ArticlePDF Available

A Review on Miscanthus Biomass Production and Composition for Bioenergy Use: Genotypic and Environmental Variability and Implications for Breeding

Authors:

Abstract and Figures

The lignocellulosic C4 perennial crop miscanthus and, more particularly, one of its species, Miscanthus × giganteus, are especially interesting for bioenergy production because they combine high biomass production with a low environmental impact. However, few varieties are available, which is risky due to disease susceptibility. Gathering worldwide references, this review shows a high genotypic and environmental variability for traits of interest related to miscanthus biomass production and composition, which may be useful in breeding programs for enhancing the availability of suitable clones for bioenergy production. The M. × giganteus species and certain clones in the Miscanthus sinensis species seem particularly interesting due to high biomass production per hectare. Although the industrial requirements for biomass composition have not been fully defined for the different bioenergy conversion processes, the M. × giganteus and Miscanthus sacchariflorus species, which show high lignin contents, appear more suitable for thermochemical conversion processes. In contrast, the M. sinensis species and certain M. × giganteus clones with low lignin contents were interesting for biochemical conversion processes. The M. sacchariflorus species is also interesting as a progenitor for breeding programs, due to its low ash content, which is suitable for the different bioenergy conversion processes. Moreover, mature miscanthus crops harvested in winter seem preferred by industry to enhance efficiency and reduce the expense of the processes. This investigation on miscanthus can be extrapolated to other monocotyledons and perennial crops, which may be proposed as feedstocks in addition to miscanthus.
Phenotypic variability and genotype contribution to the phenotypic variability in miscanthus for aboveground biomass production (a), canopy height (b), stem number per plant (c), and stem diameter (d) based on the crop age, from the first to third year of cultivation. Phenotypic variability corresponds to the mean + (environmental + genotypic variability); the genotype contribution to the phenotypic variability corresponds to genotypic variability. The aboveground biomass production was assessed for the winter harvest; the canopy height, stem number per plant, and stem diameter were obtained at the end of the growing season. The mean value (indicated by the black diamond) was calculated from the individual data collected in the literature for each year of cultivation. The ends of the vertical lines correspond to the minimum and maximum values; the black horizontal lines correspond to the upper and lower quartiles; and the white horizontal lines correspond to the median for each year of cultivation. The stars on both sides of the black horizontal lines correspond to the outlier values. The range of variation corresponds to the difference between the maximum and minimum values and is expressed as a percentage of the mean. References used—a Christian et al. [57], Clifton-Brown et al. [44], Lewandowski et al. [37], Jezowski [40], Jezowski et al. [60], Mantineo et al. [59], Amougou et al. [51], Zub et al. [41]; b Jezowski [40], Jezowski et al. [60], Zub et al. [41]; c Jezowski [40], Christian et al. [57], Jezowski et al. [60], Zub et al. [41]; d Jezowski [40], Jezowski et al. [60], Zub et al. [41]
… 
The phenotypic variability and genotype contribution to the phenotypic variability for aboveground miscanthus biomass production (a), canopy height (b), stem number per plant (c), and stem diameter (d) among and within the three Miscanthus species M. × giganteus, M. sacchariflorus, and M. sinensis for the first 3 years of cultivation. The phenotypic variability corresponds to the mean + (environmental + genotypic variability); the genotype contribution to phenotypic variability corresponds to genotypic variability. The aboveground biomass production was assessed for the winter harvest; the canopy height, stem number per plant, and stem diameter were obtained at the end of the growing season. The mean value (indicated by the black diamond) was calculated from the individual data collected in the literature for each year of cultivation. The ends of the vertical lines correspond to the minimum and maximum values; the black horizontal lines correspond to the upper and lower quartiles; and the white horizontal lines correspond to the median for each year of cultivation. The stars on both sides of the black horizontal lines correspond to the outlier values. The range of variation corresponds to the difference between the maximum and minimum values and was been expressed as a percentage of the mean. For each trait, genotypic variability in the M. sacchariflorus species during the second year was the same as the phenotypic variability; the two M. sacchariflorus genotypes were observed in a single environment. No data were available for canopy height (b), stem number per plant (c), and stem diameter (d) on the first year of cultivation for the M. sacchariflorus species in the literature. References used—a Christian et al. [57], Lewandowski et al. [37], Jezowski [40], Jezowski et al. [60], Mantineo et al. [59], Amougou et al. [51], Zub et al. [41]; b Jezowski [40], Jezowski et al. [60], Zub et al. [41]; c Christian et al. [57], Jezowski [40], Jezowski et al. [60], Zub et al. [41]; d Jezowski [40], Jezowski et al. [60], Zub et al. [41]
… 
Content may be subject to copyright.
1 23
BioEnergy Research
ISSN 1939-1234
Volume 8
Number 2
Bioenerg. Res. (2015) 8:502-526
DOI 10.1007/s12155-014-9524-7
A Review on Miscanthus Biomass
Production and Composition for Bioenergy
Use: Genotypic and Environmental
Variability and Implications for Breeding
Stéphanie Arnoult & Maryse Brancourt-
Hulmel
1 23
Your article is published under the Creative
Commons Attribution license which allows
users to read, copy, distribute and make
derivative works, as long as the author of
the original work is cited. You may self-
archive this article on your own website, an
institutional repository or funder’s repository
and make it publicly available immediately.
A Review on Miscanthus Biomass Production and Composition
for Bioenergy Use: Genotypic and Environmental Variability
and Implications for Breeding
Stéphanie Arnoult &Maryse Brancourt-Hulmel
Published online: 16 September 2014
#The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract The lignocellulosic C4 perennial crop miscanthus
and, more particularly, one of its species, Miscanthus ×
giganteus, are especially interesting for bioenergy production
because they combine high biomass production with a low
environmental impact. However, few varieties are available,
which is risky due to disease susceptibility. Gathering world-
wide references, this review shows a high genotypic and
environmental variability for traits of interest related to
miscanthus biomass production and composition, which
may be useful in breeding programs for enhancing the avail-
ability of suitable clones for bioenergy production. The M. ×
giganteus species and certain clones in the Miscanthus
sinensis species seem particularly interesting due to high
biomass production per hectare. Although the industrial re-
quirements for biomass composition have not been fully defined
for the different bioenergy conversion processes, the M. ×
giganteus and Miscanthus sacchariflorus species, which show
high lignin contents, appear more suitable for thermochemical
conversion processes. In contrast, the M. sinensis species and
certain M. ×giganteus clones with low lignin contents were
interesting for biochemical conversion processes. The
M. sacchariflorus species is also interesting as a progenitor for
breeding programs, due to its low ash content, which is suitable
for the different bioenergy conversion processes. Moreover,
mature miscanthus crops harvested in winter seem preferred
by industry to enhance efficiency and reduce the expense of
the processes. This investigation on miscanthus can be extrap-
olated to other monocotyledons and perennial crops, which may
be proposed as feedstocks in addition to miscanthus.
Keywords Miscanthus .Phenotypic and genotypic
variability .Biomass production .Biomass composition .
Breeding .Bioenergy use
Abbreviations
DM Dry matter
ha Hectare
Gig Miscanthus ×giganteus species
Sacc Miscanthus sacchariflorus species
Sin Miscanthus sinensis species
tTon
Introduction
Currently, climate change and fossil fuel resource deple-
tion are major global concerns. To limit climate change,
reduce greenhouse gas emissions, and replace fossil fuel
resources, renewable energy sources must be developed.
Many reports have shown that biomass crops are signif-
icant contributors as bioenergy sources for heat, electricity, or
biofuel production through thermochemical or biochemical
processes [15].
Currently, a wide range of crops are biomass production
candidates for bioenergy use: perennial C4 crops, such as
miscanthus (Miscanthus), switchgrass (Panicum virgatum),
or sugarcane (Saccharum officinarum L.); perennial C3 crops,
such as reed canary grass (Phalaris arundinacea L.), giant
reed (Arundo donax L.), short-rotation poplar coppices
(Populus), or willow (Salix); annual C4 crops, such as fiber
S. Arnoult (*)
INRA, UMR1281 SADV, 2 Chaussée Brunehaut, Estrées-Mons,
BP 50136, 80203 Péronne Cedex, France
e-mail: Stephanie.Arnoult@mons.inra.fr
M. Brancourt-Hulmel
INRA, UR1158 AgroImpact, Site dEstrées-Mons, 2 Chaussée
Brunehaut, Estrées-Mons, BP 50136, 80203 Péronne Cedex, France
Present Address:
S. Arnoult
INRA, UE0972 GCIE Picardie, 2 Chaussée Brunehaut,
Estrées-Mons, BP 50136, 80203 Péronne Cedex, France
Bioenerg. Res. (2015) 8:502526
DOI 10.1007/s12155-014-9524-7
sorghum (Sorghum bicolor L.) or maize (Zea mays); or annual
C3 crops, such as triticale (Triticum sativum)[615].
Among the few studies that have compared several such
crops, the C4 perennial crops have been highlighted as advan-
tageous for sustainable biomass feedstock [6].
For sustainable crops dedicated to producing bioenergy, the
following must be combined: (i) high biomass production per
hectare in various climates, (ii) suitable biomass composition
for various bioenergy conversion processes, and (iii) positive
environmental footprint (lowest water requirement, lowest N,
P, and K fertilization, low greenhouse gas emissions, low
invasiveness, etc.). For these requirements, C4 crops appear
more promising as bioenergy crops than C3 crops because
they are more productive and exhibit efficient water and
nitrogen use [16] and efficient sunlight interception [17].
Among these C4 crops, C4 perennial crops such as
miscanthus and switchgrass are particularly interesting be-
cause they yield a better environmental footprint than annual
crops [6]. Moreover, perennial crops could be used to cover
marginal lands for biomass production where annual crops are
not suitable [18]. In comparison with switchgrass, miscanthus
produces a higher biomass [16,19] and also displays a larger
solar energy conversion efficiency [20]. It is considered one of
the most promising perennial bioenergy crops [15,2123].
More particularly, Miscanthus ×giganteus, which is a triploid
sterile hybrid descendent of a cross between Miscanthus
sacchariflorus and Miscanthus sinensis [24], is particularly
interesting because it combines high biomass production per
hectare with a low environmental impact [6]. However, M. ×
giganteus also has certain disadvantages: (i) this species can-
not produce high quantities of biomass under various climates
because it is sensitive to heavy frost [25] and a lack of
water [26], and further, (ii) M×giganteus, which has
been the most commonly cultivated species until now, has
only few clones [27,28], which could be risky due to disease
susceptibility [29].
Considering these disadvantages, developing miscanthus
as suitable feedstock for bioenergy will enhance the
miscanthus varieties available for cultivation. Therefore,
miscanthus breeding programs must be developed to propose
a range of miscanthus varieties that can produce high quanti-
ties of biomass per hectare under various climatic conditions.
Other Miscanthus species could also be sought for breeding
because the genus Miscanthus contains more than 20 species
that originate from a broad geographical area [30]. Certain
species, such as M. sinensis, show high genetic diversity
compared with M. ×giganteus [27,28].
In addition to the above requirements for biomass produc-
tion, using miscanthus as a feedstock for bioenergy requires
that the biomass composition is adapted to various bioenergy
conversion processes. This suitability remains a difficult chal-
lenge and constitutes a major bottleneck to proposing suitable
lignocellulosic crops for bioenergy use.
First, the industrial requirements for lignocellulosic crop
biomass composition have not been fully defined for the
various bioenergy conversion processes. For instance, the
most favorable balance between lignins, cellulose, and hemi-
celluloses in the cell wall for producing biofuels from bio-
chemical conversion processes has not been defined [31].
In recent years, several global projects have been launched
to precisely define these requirements and develop industrial-
scale tests for several bioenergy conversion processes. The
results from these projects should precisely define the indus-
trial requirements for lignocellulosic crop biomass composi-
tion in bioenergy production.
Second, biomass composition can influence bioenergy
conversion process efficiency. For instance, high lignin con-
tent is positive for thermochemical processes [32,33], while it
is negative for biochemical processes [3436]. Regardless of
the process, ash can be problematic because it deposits on the
heat surfaces, which causes slagging and fouling [37,38].
Third, the methods used to assess the biomass components
and, more particularly, the cell wall components differ and are
not reliable. Standardized methods have not been established
for either miscanthus or other crops. This complicates the
definition of the most suitable biomass composition for each
bioenergy conversion process.
These observations demonstrate that biomass composition
must be better understood and considered in miscanthus
breeding programs to develop miscanthus as a suitable feed-
stock for bioenergy use.
Developing such breeding programs also requires consid-
eration of interesting traits related to biomass production and
composition as well as the factors of variation influencing
such traits.
For the traits of interest, aboveground biomass pro-
duction must be considered first. In addition, canopy
height, stem number, or stem diameter, which are the
main components that contribute to biomass production,
must also be investigated [3941]; cellulose, hemicellulose,
lignin, and ash contents are the main traits considered in
biomass composition.
For the factors of variation, studies on miscanthus have
reported effects of species, clone, and ploidy level on biomass
production and certain biomass production components
[3944] or biomass composition [33,37,43,4547].
Furthermore, these traits appear influenced by other factors,
such as geographical area, climate conditions, or crop man-
agement practices, which can influence the conversion pro-
cesses [33,38,44,4850].
Therefore, it is essential to consider the traits together
with the factors of variation that can influence these
traits in developing suitable miscanthus for bioenergy
production.
Breeding program development requires knowing the ge-
notypic variability related to the traits of interest [31]. As the
Bioenerg. Res. (2015) 8:502526 503
genotypic variability related to biomass production and bio-
mass composition has not been well studied for miscanthus, it
is crucial to explore the variability in this genus for biomass
production and composition.
Therefore, the present paper reviews worldwide ref-
erences about the phenotypic variability in the
Miscanthus genus for the following traits related to
biomass production and biomass composition: above-
ground biomass production, canopy height, stem number
per plant, stem diameter, cellulose content, hemicellu-
lose content, lignin content, and ash content. The phe-
notypic variability was deconstructed into environmental
variability and genotypic variability to highlight the genotypic
variability contribution that may be used in breeding pro-
grams. This paper gathered the available data for the three
most studied Miscanthus species: M. ×giganteus,M. sinensis,
and M. sacchariflorus.
This paper is presented in four sections. The first
section describes the data set used in the present review
and collected from scientific literature. A second section
investigates the phenotypic variability of the traits of
interest related to miscanthus biomass production and
biomass composition. In this section, environmental
and genotypic variability are investigated with a focus
on the factors of variation that are most studied in the
literature. Given the miscanthus genotypic variability for
biomass production and composition, in the third section, we
discuss the miscanthus breeding implications for bioenergy
use considering the industrial requirements for bioenergy
conversion processes. Finally, we provide certain proposals
in the fourth section.
Database Description and Compilation
A literature search was conducted using the Thomson Reuters
Web of Knowledge electronic database for several traits of
interest related to miscanthus biomass production and biomass
composition.
The first observation is that the studies reporting data
on these traits are relatively recent, with publication
dates mainly ranging from 1997 to 2014. Certain
miscanthus traits have not been thoroughly studied.
Traits related to biomass production studied in literature
mainly concern aboveground biomass production, cano-
py height, stem number, and stem diameter. Among
these traits, fewer studies have considered canopy
height, stem number, and stem diameter than above-
ground biomass production. Fewer studies have consid-
ered traits related to miscanthus biomass composition
than miscanthus biomass production. For the present
review, we will focus on traits related to cell wall
components (cellulose content, hemicellulose content,
and lignin content) and ash content, which are interest-
ing traits for industrial bioenergy production.
A second observation is that the studies differed.
First, the data from each publication were not comparable
for a given trait; certain publications indicate individual values
for each factor of variation (year, harvest date, nitrogen, loca-
tion, genotype, etc.), and others reported only a mean value for
a given trait. Therefore, to investigate the traits of interest
related to miscanthus biomass production and biomass com-
position in this review, we selected publications that allowed
us to collect individual data for each trait. Therefore, each
individual data has the same weight and was comparable
between publications.
Secondly, different factors of variation for such traits in the
literature were discussed. In this review, we focus on the
factors related either to crop age, climate conditions, geo-
graphical area, crop management practices (harvest date, ni-
trogen fertilization, irrigation, or plant density), species,
clones, or ploidy level, which were the most commonly stud-
ied factors in the literature. For the crop age, most studies
reported data for the first 3 years of cultivation. Therefore, we
focus on these first 3 years to investigate the influence of crop
age on these traits. In addition, only three Miscanthus species
have been investigated in the literature for traits related to
biomass production and composition: M. ×giganteus,
M. sinensis,andM. sacchariflorus. Moreover, the clone num-
ber and genetic background in the literature differed between
the three species. Therefore, to compare the publications
based on the Miscanthus species, we categorized the geno-
types into three groups: (i) interspecific M. ×giganteus-type
hybrids that correspond to M. ×giganteus clones or
M. sacchariflorus ×M. sinensis clones, (ii) M. sinensis species
that correspond to M. sinensis clones or intra-specific hybrids
between M. sinensis clones, and (iii) M. sacchariflorus species
that correspond to M. sacchariflorus clones or intra-specific
hybrids between M. sacchariflorus clones.
Third, the method used to determine the values for traits
related to biomass composition differed between studies. The
cellulose, hemicellulose, and lignin contents were determined
using several chemical analysis methods, such as the Van
Soest method, the KS M 7044 method to determine
cellulose content, or the Klason method to determine
lignin content [51,52]. Based on the method used, the
cellulose, hemicellulose, or lignin contents differed and may
be underestimated or overestimated (Chabbert, personal com-
munication). Therefore, to ensure reliable comparisons for
such contents between publications, we focused only on the
publications that used the Van Soest method as the method of
reference [53] because it was most commonly used [33,45,
5456].
Herein, Tables 1and 2gather the publications which re-
ported data on the miscanthus phenotypic variability for the
traits of interest. For each study, we provide information
504 Bioenerg. Res. (2015) 8:502526
Tab l e 1 Description of the scientific references collected for the present review reporting data on aboveground miscanthus biomass production, canopy height, stem number per plant, and stem diameter
Trait Reference Year of
publication
Factors of variation Experimental design Statistical data
Crop
age
Country
(number of sites)
Harvest
date
Nitrogen
fertilization
(k g/ha)
Irrigation Plant
density
(pl/m
2
)
Number of
Miscanthus
species (name
of species)
Number
of clones
Number
of replicated
plots
Plot
surface
(m
2
)
Sampling
area (m
2
)
Std
(min-max)
CV (%) LSD
(min-max)
(0.05)
MSD
(min-max)
SED
(min-max)
Aboveground
biomass
production
(t DM/ha)
Clifton-Brown
et al. [44]
2001 1, 2, 3 Sweden, Denmark,
England, Germany,
Portugal
(five sites)
Autumn 2 3
(Gig, Sacc, Sin)
15 3,
only 2 in
Portugal
25 2.5 0.617.1
Lewandowski
et al. [37]
2003 3 Sweden, Denmark,
England, Germany,
Portugal
(five sites)
Autumn and
winter
23
(Gig, Sacc, Sin)
15 3,
only 2 in
Portugal
49 2 2.518.0
Christian
et al. [57]
2008 1 to 14 UK
(one site)
Winter 0, 160, 120
(three levels)
41
(Gig)
1 3 100 36 0.31.0
Jezowski [40]2008 1, 2, 3 Poland
(one site)
Winter 1 2
(Gig, Sin)
63 2010
Angelini
et al. [58]
2009 1 to 12 Italy
(one site)
Autumn 2 1
(Gig)
1 4 100 10
Mantineo
et al. [59]
2009 1 to 5 Italy
(one site)
Winter 50, 100
(two levels)
75, 25 % Etm
(two levels)
21
(Gig)
124
Jezowski
et al. [60]
2011 1,2,3 Poland
(two sites)
Winter 1 2
(Gig, Sin)
63 2010
Amougou
et al. [51]
2011 2, 3 France
(one site)
Autumn and
winter
0, 120
(two levels)
1.5 1
(Gig)
1 3 360 4 4.99.6
Strullu et al. [61] 2011 2, 3 France
(one site)
Autumn and
winter
0, 120
(two levels)
1.5 1
(Gig)
1 3 360 4
Zub et al. [41]2011 2, 3 France
(one site)
Autumn and
winter
23
(Gig, Sacc, Sin)
21 3 16 16 1.24.5 19 1.8
Behnke
et al. [62]
2012 2, 3 IL, USA
(one site)
Winter 0, 60, 120
(three levels)
1
(Gig)
1 3 100 0.72.9
Dohleman
et al. [63]
2012 5, 6, 7 IL, USA
(one site)
Summer,
autumn, and
winter
11
(Gig)
1 4 100 0.38
Gauder
et al. [39]
2012 1 to 14 Germa ny
(one site)
Autumn 2 3
(Gig, Sacc, Sin)
15 3 25 1 to 4
Arundale
et al. [19]
2014 3 to 10 IL, USA
(seven sites)
Winte r 1 1
(Gig)
1 4 25 0.38
Arundale
et al. [64]
2014 3 to 9 IL, USA
(seven sites)
Winter 0, 67, 134, 202
(four levels)
11
(Gig)
1 4 25 0.19
Haines
et al. [65]
2014 1, 2, 3 NC, USA
(two sites)
Winter 0, 45, 90, 135
(four levels)
11
(Gig)
14 164
Larsen
et al. [66]
2014 1 to 20 Denmark
(two sites)
Autumn and
winter
0, 75 (NPK
and slurry),
150 (slurry)
(four levels)
0, irrigated
(two levels)
1.8, 1.1
(two levels)
2
(Gig, Sin)
2 3 160 22 and 37
Palmer
et al. [67]
2014 1 to 4 NC, USA
(two sites)
Winter 0, 34, 67, 134
(four levels)
12
(Gig, Sin)
21 259
Canopy
height (cm)
Jezowski [40]2008 1, 2, 3 Poland
(one site)
12
(Gig, Sin)
6 3 20 1446
Jezowski
et al. [60]
2011 1,2,3 Poland
(two sites)
12
(Gig, Sin)
63 2010
Zub et al. [41]2011 2, 3 France
(one site)
23
(Gig, Sacc, Sin)
21 3 15 16 1725 6 3
Stem number
per plant
Christian
et al. [57]
2008 1 to 14 UK
(one site)
0, 160, 120
(three levels)
41
(Gig)
1 3 100 1.25 24
Jezowski [40]2008 1, 2, 3 Poland
(one site)
12
(Gig, Sin)
63 20 719
Bioenerg. Res. (2015) 8:502526 505
Tab l e 1 (continued)
Trait Reference Year of
publication
Factors of variation Experimental design Statistical data
Crop
age
Country
(number of sites)
Harvest
date
Nitrogen
fertilization
(k g/ha)
Irrigation Plant
density
(pl/m
2
)
Number of
Miscanthus
species (name
of species)
Number
of clones
Number
of replicated
plots
Plot
surface
(m
2
)
Sampling
area (m
2
)
Std
(min-max)
CV (%) LSD
(min-max)
(0.05)
MSD
(min-max)
SED
(min-max)
Jezowski
et al. [60]
2011 1, 2, 3 Poland
(two sites)
12
(Gig, Sin)
63 2010
Zub
et al. [41]
2011 2, 3 France
(one site)
23
(Gig, Sacc, Sin)
21 3 16 16 1549 17 3
Gauder
et al. [39]
2012 1 to 14 Germany
(one site)
23
Gig, Sacc, Sin)
15 3 25 1 to 4
Stem diameter
(mm)
Jezowski [40]2008 1, 2, 3 Poland
(one site)
12
(Gig, Sin)
63 20
Jezowski
et al. [60]
2011 1, 2, 3 Poland
(two sites)
12
(Gig, Sin)
63 20
Zub et al. [41]2011 2, 3 France
(one site)
23
(Gig, Sacc, Sin)
21 3 16 16 1.92.4 6 1.2
Gauder
et al. [39]
2012 1 to 14 Germany
(one site)
23
Gig, Sacc, Sin)
15 3 25 1 to 4
In the study of Zub et al. [41], M. floridulus clone belongs to the M. ×giganteus species according to the authors. Data written in italics were calculated in addition to the initial paper. The LSD values from
Zub et al. [41] were calculated using Dagnelie [68]. In bold, all the references reported individual data which were used in the Figs. 1,3,4,5,6,and7
Gig M. ×giganteus species, Sacc M. sacchariflorus species, Sin M. sinensis species, Std standard deviation, CV coefficient of variation, SED standard error of difference, LSD least significant difference,
MSD minimum significant difference
506 Bioenerg. Res. (2015) 8:502526
Tab l e 2 Description of the scientific references collected for the present review reporting data on the miscanthus biomass composition for the aboveground biomass: cellulose, hemicellulose, lignin, and
ash content
Trait Reference Year of
publication
Factors of variation Experimental design Statistical data
Crop age Country
(number
of sites)
Harvest date Nitrogen
fertilization
(kg/ha)
Plant
density
(pl/m
2
)
Number of
Miscanthus
species
(name of
species)
Number
of clones
Number
of replicated
plots
Plot
surface
(m
2
)
Sampling
area (m
2
)
Std
(min-
max)
LSD
(min-
max)
(0.05)
MSD
(min-
max)
SED
(min-
max)
Cellulose
(% DM)
Hodgson et al. [33]
a
2010 3 Sweden, Denmark,
England, Germany,
Portugal
(five sites)
Autumn and
winter
2 3 (Gig, Sacc, Sin) 15 3, only 2 in
Portugal
49 2 1.26.8 0.62.6
Hemicellulose
(% DM)
Hodgson et al. [33]
a
2010 3 Sweden, Denmark,
England, Germany,
Portugal
(five sites)
Autumn and
winter
2 3 (Gig, Sacc, Sin) 15 3, only 2 in
Portugal
49 2 1.05.1 0.42.0
Lignin
(% DM)
Hodgson et al. [33]
a
2010 3 Sweden, Denmark,
England, Germany,
Portugal
(five sites)
Autumn and
winter
2 3 (Gig, Sacc, Sin) 15 3, only 2 in
Portugal
49 2 0.32.6 0.11.0
Hayes [69]
b
2013 1, 2, 3, 13 Ireland
(six sites)
Autumn, winter,
and spring
1(Gig) 1
Ash
(% DM)
Clifton-Brown and
Lewandowski [43]
2002 1, 2, 3 Germany
(one site)
Autumn and
winter
2 3 (Gig, Sacc, Sin) 15 3 25 0.84.2
Lewandowski et al. [37]2003 3 Sweden, Denmark,
England, Germany,
Portugal
(five sites)
Autumn and
winter
2 3 (Gig, Sacc, Sin) 15 3, only 2 in
Portugal
49 2 0.53.2
Baxter et al. [70] 2012 3, 4 UK
(one site)
Autumn and winter 1 (Gig) 1 3
Hayes [69] 2013 1, 2, 3, 13 Ireland
(six sites)
Autumn, winter,
and spring
0, 100, 250
(three levels)
1(Gig) 1
Meehan et al. [71] 2013 15, 16 Ireland
(one site)
Autumn, winter,
and spring
1 (Gig) 1 4 0.38
In bold, all the references reported individual data which were used in the Figs. 1,3,4,5,6,and7
Gig M. ×giganteus species, Sacc M. sacchariflorus species, Sin M. sinensis species, SED standard error of difference, LSD least significant difference, MSD minimum significant difference
a
The cellulose, hemicellulose, and lignin contents were determined using the Van Soest method as the method of reference
b
Lignin content was determined using the Klason method
Bioenerg. Res. (2015) 8:502526 507
related to the factors of variation, experimental design con-
struction, and statistical indices.
To investigate each trait, we selected in Tables 1and 2the
studies for which individual data were available (in bold in the
tables). We represented these individual data using box-plots
(Figs. 1,3,4,5,6, and 7). Two types of box-plots were
constructed for each illustration: a first type of box-plot shows
the phenotypic variability that combines both environmental
Fig. 1 Phenotypic variability and genotype contribution to the pheno-
typic variability in miscanthus for aboveground biomass production (a),
canopy height (b), stem number per plant (c), and stem diameter (d)based
on the crop age, from the first to third year of cultivation. Phenotypic
variability corresponds to the mean + (environmental + genotypic vari-
ability); the genotype contribution to the phenotypic variability corre-
sponds to genotypic variability. The aboveground biomass production
was assessed for the winter harvest; the canopy height, stem number per
plant, and stem diameter were obtained at the end of the growing season.
The mean value (indicated by the black diamond) was calculated from the
individual data collected in the literature for each year of cultivation. The
ends of the vertical lines correspond to the minimum and maximum
values; the black horizontal lines correspond to the upper and lower
quartiles; and the white horizontal lines correspond to the median for
each year of cultivation. The stars on both sides of the black horizontal
lines correspond to the outlier values. The range of variation corresponds
to the difference between the maximum and minimum values and is
expressed as a percentage of the mean. References usedaChristian
et al. [57], Clifton-Brown et al. [44], Lewandowski et al. [37], Jezowski
[40], Jezowski et al. [60], Mantineo et al. [59], Amougou et al. [51], Zub
et al. [41]; bJezowski [40], Jezowski et al. [60], Zub et al. [41]; c
Jezowski [40], Christian et al. [57], Jezowski et al. [60], Zub et al. [41];
dJezowski [40], Jezowski et al. [60], Zub et al. [41]
508 Bioenerg. Res. (2015) 8:502526
and genotypic variability, and a second type shows the part of
the phenotypic variability due only to the genotype. This
genotype effect (α
i
) was calculated in each collected environ-
ment according to the following additive model including the
studied factors:
EY
i
ðÞ¼μþαiþother factor effects
In addition, the range of variation was calculated for
each trait.
Finally, the remaining references of the Tables 1and 2are
cited in the text to support the discussion.
The Phenotypic Variability for the Traits of Interest
The Phenotypic Variability for Aboveground Biomass
Production and Biomass Components
The Influence of Crop Age
Miscanthus is a perennial crop that can be cultivated for up to
25 years [72], during which miscanthus biomass is produced
in two phases: a yield-building phase, where the biomass
gradually increases, and an adult phase often described as a
plateau phase, where the biomass production is maintained
[29,42,73].
Fig. 1 (continued)
Bioenerg. Res. (2015) 8:502526 509
Most studies in the literature have reported data on
miscanthus aboveground biomass production and compo-
nents for the first 3 years of cultivation, which correspond to
the yield-building phase (Table 1). Data on these traits for
subsequent years were less frequent in the literature and only
concern the M. ×giganteus species, which is the most com-
monly studied species [6,38,57,59].
Therefore, in the following, we describe the influence of
crop age on the aboveground biomass and its components
based on the two biomass production phases. We better detail
the first three cultivation years because more information is
available in literature.
&The first years: the yield-building phase
The first year of cultivation corresponds to the
year that the crop is established [29]. The biomass
production is generally low during the first year
with a 5.9 t dry matter/ha (DM/ha) average from
experiments using irrigated or rainfall conditions
(Fig. 1a).
During the 2 years that follow, the miscanthus biomass
gradually increases: biomass production reaches 8.3 and
13.0 t DM/ha on average during the second and third
years, respectively, under irrigated or rainfall conditions
(Fig. 1a).
0
5
10
15
20
25
30
1234567891011121314
Above- tsevrahret
n
iwninoitcudorpssamoibdnuorg
(tDM/ha)
Yea r of cu lv ao n
Gig_Chrisan et al. 2008 [57]
Gig-1_Gauder et al. 2012 [39]
Gig-2_Gauder et al. 2012 [39]
Gig-3_Gauder at al. 2012 [39]
Gig-4_Gauder et al. 2012 [39]
Gig_Cadoux et al. 2014 [6]
Fig. 2 M. ×giganteus
aboveground biomass production
during the winter harvest for
several successive years
from Christian et al. [57],
Gauder et al. [39], and
Cadoux et al. [6]
Fig. 3 The phenotypic variability and genotype contribution to the
phenotypic variability for aboveground miscanthus biomass production
based on the harvest date (autumn and winter harvests) during the second
and third years of cultivation. The phenotypic variability corresponds to
the mean + (environmental + genotypic variability); the genotype contri-
bution to the phenotypic variability corresponds to the genotypic vari-
ability. The mean value (indicated by the black diamond) was calculated
from the individual data collected in the literature for each year of
cultivation and harvest date. The ends of the vertical lines correspond to
the minimum and maximum values; the black horizontal lines correspond
to the upper and lower quartiles; and the white horizontal lines correspond
to the median for each year of cultivation and harvest date. The
stars on both sides of the black horizontal lines correspondtothe
outlier values. The range of variation corresponds to the difference
between the maximum and minimum values and is expressed as a
percentage of the mean. References usedLewandowski et al. [37],
Zub et al. [41], Amougou et al. [51]
510 Bioenerg. Res. (2015) 8:502526
Biomass components, such as canopy height and stem
number per plant, also clearly increase from the first to
third year; on average, the canopy height increases from
66 to 176 cm, and the stem number per plant increases
from 21 to 53 (Fig. 1b, c). However, with mean values
between 4.2 and 5.2 mm, the stem diameter was more
consistent than the biomass production, canopy height,
and stem number during the first 3 years (Fig. 1).
These observations confirmed that for miscanthus, the
first 3 years correspond to a yield-building phase, where
the biomass production and its components, such as can-
opy height and stem number per plant, gradually increase.
The range of variation was large within each of the first
3 years of cultivation for each trait. This range of variation
was generally smaller during the first year of cultivation
compared with the second and third years (Fig. 1). In
addition, the data scattering for biomass production, stem
number, canopy height, and stem diameter, to a lesser
extent, increased each year during the first 3 years; mini-
mum values were consistent among the years, but the
Fig. 4 The phenotypic variability and genotype contribution to the
phenotypic variability for aboveground miscanthus biomass production
(a), canopy height (b), stem number per plant (c), and stem diameter (d)
among and within the three Miscanthus species M. ×giganteus,
M. sacchariflorus,andM. sinensis for the first 3 years of cultivation.
The phenotypic variability corresponds to the mean + (environmental +
genotypic variability); the genotype contribution to phenotypic variability
corresponds to genotypic variability. The aboveground biomass produc-
tion was assessed for the winter harvest; the canopy height, stem number
per plant, and stem diameter were obtained at the end of the growing
season. The mean value (indicated by the black diamond) was calculated
from the individual data collected in the literature for each year of
cultivation. The ends of the vertical lines correspond to the minimum
and maximum values; the black horizontal lines correspond to the upper
and lower quartiles; and the white horizontal lines correspond to the
median for each year of cultivation. The stars on both sides of the black
horizontal lines correspond to the outlier values. The range of variation
corresponds to the difference between the maximum and minimum
values and was been expressed as a percentage of the mean. For each
trait, genotypic variability in the M. sacchariflorus species during the
second year was the same as the phenotypic variability; the two
M. sacchariflorus genotypes were observed in a single environment.
No data were available for canopy height (b), stem number per plant
(c), and stem diameter (d) on the first year of cultivation for the
M. sacchariflorus species in the literature. References useda
Christian et al. [57], Lewandowski et al. [37], Jezowski [40], Jezowski
et al. [60], Mantineo et al. [59], Amougou et al. [51], Zub et al. [41]; b
Jezowski [40], Jezowski et al. [60], Zub et al. [41]; cChristian et al. [57],
Jezowski [40], Jezowski et al. [60], Zub et al. [41]; dJezowski [40],
Jezowski et al. [60], Zub et al. [41]
Bioenerg. Res. (2015) 8:502526 511
maximum values increased each year (Fig. 1). Similarly,
genotypic variability was more visible in the second and
third years than the first year for each trait studied; the
higher variability in second and third years was certainly
due to the most productive genotypes, which began to
display their potential during the second year.
Finally, among the traits studied, stem number and
aboveground biomass production appeared to be the most
variable traits (301 and 242 %, respectively, in the third
year) compared with canopy height and stem diameter,
which seemed less variable (163 and 158 %, respec-
tively, in the third year) (Fig. 1). This large observed
range of variation for the traits related to biomass
production was particularly interesting in breeding pro-
grams to select the best genotypes and achieve maximum
biomass production.
&The subsequent years: the adult phase and decline
Several studies have reported data on miscanthus bio-
mass production, canopy height, stem number, or stem
diameter in long-term experiments. They mainly con-
cerned long-term biomass production which was reported
for the M. ×giganteus species in Europe (France [6], UK
[57], Germany [39], Italy [58], Denmark [66]) and in the
USA (Illinois [19,63]). The winter harvest date
corresponded to the harvest date commonly used by
farmers. From the available individual data, we assume
that the M. ×giganteus biomass production crop reached a
first peak at variable time: at 3 years in the study of Gauder
et al. [39], at 6 years in the study of Christian et al. [57],
and this first peak was not reached after 4 years of culti-
vation in the study of Cadoux et al. [6](Fig.2). Clifton-
Brown et al. [42] in the UK and Miguez et al. [74]inthe
USA showed that biomass production plateaued after 2 to
5 years for M. ×giganteus depending on the environmen-
tal conditions and crop management practices. Larsen
et al. [66] reported in Denmark an increase of the M. ×
giganteus biomass production during the first years, opti-
mum yields after 7 and 8 years, and a decrease to a lower
level which remained relatively constant from year 11 to
20. In addition, Gauder et al. [39] found even more vari-
able yields from the 3rd to the 14th years of cultivation.
In summary, based on the long-term experimental data
published, it is difficult to estimate the time of the first
peak of production and the length of time that miscanthus
maintains biomass production. It seems also that the yields
Fig. 4 (continued)
512 Bioenerg. Res. (2015) 8:502526
during the plateau phase are variable. Lastly, the time that
biomass production begins to decline is also variable. This
variability during the plateau phase and the variability of
the time of the decline depend on possible causes such as
the soil and climate conditions as well as crop manage-
ment practices [74]. In addition, the biomass production
decline depends itself on soil compaction and pest and
disease pressure [19].
The Influence of Climate Conditions
Climate conditions, such as temperature, affect
miscanthus biomass production more during the first
year of crop establishment. For instance, a severe frost
that occurs immediately after establishment can cause
plant death in M. ×giganteus and irreversibly damage
biomass production [25]. In addition, low temperatures
during the first winter after planting can damage
miscanthus biomass production. For instance, M. ×
giganteus and M. sacchariflorus clones died the first
winter after planting in Sweden and Denmark, whereas
M. sinensis clones survived [44]. The authors explained
the plant death by the winter soil temperatures, which
fell below 4.5 °C during the first year after planting in
these countries.
During subsequent years of cultivation, Jezowski et al. [60]
concluded that climate conditions significantly affected
miscanthus biomass production based on the experiment lo-
cation by comparing two sites in Poland during the first 3 years
of cultivation. Clifton-Brown et al. [44] also concluded that
biomass production varied based on the trial location; they
showed that biomass production was generally higher in
Central and Southern Europe than Northern Europe over the
first 3 years of cultivation. Moreover, Clifton-Brown et al.
[44] concluded that the clone and country significantly
interacted; clones that produced the highest yields in
Sweden and Denmark were among clones that produced the
lowest yields in Portugal and Germany. Despite this
interaction, the authors showed that certain M. sinensis
hybrids can be found for a wide range of climate
conditions in Europe. Gauder et al. [39] and Heaton et al.
[75] also reported that climate parameters, such as rainfall and
temperature, were important in miscanthus biomass accumu-
lation. More particularly, in Germany, Gauder et al. [39]
reported a strong correlation between the rainfall level
during plant growth and miscanthus biomass production
for a winter harvest.
Therefore, climate conditions can irreversibly damage bio-
mass production during the first year of cultivation, which
corresponds to the establishment year. This observation im-
plies that climate conditions should be considered during the
Fig. 5 The phenotypic variability and genotype contribution to the
phenotypic variability for aboveground miscanthus ash content based
on the crop age from the first to third year during the winter harvest based
on Clifton-Brown and Lewandowski [43]. The phenotypic variability
corresponds to the mean + (environmental + genotypic variability); the
genotype contribution to the phenotypic variability corresponds to geno-
typic variability. The mean value (indicated by the black diamond)was
calculated from the individual data collected in the literature for each year
of cultivation. The ends of the vertical lines correspond to the minimum
and maximum values; the black horizontal lines correspond to the upper
and lower quartiles; and the white horizontal lines correspond to the
median for each year of cultivation. The stars on both sides of the black
horizontal lines correspond to the outlier values. The range of variation
corresponds to the difference between the maximum and minimum
values and is as a percentage of the mean. For each of the 3 years, the
genotypic variability equaled the phenotypic variability; the 15 genotypes
were observed in a single environment
Bioenerg. Res. (2015) 8:502526 513
first year, as it was the most critical phase for biomass pro-
duction [76]. Variations in climate conditions, such as rainfall
and temperature, can also influence biomass production during
the subsequent years of cultivation. Therefore, to maximize
biomass production, Miscanthus genotypes have to be selected
based on their climate condition requirements because an inter-
action between location and genotype was observed for bio-
mass production [44].
Fig. 6 The phenotypic variability and genotype contribution to the
phenotypic variability for aboveground miscanthus cellulose content
(a), hemicellulose content (b), lignin content (c), and ash content (d)
between the autumn and winter harvests during the third year of cultiva-
tion. The phenotypic variability corresponds to the mean + (environmen-
tal + genotypic variability); the genotype contribution to phenotypic
variability corresponds to the genotypic variability. Cellulose, hemicellu-
lose, and lignin contents were determined using the Van Soest method as
the method of reference. The mean value (indicated by the black
diamond) was calculated from the individual data collected in the
literature for each year of cultivation. The ends of the vertical lines
correspond to the minimum and maximum values; the black horizontal
lines correspond to the upper and lower quartiles; and the white horizontal
lines correspond to the median for each year of cultivation. The stars on
both sides of the black horizontal lines correspond to the outlier values.
The range of variation corresponds to the difference between the maxi-
mum and minimum values and is expressed as a percentage of the mean.
References usedacHodgson et al. [33]; dClifton-Brown and
Lewandowski [43], Lewandowski et al. [37]
514 Bioenerg. Res. (2015) 8:502526
The Influence of Crop Management Practices
&The influence of harvest date
On average, 1.8 t DM/ha (29 %) and of
4.2 t DM/ha (26 %) decreases were observed be-
tween the autumn and winter harvests for the second
and third years, respectively (Fig. 3). For each grow-
ing season, maximum biomass productions were ob-
tained during autumn at flowering [26] then declined
during winter mainly due to leaf loss, senescence, and
assimilate translocation [41,43,77,78].
With variations ranging from 253 to 341 %, biomass
production was highly variable during each year and each
harvest date (Fig. 3). Interestingly, the data appeared more
scattered during the autumn harvests than winter harvests
(Fig. 3).
Finally, the genotype contribution to the phenotypic
variability was high for aboveground biomass production
and more visible during the third than second year for both
harvest dates (Fig. 3).
In addition, Strullu et al. [61] showed on M. ×giganteus
an interaction of the harvest date with the nitrogen
Fig. 6 (continued)
Bioenerg. Res. (2015) 8:502526 515
fertilization: in winter harvest, there was no effect of
the nitrogen treatment whereas in autumn harvest,
the biomass production increased with the nitrogen
fertilization.
&The influences of nitrogen fertilization and irrigation
Most studies investigated the effects of nitrogen input
levels and irrigation on M. ×giganteus biomass produc-
tion until now, excepting the study of Palmer et al. [67].
In a review, Cadoux et al. [79] concluded that the effect
of nitrogen fertilization on aboveground M. ×giganteus
biomass production varied. Certain studies reported no
effect from nitrogen fertilization on aboveground M. ×
giganteus biomass production [51,57,59,62,80]. Other
Fig. 7 The phenotypic variability and genotype contribution to the
phenotypic variability in miscanthus among and within the three
Miscanthus species M. ×giganteus,M. sacchariflorus,andM. sinensis
for cellulose content (a), hemicellulose content (b), lignin content (c), and
ash content (d) in the aboveground biomass during winter harvest in the
third year of cultivation. The phenotypic variability corresponds to mean
+ (environmental + genotypic variability); the genotype contribution to
the phenotypic variability corresponds to genotypic variability. The cel-
lulose, hemicellulose, and lignin contents were determined using the Van
Soest method as the method of reference. The mean value (indicated by
the black diamond) was calculated from the individual data collected in
the literature for each year of cultivation. The ends of the vertical lines
correspond to the minimum and maximum values; the black horizontal
lines correspond to the upper and lower quartiles; and the white horizontal
lines correspond to the median for each year of cultivation. The stars on
both sides of the black horizontal lines correspond to the outlier values.
The range of variation corresponds to the difference between the maxi-
mum and minimum values and is expressed as a percentage of the mean.
For each trait, the genotypic variability of the M. sacchariflorus species
equaled zero; only one M. sacchariflorus genotype was observed.
References usedacHodgson et al. [33]; dClifton-Brown and
Lewandowski [43], Lewandowski et al. [37]
516 Bioenerg. Res. (2015) 8:502526
studies have reported a positive effect on the aboveground
biomass for nitrogen input ranging from 0 to 200 kg N/ha
[6,26,64,81,82]. More recently, variable effects for
nitrogen fertilization were even observed on the biomass
production between years [65].
As reported by Cadoux et al. [79], the effect from
nitrogen fertilization on aboveground M. ×giganteus bio-
mass production was generally limited but the effect was
amplified with irrigation. For instance, during the third year
of cultivation, Mantineo et al. [59] observed maximum M.
×giganteus biomass production with the highest nitrogen
fertilization level (100 kg N/ha) and under the best soil
water content conditions (75 % water restoration in the soil
with maximum evapotranspiration). Moreover, Ercoli et al.
[82] concluded that the effect of irrigation on M. ×
giganteus biomass production increased with the nitrogen
level, whereas irrigation did not affect biomass production
without nitrogen fertilization. With 100 kg N/ha, irrigation
increased the biomass production by 3.7 t DM/ha, and with
200 kg N/ha, irrigation increased the biomass production
by 9.8 t DM/ha compared with the rainfed treatment over
the first 4 years of cultivation.
These observations suggest that high M. ×giganteus
yields could be obtained with low nitrogen input levels
because biomass production increases recorded with 0
and 200 kg N/ha were moderate or insignificant.
However, because the irrigation and nitrogen level influ-
enced M. ×giganteus biomass production in the interaction,
Fig. 7 (continued)
Bioenerg. Res. (2015) 8:502526 517
it would be interesting to perform a more detailed investi-
gation of the miscanthus capacity to produce high biomass
under low nitrogen input and low irrigation levels with
other Miscanthus species than M. ×giganteus. Identifying
clones that can produce a high biomass with the lowest
environmental footprint (lowest water requirement and ni-
trogen fertilization levels) is necessary for improving
miscanthus growth for bioenergy use.
&The influence of plant density
Different plant densities were used in miscanthus ex-
periments (0.6 to 4 plants/m
2
,seeTables1and 2), but only
a few studies reported the effect of plant density on
miscanthus biomass production. Danalatos et al. [80]ob-
served that a 1- and 2-plants/m
2
density improved
miscanthus biomass production compared with 0.67
plants/m
2
in central Greece. Based on this result, we
suggest that the higher plant density increased biomass
production. Based on this observation, two hypotheses
can be formulated for the evolution of miscanthus biomass
production over several years: (i) at higher plant density,
biomass production will plateau more rapidly, and (ii) a
higher plant density will yield higher biomass production
levels during the plateau phase. More investigation is
necessary to confirm these hypotheses and select an opti-
mal plant density to maximize aboveground miscanthus
biomass production. In addition, this choice should also
depend on the cost of the miscanthus rhizomes because
the expense of establishing the rhizome remains high.
Producing plantlets through vitro culture or producing
seed-propagated varieties may be good alternatives for
combining high plant densities, high biomass production,
and lower establishment costs. Nevertheless, the environ-
mental footprint, such as non-invasiveness, must also be
preserved. For example, sterility must be obtained for
seed-propagated varieties.
The Influence of Species and Ploidy Level
First, the Miscanthus species differed in biomass production and
biomass components. With the average values 12 and 18 t DM/
ha for the winter harvest during the second and third years,
respectively, M. ×giganteus seemed the best biomass producer
compared with the M. sacchariflorus and M. sinensis species
(Fig. 4a). The M. ×giganteus species also seemed better than
the M. sacchariflorus and M. sinensis species. Moreover, the
M. sinensis species seemed smaller than the M. ×giganteus and
M. sacchariflorus species (Fig. 4b). During the third year, the M.
×giganteus species reached 231 cm on average, whereas the
M. sacchariflorus and M. sinensis species reached 185 and
132 cm, respectively (Fig. 4b). For stem number per plant, the
M. sinensis and M. ×giganteus species appeared to produce
more stems than the M. sacchariflorus species. For example, the
M. sinensis and M. ×giganteus species produced 49 and 58
stems per plant on average, respectively, during the third year
compared with the M. sacchariflorus species, which produced
26 stems (Fig. 4c). Finally, for stem diameter, the
M. sacchariflorus and M. ×giganteus species showed the
highest values compared with the M. sinensis species for the
second and third years of cultivation (Fig. 4d). For example,
during the third year, the M. sacchariflorus and M. ×giganteus
species showed the mean stem diameters 7.2 and 5.6 mm on
average, respectively, compared with M. sacchariflorus species,
which exhibited the stem diameter 4.4 mm (Fig. 4d).
The range of variation within species was higher for
M. sinensis than both the M. ×giganteus and
M. sacchariflorus species, particularly for biomass production
and stem number per plant (Fig. 4a, c). For example, during the
third year, the range of variation for aboveground biomass
production and stem number per plant in the M. sinensis species
reached 324 and 276 % on average, respectively (Fig. 4a, c). In
contrast, the range of variation for canopy height and stem
diameter appeared similar among the three Miscanthus species
(Fig. 4b, d).
For each trait, the genotype contribution to the phenotypic
variability was high. This genotypic variability was more
visible in the second and third years than the first year
(Fig. 4). Moreover, this genotypic variability was generally
more visible for the M. sinensis species than the M. ×
giganteus and M. sacchariflorus species (Fig. 4). This
trend confirmed that the genotypic diversity among miscanthus
is relatively high, particularly among the M. sinensis
species [28].
Second, certain Miscanthus clones exhibited high biomass
production and biomass component values and, thus, ap-
peared suitable for biomass production in contrast to other
clones that exhibited low values.
Interestingly, certain M. sinensis clones produced more
biomass than certain M. ×giganteus clones. Based on the
studies shown in Fig. 4a, the maximum biomass production
31.9 t DM/ha was recorded for a hybrid composed of two
M. sinensis clones (EMI no. 7) in Portugal with irrigation
compared with a 30.6 t DM/ha maximum for a M. ×giganteus
clone (Greef et Deu.) in Italy with irrigation, as well as
100 kg N/ha from a winter harvest during the third year of
cultivation. Although a higher aboveground biomass produc-
tion of 49 t DM/ha was recorded for a M. ×giganteus clone in
France under irrigated conditions [83], based on these obser-
vations, we suggest that specific M. sinensis clones can pro-
duce more biomass than certain M. ×giganteus clones under
specific climate conditions and locations. This suggestion was
confirmed by the results from Clifton-Brown et al. [44]. In
addition, certain studies have reported the superiority of cer-
tain Miscanthus hybrids for biomass production compared
with non-hybrids. For example, Clifton-Brown et al. [44]
518 Bioenerg. Res. (2015) 8:502526
observed that M. sinensis hybrids (EMI nos. 610) produced
31 % more biomass on average than non-hybrid M. sinensis
clones (EMI nos. 1115) during a winter harvest in the third
year of cultivation.
Considering the biomass components, two M. sinensis
clones tested in Northern France [41] showed a high stem
number per plant: M. sinensis Herman Müssel reached 85
stems during the second year of cultivation, and M. sinensis
Silberspinne reached 176 stems during the third year of culti-
vation (Fig. 4c). One M. ×giganteus clone (Flo) that was also
tested in Northern France [41] showed a high stem diameter,
reaching 9.8 mm in the third year (Fig. 4d).
Finally, ploidy level influenced the biomass production and
its components. Glowaka et al. [84] observed significant but
contrasting effects on biomass production and its components
from increasing the ploidy level, depending on the genotype.
Interestingly, the authors generated polyploid forms for certain
genotypes, which showed more dry matter, higher plants, or a
greater stem diameter compared with the corresponding con-
trols. The results from Zub et al. [41] suggest that the
miscanthus tetraploid or triploid forms were more productive
than the diploid forms, and although the miscanthus forms
studied were from distinct backgrounds, they reinforced the
results from Glowaka et al. [84]. These results show that a
greater ploidy level can improve biomass production.
Despite these encouraging results, this investigation must
be considered in its context for several reasons. First, the
species compared from various environments did not have
the same genetic background. Second, the number of clones
used differed between the three Miscanthus species studied:
fewer clones were observed for the M. ×giganteus and
M. sacchariflorus species than the M. sinensis species; this
could partially explain the lower variability in the M. ×
giganteus and M. sacchariflorus species. Finally, the effect
of ploidy level on biomass production and its components
seemed to differ based on the genotype.
The Phenotypic Variability for the Traits Related to Biomass
Composition
The Influence of Crop Age
Allison et al. [45]studiedtheeffectofyearofcultivationon
cellulose, hemicellulose, and lignin contents in miscanthus.
Based on their experiments, the authors suggested that
miscanthus biomass composition remained relatively consis-
tent over the successive second, third, and fourth years.
Clifton-Brown and Lewandowski [43]comparedseveral
miscanthus clones for ash content in aboveground biomass
during the first 3 years of cultivation. They observed that this
content decreased from 8.9 to 2.3 % DM on average between
the first and third years during the winter harvest (Fig. 5).
Moreover, with a 79 % range of variation during the first year,
the ash content appeared more variable during the first year
compared with the second and third years (Fig. 5).
The Influence of Geographical Area
By studying the biomass compositions of 15 Miscanthus
genotypes in five European locations during the third year of
cultivation, Lewandowski et al. [37] showed that the ash
content in the aboveground biomass varied based on the
experiment location. Hodgson et al. [33] also highlighted a
significant effect from location on cellulose and hemicellulose
contents in the aboveground biomass. In contrast, these au-
thors showed that this effect was not significant for lignin
content in the aboveground biomass.
The influence of location on biomass composition could be
partly explained by climatic conditions. The rainfall differ-
ences between locations could cause mineral leaching. Wind
force differences between locations could affect the composi-
tion due to leaf loss [38].
These observations demonstrated that miscanthus bio-
mass composition differed based on location. However,
in contrast to biomass production, for which clear trends
between Northern Europe countries and Central/Southern
Europe countries have been highlighted [37], no clear
trends identified an area that is most suitable for miscanthus
biomass composition.
The Influence of Crop Management Practices
&The influence of harvest date
The cellulose and lignin contents in the aboveground
biomass increased from 40.6 to 46.4 % DM and 8.0 to
9.4 % DM on average between the autumn and winter
harvests, respectively (Fig. 6a, c). In contrast, the hemi-
cellulose content tended decreased between the two har-
vest dates with the averages 29.4 and 28.8 % DM during
the autumn and winter harvests, respectively (Fig. 6b).
These cellulose, hemicellulose, and lignin content differ-
ences between the autumn and winter harvests could be
due to leaf loss between the two harvest dates [69].
Delaying the harvest time also significantly decreased
the ash content [37,43,70]aswellascuttingthebiomass
and leaving it in the field [71]. For example, the ash
content decreased from 3.9 to 2.5 % DM on average
between the autumn and winter harvests during the third
year of cultivation (Fig. 6d).
In addition, the genotype contribution to phenotypic
variability was high for cellulose, hemicellulose, lignin,
and ash contents in the aboveground biomass (Fig. 6).
Finally, with variations ranging from 25 to 213 %
during the third year of cultivation, the cellulose, hemicel-
lulose, lignin, and ash contents were highly variable traits
(Fig. 6). Interestingly, the highest ranges of variation for
Bioenerg. Res. (2015) 8:502526 519
ash content (145 and 213 %) showed that this trait was
more variable than cellulose, hemicellulose, and lignin
contents regardless of the harvest date (Fig. 6).
&The influence of nitrogen fertilization
Hodgson et al. [32] showed that the nitrogen input
negatively impacted the M. ×giganteus biomass quality;
when nitrogen input increased, the cellulose, hemicellulose,
and lignin contents in the aboveground biomass decreased,
while ash increased. The authors concluded that low nitro-
gen fertilization yielded better quality biomass for thermo-
chemical processes. These observations were encouraging
as environmentally and economically beneficial and as
contributor to cropping system sustainability.
The Influence of Species
Differences among Miscanthus species were highlighted for
cellulose, hemicellulose, lignin, and ash contents during the
winter harvest of the third year of cultivation (Fig. 7). With the
average values of 47.3 and 49.4 % DM, respectively, the M. ×
giganteus and M. sacchariflorus species appeared to contain
more cellulose than the M. sinensis species (Fig. 7a).
Moreover, with the average values of 10.0 and 10.6 % DM,
respectively, the M. ×giganteus and M. sacchariflorus species
also seemed to exhibited higher lignin content than the
M. sinensis species (Fig. 7c). In contrast, with the average
value 30.2 % DM, the M. sinensis species exhibited higher
hemicellulose content than the M. ×giganteus and
M. sacchariflorus species (Fig. 7b). Finally, with the average
value of 1.6 % DM, the M. sacchariflorus species exhibited
lower ash content in the aboveground biomass than the M. ×
giganteus and M. sinensis species (Fig. 7d).
In addition, range of variation seemed variable based on the
Miscanthus species and its traits. The range of variation for the
cellulose and hemicellulose contents seemed to be the same as
for the M. ×giganteus,M. sinensis,andM. sacchariflorus
species. In contrast, the range of variation for the lignin and
ash contents appeared significantly lower for the
M. sacchariflorus species than for the M. ×giganteus and
M. sinensis species. Among the traits studied, the ash content
seemed the most variable trait with a range of variation of up to
178 % compared to the cellulose, hemicellulose, and lignin
contents in the aboveground biomass (Fig. 7).
Finally, for each trait, the portion of phenotypic variability
due to the genotype was high, except for the M. sacchariflorus
species, for which a single clone has been studied (Fig. 7).
The biomass composition observations among the
Miscanthus species can only be considered trends because
only few clones were used in the studies reported in Fig. 7,
wherein only a single M. sacchariflorus clone, six M. ×
giganteus clones, and eight M. sinensis clones were used.
Interestingly, certain Miscanthus clones showed high cel-
lulose and hemicellulose contents, and low lignin and ash
contents. Certain M. sinensis clones presented lower minimal
ash content than M. sacchariflorus clones (Fig. 7d):
Clifton-Brown and Lewandowski [43] and Lewandowski
et al. [37] identified two M. sinensis clones (Sin-11
and Sin-15) with the lowest ash content. Moreover,
one M. ×giganteus clone (EMI08) studied in England by
Hodgson et al. [33] showed lower minimum lignin content
than M. sinensis clones (Fig. 7c).
These observations suggest that certain Miscanthus species
or clones are useful for breeding programs considering bio-
mass composition. In addition, two papers reinforced the
suggestions based on these data: the study by Allison et al.
[45] which did not indicate individual values, and the study by
Hodgson et al. [46] which concerned older crops that were
9 years old.
Key Points and Conclusions on Phenotypic Variability
for Miscanthus Biomass Production and Biomass
Composition
The present review shows that the traits of interest
related to biomass production and biomass composition
were generally highly variable for miscanthus. Among
these traits, aboveground biomass production and stem
number per plant appeared more variable than canopy
height and stem diameter with values up to 300 %
during the third year, the range of variation for the
aboveground biomass production and stem number per
plant were approximately 2-fold greater than the range
of variation for the canopy height and stem diameter.
Interestingly, for the biomass composition, the ash con-
tent in the aboveground biomass appeared to be the
most variable trait; with values that could reach over
200 %, the range of variation for ash content reached
levels 5-fold greater than the range of variation for cellulose,
hemicellulose, and lignin contents. Moreover, the genotype
contribution to the phenotypic variability was high for each
trait studied.
Therefore, the high phenotypic variability for the traits
studied and high genotype contribution to the variability were
important for enhancing miscanthus breeding for bioenergy.
In addition, most of the factors of variation investigated in
this review influenced the traits related to biomass production
and biomass composition and, therefore, must be considered
in breeding programs.
Miscanthus biomass production and canopy height ap-
peared to mainly be influenced by crop age. With the maxi-
mum mean up to 13.0 t DM/ha and 176 cm during the third
year of cultivation, the biomass production and canopy height
may be approximately 2.2- and 2.7-fold greater for a 3-year-
old crop than a 1-year-old crop, respectively.
520 Bioenerg. Res. (2015) 8:502526
Similarly, the stem number per plant seemed mainly affect-
ed by the crop age and species: (i) With the maximum mean at
53 stems per plant in a 3-year-old crop, the stem number per
plant increased 2.5-fold from the first to third year, and (ii) in
comparison, with the maximum mean 58 stems in the
M. sinensis species, the stem number per plant was 2.2-fold
greater in the M. sinensis species than in the M. sacchariflorus
species, which showed the lowest mean in the third year of
cultivation.
The stem diameter appeared to mainly be influenced
by species; with the maximum mean 7.2 mm for the
M. sacchariflorus species, the stem diameter was 1.6-
fold greater than for the M. sinensis species, which
showed the lowest mean during the third year of
cultivation.
For biomass composition, the cellulose, hemicellulose, and
lignin contents in the aboveground biomass appeared to be, at
a same level, most affected by the harvest date and species,
while it was not affected by crop age.
With a 46.4 % DM average during the winter harvest, the
cellulose content was 1.1-fold greater during the winter har-
vest than the autumn harvest during the third year of cultiva-
tion. Similarly, with a 9.4 % DM average during the winter
harvest, the lignin content was 1.2-fold higher during the
winter harvest than the autumn harvest during the third year
of cultivation. In contrast, with the mean 28.8 % DM
during the winter harvest, the hemicellulose content was
similar during the winter and autumn harvests in the
third year of cultivation. In comparison, with means up
to 49.4 and 10.6 % DM in M. sacchariflorus species,
the cellulose and lignin contents were, respectively, 1.1- and
1.2-fold greater in this species than in the M. sinensis species,
which exhibited the lowest average values. With the maxi-
mum mean 30.2 % DM in the M. sinensis species, the hemi-
cellulose content was 1.2-fold greater in this species than in
the M. sacchariflorus species, which exhibited the lowest
average value.
The ash content in the aboveground miscanthus biomass
appeared mainly affected by the crop age. With the average
2.3 % DM in a 3-year-old crop, the ash content can decrease
by 3.9 from the first to the third year.
Therefore, developing miscanthus as sustainable feed-
stock for bioenergy required tackling such factors that
influence traits related to biomass production and
composition.
With bioenergy use as the breeding objective, the pheno-
typic variability, the genotype contribution to this variability,
and the factors of variation investigated in the present review
yielded guidelines (i) to select the traits for consideration in
breeding programs, (ii) to propose the miscanthus species or
clones that appear interesting as progenitors for breeding, and
(iii) to select the optimal crop age, crop management practices,
or geographical areas.
Implications for Miscanthus Breeding Considering
the Industrial Requirements for Bioenergy Use
Few studies have investigated the ability to transform ligno-
cellulosic crops through the different bioenergy conversion
processes on an industrial scale. However, the researchers and
industrial companies involved in recent bioenergy projects
have begun to generate more precise requirements for opti-
mizing lignocellulosic crop biomass production and
composition.
First, for biomass production, the industry supports the
notion that biomass production per hectare must be maxi-
mized under various climates with a lower environmental
footprint for both biomass conversion processes: biochemical
and thermochemical processes (Table 3(A)).
Second, although the lignocellulosic crop biomass compo-
sition requirements have not been fully defined by the indus-
try, data are available from several private partners or publi-
cations. These data concern cellulose, hemicellulose, lignin,
and ash contents in aboveground biomass, which are consid-
ered the main traits that impact biomass conversion efficiency.
Lignin and ash content appear to be the main traits that
affect thermochemical process efficiency. High lignin content
is preferable for this process because high lignin content aids
in improving calorific value [32,33,85] (Table 3(A)). In
contrast, low ash content is preferred, especially for combus-
tion, as it implies a more efficient combustion processes,
higher heating values, lower NO
x
emissions and, thus, lower
combustion costs [37,38](Table3(A)).
Cell wall composition and, more particularly, cellulose,
hemicelluloses, and lignins appear to be the main traits that
affect biochemical process efficiency. The cell wall composi-
tion and its digestibility must be optimized to ensure high
sugar extraction efficiency in biochemical processes [31,86].
Among cellulose, hemicelluloses, and lignins, lignins seem
the main restrictive factors because they apparently limit
microbial accessibility of fermentescible sugars stored in the
cell wall during fermentation [3436]. Cellulose and hemicel-
lulose contents also affect the efficiency of such processes
during hydrolysis or fermentation [31,87]. Based on these
observations and the initial results from recent projects, the
industry expects high cellulose and hemicellulose contents but
low lignin and ash contents to enhance the efficiency of
biochemical processes [Colonna, Dumas, personal communi-
cation, 88](Table3(A)). However, a favorable balance be-
tween lignin, cellulose, and hemicellulose for this process has
not been fully defined because (i) data for the influence of
biomass composition on this processefficiency have not been
published, and (ii) the methods used to assess biomass cell
wall components differ, are not standardized, and yield vari-
able results [31].
All these industrial requirements regarding biomass pro-
duction and composition as well as the high genotypic
Bioenerg. Res. (2015) 8:502526 521
variability described for these miscanthus traits herein aid in
proposing guidelines for developing miscanthus breeding
programs to produce bioenergy.
Firstly, the industry must maximize lignocellulosic crop
biomass production for each bioenergy conversion process
(Table 3(A)). Based on the current investigation, breeding
programs must include developing varieties that (i) reach their
maximum production potential per hectare in a specific envi-
ronment and (ii) are adapted to a wide range of climates.
Among the three Miscanthus species studied in this review,
the M. ×giganteus species and certain M. sinensis species
clones appear particularly interesting because they produce a
high quantity of biomass per hectare (Table 3(B)). Although
the M. sacchariflorus species and M. sinensis species produce
lower biomass per hectare on average than the M. ×giganteus
species (Table 3(B)), these species can offer progenitors
for breeding programs because they are adapted to a
wider range of climates than M. ×giganteus.Moreover,
breeding programs must consider nitrogen and water use
efficiencies for a low environmental impact. In addition,
traits such as canopy height, stem number per plant, or
stem diameter must be considered in miscanthus breed-
ing programs because these traits correlate with biomass
production [40,41].
Secondly, although the industrial requirements for ligno-
cellulosic crop biomass composition have not been fully de-
fined, these requirements appear to contrast based on the
conversion processes (Table 3(A)). Therefore, currently, im-
proving biomass composition is the most important challenge
for bioenergy breeding programs to improve the efficiency
and economic expense of conversion processes [31]. Based on
the present review, we conclude that the species to consider
may differ based on the conversion process. A given
Miscanthus species may be suitable for one conversion pro-
cess but not another. For thermochemical processes, such as
combustion, pyrolysis, or gasification, the M. ×giganteus and
M. sacchariflorus species seemed better adapted than the
M. sinensis species due to their high lignin content
(Table 3(B)). For biochemical processes, such as hydrolysis,
fermentation, or methanation, the M. ×giganteus and
M. sacchariflorus species were also interesting because they
had high cellulose content. However, these species also had
high lignin content, which can reduce efficiency for these
processes (Table 3(B)); therefore, they were not entirely suit-
able. One M. ×giganteus clone (EMI08) and the M. sinensis
species showed lower lignin content and, therefore, may be
particularly interesting for improving lignin content from bio-
chemical processes in breeding programs (Table 3(B)). As low
ash content is more suitable for all of these processes
(Table (3A)), it may also be interesting to consider the
M. sacchariflorus species for improving this trait because this
species contained less ash than the M. ×giganteus and
M. sinensis species (Table 3(B)). The two M. sinensis clones
Tab l e 3 (A) Summary of the requirements expected in the industry for lignocellulosic crop biomass production and composition for each biomass conversion process to produce bioenergy and (B)
Miscanthus species characteristics relevant to biomass production and composition based on the observations in this review
Trait
Biomass conversion
process
Type of conversion Bioenergy end-use Aboveground biomass
production per hectare
Cellulose content Hemicellulose
content
Lignin content Ash content
(A)
Thermochemical Combustion, pyrolysis,
or gasification (syngas)
Heat, electricity,
cogeneration,
or biofuels
+NANA+
Biochemical Fermentation, hydrolysis,
or methanation (biogas)
Heat, electricity,
cogeneration,
or biofuels
+++−−
(B)
Thermochemical Combustion, pyrolysis,
or gasification (syngas)
Heat, electricity,
cogeneration,
or biofuels
+, Gig, Sin (some clones);
, Sacc, Sin
+, Gig, Sacc;
, Sin, Gig (some clones)
+, Gig, Sin;
,Sacc,Sin(someclones)
Biochemical Fermentation, hydrolysis,
or methanation (biogas)
Heat, electricity,
cogeneration,
or biofuels
+, Gig, Sin (some clones);
, Sacc, Sin
+, Gig, Sacc;
,Sin
+, Sin;
, Gig, Sacc
+, Gig, Sacc;
, Sin, Gig (some clones)
+, Gig, Sin;
,Sacc,Sin(someclones)
+corresponded to high value, corresponded to low value, NA data not available
522 Bioenerg. Res. (2015) 8:502526
(Sin-11 and Sin-15) also appeared interesting because they
had the lowest ash content.
In addition to these guidelines, other factors that influence
miscanthus biomass production and composition must be
considered. The variability described in this reviews allow
us to suggest that mature miscanthus crops are more suitable
than younger crops for these processes due to (i) the maximum
canopy height, stem number, and biomass production, as well
as (ii) the lower ash content. Moreover, for thermochemical
processes and, more particularly, combustion, delaying the
miscanthus harvest date from autumn to winter yielded a
better biomass composition, which leads to better combustion
efficiency [38]. Hodgson et al. [33] also predicted that the
winter harvest is more suitable for thermochemical processes
because the calorific value of the winter-harvested biomass
should increase. Finally, delaying the harvest date decreased
the drying and transport expenses because the moisture con-
tent was lower for the winter harvest. Therefore, the winter
miscanthus harvest appeared more economical than the au-
tumn harvest for such conversion processes [89,90].
Interestingly, the crop age and harvest date recommendations
are consistent with recommendations from agronomical and
environmental perspectives [91].
Prospects
The present review provides relevant information on genotypic
and environmental variability for the biomass production and
composition of the three Miscanthus species which gained most
of the attention today. The broad genotypic variability in these
traits shows that the potential for improving miscanthus as a
bioenergy feedstock is great. However, miscanthus is a recent
undomesticated crop; this review shows that little information
on miscanthus biomass production and composition has been
reported compared with major crops, and it must be genetically
improved. Therefore, more information is necessary.
The relationships between the traits related to biomass
production and composition that have not been studied for
miscanthus until now must be investigated. It is important to
improve the biomass composition while at least maintaining
the biomass production. Moreover, little information about the
heritability has been reported up to now. Heritability for these
traits must be thoroughly investigated in the future. The pres-
ent review showed extreme clones for many traits of interest,
which can guide the breeder to create the relevant populations
for assessing such estimates of heritability. In addition,
miscanthus germplasm collections must be developed because
the current collections do not cover the broad genetic diversity
in the Miscanthus genus. Exploring miscanthus species other
than the three Miscanthus species discussed herein may be
promising for assessing new and interesting variables for
miscanthus breeding. In addition, the breeding considerations
for biomass composition should be accompanied by a com-
plete definition of the industrial requirements for the different
bioenergy conversion processes.
Finally, the results from the present investigation on
miscanthus can be extrapolated to other monocotyledons or
perennial crops that are close to miscanthus from a taxonomic
perspective; sugarcane, sorghum, or maize are also promising
feedstock candidates for bioenergy production. Enhancing oth-
er lignocellulosic crops is particularly interesting for the indus-
try. The most sustainable scenario currently proposed includes
developing crop areas close to local facilities. The most suitable
feedstock for each facility will have to fulfill a continuous
supply of the facility and will depend on the areas climatic
and soil conditions. Therefore, this feedstock will be composed
not only of a single crop but a set of several crops specifically
adapted to the production area and harvested at different times
throughout the year to continuously supply the facilities.
Acknowledgments The authors acknowledge the FUTUROL project,
which supported this work. We particularly thank Catherine Bastien for her
helpful comments on the manuscript and for asking the relevant question
that initiated this manuscript. We also thank Isabelle Lejeune-Hénaut,
Catherine Giauffret, Marion Zapater, Jean Carpentier, and the anonymous
reviewers for their valuable recommendations.
Open Access This article is distributed under the terms of the Creative
Commons Attribution License which permits any use, distribution, and
reproduction in any medium, provided the original author(s) and the
source are credited.
References
1. Greenpeace (2013) Scénario de transition énergétique, 27 p
2. Eurelectric (2011) Power choices pathways to carbon-neutral electricity
in Europe by 2050, Full Report, Union of the Electricity Industry, 100 p
3. EREC (2010) RE-thinking 2050, a 100 % renewable energy vision
for the European Union, 76 p
4. ECF (2010) Roadmap 2050: a practical guide to a prosperous, low-
carbon Europe, Technical Analysis, 100 p
5. IEA/OECD (2006) Perspectives des Technologies de lEnergie,
Scénarios et Stratégies à lhorizon 2050, Synthèse et Implications
Stratégiques. International Energy Agency (IEA), 15 p
6. Cadoux S, Ferchaud F, Demay C, Boizard H, Machet J-M, Fourdinier
E, Preudhomme M, Chabbert B, Gosse G, Mary B (2014)
Implications of productivity and nutrient requirements ongreenhouse
gas balance of annual and perennial bioenergy crops. GCB
Bioenergy 6:425438
7. Karp A, Shield I (2008) Bioenergy from plants and the sustainable
yield challenge. New Phytol 179:1532
8. Sanderson MA, Adler PR (2008) Perennial forages as second gener-
ation bioenergy crops. Int J Mol Sci 9:768788
9. Zegada-Lizarazu W, Parrish D, Berti M, Monti A (2013) Dedicated
crops for advanced biofuels: consistent and diverging agronomic
points of view between the USA and the EU-27. Biofuels Bioprod
Biorefin 7:715731
10. Rabelo SC, Carrere H, Filho RM, Costa AC (2011) Production of
bioethanol, methane and heat from sugarcane bagasse in a biorefinery
concept. Bioresour Technol 102:78877895
Bioenerg. Res. (2015) 8:502526 523
11. Dillen SY, Djomo SN, Al Afas N, Vanbeveren S, Ceulemans R
(2013) Biomass yield and energy balance of a short-rotation poplar
coppice with multiple clones on degraded land during 16 years.
Biomass Bioenergy 56:157165
12. Nissim WG, Pitre FE, Teodorescu TI, Labrecque M (2013) Long-
term biomass productivity of willow bioenergy plantations main-
tained in southern Quebec, Canada. Biomass Bioenergy 56:361369
13. Vermerris W (2011) Survey of genomics approaches to improve
bioenergy traits in maize, sorghum and sugarcane free access. J
Integr Plant Biol 53:105119
14. Souza A, Grandis A, Leite DC, Buckeridge M (2014) Sugarcane as a
bioenergy source: history, performance, and perspectives for second-
generation bioethanol. Bioenergy Res 7:2435
15. Hastings A, Clifton-Brown J, Wattenbach M, Stampfl P, Mitchell CP,
Smith P (2008) Potential of Miscanthus grasses to provide energy and
hence reduce greenhouse gas emissions. Agron Sustain Dev 28:465472
16. Heaton EA, Dohleman FG, Long SP (2008) Meeting US biofuel
goals with less land: the potential of Miscanthus. Glob Chang Biol
14:20002014
17. Dohleman FG, Long SP (2009) More productive than maize in the
Midwest: how does miscanthus do it? Plant Physiol 150:21042115
18. Wagoner P (1990) Perennial grain development - past efforts and
potential for the future. Crit Rev Plant Sci 9:381408
19. Arundale RA, Dohleman FG, Heaton EA, McGrath JM, Voigt TB,
Long SP (2014) Yields of Miscanthus ×giganteus and Panicum
virgatum decline with stand age in the Midwestern USA. Glob
Chang Biol Bioenergy 6:113
20. Dohleman FG, Heaton EA, Leakey ADB, Long SP (2009) Does
greater leaf-level photosynthesis explain the larger solar energy con-
version efficiency of Miscanthus relative to switchgrass? Plant Cell
Environ 32:15251537
21. Cadoux S, Ferchaud F, Preudhomme M, Demay C, Fourdinier E,
Strullu L, Mary B, Machet JM, Boizard H, Gosse G (2010)
Production de biomasse et impacts environnementaux des cultures
énergétiques, Colloque final du programme REGIX, 37 mai 2010,
Lyon, France, pp 2739
22. Clifton-Brown JC, Stampfl PF, Jones MB (2004) Miscanthus biomass
production for energy in Europe and its potential contribution to
decreasing fossil fuel carbon emissions. Glob Chang Biol 10:509518
23. Lewandowski I, Schmidt U (2006) Nitrogen, energy and land use
efficiencies of Miscanthus, reed canary grass and triticale as deter-
mined by the boundary line approach. Agric Ecosyst Environ 112:
335346
24. Hodkinson TR, Chase MW, Takahashi C, Leitch IJ, Bennett MD,
Renvoize SA (2002) The use of DNA sequencing (ITS and trnL-F),
AFLP, and fluorescent in situ hybridization to study allopolyploid
Miscanthus (Poaceae). Am J Bot 89:279286
25. Zub HW, Arnoult S, Younous J, Lejeune-Hénaut I, Brancourt-
Hulmel M (2012) The frost tolerance of Miscanthus at the juvenile
stage: differences between clones are influenced by leaf-stage and
acclimation. Eur J Agron 36:3240
26. Cosentino SL, Patane C, Sanzone E, Copani V, Foti S (2007) Effects
of soil water content and nitrogen supply on the productivity of
Miscanthus ×giganteus Greef et Deu. in a Mediterranean environ-
ment. Ind Crop Prod 25:7588
27. de Cesare M, Hodkinson TR, Barth S (2010) Chloroplast DNA
markers (cpSSRs, SNPs) for Miscanthus,Saccharum and related
grasses (Panicoideae, Poaceae). Mol Breed 26:539544
28. Greef JM, Deuter M, Jung C, Schondelmaier J (1997) Genetic
diversity of European Miscanthus species revealed by AFLP finger-
printing. Genet Resour Crop Evol 44:185195
29. Zub HW, Brancourt-Hulmel M (2010) Agronomic and physiological
performances of different species of Miscanthus, a major energy
crop. A review. Agron Sustain Dev 30:201214
30. Greef JM, Deuter M (1993) Syntaxonomy of Miscanthus ×giganteus
Greef-et-Deu. Angew Bot 67:8790
31. Jakob K, Zhou FS, Paterson A (2009) Genetic improvement of C4
grasses as cellulosic biofuel feedstocks. In Vitro Cell Dev Biol Plant
45:291305
32. Hodgson EM, Fahmi R, Yates N, Barraclough T, Shield I, Allison G,
Bridgwater AV, Donnison IS (2010) Miscanthus as a feedstock for
fast-pyrolysis: does agronomic treatment affect quality? Bioresour
Technol 101:61856191
33. Hodgson EM, Lister SJ, Bridgwater AV, Clifton-Brown J, Donnison
IS (2010) Genotypic and environmentally derived variation in the cell
wall composition of Miscanthus in relation to its use as a biomass
feedstock. Biomass Bioenergy 34:652660
34. Klimiuk E, Pokoj T, Budzynski W, Dubis B (2010) Theoretical and
observed biogas production from plant biomass of different fibre
contents. Bioresour Technol 101:95279535
35. Monlau F, Barakat A, Trably E, Dumas C, Steyer JP, Carrere H
(2013) Lignocellulosic materials into biohydrogen and biomethane:
impact of structural features and pretreatment. Crit Rev Environ Sci
Technol 43:260322
36. Boudet AM, Kajita S, Grima-Pettenati J, Goffner D (2003) Lignins
and lignocellulosics: a better control of synthesis for new and im-
proved uses. Trends Plant Sci 8:576581
37. Lewandowski I, Clifton-Brown JC, Andersson B, Basch G, Christian
DG, Jorgensen U, Jones MB, Riche AB, Schwarz KU, Tayebi K,
Teixeira F (2003) Environment and harvest time affects the combus-
tion qualities of Miscanthus genotypes. Agron J 95:12741280
38. Lewandowski I, Kicherer A (1997) Combustion quality of biomass:
practical relevance and experiments to modify the biomass quality of
Miscanthus × giganteus. Eur J Agron 6:163177
39. Gauder M, Graeff-Hönninger S, Lewandowski I, Claupein W (2012)
Long-term yield and performance of 15 different Miscanthus geno-
types in southwest Germany. Ann Appl Biol 160:126136
40. Jezowski S (2008) Yield traits of six clones of Miscanthus in the first
3 years following planting in Poland. Ind Crop Prod 27:6568
41. Zub HW, Arnoult S, Brancourt-Hulmel M (2011) Key traits for
biomass production identified in different Miscanthus species at
two harvest dates. Biomass Bioenergy 35:637651
42. Clifton-Brown JC, Chiang YC, Hodkinson TR (2008) Miscanthus:
genetic resources and breeding potential to enhance bioenergy pro-
duction. In: Vermerris W (ed) Genetic improvement of bioenergy
crops. Springer, USA, pp 273294
43. Clifton-Brown JC, Lewandowski I (2002) Screening Miscanthus
genotypes in field trials to optimise biomass yield and quality in
Southern Germany. Eur J Agron 16:97110
44. Clifton-Brown JC, Lewandowski I, Andersson B, BaschG, Christian
DG, Kjeldsen JB, Jorgensen U, Mortensen JV, Riche AB, Schwarz
KU, Tayebi K, Teixeira F (2001) Performance of 15 Miscanthus
genotypes at five sites in Europe. Agron J 93:10131019
45. Allison GG, Morris C, Clifton-Brown J, Lister SJ, Donnison IS
(2011) Genotypic variation in cell wall composition in a diverse set
of 244 accessions of Miscanthus. Biomass Bioenergy 35:47404747
46. Hodgson EM, Nowakowski DJ, Shield I, Riche A, Bridgwater AV,
Clifton-Brown JC, Donnisona IS (2011) Variation in Miscanthus
chemical composition and implications for conversion by pyrolysis
and thermo-chemical bio-refining for fuels and chemicals. Bioresour
Technol 102:34113418
47. Kalembasa D, Jezowski S, Pude R, Malinowska E (2005) The
content of carbon, hydrogen and nitrogen in different development
stage of some clones of Miscanthus. Pol J Soil Sci 38:169177
48. Lewandowski I, Heinz A (2003) Delayed harvest of miscanthus -
influences on biomass quantity and quality and environmental im-
pacts of energy production. Eur J Agron 19:4563
49. Le Ngoc Huyen T, Rémond C, Dheilly RM, Chabbert B (2010)
Effect of harvesting date on the composition and saccharification of
Miscanthus ×giganteus. Bioresour Technol 101:82248231
50. Mos M, Banks SW, Nowakowski DJ, Robson PRH, Bridgwater AV,
Donnison IS (2013) Impact of Miscanthus ×giganteus senescence
524 Bioenerg. Res. (2015) 8:502526
times on fast pyrolysis bio-oil quality. Bioresour Technol 129:335
342
51. Amougou N, Bertrand I, Machet JM, Recous S (2011) Quality and
decomposition in soil of rhizome, root and senescent leaf from
Miscanthus ×giganteus, as affected by harvest date and N fertiliza-
tion. Plant Soil 338:8397
52. Kim SJ, Kim MY, Jeong SJ, Jang MS, Chung IM (2012) Analysis of
the biomass content of various Miscanthus genotypes for biofuel
production in Korea. Ind Crop Prod 38:4649
53. Van Soest PJ, Wine RH (1967) Use of detergents in the analysis of
fibrous feeds. IV. Determination of plant cell-wall constituents. J
Assoc Off Anal Chem 50:5055
54. Hulle S, Waes C, Vliegher A, Baert J, Muylle H (2012) Comparison
of dry matter yield of lignocellulosic perennial energy crops in a long-
term Belgian field experiment, Grasslanda European resource?
Proceedings of the 24th General Meeting of the European
Grassland Federation, Lublin, Poland, 37June2012
55. Qin J, Yang Y, Jiang J, Yi Z, Xiao L, Ai X, Chen Z (2012)
Comparison of lignocellulose composition in four major species of
Miscanthus. Afr J Biotechnol 11:1252912537
56. Van Hulle S, Roldan-Ruiz I, Van Bockstaele E, Muylle H (2010)
Comparison of different low-input lignocellulosic crops as feedstock
for bio-ethanol production. In: C Huyghe (ed) Proceedings of
the conference of the Eucarpia Fodder and Amenity Species
Section: sustainable use of genetic diversity in forage and turf breed-
ing, 2010
57. Christian DG,Riche AB, Yates NE (2008) Growth, yield andmineral
content of Miscanthus ×giganteus grown as a biofuel for 14 succes-
sive harvests. Ind Crop Prod 28:320327
58. Angelini LG, Ceccarini L, Di Nassa NNO, Bonari E (2009)
Comparison of Arundo donax L. and Miscanthus × giganteus in a
long-term field experiment in Central Italy: analysis of productive
characteristics and energy balance. Biomass Bioenergy 33:635643
59. Mantineo M, DAgosta GM, Copani V, Patane C, Cosentino SL
(2009) Biomass yield and energy balance of three perennial crops
for energy use in the semi-arid Mediterranean environment. Field
Crop Res 114:204213
60. Jezowski S, Glowacka K, Kaczmarek Z (2011) Variation on biomass
yield and morphological traits of energy grasses from the genus
Miscanthus during the first years of crop establishment. Biomass
Bioenergy 35:814821
61. Strullu L, Cadoux S, Preudhomme M, Jeuffroy MH, Beaudoin N
(2011) Biomass production and nitrogen accumulation and
remobilisation by Miscanthus ×giganteus as influenced by nitrogen
stocks in belowground organs. Field Crop Res 121:381391
62. Behnke GD, David MB, Voigt TB (2012) Greenhouse gas emissions,
nitrate leaching, and biomass yields from production of Miscanthus ×
giganteus in Illinois, USA. Bioenergy Res 5:801813
63. Dohleman FG, Heaton EA, Arundale RA, Long SP (2012) Seasonal
dynamics of above- and below-ground biomass and nitrogen
partitioning in Miscanthus × giganteus and Panicum virgatum across
three growing seasons. GCB Bioenergy 4:534544
64. Arundale R, Dohleman F, Voigt T, Long S (2014) Nitrogen fertiliza-
tion does significantly increase yields of stands of Miscanthus ×
giganteus and Panicum virgatum in multiyear trials in Illinois.
Bioenergy Res. doi:10.1007/s12155-013-9385-5:1-9
65. Haines SA, Gehl RJ, Havlin JL, Ranney TG (2014) Nitrogen and
phosphorus fertilizer effects on establishment of giant Miscanthus.
Bioenergy Res. doi:10.1007/s12155-014-9499-4:1-11
66. Larsen SU, Jorgensen U, Kjeldsen JB, Laerke PE (2014) Long-term
Miscanthus yields influenced by location, genotype, row distance,
fertilization and harvest season. Bioenergy Res 7:620635
67. Palmer IE, Gehl RJ, Ranney TG, Touchell D, George N
(2014) Biomass yield, nitrogen response, and nutrient uptake
of perennial bioenergy grasses in North Carolina. Biomass Bioenergy
63:218228
68. Dagnelie P (1969) Théorie et méthodes statistiques. Presses
agronomiques, Gembloux
69. Hayes DJM (2013) Mass and compositional changes, relevant to
biorefining, in Miscanthus × giganteus plants over the harvest win-
dow. Bioresour Technol 142:591602
70. Baxter XC, Darvell LI, Jones JM, Barraclough T, Yates NE, Shield I
(2012) Study of Miscanthus ×giganteus ash compositionvariation
with agronomy and assessment method. Fuel 95:5062
71. Meehan PG, Finnan JM, Mc Donnell KP (2013) The effect of harvest
date and harvest method on the combustion characteristics of
Miscanthus ×giganteus. Glob Chang Biol Bioenergy 5:487496
72. Lewandowski I, Scurlock JMO, Lindvall E, Christou M (2003) The
development and current status of perennial rhizomatous grasses as
energy crops in the US and Europe. Biomass Bioenergy 25:335361
73. Clifton-Brown J, Long SP, Jorgensen U (2001) Miscanthus produc-
tivity. In: Jones M, Walsh M (eds) Miscanthus for energy and fibre.
James and james, London, pp 4667
74. Miguez FE, Villamil MB, Long SP, Bollero GA (2008) Meta-analysis
of the effects of management factors on Miscanthus ×giganteus
growth and biomass production. Agric For Meteorol 148:12801292
75. Heaton E, Voigt T, Long SP (2004) A quantitative review comparing
the yields of two candidate C-4 perennial biomass crops in relation to
nitrogen, temperature and water. Biomass Bioenergy 27:2130
76. Christian DG, Haase E (2001) Agronomy of miscanthus. In: Jones
MB, Walsh M (eds) Miscanthus for energy and fibre. James and
james, London, pp 2145
77. Beale CV, Bint DA, Long SP (1996) Leaf photosynthesis in the C-4-
grass Miscanthus ×giganteus, growing in the cool temperate climate
of southern England. J Exp Bot 47:267273
78. Himken M, Lammel J, Neukirchen D, CzypionkaKrause U, Olfs HW
(1997) Cultivation of Miscanthus under west European conditions:
seasonal changes in dry matter production, nutrient uptake and re-
mobilization. Plant Soil 189:117126
79. Cadoux S, Riche AB, Yates NE, Machet J-M (2012) Nutrient re-
quirements of Miscanthus ×giganteus: conclusions from a review of
published studies. Biomass Bioenergy 38:1422
80. Danalatos NG, Archontoulis SV, Mitsios I (2007) Potential growth
and biomass productivity of Miscanthus ×giganteus as affected by
plant density and N-fertilization in central Greece. Biomass
Bioenergy 31:145152
81. Acaroglu M, Aksoy AS (2005) The cultivation and energy balance of
Miscanthus ×giganteus production in Turkey. Biomass Bioenergy
29:4248
82. Ercoli L, Mariotti M, Masoni A, Bonari E (1999) Effect of
irrigation and nitrogen fertilization on biomass yield and effi-
ciency of energy use in crop production of Miscanthus. Field Crop
Res 63:311
83. Tayot X, Chartier M, Varlet-Grancher C, Lemaire G (1995) Potential
above-ground dry matter production of miscanthus in north-central
France compared to sweet sorghum. In: Chartier P, Beenackers A,
Grassi G (eds) Biomass for energy, environment, agriculture and
industry. Elsevier, Oxford, pp 556564
84. Glowacka K, Jezowski S, Kaczmarek Z (2010) In vitro induction of
polyploidy by colchicine treatment of shoots and preliminary char-
acterisation of induced polyploids in two Miscanthus species. Ind
Crop Prod 32:8896
85. Fahmi R, Bridgwater A, Donnison I, Yates N, Jones JM (2008) The
effect of lignin and inorganic species in biomass on pyrolysis oil
yields, quality and stability. Fuel 87:12301240
86. Zhao X, Zhang L, Liu D (2012) Biomass recalcitrance. Part I: the
chemical compositions and physical structures affectingthe enzymat-
ic hydrolysis of lignocellulose. Biofuels Bioprod Bioref Biofpr 6:
465482
87. Slavov G, Allison G, Bosch M (2013) Advances in the genetic
dissection of plant cell walls: tools and resources available in
Miscanthus. Front Plant Sci 4:217. doi:10.3389/fpls.2013.00217
Bioenerg. Res. (2015) 8:502526 525