Application of molecular markers in cereals breeding

Dr. Viktor Korzun – KWS LOCHOW GMBH

2

Worldwide development of food supply

0 2 4 6 8 10

2050

2025

2000

1975

1950

World population(in Mrd)

0 0,3 0,6 0,9 1,2 1,5

2050

2025

2000

1975

1950

Cultivated area(in Mrd ha)

Arable land per-capita

( in ha)

Ressource: UNO, 2007.

0 0,2 0,4 0,6

2050

2025

2000

1975

1950

3

Possible solution

Breeding of high yielding and resistance to abiotic

and biotic stress cultivars will play a key role

4

Agronomic traits� Yield StabilityQuality

� Nutrition

� Processing

Resistance� Diseases

Yield� Grain yield

Nutrient use efficiency

� Nitrogen

Conventional Plant Breeding

5

Conventional Plant Breeding

GENETIC RESOURCES

Evaluation of plants

Crossing

Evaluation

Selection

Official testing

Variety

years, places,

replications

resistances,

yield, quality

2 to 4 years

6 to >8 years

8 to >12 years, 5 to 6 mn €, release rate < 10%

V a

r i a

t i o

nS e

l e

c t

i o

n

6

State of the art in plant breeding technologies

Breeding speed-up and add-on technologies

Yield improvement

Gene technology

DNA diagnostics

Cell- and tissue culture

Hybrid breeding

Cross breeding and selection

Genomic research

1856 1910 1940 1970 Today

7

Advantages of molecular markers:

• environment independent detection of trait(s)

• application in early generations

• possibility for HT automatisation ☺☺☺☺

����

Molecular markers

☺☺☺☺

Disadvantages of molecular markers:

� relatively high cost of application

� limited application on quantitative trait(s)

RYE

9

Pre-basic seed (1st year)

Basic seed( 2nd year)

Certified seed

(3rd year)

Hybrid rye,commercially grown (4th year)

A line B line Restorer synthetic C

general formula of a hybrid rye variety: (A-cms x B-N) x R-Syn

Seed Production of Hybrid Rye

10

Ergot

Germinating sclerotia

Ergot intoxification in the middle

ages (fire of St. Anton)

Altar of Isenheim (Alsassia, France)

Pollen grains

germinating on

pistils

Fertility restoration in rye

11

4RS26.4

0.9

1.8

2.7

0.1

0.1

1.7

21.4

11.3

19.6

0.1

10.2

4.5

6.6

1.4

10.9

11.1

4.1

5.1

3.3

Xmwg77

Xscb230Xmwg2299

Xiag120

Xpsr150

Xscb258a

Xpsr584Xiag115

Xmwg89

Px(en)

Xscb52Xmwg530

Xmwg539

Xpsr119

Xpsr392

Xscb21

Xcmwg652

Xmwg2053b

Xpsr899

Xmwg2202

Xpsr167

4RL

Xpsr8

S4R09

AFLP1555

RfS4R04

AFLP1455

PSR167

4RL

PSR8

STSR022

STS4R014

SNP4R015

SNP4R028

Fertility restoration in rye

12

Marker analysis

Fertility restoration in rye

13

„old“ assay

„modern“ assay

Fertility restoration in rye

14

B = 64,5 % RP

A = 87,5 % RP

BC

1 =

75

% R

P

BC

2 =

87

,5%

RP

15

Hybrid Rye with PollenPlus

increased pollen shedding

= less ergot

tolerant of stress

medium hybrid

rye

medium

conventional rye

PollenPlus

hybrid rye

high · ergot infection level · low

An additional information about POLLEN+ strategy see on www. pollenplus.de

Fertility restoration in rye

WHEAT

17

Foto: Prof. Ruckenbauer (IFA, Tulln)

�Winter wheat is most prominent in Germany, followed by barley, rye and triticale

�Complex of Fusarium species: Fusarium

graminearum, F. culmorum, F. avenaceum, F. poae

�Main focus on deoxynivalenol (DON), but nivalenol (NIV) occurs also

�Epidemics do not occur regularly (1998 and 2002 in Germany), however, mycotoxins can be found every year

�Mycotoxin contamination is of greater concern than yield loss (food safety)

Fusarium Head Blight (FHB) in cereals

18

Commission Regulation (EC) No 856/2005

DON in unprocessed cereals: 1250 µg kg-1

DON in bread and bakeries: 500 µg kg-1

DON in baby food: 200 µg kg-1

Durum products

Cereals for human consumption

except Bread, pastries, beer

Baby food

-

500

350

100

Germany1

1 Since February 2004 valid

Mycotoxins in cereals

19

1

2

3

4

5

6

1986 1988 1990 1992 1994 1996 1998 2000 2002 2004

Year

Note 1-9, (1 is full resistant)

Powdery

mildew

FHB

Resistance to

BSL 1986-2005

Selection success by resistances breeding

20

Molecular breeding Geneticengineering

HOH/LP/LfLLfL/HOH/LPLfL/IFA PLT/MPI/LP

Phenotypic

selection

Marker-

based

selection

Recurrent selection

QTL-

Mapping

QTL-

Validation

Marker development

Strategies towards improving resistance to Fusarium Head Blight

21

Benefit of exotic resistance sources

0

10

20

30

40

50

60

Sumai

3

Arin

aPet

rus

Dre

am

His

tory

G16

_92

Spark

Ren

an

Arc

heA

pach

eF20

1_R

Bra

ndo

Cha

rger

G

CH

GB

C

10 environments, 2003-2004% FHB rating

LSD5%

FR

1. Resistance level of Sumai 3 is much higher

2. Low number of QTL with high effects that are less

dependent on environment

22

CM 82036

(CHINA)

Munk

(D)

Frontana

(BRAS)

G16-92 Brando Dream LP 235.1

Spring wheat Winter wheat

FHB resistance of parents

23

(ER • F) x (GR • H)

S+MBS

CM

Phenotyp. Sel. Marker-based Sel.

C0

B. Winter wheat

S0

Recomb.

xS

C1

Test 1

MBS

(AR • B) x (CR • D)

S0

Test 1 xS

Recomb.

2000

2001

Year

Test 2

S0

Test 1‘ xS

CP1 CP1‘

unsel. sel.

2003

2004

2005

Phenotyp.Sel.

MBS

S+MBS

CM

S+MBS

Marker-based Sel.

2002

C0

A. Spring wheat

Data analysis, use of selected lines in breeding programmes

Phenotypic vs. marker-based selection

24

0

5

10

15

20

25

30

35

40

45

3B+5

A+3

A

3B+5

A

3B+3

A

3A+5

A 3B 5A 3AS

usce

pt.

a

e

dcd

bcbcb

a

DON content [mg/kg]

Donor-QTL alleles

-80%

Effect of exotic QTL on DON content in spring wheat

25

Variant

N Total Per year

Phenotypic

selection

1000 6.2 (2) 3.1

Marker-based

selection CM 35 3.3 (1) 3.326 8.8 (1) 8.8

40 9.6 (2) 4.8

N Total Per year

Spring wheat Winter wheat

135 12.6 (3) 4.2

CP1

CP1‘

FHB rating (%)

EUREKA – 2004, 4 locations

Mean realised selection gain

26

GABI-Canada

Canadian-German Collaboration on Plant Genomics Research

Reducing Fusarium Toxins in Wheat Through Genomics - Guided Strategies

Duration: 4 Years 1.04.2006 - 31.03.2010

Project coordination (Canada): Project coordination (Germany):Dr. Daryl Somers /Christof Rampitsch scientific: Prof. Dr. Gerhard Wenzel / Dr. M. Schmolke (TUM)

industrial: Dr. E. Ebmeyer (KWS-L)

Cooperating Partners in Germany:

University of Hohenheim (UHOH)Bavarian State Research Center for Agriculture (LfL)Technical University of Munich (TUM)Federal Centre for Breeding Research on Cultivated Plants (JKI)Institute of Plant Genetics and Crop Plant Research (IPK)

KWS LOCHOW GMBHSaaten-Union Resistenzlabor GmbH

27

GABI Canada : German subprojets

28

FHB resistance from exotic sources

29

FHB resistance tests – spray inoculation

30

1. Marker based transfer of the exotic resistance QTL 3B und 5A into elitewinter wheat

2. Linkage drag: Evaluation of possible negative effects of the exoticresistance loci on yield and other characters

3. Dissection of QTL: Diminution of the exotic chromosome segment byretaining the FHB resistance

GABI Canada: Aims of our part (module 2) of the project

31

Development of material for Dissection and Linkage Drag

Generation Sowing Marker Analysis Dissection (Opus) Linkage Drag (Opus/Anthus)

Nov. 2002

F1 x Opus/Anthus Aug. 2003 1.940 GT

BC1 x Opus/Anthus Feb. 2004 2.500 GT

BC2 x Opus/Anthus Oct. 2004 3.300 GT

BC3 F1 July 2005 3.600 GT Selection: Bbaa, bbAa Selection: aaBb, bbAa, AaBb

Jan. 2006BC3 F2 3.076 GT Selection: Bbaa, bbAa Selection: aabb, aaBB, bbAA, AABB

Aug. 2006BC3 F3 7.700 GT B_aa: 402 GT with recomb. BC3 F2:3

(3.000 GT) bbA_: 300 GT with recomb.

Jan. 2007BC3 F4 BC3 F2:4

BC3 F4:5 BC3 F2:5Yield and resistance trial 2008

T=180, P=5, R=2

Yield and resistance trial 2009

3B: 165 GT, 5A: 183 GT T=180, P=5, R=2

GT: Genotypes Resistance Donor: SW1-91 (CM 82036 x Nandu) x (Frontana x Munk)

Marker assisted background selection

Propagation 2008

FHB resistance Trial 2009 (P=4)

Homozygous sub-NILs

Opus x Donor Anthus x Donor

Marker assisted background selection

Marker assisted background selection

32

Exotic resistance loci in elite winter wheatSusceptibility for FHB in different marker classes

0

5

10

15

20

25

30

35

40

45

50

AABB AAbb aaBB aabb Opus AABB AAbb aaBB aabb Anthus

c

a

b

d

cbb

a

FHB rating (%)

33

TraitGenetics GmbH

Syngenta Seeds GmbH

KWS LOCHOW GmbH

Leibniz Institute of Plant Genetics and Crop Plant Research (IPK)

GABI-Wheat: Establishment of a platform for genome-wide association mapping in wheat

Poster ITMI2009_036

BARLEY

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Malting Quality

Malting quality

36

--ym4/ym5ym4/ym5

++ ym4/ym5ym4/ym5

Yield

+ ym4/ym5- ym4/ym5

Virus resistance in barley

37

TriGen COST action

ITMI

Magic triangle

Thank you for your attention!