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Carbon and greenhouse gas dynamics in
Japanese Miscanthus grasslands and
prospects of genetic improvement of
Miscanthus
Toshihiko Yamada
Field Science Center for Northern Biosphere,
Hokkaido University, Japan
Climate of Sapporo
40,0
600,0
30,0
500,0
20,0
400,0
10,0
300,0
0,0
200,0
-10,0
100,0
0,0
-20,0
Jan
Feb
Mar
Precipitation
Apr
May
Jun
Max. Temperature
Jul
Aug
Sep
Mean Temperature
Average snow cover period: 4th Dec-3rd Apr
Total amount of snowfall per year 5.0m
Oct
Nov
Dec
Min. Temperature
Hokkaido
Sapporo
Osaka
Tokyo
In July 1876 , Dr. William S. Clark, then the
President of the Massachusetts Agricultural
College was invited to be the President of Sapporo
Agricultural College (now Hokkaido University)
Some pictures of Hokkaido University
Main Entrance
Central Lawn
Furukawa Memorial Hall
Roadside with Ginnkgo
Experiment Farm
The 2011 Eastern Japan Earthquake and Tsunami on
11th March 2011
Failure of Cooling Systems at Fukushima I Nuclear
Power Plant after Earthquake and Tsunami 2011
Renewable energy
Solar energy
Wind power
Geothermal energy
Wave energy
Hydroelectricity
Biomass energy
Biorefinery
-From petroleum to biomass
Fossil resource use type
CO2
CO2
Fractional
distillation
Crude
CO2
Gasoline
Naphtha
Kerosene
Light oil
Others
Fuels
Chemicals
Petrochemical
process
Materials
CO2
Biomass utilization type
CO2
Extraction
Biomass
CO2
Synthesis
gas
Sugar
Fuels
Chemicals
Bioprocess
Oil
etc.
・Stable supply of good quality biomass feedstock
・Development of dedicated biomass crops
Biomaterials
Practical
Implementation
Demonstration
Laboratory
Basic research
Phase1
(2005）
Phase2
(2010）
Phase3
(2020）
Phase4
(2050）
Waste
Biomass
Technological
Development
Infrastructure
Improvement
Unused
Biomass
Energy Crops
Novel Energy
Crops
Motivation
Summary of Biomass Targets and Scenario in National Strategy
for Biomass Use in Japan(2006)
preparatory phase
practical phase
Miscanthus spp. is
a potential energy crop
Burning
Cutting for forage
Biotic pressures involved maintenance
of semi-natural Miscanthus grassland
Grazing
Famous Miscanthus grasslands in Japan
Aso (Kumamoto)
Kawatabi (Miyagi）
Soni (Nara)
Sugadaira (Nagano)
Advantage of perennial grasses for biomass production
A low demand for nutrient inputs
Higher yields on relatively poor quality land
Longer persistency
Increase in soil carbon content
Effect on stability and cover value for wildlife
Mineral Nutrients Use Efficiency
Theory for Perennials
Fall
Winter
Mineral
nutrients
Mineral
nutrients
Spring and
Summer
Translocation
from
rhizomes to
growing shoot
Translocation to Dry shoots harvested,
rhizomes as shoot nutrients stay in
rhizomes
senesces
Miscanthus ×giganteus
(Giant Miscanthus)
Triploid natural hybrid: M. sinensis x M. sacchariflorus
Introduction to Denmark in
1935 from Japan as an
ornamental variety
Potential energy crop
since oil crisis happened
in 1970.
Geographical Distribution of
the Miscanthus spp.
Ogi（荻、オギ）
Use of Miscanthus plants
in Japan
 Roof materials of traditional houses
 Animal feeds
 Component of manure
Susuki（薄、ススキ）
Traditional house with a thatched roof
(Clifton-Brown et al. 2008)
Carbon and greenhouse gas dynamics in semi-natural
Miscanthus sinensis grassland in Aso, Southern Japan
Aso in southern Japan
Japan
Sapporo
40°N
Annual Precipitation : 3250 mm yr-1
Mean Annual Air Temperature : 9.6°C
Altitude : 794 m
(30 years average)
Tokyo
Aso
42ﾟ40’15.3” N
141ﾟ45’49.8” E
30°N
130°E
140°E
Long history of M. sinensis utilization for people in Aso
天皇（景行天皇）は九州各地を経て阿蘇の国に来ら
れたが、野は広く遠く、人家が見えなかった。天皇
が「この国に人はいるのか」と仰せられたところ、
阿蘇都彦、阿蘇都媛の二神が人の姿で現れ、「我ら
二人がいます。どうして人がいないものですか」と
述べた。
西暦７２０年頃
日本書紀－Nihon shoki－
（Japanese oldest official histories）
[The Emperor (Keiko) came to Aso. In Aso, the field was far
and large, and he didn’t find a House]
(around 720 CE)
Evidence for long-term existence
of M. sinensis grassland
By analyzing plant phytlith,
vegetation succession can
be estimated.
→ M. sinensis has been
dominated for more than
10,000 years in Aso region.
13,000yr ago
Miyabuchi & Sugiyama (2006), The Quaternary Research, 45, 15-28
Burning is essential for keeping Miscanthus vegetation in grassland
Burning is essential for keeping Miscanthus vegetation in grassland
Burning is essential for keeping Miscanthus vegetation in grassland
After 1 min.
After burning
1. Carbon dynamics as affected by burning
in semi-natural grassland
Toma et al. (2010) GCB Bioenergy, 2, 52-62
Most of the aboveground biomass was burned.
1. Carbon dynamics as affected by burning
in semi-natural grassland
Toma et al. (2010) GCB Bioenergy, 2, 52-62
・Soil C was not burned.
・Soil CH4 and N2O flux did not increased after the burning.
2. How much C has been sequestrated in soil in Aso?
Soil type
Typic Melanudans
Much C is stored in semi-natural
grassland in Aso.
(Soil C content is sometime more
than 20%)
→ How much soil C has been
ANNUALLY accumulated in the
grassland?
K-Ah horizon (7,300yr ago)
2. How much C has been sequestrated in soil in Aso?
Select 6 sites semi-natural
grassland in Aso, and measure
soil C content, δ13C, 14C dating,
plant phytolith, and bulk density
→ Estimate…
・C4 plant contribution to soil C
・Soil C accumulation rate using
mass soil C and 14C dating
・Dominant species in the
grassland
2. How much C has been sequestrated in soil in Aso?
・C from C4 plant contributed to
more than 50% in all sites.
・Miscanthus was dominant
species (50-100%) in many
depth and sites.
Toma et al. (2013) GCB, 19, 1676-1687
2. How much C has been sequestrated in soil in Aso?
・Soil C sequestration rate decreased
with increasing soil C accumulation
interval due to the decomposition of
soil organic C as year passes.
→ 34yrs average: 618 (kg C ha-1 yr-1)
50yrs
: 483 (kg C ha-1 yr-1)
100yrs
: 332 (kg C ha-1 yr-1)
2yrs average:1788 (kg C ha-1 yr-1)
→ This value was used for the
assessment study of the impact of
the semi-natural grassland on
Global Warming.
Toma et al. (2013) GCB, 19, 1676-1687
3. Does semi-natural grassland work for mitigating
Global Warming?
Input
Atmosphere
Biomass utilization
For agriculture
Root
Respiration
Output
Photosynthesis
Soil organic matter
decomposition
CO2, CH4, N2O
Emission
by burning
CH4 uptake
Charcoal
Soil
Dead plant material
3. Does semi-natural grassland work for mitigating
Global Warming?
Input
Atmosphere
Biomass utilization
For agriculture
Output
Photosynthesis
Soil organic matter
decomposition
Carbon budget: - Soil C sequestration rateCO
+ CH
4 emission/uptake
2, CH4, N2O
(Soil C sequestration rate
Emission
= C fix by plant – C loss by burning andbySOM
decomposition)
burning
Root
Respiration
→ Global
Warming Potential
CH4 uptake
= - Soil C sequestration rate + CH4 emission/uptake + N2O emission
Charcoal
Soil
Dead plant material
3. Does semi-natural grassland work for mitigating
Global Warming?
→ Global Warming Potential
= - Soil C sequestration rate + CH4 emission/uptake + N2O emission
・Soil C sequestration rate: 1788 kg C ha-1 yr-1 (Toma et al. 2013)
・Soil CH4, N2O emissions：2 year field monitoring
・CH4, N2O emissions by burning：Burned biomass × Emission Factor
Biomass burning: 2 year field monitoring
Emission factor: CH4 0.66, N2O 1.33 g/kg
(Miura & Kanno (1997) Soil Sci. Plant Nutri., 43, 849-854.)
3. Does semi-natural grassland work for mitigating
Global Warming?
GWP
(MgCO2eq ha-1 yr-1)
Accelerate
3
N2O by burning
CH4 by burning
Soil N2O
emission
Global Warming
0
Soil C
sequestration
-3
-6
Mitigate
-9
Soil CH4
uptake
Total -4.9
Toma et al., Grass. Sci., Submitted
・Negative GWP indicated
semi-natural grassland
mitigated global
warming, even though
CH4 and N2O were
emitted by burning
・Soil C accumulation is
important factor for
mitigating global
warming.
4. Comparison of soil C sequestration rate between
Miscanthus semi-natural grassland and Japanese Cedar
forest plantation
・Two adjusted sites, M. sinensis grassland and Criptomeria japonica forest
plantation (47 years), were investigated.
・Measurement: Soil C content, δ13C, and bulk density and δ13C of plant materials
and root.
←M. sinensis grassland
Japanese Cedar forest
plantation →
Tree seedlings were
transplanted to M. sinensis
grassland 47 years ago.
4. Comparison of soil C sequestration rate between
Miscanthus semi-natural grassland and Japanese Cedar
forest plantation
・Soil mass C in the
Miscanthus grassland
was significantly higher
than that in the forest.
・Soil δ13C was higher in
the grassland than the
forest.
Toma et al. (2012) GCB Bioenergy, 4, 566-575
4. Comparison of soil C sequestration rate between
Miscanthus semi-natural grassland and Japanese Cedar
forest plantation
M. sinensis grassland
Soil C sequestration rate
503 kg/ha
Japanese Cedar forest
Soil C sequestration rate
284 kg/ha
Toma et al. (2012), GCB Bioenergy, 4, 566-575
Conclusion
Higher soil C accumulation in semi-natural grassland than
that in Japanese Cedar forest significantly contribute for
mitigating global warming even thought CH4 and N2O was
emitted by burning, which is essential for maintaining
semi-natural grassland.
Breeding and cultivation programs of Miscanthus in
Hokkaido University
Discovery of new natural Miscanthus triploid plants in sympatric
populations of M. sacchariflorus and M. sinensis
Distribution map of the sympatric population of M.
sacchariflorus and M. sinensis plants in Kushima,
Miyazaki Prefecture, Japan.
Nishiwaki et al. (2011) Amer. J. Bot. 98, 154-159
Confirmation of hybridty by nuclear ribosomal
DNA ITS and chloroplast DNA sequences.
Dwiyanti et al. (2013) BioEnergy Res. 6:486–493
3x=57
Ogi79 Ogi63 Ogi80 M.×g
New natural Miscanthus triploid plants
DNA markers for identifying interspecific hybrids between
Miscanthus sacchariflorus and Miscanthus sinensis
Miscanthus sacchariflorus Miscanthus sinensis
Miscanthus ×giganteus ‘Illinois’
M 1 2 3 4 5 6 7 8 9 10 11 12 13
Sa_IP_139
Sa_IP_151
Sa_IP_303
Intron-flanking primer
pairs developed using
genomic information
from sugarcane
(Saccharum hybrid)
and Sorghum bicolor
Sa_IP_537
Sa_IP_728
Sa_IP_817
Sa_IP_877
Tamura et al., in press,
Grassland Science
Evaluation of biomass potential in wild Japanese Miscanthus
sinensis populations
Northern Hokkaido
Eastern
Hokkaido
Central Hokkaido
Collection sites of wild
Miscanthus sinensis
accessions
Southern Hokkaido
Kanto
Tohoku
Hachijojima
Anzoua et al. in press, Grassland Science
Northern-H Eastern-H Central-H Southern-H Tohoku Kanto
Chubu Shikoku Kyushu
Origin
Days to heading in each area of wild Miscanthus sinensis populations
Northern-H Eastern-H Central-H Southern-H Tohoku Kanto
Chubu
Shikoku Kyushu
Biomass productivity of 41 wild Miscanthus sinensis
populations divided into nine areas at Sapporo over three
seasons (2008–2010)
Correlations of biomass productivity with plant height of 41
wild Miscanthus sinensis populations at Sapporo over two
seasons (2008–2009)
Correlations of biomass productivity with tiller number of 41
wild Miscanthus sinensis populations at Sapporo over two
seasons (2008–2009)
Change of biomass production in superior strains
(2010～2014)
Change of biomass production among 5 breeding strains (Zero fertilizer)
Cultivar
Akeno
Gunma
Matsumae
Morioka
Shiozuka
2010
11.5
15.6
17.3
13.5
20.5
Established in 2009
Biomass potential (t ha-1 yr-1)
2011
2012
24.4
44.1
28.3
37.7
17.6
30.3
22.9
38.7
25.2
46.3
2013
37.9
27.8
24.4
27.8
35.7
2014
29.7
29.3
26.4
27.3
31.9
Miscanthus sinensis ‘Shiozuka’
High biomass potential
High number of tillers
Lower lignin content
First genetically modified Miscanthus grass developed
Particle bombardment-mediated transformation systems of Miscanthus sinensis
by reporter gfp (green fluorescence protein) gene
GFP-illuminated genetically
transformed Miscanthus callus
GFP-illuminated genetically transformed
Miscanthus leaf
Wang et al. (2011) GCB Bioenergy 3: 322-332
Fructans are linear or branched forms of
fructose polymers
O
O
G
F
O
O
F
F
levan series
G
O
F
O
F
O
F
1-SST
1-SST
F
Sucrose
inulin and6-SFT
mixed levan
series
O
F
O
G
Glucose
O
O
O
G
F
6-SFT
F
O
O
O
F
Fructose
O
F
O
F
O
F
O
F
O
F
O
F
O
F
Outward symptoms of prft4 transgenic Miscanthus
plants after cold treatment
Wang et al.
submitted
The flow from harvest to storage in demonstration tests
: flow
Round bales
machineries used
/feed stocks
Harvester
Storage
Baling
Harvest
Farm truck
（transport）
Wheel loader
（ loading ）
Wheel loader
（ loading ）
Roll Baler
Miscanthus was secured in nets first, then wrapped in films.
※ Films are usually used for securing, but nets were suitable
for securing Miscanthus. Costs： Films＞Nets
Bunker silo
Seal
Harvest
machineries used
/feed stocks
Harvester
Farm truck
（transport）
Wheel loader
（ loading ）
1.2m
Diameter
Storage
sealing sheets
Funds
•
•
•
•
New Energy and Industrial Technology Development Organization(NEDO)
Ministry of Agriculture, Forestry and Fisheries, Japan (MAFF)
United States Department of Energy(DOE)
Energy Biosciences Institute , USA (EBI)