Challenges for Improving Wheat
Quality
Peter Shewry
(Rothamsted Research and the University of Reading)
The major grain components
determine end use quality
starch
UK usage of wheat
2013
Total= 14 m tonnes
feed
milling
seed
other
gluten
proteins
cell walls (fibre)
vitamins
minerals
phytochemicals
distilling
biofuels
livestock
feed
food
processing
Human
health
Challenges for Wheat Quality in Europe
• Reduce N requirement for growing high yielding
bread making wheats
• Increase STABILITY of yield and quality
• Improve health benefits
The high grain protein requirement for bread
making results in N applications above the
optimum for yield
16
14
12
Grain yield
10
(tonnes/Ha)
8
Grain protein %
6
(%N x 5.7)
4
2
0
48
144
192
240
288
N application (kg/Ha)
cost of N fertiliser
Improved quality with reduced N
• Exploit grain protein deviation (GPD)
• Improve quality at low protein
• Exploit non-protein sources of quality
Grain protein deviation (GPD):
Mean grain N contents and yields for 47 wheat
cultivars grown at Rothamsted Research between
2004 and 2012.
Exploiting GPD: project components
Six cultivars
• Trials
•
11 environments (sites or years)
• Analyses
•
Yield
•
Grain protein
•
Milling and baking
• Transcriptomics (21 dpa)
WITH GPD
Hereward: Group 1, stable high
protein
Cordiale: Group 2, medium protein,
Marksman: Group 2, medium protein,
WITHOUT GPD
Xi 19: Group 1, medium protein
Malacca: Group 1, medium protein
Istabraq: Group 4, low protein,
suitable for feed and biofuel
Transcriptomics
2009 and 2010
2011
Expression of 8770 genes varied with varietyand/or environment
(out of 60000 total)
CO
He
IS
Ma
MK
XI
Co Co
Ma
MK
IS
He
Xi
Expression data processed to remove effects of
1. Nitrogen fertilisation
2. Yield
Identified 136 transcripts related to cultivar differences in GPD
Expression of genes positively related to GDP in 2009 (open
circles), 2010 (closed circles) and 2011 (open squares)
The samples are
sorted along the xaxis according to
the cultivar
With GPD
Red:
Hereward
Green: Cordiale
Yellow: Marksman
Without GPD
Blue:
Istabraq
Black: Malacca
Purple: Xi19
The challenge is to confirm gene function
- genetic analysis
- transgenesis
Can we identify new sources of
improved and STABLE quality?
• Lipids by MS/MS
• LMW metabolites by NMR and ESI-MS
The role of lipids in determining gas bubble
stability in wheat dough
• Formation of visco-elastic gluten network
during mixing
• Expansion of the network by entrapment of
CO2 during proofing
• Retention of gas bubble structure through to
baking
These are determined by:
•
Gluten viscoelasticity
•
Lipids at the gas bubble interface
Surface active lipids can be studied
in dough liquor
Centrifuge
dough
Foam stability
Electrode
Foam
Over -foaming to
separation and collect
active lipids.
Jet
Surface composition and structure
Surface tension
& surface
rheology
Dough liquor
zoom
microscope
digital
camera
Foam microconductivity to
measure stability.
Dough liquor
Pendant drop
Langmuir trough
allows sampling of
surface lipids for MS
analysis
Lipidomics
Extraction
MS analysis
Data
Processing
Direct infusion:
CTC PAL
+NL (341.00): 40 MCA scans from Sample 5 (BD_5_DGDG) of 2013-04-03LOUISE-FLOUR_DGDG.wiff (Nanospray)
Max. 2.5e6 cps.
2.5e6
2.4e6
2.3e6
2.2e6
2.1e6
2.0e6
1.9e6
1.8e6
1.7e6
1.6e6
1.5e6
Intensity, cps
MS/MS
4000 QTRAP
1.4e6
1.3e6
1.2e6
1.1e6
1.0e6
9.0e5
8.0e5
7.0e5
6.0e5
5.0e5
4.0e5
3.0e5
2.0e5
1.0e5
0.0
900
905
910
915
920
925
930
935
940
945
950
955
m/z, Da
960
965
970
975
980
985
990
995
1000
Separates, identifies and and quantifies over150 molecular species
Lipids in Foaming of dough liquor
Foam stability
Drainage volume (ml)
20
15
10
5
0
0
20
40
60
80 100 120 140 160 180
Time (min)
Comparison of phospholipids in flour
and dough liquor of cv Hereward
70
60
% mol
50
DL HEREWARD
40
30
HEREWARD FLOUR
20
10
0
LPC
PA
PC
PE
PG
PI
Will allow
• Selection of improved lines
• Innovative milling to concentrate lipids
High stable quality at lower N can be
achieved by combining
• Exploit GPD
• High dough strength at lower protein
• Novel non-protein sources of quality
This will be facilitated by new technologies
Improving wheat quality for diet and
health- major components in grain
• Dietary fibre
• Minerals (Fe, Zn, Se)
• B vitamins (folates)
• Methyl donors (betaine)
Bread contributes significantly to the daily intake
of dietary fibre in the UK (% adult intake)
Energy
Protein
Carbohydrate
Fibre
13
12
21
20
White bread
8
8
14
11
Wholemeal
Bread
2
2
3
5
All bread
Fibre is deficient in EU diets
• UK intake 14g/day
• EFSA recommended 25g/day
Source: Steer et al. Proc Nutr Soc/ 2008, 67, E363.
Some potential mechanisms for benefits of DF
Improved bowel function
Reduce cholesterol
Slow glucose absorption,
improve insulin
sensitivity
Reduce colorectal cancer
• Decreased transit time
• Increased stool bulk
• Reduce absorption
• Fermentation in colon to short chain fatty acids
• Slow digestion and absorption due to increased
viscosity
• Inhibit amylase and absorption of sugars
• Beneficial effects of butyric acid on colonocytes
• Dilute and reduced exposure to carcinogens
Based on slide provided by Janet Cade, Leeds
Some take home messages from metaanalyses of fibre and health
• 10% reduction in risk of colon cancer with10g/day
extra cereal/wholegrain fibre
• 5% reduction in risk of breast cancer with 10g/day
extra soluble fibre
• 7% reduction in risk of stroke with 7g/day extra
total fibre
Thanks to Janet Cade, University of Leeds
Diversity screen of 150 bread wheat lines and 50 other cereals
grown at Martonvasar in 2004/5
Types
Geographical origins
Western Europe
France, UK,
Netherlands
34
West-Central Europe
Germany, Austria, Switzerland, Czech
Republic,Poland
22
South Europe
Italy, Bulgaria, Serbia, Croatia
28
South Continental Europe
Hungary, Romania
14
Steppe
Russia, Ukraine
Land races and old variety populations
14
Old and transitional varieties
13
Americas
USA, Canada, Mexico, Argentina
30
Asia, Near East, Australia
China, Korea, Australia, Turkey
15 l
Aims: identify variation
determine heritability
Modern varieties
Germplasm accessions
Winter type
Spring type
Spelt
durum wheat
T. monococcum
T. dicoccum
Rye
Barley
Oats
64
117
26
184
37
5
10
5
5
10
10
5
Arabinoxylan (AX) is the major dietary
fibre component in wheat grain
White flour
Bran
2-3% fibre
45-50 % fibre
70% AX
50% AX
The contents of soluble and total AX fibre vary widely in
bran and white flour of 150 wheat lines
25
15
150.00
250.00
200.00
10
50.00
5
0.00
0
THESEE
CHINESE-…
CF99007
SAVA
MANITAL
KEY
SULTAN95
MALACCA
BILANCIA
KUKRI
ALBA
RIBAND
GALAHAD
THATCHER
KANZLER
CUBUS
CHARA
ISENGRAIN
KRASNODA…
BISCAY
TREMIE
JUBILEJNAJ…
CF99102
LIBELLULA
ATLAS-66
PRODUTTORE
GRANBEL
SARATOV-29
BUCK-…
SUNSTAR
AUTONOMIA
COURTOT
KARL-92
BARANJKA
CF99105
LONA
GENE
AMADEUS
VALORIS
DEKAN
SEU-SEUN-27
CATBIRD
TOMMI
STEPHENS
MANITOBA
AGRON
ALABASSKA…
PROBSTDO…
APACHE
NOMADE
MIETI
AKTEUR
SCOUT66
POBEDA
TAMARO
HEREW ARD
FERTODI-293
BANKUTI-1201
MILAN
BALKAN
MOMTCHIL
NAP-HAL
VONA
SAGITTARIO
CARDINAL
SU321
ZVEZDA
ARINA
SADOVO-1
BEGRA
UKRAINKA
HANA
ETOILE-DE-…
BEZOSTAJA-1
ALBATROS-…
SOLUBLE: 0.3-0.85%
BEGRA
BARANJ KA
STEPHENS
ALBA
KANZLER
PLAINSMAN-V
CHARA
GERONIMO
KEY
ALABASSKAJ A
MONOPOL
BUCK-CATRIEL
HEREW ARD
AURORA
RAVENNA
SOISSONS
NAP-HAL
SCOUT66
CLAIRE
AUGUSTA
RED-RIVER
QUALITAL
MV-PALOTAS
PRODUTTORE
RIBAND
GENE
GLENLEA
PASTOR
KORW ETA
DISPONENT
AVALON
J UBILEJ NAJ A-50
KIRKPINAR-79
FLAMURA-85
THATCHER
RUSALKA
MEXIQUE-50
MALACCA
GALAHAD
J ANZ
SU321
B16
BALKAN
FUNDULEA-29
AGRON
CARMEN
APACHE
KLEIN-…
CF99007
KRASNODARS…
MANITAL
MARIS-…
CAPHORN
HANA
COURTOT
LIBELLULA
MV-SUBA
SUNSTAR
SPARK
ATAY-85
DEKAN
VALORIS
G K-TISZATAJ
MOMTCHIL
SEU-SEUN-27
BISCAY
RENAN
TOMMI
TAM200
RECITAL
LYNX
SARATOV-29
AMADEUS
CUBUS
YUMAI-34
SOLUBLE: 0.3-1.4%
100.00
Yumai 34
30
GLENLEA
CAMP-REMY
AKTEUR
KORW ETA
RUSALKA
ATAY-85
COURTOT
UKRAINKA
DEKAN
GERONIMO
BISCAY
SPARTANKA
KIRKPINAR-79
TALDOR
PALESIO
MIETI
SULTAN95
CHARA
TOMMI
JANZ
ALBATROS-…
AGRON
CLAIRE
KANZLER
APACHE
BLUE/AG
KUKRI
SUNSTAR
ETOILE-DE-…
VONA
MILLENNIUM
GUARNI
THESEE
CADENZA
BLASCO
GEREK-79
ARINA
HANA
RAVENNA
KIRAC66
RENAN
CF99102
KARL-92
FLAMURA-85
SAVA
KRASNODAR…
GRANBEL
MV-PALOTAS
SARATOV-29
KEY
CHINESE-…
AUTONOMIA
PAN
MANITAL
RECITAL
MOMTCHIL
SAGITTARIO
MEXIQUE-50
GALAHAD
ATLAS-66
AMADEUS
CAMPARI
RIALTO
B16
JUBILEJNAJA…
BALKAN
SU321
CARMEN
LASTA
CF99007
LYNX
SEU-SEUN-27
MILAN
BILANCIA
YUMAI-34
CHINESE-SPRING
SOISSONS
SAVA
QUALITAL
TALDOR
GLENLEA
AUTONOMIA
LONA
FREDRICK
FLEISCHMANN-…
ORNICAR
OBRIY
BISCAY
CAPO
POBEDA
RAVENNA
BEGRA
THATCHER
ARTHUR-71
SPARTANKA
JUBILEJNAJA-50
GLORIA
MONOPOL
FLAMURA-85
AKTEUR
CAMP-REMY
FUNDULEA-29
AUGUSTA
HANA
NS-RANA-1
BLASCO
SAGITTARIO
ATAY-85
BLUE/AG
RED-FIFE
ALBA
SCOUT66
HEREW ARD
AGRON
ARINA
DEKAN
HERZOG
ALBATROS-…
JANZ
B16
ESTICA
ELLVIS
RENAN
CLAIRE
PROBSTDORFE…
BLE-DES-DOMES
TOMMI
SKOROSPELKA-…
CHARA
MALACCA
BALKAN
CF99102
SU321
AMADEUS
CATBIRD
KIRKPINAR-79
AVALON
MILLENNIUM
UKRAINKA
SEU-SEUN-27
MOULIN
MAGDALENA-FR
CUBUS
GRANBEL
AZTECA67
MANITAL
CF99007
CAMPARI
MIETI
YUMAI-34
% SOLUBLE: 20-50%
% SOLUBLE: 2-5%
TOTAL: 1.35-2.75%
TOTAL: 12.7-22.1%
Yumai 34
1.60
1.40
1.20
1.00
0.80
0.60
0.40
0.20
0.00
0.90
0.80
0.70
0.60
0.50
0.40
0.30
0.20
0.10
0.00
FLOUR
BRAN
20
Data of Kurt Gebruers, Christophe Courtin and Jan Delcour (KU Leuven)
AX content of flour and bran is highly
heritable
Exploiting natural variation in AX fibre: Yumai 34
 A Chinese wheat variety released
in 1998 in Henan province
 High fibre content (total and
soluble)
 High viscosity of aqueous
extracts
 Good breadmaking quality
3
2.5
2
TOT-AX
1.5
WE-AX
1
0.5
0
Yumai 34
Mean 151 lines
Healthgrain Diversity screen data from Gebruers et al
(2008) J. Ag. Food Chem., 56, 9740-9749.
Relative Viscosity
3.00
2.50
2.00
1.50
1.00
Yumai 34
Control
Yumai 34
Control
Exploiting natural variation in AX fibre:
Yumai 34 x Ukrainka
AX
content
2.500
TOT
WE
Relative
viscosity
2.000
1.500
1.000
Ukrainka
Yumai 34
0.500
2.400
2.200
Ukrainka
Yumai 34
2.000
1.800
1.600
1.400
1.200
1.000
0.000
96 F6 lines + parents
 Fibre analysis
 Genotyping
 Transcriptome analysis
of bulk segregants
Ukrainka
Yumai 34
New sources of variation: the Watkins collection
Collected by Board of Trade
for A E Watkins (University of
Cambridge)
From markets and farms
in1920s and 30s
Initially several thousand but
now down to1300
From 34 countries
Held at JIC
Core collection 119 lines
Total and WE-AX in wholemeal flours of the Watkins lines
WE-AX
Yumai 34
TOT-AX
Yumai 34
40%
WE-AX as % of TOT-AX
Yumai 34
0%
Exploring the role of AX fibre in the human GI tract
SMALL
INTESTINE
effects of AX on
digestion,
viscosity and
mass transfer
prebiotic
effects of AX:
SCFAs, “good
bacteria”
COLON
ferulate and
vascular
function/blood
pressure
dough
fermentation
and IBS
Conclusions- Priorities
• Develop N efficient wheat with good quality at
low grain N
• New protein and non-protein sources of qualitygreater stability?
• Improve health benefits
DF is an important target with established health
benefits and high heritability
Thanks
GPD
Malcolm Hawkesford
Yongfang Wan
Gemma Chope (Campden BRI)
Simon Penson (Campden BRI)
Ellen Mosleth Faergestad
(Nofima/Oslo)
Millers and bakers
Breeders
DIETARY FIBRE
Rowan Mitchell
Alison Lovegrove
Mark Wilkinson
Till Pellny
Jackie Freeman
Ondrej Kosik
Zoltan Bedo (MTA)
Mariann Rakszegi (MTA)
GRAIN LIPIDS
Richard Haslam (RRes)
Peter Wilde (IFR)
Louise Salt (IFR)
Irene Gonzalez-Thuillier
Paola Tosi (Reading)
Simon Penson (Campden BRI)
Peter Skeggs (Hovis)