Breeding resistant cultivars to reduce mycotoxin risks in oatsoats2016.org/files/present/1/4/3.pdfSt...

Breeding resistant cultivars to

reduce mycotoxin risks in oats

International Oat Conference

St Petersburg July 13, 2016

A collaborative research effort

Norwegian University of Life Sciences Choice of resistant cultivars 2

• Trond Buraas

• Lars Reitan

• Åsmund Bjørnstad

• Selamawit Tekle

• Helge Skinnes

• Morten Lillemo

FHB in Norway (1)

• Severe FHB epidemics in Norway since 2000

• Increased F. graminearum prevalence; major

DON producer in Norway

• Reduced germination in certified seeds 2006-

• Cultivars Gere, Nes and Lena (Graminor) and

Bessin (Norsaat) withdrawn from the market

DEPARTMENT OF PLANT SCIENCES 3

FHB in Norway (2)

• Oat cannot be used as feed for monogastric

animals in “FHB-years”

• For food 10% of the crop, but almost 1/3

imported every year due to FHB

• Due to FHB oat acreage should be reduced

from 30 to 20%, but a key rotation crop in a

specialized cereal production

• 22% in 2015, but no FHB!

•


Two types of trials

• Breeding trials:

–Designed by Graminor primarily to identify new

varieties that are resistant to FHB and DON

• Genetic trials

–To study the resistance genetics of FHB, germination

and DON in oats


Reliable field testing • Spawn inoculation

• Mist irrigation in the evenings

• Field data (Flowering time, height, disease...)

• Parameters: FHB, Mycotoxin (DON), germination, seed

infection


Parameter 1. FHB: Unreliable, confused

with maturation- abandoned

(Left cv. ‘Hurdal’, right ‘Z615’)

Parameter 2. Degree of fungal infection

left ’Belinda’, right ’615-4’: Less infected

Poor correlation with DON- abandoned

Parameter 3. Germination percentage

used in breeding - but a composite

problem:

• DON kills the seed at early infections

• Does not kill the healthy seed, but inhibits growth:


• Seedling blight:

Reversible by dehulling

and fungicides

Infection kills the seed.

Effects on the ultrastructure

10

Surface infections:

seedling blight

Parameter 4. Mycotoxin content:

used in breeding

• DON measured by GC-MS: direct and efficient

• Used since 2008

• NECESSARY to have strong infection levels

• DON-levels preferably between 3-30 ppm, but 1-10 may

be significant

• Need several environments (two locations since 2011)

• Need repeated checks from the best to the worst

• Progress is possible: Recent varieties have 40% less

DON


Norwegian University of Life Sciences Wheat breeding in Norway 13

Varietal differences in DON 2007-2015

LSD=2.0 ppm

5,6

8,1

7,3

6,3

8,0

5,9

8,0

6,0

9,7

7,1 6,6

7,1 7,3

7,0

10,1

7,0 6,6

7,1

7,8

10,2

6,0

8,6

9,5

7,7

12,7

12,1

12,9

11,1

8,8

0

2

4

6

8

10

12

14G

N0

91

11

Vål

er

Hu

rdal

GN

09

18

0

Aks

eli

GN

12

23

0

GN

08

20

7

Od

al

Be

lind

a

GN

12

15

0

Gim

se

Hu

rum

Skar

nes

Vin

ger

Hag

a

Rin

gsak

er

GN

12

14

3

GN

11

13

5

Do

vre

Sym

ph

on

y

Nu

dis

t

GN

12

14

2

NO

RD

09

/12

7

NO

RD

11

/12

8

Be

ssin

Nik

e

Po

seid

on

Ava

nti

Sco

rpio

n

DO

N (

pp

m)

Oats DON mean (2008-2015) LSD 5% = 2.0 (8 years vs 8 years) LSD 5% = 3.0 (2 years vs 8 years)


Varietal differences in germination 2007-

2015 (LSD = 4.6%)

81,4

79,1 78,1 78,0 78,0

77,0 76,1 76,0

74,4 74,4 74,1 73,8 72,9

72,4 72,3 72,1 71,6 71,6 71,4 70,3 69,9

69,0

65,8

64,0 63,7 63,6 63,4 63,3

60,6

50

55

60

65

70

75

80

85

GN

09

11

1

Vål

er

Hu

rdal

GN

09

18

0

Aks

eli

GN

12

23

0

GN

08

20

7

Od

al

Be

lind

a

GN

12

15

0

Gim

se

Hu

rum

Skar

ne

s

Vin

ger

Hag

a

Rin

gsak

er

GN

12

14

3

GN

11

13

5

Do

vre

Sym

ph

on

y

Nu

dis

t

GN

12

14

2

NO

RD

09

/12

7

NO

RD

11/

12

8

Be

ssin

Nik

e

Po

seid

on

Ava

nti

Sco

rpio

n

Ger

min

atio

n (

%)

Oats Germination (2007-2015) LSD 5% = 4.6 (8 years vs 8 years) LSD 5% =6.8 (2 years vs 8 years)


Relationship between DON and germination:

R2= 0.45, but outliers exist

[CELLEOMRÅDE]

[CELLEOMRÅDE] [CELLEOMRÅDE] [CELLEOMRÅDE] [CELLEOMRÅDE]

[CELLEOMRÅDE] [CELLEOMRÅDE] [CELLEOMRÅDE]

[CELLEOMRÅDE] [CELLEOMRÅDE] [CELLEOMRÅDE] [CELLEOMRÅDE] [CELLEOMRÅDE]

[CELLEOMRÅDE] [CELLEOMRÅDE] [CELLEOMRÅDE] [CELLEOMRÅDE] [CELLEOMRÅDE] [CELLEOMRÅDE]

[CELLEOMRÅDE] [CELLEOMRÅDE] [CELLEOMRÅDE]

[CELLEOMRÅDE]

[CELLEOMRÅDE] [CELLEOMRÅDE] [CELLEOMRÅDE] [CELLEOMRÅDE] [CELLEOMRÅDE]

[CELLEOMRÅDE]

R² = 0,4533

58

60

62

64

66

68

70

72

74

76

78

80

82

5 6 7 8 9 10 11 12 13 14

Ger

mn

inat

ion

(%

)

DON (ppm)

Germination vs DON 2007-2015

Part II. NIR calibration of DON and

germination: useful in breeding?


10090807060504030

100

90

80

70

60

50

40

30

Actual Germination %

Pre

dic

ted

Ge

rmin

ati

on

%

Prediction using SNV and De-trended spectra

Correlation between Predicted and Actual = 0.85 (P = 0,000).

NIR calibration and prediction also possible for

germination


403020100

90

80

70

60

50

40

30

DON (ppm)

Ge

rmin

ati

on

(%

)

Germination vs DON (GC/MS)

2015105

90

80

70

60

50

40

30

DON (ppm)

Ge

rmin

ati

on

(%

)

Germination vs DON (NIR)

2015:

Actual DON values and

germination: -0.67 (P=0.000)

Predicted DON values and

germination was -0.43

(P=0.000).

Limitation: to include enough environments in the

calibration


Green and red= Staur

Blue and grey= Vollebekk

Year and location effects dominate spectral signals

Part III. Genetic studies of resistance

to DON and FHB

• QTL mapping populations

– Hurdal x Z595-7(184 lines) and Hurdal x Z615-4 (91 lines)

– 3 years field testing

– QTL mapping

• CORE spring oats from North America and Europe

– 2 years field testing, N=432

– GWAS

• Anther extrusion in relation to infection


He, et al. , 2013

Bjørnstad et al, submitted


Example 6

GMI_ES01_c25270_71oPt-17300

0

P32M66-80GMI_ES01_c17183_318

1

oPt-17991oPt-7733oPt-17172

2

GMI_ES17_c20752_1084GMI_ES17_c2826_360GMI_ES15_c5289_317GMI_ES17_c15844_381oPt-16301oPt-10253oPt-10427oPt-7687oPt-1879oPt-17156oPt-6375oPt-8654oPt-17489

3

oPt-9631oPt-13459oPt-15501GMI_ES02_c11747_563GMI_ES01_c8777_202P32M66-114

4

oPt-9004GMI_ES17_c18122_180

5

P31M72-191GMI_ES17_c1315_660

6

DO

N0

8

DO

N0

9

DO

N1

0

0

5

10

Chr_17A

DON08

DON09

DON10

FHB

PH

DH

DM

• Qdon.umb-17A/7C

– Detected in all experimental years

– independent of phenology

– Verified in HZ615 population

• Other QTL were found on 5C, 9D, 13A,

and 14D

– <10% of DON variation

– Not in all years

– QTL on 9D and 13A co-localized

with QTLs for agronomic traits

Hurdal x Z595-7: a strong QTL reduces

DON by up to 25%

He, et al. , 2013

The Spring CORE panel

• 418 spring oats tested and analyzed for DON in 2011 and

2012

• SNPs using the 6k chip and GBS, 2974 polymorphic

markers

• Analyzed for structure and trait relationships


1. Possible to score FHB if the variation is wide

(bad) enough

DEP. OF PLANT- AND ENVIRONMENTAL SCIENCES 23

2. Relationship between DON and FHB: Large

variation - Midwest oats susceptible

5

10

15

20

25

30

35

2 7 12 17 22 27 32

Mea

n D

ON

co

nte

nt

(pp

m)

Mean FHB (AUDPC)

Canada

Midwest

Nordic

US_other

Check

Horizon270

Leggett



Does less FHB and DON mean late and

tall?


DTF DTM PH

FHB_M -0.81*** -0.62*** -0.52***

DON_M -0.28*** -0.14* -0.24***


Klos et al 2016: Many QTLs for DTF coincide with FHB and DON

But low DON need not mean “late

and tall”

• Mrg11/1C for FHB

–The previously identified Qdon-umb-17A/7C

• The CORE correlations reflect the very susceptible, tall

and daylength sensitive Midwest lines

• They do not represent the “truth” about genetics of DON

resistance in oats

• Progress in Norwegian oats despite limited variation in

earliness and height!

• Resistant cv. Leggett is early and not so tall



Anther extrusion (AE): a way to avoid FHB

in oats, like in wheat and barley?

Wheat Barley

• Wheat


Variation of AE exists, but high AE rare in modern

varieties

cv. Typhon: Closed cv. Norum: Wide open

Transgressive segregation for AE and as

heritable as days to flowering

Stormogul x Fia

Stormogul x Svea

29

P2 P1 P2 P1

Wald n.d.f F d.d.f P SE

Genotype (g) 3179.84 152 20.92 18 <0.001 0.63

Year (y) 0.24 1 0.20 18 0.658 -

g×y 719.59 147 4.90 18 <0.001 0.89

Sources of error: drought, - and anthers

remaining inside

Anthers may remain

inside, even at high AE!

Beware of drought when

scoring AE

• Sensitive to drought

(and cloudy weather)

Does AE protect agains Fusarium? (1) Yes. In the

field with spawn inoculation

The higher the AE, the less FHB and DON


FXS DTF Height FHB AE*

Height ns

FHB -0.54 (P=0.00) -0.48 (P=0.00)

AE 0.33 (P=0.05) 0.53 (P=0.00) -0.62 (P=0.00)

DON -0.36 (P=0.02) -0.34 (P=0.03) 0.64 (P=0.00) -0.36 (P=0.03)

*16 RILs from each cross spawn inoculated in field 2015

Anther extrusion data was scored in previous greenhouse experiment. ns-non

significant.

Does AE protect agains Fusarium? (2)

No. The higher the AE, the more FHB and

DON in the greenhouse: spray inoculated

DTF FHB DON

FHB ns

DON ns 0.848 (P=0.000)

AE ns 0.353 (P=0.022) ns

Conclusions

• Significant breeding progress possible in oats if testing

is accurate and adequate

• NIR may be possible to supplement or replace expensive

measurements

• Anther extrusion may have a role in infection, but

confused with open flowering.

• Anther retention more accurate, but laborious to score

• Wide open flowers very susceptible to spray


Thank you

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