Breeding resistant cultivars to reduce mycotoxin risks in oats. International Oat Conference St Petersburg July 13, 2016
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1 Breeding resistant cultivars to reduce mycotoxin risks in oats International Oat Conference St Petersburg July 13, 2016
2 A collaborative research effort Trond Buraas Lars Reitan Åsmund Bjørnstad Selamawit Tekle Helge Skinnes Morten Lillemo Choice of resistant cultivars Norwegian University of Life Sciences 2
3 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producer in Norway Reduced germination in certified seeds Cultivars Gere, Nes and Lena (Graminor) and Bessin (Norsaat) withdrawn from the market DEPARTMENT OF PLANT SCIENCES 3
4 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! DEPARTMENT OF PLANT SCIENCES 4
5 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 Choice of resistant cultivars Norwegian University of Life Sciences 5
6 Reliable field testing Spawn inoculation Mist irrigation in the evenings Field data (Flowering time, height, disease...) Parameters: FHB, Mycotoxin (DON), germination, seed infection Choice of resistant cultivars Norwegian University of Life Sciences 6
7 Parameter 1. FHB: Unreliable, confused with maturation- abandoned (Left cv. Hurdal, right Z615 )
8 Parameter 2. Degree of fungal infection left Belinda, right : Less infected Poor correlation with DON- abandoned
9 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 Choice of resistant cultivars Norwegian University of Life Sciences 9
10 Infection kills the seed. Effects on the ultrastructure 10
11 Surface infections: seedling blight
12 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 DEPARTMENT OF PLANT SCIENCES 12
13 GN09111 Våler Hurdal GN09180 Akseli GN12230 GN08207 Odal Belinda GN12150 Gimse Hurum Skarnes Vinger Haga ingsaker GN12143 GN11135 Dovre ymphony Nudist GN12142 D 09/127 D 11/128 Bessin Nike Poseidon Avanti Scorpion DON (ppm) Varietal differences in DON LSD=2.0 ppm Oats DON mean ( ) LSD 5% = 2.0 (8 years vs 8 years) LSD 5% = 3.0 (2 years vs 8 years) 12,7 12,1 12,9 11,1 10 9,7 10,1 10,2 9, ,6 8,1 7,3 6,3 8,0 5,9 8,0 6,0 7,1 6,6 7,1 7,3 7,0 7,0 6,6 7,1 7,8 6,0 8,6 7,7 8, Wheat breeding in Norway Norwegian University of Life Sciences 13
14 GN09111 Våler Hurdal GN09180 Akseli GN12230 GN08207 Odal Belinda GN12150 Gimse Hurum Skarnes Vinger Haga Ringsaker GN12143 GN11135 Dovre Symphony Nudist GN12142 NORD 09/127 NORD 11/128 Bessin Nike Poseidon Avanti Scorpion Germination (%) Varietal differences in germination (LSD = 4.6%) ,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 Oats Germination ( ) LSD 5% = 4.6 (8 years vs 8 years) LSD 5% =6.8 (2 years vs 8 years) 65,8 64,0 63,7 63,6 63,4 63, , Wheat breeding in Norway Norwegian University of Life Sciences 14
15 Germnination (%) 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] Germination vs DON [CELLEOMRÅDE] [CELLEOMRÅDE] [CELLEOMRÅDE] [CELLEOMRÅDE] R² = 0,4533 [CELLEOMRÅDE] [CELLEOMRÅDE] [CELLEOMRÅDE] [CELLEOMRÅDE] DON 9 (ppm) Wheat breeding in Norway Norwegian University of Life Sciences 15
16 Part II. NIR calibration of DON and germination: useful in breeding?
17 NIR calibration and prediction also possible for germination 100 Prediction using SNV and De-trended spectra 90 Predicted Germination % Actual Germination % Correlation between Predicted and Actual = 0.85 (P = 0,000). DEPARTMENT OF PLANT SCIENCES 17
18 90 Germination vs DON (GC/MS) Germination (%) : Actual DON values and germination: (P=0.000) DON (ppm) Germination (%) Germination vs DON (NIR) Predicted DON values and germination was (P=0.000) DON (ppm) DEPARTMENT OF PLANT SCIENCES 18
19 Limitation: to include enough environments in the calibration Green and red= Staur Blue and grey= Vollebekk Year and location effects dominate spectral signals DEPARTMENT OF PLANT SCIENCES 19
20 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 DEPARTMENT OF PLANT SCIENCES 20
21 DON10 DON09 DON08 Hurdal x Z595-7: a strong QTL reduces DON by up to 25% Chr_17A 0 1 GMI_ES01_c25270_71 opt P32M66-80 GMI_ES01_c17183_318 Example 6 Qdon.umb-17A/7C Detected in all experimental years independent of phenology Verified in HZ615 population opt opt-7733 opt GMI_ES17_c20752_1084 GMI_ES17_c2826_360 GMI_ES15_c5289_317 GMI_ES17_c15844_381 opt opt opt opt-7687 opt-1879 opt opt-6375 opt-8654 opt opt-9631 opt opt GMI_ES02_c11747_563 GMI_ES01_c8777_202 P32M opt-9004 GMI_ES17_c18122_180 P31M GMI_ES17_c1315_660 DON08 DON09 DON10 FHB PH DH DM 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 He, et al., 2013 DEPARTMENT OF PLANT SCIENCES 21
22 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 DEPARTMENT OF PLANT SCIENCES 22
23 1. Possible to score FHB if the variation is wide (bad) enough DEP. OF PLANT- AND ENVIRONMENTAL SCIENCES 23
24 Mean DON content (ppm) 2. Relationship between DON and FHB: Large variation - Midwest oats susceptible 35 Canada Midwest 30 Nordic US_other Check Horizon Leggett Mean FHB (AUDPC) Bjørnstad et al, submitted DEPARTMENT OF PLANT SCIENCES 24
25 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 Bjørnstad et al, submitted DEPARTMENT OF PLANT SCIENCES 25
26 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 Bjørnstad et al, submitted DEPARTMENT OF PLANT SCIENCES 26
27 Anther extrusion (AE): a way to avoid FHB in oats, like in wheat and barley? Wheat Barley Wheat DEPARTMENT OF PLANT SCIENCES 27
28 Variation of AE exists, but high AE rare in modern varieties cv. Typhon: Closed cv. Norum: Wide open
29 Transgressive segregation for AE and as heritable as days to flowering Stormogul x Fia Stormogul x Svea P2 P1 P2 P1 Wald n.d.f F d.d.f P SE Genotype (g) < Year (y) g y <
30 Sources of error: drought, - and anthers remaining inside Anthers may remain inside, even at high AE! Sensitive to drought (and cloudy weather) Beware of drought when scoring AE
31 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 (P=0.00) (P=0.00) AE 0.33 (P=0.05) 0.53 (P=0.00) (P=0.00) DON (P=0.02) (P=0.03) 0.64 (P=0.00) (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. DEPARTMENT OF PLANT SCIENCES 31
32 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 (P=0.000) AE ns (P=0.022) ns
33 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 DEPARTMENT OF PLANT SCIENCES 33
34 Thank you
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