Challenges for Improving Wheat Quality

Peter Shewry (Rothamsted Research and the University of Reading)

starch

cell walls (fibre)

vitamins minerals

phytochemicals

The major grain components determine end use quality

biofuels

distilling

food processing

livestockfeed

Human health

milling

feed

otherseed

UK usage of wheat 2013

Total= 14 m tonnes

gluten proteins

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

48 144 192 240 2880

2

4

6

8

10

12

14

16

Grain yield (tonnes/Ha)

Grain protein % (%N x 5.7)

N application (kg/Ha)

The high grain protein requirement for bread making results in N applications

above the optimum for yield

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

• Trials• 11 environments (sites or years)

• Analyses• Yield

• Grain protein

• Milling and baking

• Transcriptomics (21 dpa)

Six cultivars

WITH GPDHereward: Group 1, stable high protein

Cordiale: Group 2, medium protein,

Marksman: Group 2, medium protein,

WITHOUT GPDXi 19: Group 1, medium protein

Malacca: Group 1, medium protein

Istabraq: Group 4, low protein, suitable for feed and biofuel

Transcriptomics

CO He IS

20112009 and 2010

Ma MK XI

Expression of 8770 genes varied with variety-and/or environment (out of 60000 total)

Ma MK IS He XiCoCo

Expression data processed to remove effects of 1. Nitrogen fertilisation2. YieldIdentified 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 x-axis according to the cultivar

With GPDRed: HerewardGreen: Cordiale Yellow: Marksman

Without GPDBlue: Istabraq Black: MalaccaPurple: 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

• 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

The role of lipids in determining gas bubble stability in wheat dough

Centrifuge dough

Surface active lipids can be studied in dough liquor

Dough liquor

zoom microscope

digital camera

Pendant drop

Surface tension & surface rheology

Foam stability

Surface composition and structure

Dough liquorLangmuir trough allows sampling of surface lipids for MS analysis

Foam micro-conductivity to measure stability.

Over -foaming to separation and collect active lipids.

FoamElectrode

Jet

Lipidomics

MS/MS4000 QTRAP

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.

900 905 910 915 920 925 930 935 940 945 950 955 960 965 970 975 980 985 990 995 1000m/z, Da

0.0

1.0e5

2.0e5

3.0e5

4.0e5

5.0e5

6.0e5

7.0e5

8.0e5

9.0e5

1.0e6

1.1e6

1.2e6

1.3e6

1.4e6

1.5e6

1.6e6

1.7e6

1.8e6

1.9e6

2.0e6

2.1e6

2.2e6

2.3e6

2.4e6

2.5e6

Intensity, cps

Extraction Data Processing

MS analysis

Separates, identifies and and quantifies over150 molecular species

0 20 40 60 80 100 120 140 160 18002468

101214161820

Foam stability

Time (min)

Dra

inag

e vo

lum

e (m

l)

Lipids in Foaming of dough liquor

Comparison of phospholipids in flour and dough liquor of cv Hereward

LPC PA PC PE PG PI0

10

20

30

40

50

60

70

DL HEREWARDHEREWARD FLOUR%

mo

l

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

All bread 13 12 21 20

White bread

8 8 14 11

WholemealBread

2 2 3 5

Source: Steer et al. Proc Nutr Soc/ 2008, 67, E363.

Fibre is deficient in EU diets• UK intake 14g/day• EFSA recommended 25g/day

Some potential mechanisms for benefits of DF

• Decreased transit time• Increased stool bulkImproved bowel function

• Reduce absorption• Fermentation in colon to short chain fatty acidsReduce cholesterol

• Slow digestion and absorption due to increased viscosity

• Inhibit amylase and absorption of sugars

Slow glucose absorption, improve insulin

sensitivity

• Beneficial effects of butyric acid on colonocytes• Dilute and reduced exposure to carcinogensReduce colorectal cancer

Based on slide provided by Janet Cade, Leeds

Some take home messages from meta-analyses 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

Geographical origins

Western Europe France, UK, Netherlands 34

West-Central EuropeGermany, Austria, Switzerland, Czech

Republic,Poland 22 South Europe

Italy, Bulgaria, Serbia, Croatia 28

South Continental EuropeHungary, Romania 14

SteppeRussia, Ukraine 13

AmericasUSA, Canada, Mexico, Argentina 30

Asia, Near East, AustraliaChina, Korea, Australia, Turkey 15 l

Types

Land races and old variety populations 14

Old and transitional varieties 64

Modern varieties 117

Germplasm accessions 26

Winter type 184 Spring type 37

Spelt 5durum wheat 10T. monococcum 5T. dicoccum 5Rye 10Barley 10Oats 5

Aims: identify variation determine heritability

Bran

45-50 % fibre

50% AX

Arabinoxylan (AX) is the major dietary fibre component in wheat grain

White flour

2-3% fibre

70% AX

The contents of soluble and total AX fibre vary widely in bran and white flour of 150 wheat lines

Data of Kurt Gebruers, Christophe Courtin and Jan Delcour (KU Leuven)

0.00

50.00

100.00

150.00

200.00

250.00

ALBATRO

S-…

BEZOSTAJA

-1ETO

ILE-D

E-…

HAN

AU

KRAIN

KABEG

RA

SADO

VO-1

ARIN

AZVEZD

ASU

321C

ARD

INAL

SAGITTAR

IOVO

NA

NAP

-HAL

MO

MTC

HIL

BALKANM

ILANBAN

KUTI

-1201FER

TOD

I-293

HER

EWAR

DTAM

ARO

POBED

ASC

OU

T66AKTEU

RM

IETIN

OM

ADE

APACH

EPR

OBSTD

O…

ALABASSKA…

AGR

ON

MAN

ITOBA

STEPHEN

STO

MM

IC

ATBIRD

SEU-SEU

N-27

DEKAN

VALOR

ISAM

ADEU

SG

ENE

LON

AC

F99105BAR

ANJKA

KARL

-92C

OU

RTO

TAU

TON

OM

IASU

NSTAR

BUC

K-…

SARATO

V-29

GR

ANBEL

PRO

DU

TTORE

ATLAS-66

LIBELLULA

CF99102

JUBILEJN

AJ…

TREM

IEBISC

AYKR

ASNO

DA

…ISEN

GR

AINC

HAR

AC

UBU

SKAN

ZLERTH

ATCH

ERG

ALAHAD

RIBAN

DALBAKU

KRI

BILANC

IAM

ALACC

ASU

LTAN95

KEYM

ANITAL

SAVAC

F99007C

HIN

ESE-…

THESEE

0.000.100.200.300.400.500.600.700.800.90

YU

MA

I-34

MIE

TI

CA

MP

AR

IC

F99007

MA

NIT

AL

AZ

TE

CA

67G

RA

NB

EL

CU

BU

SM

AG

DA

LEN

A-F

RM

OU

LINS

EU

-SE

UN

-27U

KR

AIN

KA

MILLE

NN

IUM

AV

ALO

NK

IRK

PIN

AR

-79C

AT

BIR

DA

MA

DE

US

SU

321C

F99102

BA

LKA

NM

ALA

CC

AC

HA

RA

SK

OR

OS

PE

LKA

-…T

OM

MI

BLE

-DE

S-D

OM

ES

PR

OB

ST

DO

RF

E…

CLA

IRE

RE

NA

NE

LLVIS

ES

TIC

AB

16JA

NZ

ALB

AT

RO

S-…

HE

RZ

OG

DE

KA

NA

RIN

AA

GR

ON

HE

RE

WA

RD

SC

OU

T66

ALB

AR

ED

-FIF

EB

LUE

/AG

AT

AY

-85S

AG

ITT

AR

IOB

LAS

CO

NS

-RA

NA

-1H

AN

AA

UG

US

TA

FU

ND

ULE

A-29

CA

MP

-RE

MY

AK

TE

UR

FLA

MU

RA

-85M

ON

OP

OL

GLO

RIA

JUB

ILEJN

AJA

-50S

PA

RT

AN

KA

AR

TH

UR

-71T

HA

TC

HE

RB

EG

RA

RA

VE

NN

AP

OB

ED

AC

AP

OB

ISC

AY

OB

RIY

OR

NIC

AR

FLE

ISC

HM

AN

N-…

FR

ED

RIC

KLO

NA

AU

TO

NO

MIA

GLE

NLE

AT

ALD

OR

QU

ALIT

AL

SA

VA

SO

ISS

ON

SC

HIN

ES

E-S

PR

ING

0

5

10

15

20

25

30

YUM

AI-34

CU

BUS

AMAD

EUS

SARATO

V-29

LYNX

REC

ITALTAM

200TO

MM

IR

ENAN

BISCAY

SEU-SEU

N-27

MO

MTC

HIL

GK

-TISZATAJVALO

RIS

DEKAN

ATAY-85

SPARK

SUN

STARM

V-SU

BALIBELLU

LAC

OU

RTO

TH

ANA

CAPH

OR

NM

ARIS

-…M

ANITAL

KRASN

OD

ARS

…C

F99007KLEIN

-…APAC

HE

CAR

MEN

AGR

ON

FUN

DU

LEA-29

BALKANB16SU

321JAN

ZG

ALAHAD

MALAC

CA

MEXIQ

UE

-50R

USALKA

THATC

HER

FLAMU

RA

-85KIR

KPINAR

-79JU

BILEJNAJA

-50AVALO

ND

ISPON

ENT

KOR

WETA

PASTOR

GLEN

LEAG

ENE

RIBAN

DPR

OD

UTTO

RE

MV

-PALOTAS

QU

ALITALR

ED-R

IVERAU

GU

STAC

LAIRE

SCO

UT66

NAP

-HAL

SOISSO

NS

RAVEN

NA

AUR

OR

AH

EREW

ARD

BUC

K-C

ATRIEL

MO

NO

POL

ALABASSKAJAKEYG

ERO

NIM

OC

HAR

APLAIN

SMAN

-VKAN

ZLERALBASTEPH

ENS

BARAN

JKABEG

RA

0.000.200.400.600.801.001.201.401.60

YU

MA

I-34

BILA

NC

IAM

ILANS

EU

-SE

UN

-27LY

NX

CF99007

LAS

TAC

ARM

EN

SU

321B

ALKA

NJU

BILEJN

AJA

…B

16R

IALT

OC

AMP

AR

IA

MA

DE

US

AT

LAS-66

GALA

HA

DM

EXIQ

UE

-50S

AG

ITT

ARIO

MO

MT

CH

ILR

ECIT

AL

MAN

ITA

LP

AN

AU

TO

NO

MIA

CH

INE

SE

-…K

EY

SA

RA

TOV

-29M

V-P

ALO

TAS

GR

ANB

EL

KR

AS

NO

DA

R…

SA

VA

FLA

MU

RA

-85K

AR

L-92

CF99102

REN

AN

KIR

AC66

RAV

EN

NA

HAN

AA

RIN

AG

ERE

K-79

BLA

SCO

CAD

EN

ZA

TH

ES

EEG

UAR

NI

MILLE

NN

IUM

VO

NA

ET

OILE

-DE

-…S

UN

STA

RK

UK

RI

BLU

E/A

GA

PA

CH

EK

AN

ZLE

RC

LAIR

EA

GR

ON

ALB

ATR

OS

-…JA

NZ

TO

MM

IC

HA

RA

SU

LTAN

95M

IET

IP

ALES

IOT

ALD

OR

KIR

KPIN

AR

-79S

PA

RTA

NK

AB

ISC

AY

GER

ON

IMO

DEK

AN

UKR

AIN

KAC

OU

RT

OT

AT

AY

-85R

US

ALK

AK

OR

WE

TAA

KT

EU

RC

AMP

-REM

YG

LEN

LEA

FLOUR TOTAL: 1.35-2.75%

SOLUBLE: 0.3-1.4%

BRAN TOTAL: 12.7-22.1%

SOLUBLE: 0.3-0.85%

Yumai 34

Yumai 34

% SOLUBLE: 2-5% % SOLUBLE: 20-50%

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

Yumai 34 Control1.00

1.50

2.00

2.50

3.00

Relative Viscosity

High fibre content (total and soluble)

High viscosity of aqueous extracts

Good breadmaking quality Healthgrain Diversity screen data from Gebruers et al (2008) J. Ag. Food Chem., 56, 9740-9749.

Yumai 34 Mean 151 lines

Yumai 34 Mean 26 lines

0

0.5

1

1.5

2

2.5

3

TOT-AXWE-AX

Yumai 34 Control

Exploiting natural variation in AX fibre:Yumai 34 x Ukrainka

0.000

0.500

1.000

1.500

2.000

2.500

UkrainkaYumai 34

TOT WE

Ukrainka Yumai 341.000

1.200

1.400

1.600

1.800

2.000

2.200

2.400Ukrainka Yumai 34

96 F6 lines + parents Fibre analysis Genotyping Transcriptome analysis

of bulk segregants

AX content

Relativeviscosity

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

New sources of variation: the Watkins collection

WE-AX

TOT-AX

Yumai 34

Yumai 34

Total and WE-AX in wholemeal flours of the Watkins lines

40%

0%

Yumai 34

WE-AX as % of TOT-AX

COLON

SMALL INTESTINE

Exploring the role of AX fibre in the human GI tract

prebiotic effects of AX: SCFAs, “good bacteria”

ferulate and vascular function/blood pressure

dough fermentation and IBS

effects of AX on digestion, viscosity and mass transfer

Conclusions- Priorities

• Develop N efficient wheat with good quality at low grain N

• New protein and non-protein sources of quality- greater stability?

• Improve health benefits

DF is an important target with established health benefits and high heritability

Thanks

GPDMalcolm Hawkesford Yongfang Wan Gemma Chope (Campden BRI)Simon Penson (Campden BRI)Ellen Mosleth Faergestad (Nofima/Oslo)

Millers and bakersBreeders

DIETARY FIBRERowan Mitchell Alison LovegroveMark WilkinsonTill PellnyJackie FreemanOndrej KosikZoltan Bedo (MTA)Mariann Rakszegi (MTA)

GRAIN LIPIDSRichard Haslam (RRes)Peter Wilde (IFR)Louise Salt (IFR)Irene Gonzalez-Thuillier Paola Tosi (Reading)Simon Penson (Campden BRI)

Peter Skeggs (Hovis)