Upload
others
View
2
Download
0
Embed Size (px)
Citation preview
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
Tailoring Grain Protein Composition for Wheat Using an Ecophysiological
Modeling Approach
Pierre Martre
INRA – Blaise Pascal UniversityUMR1095 Genetic, Diversity, and Ecophysiology of Cereals
Clermont-Ferrand, France
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
6
8
10
12
14
16
GrainProtein
Concentration(% DM)
CookiesCakesPastriesJapanese noodlesFlat breadsChinese noodlesCrackers
Leavened breadPastaGluten and starch extraction
The Importance of Wheat Storage ProteinsLo
afVolu
me
(cm
3)
Flour protein (%)(Finney et al., 1948)
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
Storage proteins (prolamins)
Gliadins Glutenins
ω-gliadins α,β-gliadins γ-gliadins LMW-GS HMW-GS
Low Optimum High
Loaf volumeGliadin to glutenin ratio
The Importance of Wheat Storage Proteins
% Protein 17%
HMW‐GS 19%
LMW‐GS8%Gliadins
11%
Hardness31%
Unexplained 14%
Allelic polymorphism of storage proteins account for 38% of the genetic variability
of dough strength
(Branlard et al., 2001)
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
49 106 250 359 393 503 746 950
Division Remplissage Maturation
°Cdays49 106 250 359 393 503 746 950
DivisionDivision Storage (filling)Storage (filling) MaturationMaturation
°
Kinetics of Grain Protein AccumulationP
rote
in fr
actio
n(m
g N
gra
in-1)
0.0
0.1
0.2
0.3
0.4
0.5
Thermal time after anthesis (°Cd)
0 200 400 600 800 1000
Rat
e of
pro
tein
frac
tion
accu
mul
atio
n(m
g N
gra
in-1 °
Cd-1
)
0.000
0.002
0.004
0.006 Albumins-GlobulinsGlutenins
Gliadines
How could we model these kinetics and E and G effects on their parameters?
Could we find stable response curves to environmental and/or endogenous plant variables?
Can response curves help us better understand the environmental (E) and genetic (G) bases of the variations of grain protein composition?
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
Outdoor controlledOutdoor controlled--environment conditionsenvironment conditions
2 day time post-anthesis temperatures(19°C and 28°C)
2 post-anthesis watering regimes(100% or 15% of ETP)
Materials and Growing Conditions
3 rates of N fertilisation
(0, 70, and 100 kg N ha-1)
FieldField
2 ear halving treatments
(at anthesis or 250 °Cdays later)
Yield (Mg ha-1) 7.8 6.2 6.8 4.1
Proteins (% DM) 13.1 12.7 15.0 15.4
Grain (×103 grains m-2) 22.4 18.4 16.1 10.8
Arche Récital Renan TamaroCultivars
E and G Effects on Grain N Allocation (1/2)
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
Environmental effects (cv. Récital)
Genetic effects (optimal conditions)
Pro
tein
fra
ctio
ns
(mg
N g
rain
-1)
0.0
0.1
0.2
0.3
0.4
0.5
0.6
ArcheRécitalRenanTamaro
Non-prolamins Gliadins
Glutenins Gliadins / glutenin
Days after anthesis0 10 20 30 40 50
0.0
0.1
0.2
0.3
0.4
0.5
Glia
din
to g
lute
nin
ratio
0.0
0.2
0.4
0.6
0.8
1.0
N allocation(mature grains)
Grain N (mg N grain-1)
0.0
0.1
0.2
0.3
0.4
0.5
0.6
Non-prolamins
Gliadins
Glutenins
Prot
ein
frac
tions
(mg
N g
rain
-1)
0.0
0.1
0.2
0.3
0.4
0.5
Y = 2.99 X0.684
r2 = 0.842
Y = 0.041 X1.295
r2 = 0.931
Y = 0.2502 X1.05
r2 = 0.952
0.4 0.6 0.8 1.0 1.2 1.40.0
0.1
0.2
0.3
0.4
0.5
Pro
tein
fra
ctio
ns
(mg
N g
rain
-1)
0.0
0.1
0.2
0.3
0.4
0.5
0.6
ControlLow NHigh TemperatureDrought
Non-prolamins Gliadins
Glutenins Gliadins / glutenin
Days after anthesis0 10 20 30 40 50 60
0
100
200
300
400
500
600
Glia
din
to g
lute
nin
ratio
0.00.20.40.60.81.01.2
E and G Effects on Grain N Allocation (2/2)
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
Crop simulation model ((SiriusSirius))
N, C Fluxes
Grain Number
Environmental var.(Rg, N, θ, H20, soil,…)
Single grain massSingle grain mass
Protein quantityProtein quantity
Protein compositionProtein composition
Protein Protein concentrationconcentration
Modeling Grain N Accumulation and Allocation (1/5)
(Eur. J. Agro. 2006, 25: 138-154)
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
Gra
in C
or
N(m
g gr
ain-
1 )
Thermal time (°Cdays)
1. The accumulation of structural C is a function of thermal time and grain development.
Accumulation of Structural and Storage C and N
Main Hypotheses
(Plant Physiol. 2003, 133: 1959-1967)
Dcd Der
Structural C
Total NStorage N
Modeling Grain N Accumulation and Allocation (2/5)
2. structural C : N ratio is steady.
3. The accumulation of storage C (starch) and N (gliadins and glutenins) is source driven.
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
(Plant Physiol. 2003, 133: 1959-1967)
Modeling Grain N Accumulation and Allocation (3/5)
Glia
dins
(mg
N g
rain
-1)
0.00.10.20.30.40.50.6
Total grain N(mg N grain-1)
0.0 0.3 0.6 0.9 1.2 1.5
Glu
teni
ns(m
g N
gra
in-1
)
0.00.10.20.30.40.50.6
Gliadins
Glutenins
Partitioning of Grain N
Main Hypotheses
4. The accumulation of grain storage proteins scales with total grain N
1. The accumulation of structural C is a function of thermal time and grain development stage.
2. structural C : N ratio is steady during grain growth. (Dreccer et al., 1997).
3. The accumulation of storage C (starch) and N (gliadins and glutenins) is source regulated.
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
Pro
tein
fra
ctio
ns
(mg
N g
rain
-1)
0.00.10.20.30.40.50.6
0 10 20 30 40 50 60
Time after anthesis (days)0 10 20 30 40 50
0.00.10.20.30.40.5
Arche Récital
Renan Tamaro
Modeling Grain N Accumulation and Allocation (4/5)
Simulation of genetic differences in grain protein fraction accumulation under optimal growing conditions
AmphiphilicGliadinGluténin
Albumin-globulin
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
Genetic and environmental variability of grain protein composition is mainly due to differences in N flux per grain
y = 0.94x + 21r2 = 0.87, MEP = 0.035
y = 1.25x + 82r2 = 0.93, MEP = 0.039
Protein fractions, observed (mg N grain-1)
0.0 0.2 0.4 0.6Prot
ein
frac
tions
, sim
ulat
ed(m
g N
gra
in-1
)
0.0
0.2
0.4
0.6
0.0 0.2 0.4 0.6
GluteninsGliadins
ArcheRécitalRenanTamaro
N0 L N1 19°C/14°CIrr
19°C/14°CDry
Field semi-controled conditions
28°C/15°C Irr
Modeling Grain N Accumulation and Allocation (5/5)
Simulation of genetic differences in grain protein fraction accumulation under optimal growing conditions
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
Long arm Short arm
1A
1B
1D
6A
6B
6D
c
c
c
c
c
c
Gli-A2
Gli-B2
Gli-D2
α,β,γ-Gliadins
Glu-A3
Glu-B3
LMW-GS
Glu-D3
Gli-A5Gli-A3
Gli-A1 Gli-A6
Gli-B3
Gli-B1Gli-B5
Gli-D1
ω-gliadins
Glu-A1
Glu-B1
HMW-GS
Glu-D1
Major Wheat Storage Protein Loci
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
Grain N (mg N grain-1)0.8 1.0 1.2 1.4 1.6
0.10.20.30.40.50.60.70.8
0.8 1.0 1.2 1.4 1.6 1.8
Prot
ein
frac
tions
(m
g N
gra
in-1
)
NullNullxxyGlu-D1
xyxyxxyGlu-B1
NullxyNullxGlu-A1
Null-1A1DNull-1DNull-1AParental line
(LP)
LinesHMW-GS Loci
HMW-GS Gene Dosage Effect on Grain N Allocation
Near-isogenic lines for the number of HMW-GS genes in 7 spring wheat backgrounds
LPNull-1A Null-1DNull-1A1D
Gliadins Glutenins
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
The Origins of Cultivated Wheat
Triticum monococcum Aegylops sp. Triticum tauschii
AA BB DD
X
Wild durum wheat
Triticum durum : AABB
Cultivated durum wheatCultivated durum wheat
X
Wild hexaploid wheat
Triticum aestivum : AABBDD
Cultivated bread wheatCultivated bread wheat
NaturalNaturalselectionselection
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
2 Triticum monococcum(2n = 2x = 14, AA)
3 Aegilops spp.(2n = 2x = 14, BB)
2 Triticum tauschii(2n = 2x = 14, DD) P
oly
plo
ids
Dip
loid
s
Effects of Ploidy Level on Grain N Allocation (1/3)
14 Triticum aestivum(2n = 6x = 42, AABBDD)
Courtot: 1 Parental Line
4 Isohomeoallelic lines
3 Triticum durum(2n = 4x = 28, AABB)
Gliadins Glutenins
Grain N (mg N grain-1)
0.0 0.3 0.6 0.9 1.2
Pro
tein
fra
ctio
ns
(mg
N g
rain
-1)
0.00.10.20.30.40.50.6
0.0 0.3 0.6 0.9 1.2 1.5
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
2 Triticum monococcum(2n = 2x = 14, AA)
3 Aegilops spp.(2n = 2x = 14, BB)
2 Triticum tauschii(2n = 2x = 14, DD) P
oly
plo
ids
Dip
loid
s
Effects of Ploidy Level on Grain N Allocation (2/3)
14 Triticum aestivum(2n = 6x = 42, AABBDD)
Courtot: 1 Parental Line
4 Isohomeoallelic lines
3 Triticum durum(2n = 4x = 28, AABB)
Total gliadins (mg N grain-1)
0.0 0.1 0.2 0.3 0.4
Glia
din
su
bu
nit
s(m
g N
gra
in-1
)
0.00
0.05
0.10
0.15
0.20
0.25α-, β-Gli γ-Gli ω-Gli
0.0 0.1 0.2 0.3 0.4 0.0 0.1 0.2 0.3 0.4 0.5
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
2 Triticum monococcum(2n = 2x = 14, AA)
3 Aegilops spp.(2n = 2x = 14, BB)
2 Triticum tauschii(2n = 2x = 14, DD) P
oly
plo
ids
Dip
loid
s
Effects of Ploidy Level on Grain N Allocation (3/3)
14 Triticum aestivum(2n = 6x = 42, AABBDD)
Courtot: 1 Parental Line
4 Isohomeoallelic lines
3 Triticum durum(2n = 4x = 28, AABB)
Total glutenins (mg N grain-1)
0.0 0.1 0.2 0.3 0.4 0.5Glu
ten
in s
ub
un
its
(mg
N g
rain
-1)
0.0
0.1
0.2
0.3
0.4LMW-GS HMW-GS
0.0 0.1 0.2 0.3 0.4 0.5
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
ω-Gliadins α,β,γ -Gliadins Gli-A1 Gli-B1 Gli-D1 Gli-A2 Gli-B2 Gli-D2
Récital o f b j p n
Renan f b g k m e
Cultivars Quality index
HMW-GS LMW-GS
Glu-A1 Glu-B1 Glu-D1 Glu-A3 Glu-B3 Glu-D3 Récital 62 2* 6 + 8 5 + 10 d g c
Renan 78 2* 7 + 8 5 + 10 a c b
Allelic Composition of the Parental Lines
Materials194 Double haploid lines (Récital × Renan)2 sites (Clermont-Ferrand, Rennes)
Genetic Bases of the Allometric Coefficients (1/2)
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
Y = a × Nb
b (dimensionless)0.6 0.8 1.0 1.2 1.4 1.6
0
20
40
60
80
100
a (mg N grain-1)0.10 0.12 0.14 0.16
Num
ber o
f lin
es
0
10
20
30
40
Genetic Bases of the Allometric Coefficients (2/2)
Protein Parameter h2 Chromosome Collocationfraction (%)
Gliadins a 52 1A Glu-A3 Gli-A1
Glutenins a 12 1A Glu-A3 Gli-A1
Grain N (mg N grain-1)0.3 0.6 0.9 1.2 1.5
Glu
tein
s(m
g N
gra
in-1
)
0.000.020.040.060.080.100.120.14
Glutenins (F1)
Grain N (mg N grain-1)0.3 0.6 0.9 1.2 1.5
Glia
dins
(mg
N g
rain
-1)
0.00
0.05
0.10
0.15
0.20
0.25Gliadins (F4)
Clermont-FerrandRennes
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
The Synthesis of Grain Storage Proteins is Regulated at the Transcriptional Level
TFs regulating the synthesis of grain storage proteins are well conserved among cereal species
916
963
556
690
711
982
738
MAIZE O2
SHORGUME O2RICE
WHEAT SPA
BARLEY BLZ2
MAIZE OHP
BARLEY BLZ1WHEAT SPA2
Arabidoposis
RICE
916
963
556
690
711
982
738
MAIZE O2
SHORGUME O2
MAIZE O2
SHORGUME O2RICE
WHEAT SPA
BARLEY BLZ2
MAIZE OHP
BARLEY BLZ1WHEAT SPA2
Arabidoposis
RICE
SPA
SPA2
(Rubio-Somoza et al., 2006)
Barley
Can the trans-regulation of grain storage proteins by TFs explain the grain N scaling laws?
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
Nucleotide Polymorphism in SPA Influences Grain Protein Allocation (1/2)
(Plant Physiol., 2009, 151, 2133-2144)
5478 bp
41 mutations - 6 indels
1 mutation/136 bp
Mean r2 (LD)= 0.73
Identification of 2 major haplotypesfor each homeologous gene
Number of base pairs fromthe transcription start site
-2000-1000 0 1000 2000 3000 400005
101520253035N
ucle
otid
e di
vers
ity, π
(x
10-3
)
05
10152025303505
101520253035
Spa-D
Spa-B
Spa-A
Spa-A
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
Gli / Glu
Haplotype 1 0.49 + 0.04
Haplotype 2 0.67 + 0.03
P-value 0.0006
Glutenins (mg N grain-1)0.1 0.2 0.3 0.4
Glia
dins
(mg
N g
rain
-1)
0.1
0.2
0.3
0.4
Dough viscoelasticityTenacity Extensibility
Haplotype 1 43.7 106.1Haplotype 2 58.9 77.0P-value 0.005 0.006
(Plant Physiol. 2009, 151: 2133-2144)
Nucleotide Polymorphism in SPA Influences Grain Protein Allocation (2/2)
Thermal time after anthesis(°Cd above 0°C)
0 100 200 300 4000
1
2
3
Nor
mal
ized
exp
ress
ion
x 10
3
0
1
2
3
0
1
2
3
Spa-D
Spa-B
Spa-A
0 100 200 300 4000.00
0.05
0.10
0.15
0.20
0.25
0 100 200 300 4000.0
0.1
0.2
0.3
0.4
0.5
Haplotype 1Haplotype 2
Prot
ein
fract
ions
(mg
N g
rain
-1)
0.0
0.1
0.2
0.3
0.4
Total grain N (mg N grain-1)0.5 0.6 0.7 0.8 0.9 1.0 1.1
0.0
0.1
0.2
0.3
0.4
0.0
0.1
0.2
0.3
0.4
0.5
Glutenins
Gliadins
Non-prolamins
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
MCB1
HMWLMW
Chimericgliadins
SAD GAMYB SPA2
SPA
MYBS3 PBF
Transcriptional Regulatory Network of Grain Storage Protein Regulations
Grain N scaling laws are emergent properties of a transcriptional regulatory network
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
Concluding Remarks
Grain protein composition shows high G x (E x M) interactions
These interactions can be explained by simple scaling laws
As a consequence of these scaling laws grain protein concentration is primarily determined by the rate and duration of grain N accumulation => determined at the whole plant (canopy) level
But we could identify both natural and induced genetic variations in the parameters of the scaling laws of grain N allocations => “decorrelate” grain protein composition from total grain N => allows manipulating grain protein composition and grain N (concentration) independently (low input systems)
Scaling laws of grain N allocation are emergent properties of a regulatory transcriptional network
Analysing and modelling these regulatory networks in the frame of the scaling laws of grain N allocation can helps us “decorrelate” grain protein composition from total grain N and develop new ideoptypewith less total grain protein by with the right balance
N U T R I T I O N A G R I C U L T U R E
E N V I R O N M E N T
U M R 1 0 9 5 G D E C
A G R O 2 0 1 0 – 31 Sept. 2010
UMR1095 GDEC, Clermont-Ferrand
François BalfourierGérard BranlardGilles CharmetCatherine RavelEugène Triboi
Vitalie SamoilZhanwu DaiAnne Plessis
Sibille PerrochonMireille DardevetNathalie Duchateau
Founding
Marie Agier
Gene network modelling
TransgenesisStéphane LafargeFrançois Torney
Acknowledgements