005) 515–535.
[36] J.J. Brejda, Fertilization of native warm-season grasses, in: K.J. Moore, B.
E. Anderson (Eds.), Native Warm-Season Grasses: Research Trends and Issues, Crop
Science Society of America, Madison, 2000, pp. 177–200.
[37] B. Eghball, J.F. Power, Composted and noncomposted manure application to
conventional and no-tillage systems: corn yield and nitrogen uptake, Agron. J. 91
(1999) 819–825.
[38] D.D. Warnock, J. Lehmann, T.W. Kuyper, M.C. Rillig, Mycorrhizal responses to
biochar in soil–concepts and mechanisms, Plant Soil 300 (2007) 9–20.
[39] E.H. Tryon, Effect of charcoal on certain physical, chemical, and biological
properties of forest soils, Ecol. Monogr. 18 (1948) 81–115.
[40] R.E. Lucas, J. Davis, Relationships between pH values of organic soils and
availabilities of 12 plant nutrients, Soil Sci. 92 (1961) 177–182.
[41] B. Glaser, J. Lehmann, W. Zech, Ameliorating physical and chemical properties of
highly weathered soils in the tropics with charcoal–a review, Biol. Fertil. Soils 35
(2002) 219–230.
[42] J. Lehmann, J.P. da Silva, C. Steiner, T. Nehls, W. Zech, B. Glaser, Nutrient
availability and leaching in an archaeological Anthrosol and a Ferralsol of the
Central Amazon basin: fertilizer, manure and charcoal amendments, Plant Soil 249
(2003) 343–357.
[43] M.J. Gundale, T.H. DeLuca, Temperature and source material influence ecological
attributes of ponderosa pine and Douglas-fir charcoal, For. Ecol. Manag. 231
(2006) 86–93.
[44] T.H. DeLuca, M.D. MacKenzie, M.J. Gundale, W.E. Holben, Wildfire-produced
charcoal directly influences nitrogen cycling in ponderosa pine forests, Soil Sci.
Soc. Am. J. 70 (2006) 448–453.
[45] T.F. Ducey, J.A. Ippolito, K.B. Cantrell, J.M. Novak, R.D. Lentz, Addition of
activated switchgrass biochar to an aridic subsoil increases microbial nitrogen
cycling gene abundances, Appl. Soil Ecol. 65 (2013) 65–72.
[46] K. Ker, P. Seguin, B.T. Driscoll, J.W. Fyles, D.L. Smith, Switchgrass establishment
and seeding year production can be improved by inoculation with rhizosphere
endophytes, Biomass Bioenergy 47 (2012) 295–301.
[47] J.A. Alburquerque, C. De la Fuente, M. Campoy, L. Carrasco, I. Nájera, C. Baixauli,
F. Caravaca, A. Roldán, J. Cegarra, M.P. Bernal, Agricultural use of digestate for
horticultural crop production and improvement of soil properties, Eur. J. Agron. 43
(2012) 119–128.
[48] J.A. Alburquerque, C. de la Fuente, A. Ferrer-Costa, L. Carrasco, J. Cegarra,
M. Abad, M.P. Bernal, Assessment of the fertiliser potential of digestates from farm
and agroindustrial residues, Biomass Bioenergy 40 (2012) 181–189.
[49] C.T. Lukehurst, P. Frost, T. Al Seadi, Utilisation of digestate from biogas plants as
biofertiliser, IEA bioenergy, 2010. http://www.centri-force.co.uk/wp-content
/uploads/2014/07/Utilisation-of-Digestate-as-Biofertiliser-V2.0.pdf.
[50] S. Kizito, H. Luo, J. Lu, H. Bah, R. Dong, S. Wu, Role of nutrient-enriched biochar as
a soil amendment during maize growth: exploring practical alternatives to recycle
agricultural residuals and to reduce chemical fertilizer demand, Sustainability 11
(2019) 3211.
[51] D.J. Parrish, J.H. Fike, The biology and agronomy of switchgrass for biofuels, Crit.
Rev. Plant Sci. 24 (2005) 423–459.
[52] W.E. McMurphy, C.E. Denman, B.B. Tucker, Fertilization of native grass and
weeping lovegrass, Agron. J. 67 (1975) 233–236.
[53] R.W. Taylor, D.W. Allinson, Response of three warm-season grasses to varying
fertility levels on five soils, Can. J. Plant Sci. 62 (1982) 657–665.
[54] M.A. Sanderson, J.C. Read, R.L. Reed, Harvest management of switchgrass for
biomass feedstock and forage production, Agron. J. 91 (1999) 5–10.
[55] J.R. George, D.J. Obermann, D.D. Wolf, Seasonal trends for nonstructural
carbohydrates in stem bases of defoliated switchgrass, Crop Sci. 29 (1989)
1282–1287.
[56] M.K. Kering, J.T. Biermacher, T.J. Butler J, J.A. Mosali, Guretzky, Biomass yield
and nutrient responses of switchgrass to phosphorus application, Bioenergy Res 5
(2012) 71–78.
[57] A. Demirbas, Combustion characteristics of different biomass fuels, Prog. Energy
Combust. Sci. 30 (2004) 219–230.
[58] R. Lemus, E.C. Brummer, C.L. Burras, K.J. Moore, M.F. Barker, N.E. Molstad,
Effects of nitrogen fertilization on biomass yield and quality in large fields of
established switchgrass in southern Iowa, USA, Biomass Bioenergy 32 (2008)
1187–1194.
[59] N. Waramit, K.J. Moore, A.H. Heggenstaller, Composition of native warm-season
grasses for bioenergy production in response to nitrogen fertilization rate and
harvest date, Agron. J. 103 (2011) 655–662.
[60] C.O. Hong, V.N. Owens, D. Bransby, R. Farris, J. Fike, E. Heaton, S. Kim,
H. Mayton, R. Mitchell, D. Viands, Switchgrass response to nitrogen fertilizer
across diverse environments in the USA: a regional feedstock partnership report,
Bioenergy Res 7 (2014) 777–788.
[61] P.R. Bakker, H.W. Elbersen, Managing ash content and quality in herbaceous
biomass: an analysis from plant to product, in: 14th European Biomass Conference,
2005, pp. 210–213.
[62] F.C. Lanning, L.N. Eleuterius, Silica and ash in native plants of the central and
southeastern regions of the United States, Ann. Bot. 60 (1987) 361–375.
M.-S. Lee et al. Biomass and Bioenergy 144 (2021) 105928

[63] L.J. Perry, D. Baltensperger, Leaf and stem yields and forage quality of three N- [65] C.W. Wood, H.A. Torbert, D.P. Delaney, Poultry litter as a fertilizer for
fertilized warm-season grasses, Agron. J. 71 (1979) 355–358. bermudagrass: effects on yield and quality, J. Sustain. Agric. 3 (1993) 21–36.
[64] P.R. Adler, M.A. Sanderson, A.A. Boateng, P.J. Weimer, H.J.G. Jung, Biomass yield
and biofuel quality of switchgrass harvested in fall or spring, Agron. J. 98 (2006)
1518–1525.