improving the nitrogen responses of uk wheat varieties
DESCRIPTION
International Gluten Workshop, 11th; Beijing (China); 12-15 Aug 2012TRANSCRIPT
Yongfang Wan, Malcolm Hawkesford, Rowan Mitchell and Peter Shewry
Rothamsted Research, UK
Improving the nitrogen responses of UK wheat varieties
Background
• Most varieties exhibit a negative relationship between grain protein and yield.
• Few varieties show reproducible deviations, such as
Cordiale and Marksman with high yield and grain protein.
• These varieties can make most efficient use of nitrogen inputs, have better economic costs and minimise environmental pollution.
• Little is known about the molecular basis for the protein deviation resulting from:
• nitrogen uptake • source metabolism • sink activity
This project will: • identify some candidate transcripts responding
to nitrogen application and /or related to the protein deviation;
• determine the effect of nitrogen levels on protein
composition; • examine interaction between nitrogen, protein
deviation and bread-making quality.
Strategy
Six cultivars (He, Co, Mk, Is,
Ma, Xi)
Three N levels (N100, N200, N350kg/ha)
AA 14 DAA AA 21DAA AA 28 DAA AA 35DAA AA 42DAA mature
Transcriptome Affymetrix and qPCR
Protein composition SE-HPLC;SDS-PAGE
Baking quality
Agronomic traits Grain yield and protein
Field trials
Bioinformatics Hypothesis, Genes, markers
integrated
Grain protein responses to nitrogen in 2009 and 2010 at Rothamsted.
0
0.5
1
1.5
2
2.5
3
Co He Is Ma Mk Xi Co He Is Ma Mk Xi
Gra
in %
N
2009 I 2010
N100
N200
N350
Grain yield responses to nitrogen in 2009 and 2010 at Rothamsted
0
2
4
6
8
10
12
14
Co He Is Ma Mk Xi Co He Is Ma Mk Xi
Gra
in y
ield
(t/
ha@
85
% D
M)
2009 I 2010
N100
N200
N350
105 transcripts are significantly up-regulated by nitrogen.
Transcriptome analysis at 21D in 2009 and 2010 by Affymetrix .
N100 N200 N350 N100 N200 N350
2009 2010 Storage
proteins(13)
Defence genes (6)
photosynthesis genes(5)
Unknown genes (81)
10000
11000
12000
13000
14000
15000
N1
00
N2
00
N3
50
N1
00
N2
00
N3
50
N1
00
N2
00
N3
50
N1
00
N2
00
N3
50
N1
00
N2
00
N3
50
N1
00
N2
00
N3
50Co
HeIs
MaMk
Xi
Raw
exp
ress
ion
Gliadin transcripts
2009
2010
2009
10000
12000
14000
16000
18000
20000
22000
N1
00
N2
00
N3
50
N1
00
N2
00
N3
50
N1
00
N2
00
N3
50
N1
00
N2
00
N3
50
N1
00
N2
00
N3
50
N1
00
N2
00
N3
50Co
He IsMa
MkXi
raw
exp
ress
ion
HMW-glutenin subunit transcripts
2009
2010
2009
10000
15000
20000
25000
30000
N1
00
N3
50
N2
00
N1
00
N3
50
N2
00
N1
00
N3
50
N2
00
Co He Is Ma Mk XiR
aw e
xpre
ssio
n
LMW-glutenin subunit transcripts
2009
2010
Response of storage protein transcripts to nitrogen
0
1
2
3
4
5
6
7
8
9
10
14 21 28 35
REL
ATI
VE
EXP
RES
SIO
N
days after anthesis
N100
N200
N350
0
2000
4000
6000
8000
10000
12000
14000
CO HE IS MA MK XI
N100
N200
N350
Exp
ress
ion
inte
nsi
ty
0
2000
4000
6000
8000
10000
12000
14000
CO HE IS MA MK XI
N100
N200
N350
2009 2010
A novel γ-gliadin gene is up-regulated by nitrogen.
Expression on Affymetrix
Expression by realtime RT-PCR
1AS
CS-typical γ-gliadin
γ-3 hordein (Hordeum chilense) CS-3
CS-3
CS-1
He-1
T. monococcum
Ae. tauchii
CS-2
He-2
1DS
1BS
Comparison of novel γ-gliadin genes
Xi IS Co Ma MK He
HMW-glutenins
ω- gliadins
LMW-glutenins & α,β,γ gliadins
By
Dx Bx
Ax
Dy
Globulin and Albumin
Effect of nitrogen on protein composition
N100 N200 N350
HMW glutenins ω - gliadins LMW-glutenin & gliadins
Dx2%=Dx2 volume/total HMW glutenins
ω- 5 Dx
Bx By
Dy ω- (1+2)
7.5
12.5
17.5
22.5
27.5
32.5
1Ax1 1Dx5 1Bx17 1By18 1Dy10
N100
N200
N350
0.0
10.0
20.0
30.0
40.0
50.0
60.0
70.0
ω-5a ω-(1+2)a ω-(1+2)b
N100
N200
N350
7.5
12.5
17.5
22.5
27.5
32.5
1Ax1 1Dx5 1Bx17 1By18 1Dy10
N100
N200
N350
0.0
10.0
20.0
30.0
40.0
50.0
60.0
70.0
ω-5a ω-(1+2)a ω-(1+2)b
N100
N200
N350
0.0
10.0
20.0
30.0
40.0
50.0
60.0
70.0
80.0
HMW-GS% ω-GLIADIN% LMW-GS+GLIADINS(%)
N100
N200
N350
0.00
10.00
20.00
30.00
40.00
50.00
60.00
70.00
80.00
90.00
HMW-GS% ω-GLIADIN% LMW-GS+GLIADINS (%)
N100
N200
N350
HMW-glutenin
Omega-gliadin
Group protein
2009 2010
Cordiale
IS IS
32.0
33.0
34.0
35.0
36.0
37.0
38.0
39.0
40.0
40.0 42.0 44.0 46.0 48.0 50.0 52.0
Po
lym
eri
c (F
1+
F2)
Monomeric (F3+F4)
Linear (N100)
Linear (N200)
Linear (N350)
HE
Ma
CO MKMK
IS
HE HE
IS ISXI
N100 N200 N350
Relationship of polymeric and monomeric proteins by SE-HPLC in 2009 mature seeds
2010 baking quality from Rothamsted site
Summary
1: Nitrogen application can increase both grain protein and yield.
2: The storage protein transcripts are significantly up-regulated by nitrogen. 3: The protein compositions respond to nitrogen disproportionally. 4: The baking quality is improved by higher nitrogen levels, but is affected by different sites, and years .
Gene expression Protein composition
Grain protein Grain yield
Nitrogen
Baking quality
environments
Protein deviation
Future
Acknowledgment
Funding:
Partners: Campden BRI, Gemma Chope
HGCA, Dhan Bhandari
Nickerson Seeds RAGT seeds Syngenta seeds KWS-UK
ADM milling Ltd Heygates ltd Warbourtons Rank Hovis McDougal