International collaboration to improve wheat quality. Tatsuya M. Ikeda NARO Western Agricultural Research Center, Fukuyama, Japan

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1 International collaboration to improve wheat quality Tatsuya M. Ikeda NARO Western Agricultural Research Center, Fukuyama, Japan

2 Wheat quality Cultivars Seed shape Flour N content Product Taste N content Milling Ash content, color Processing Flavor, color Grain hardness Gluten properties Texture Vitreousness Ash content Fusarium toxin Starch properties Amylase activity Staling Nutrient, Health Safety Agricultural products Standardized products Commercial products 2

3 Analysis of gluten proteins kda High molecular weight glutenin subunits (HMW-GS) ω-gliadin Low molecular weight glutenin subunits LMW-GS), α/β/γ-gliadin

4 Classification of Glu-1 alleles 2 1 2* 7 Glu-A null a b c a b c d e f g h i j k 22 Glu-B Glu-D a b c d e f 12 Payne & Lawrence 1983

5 Classification of Glu-3 alleles 1. Bungulla, 2. Gabo, 3. Insignia, 4. Kharkof, 5. Orca, 6. Rescue, 7. Grandin, 8. Hope, 9. Thatcher, 10. Timstein, 11. Arrino, 12. Cadoux, 13. Corrigin, 14. Cranbrook. 15. Eradu. 16. Chinese Spring Glu-A3 Glu-B3 Glu-D3 by Gupta and Shepherd (1990) based on 2-step SDS-PAGE

6 Identifying Glutenin subunits by 2DE and MS We still need to optimize these conditions.

7 No one method can identify all alleles

8 Effects of allelic variation of glutenin subunits SS bond subunit Increasing glutenin polymer (Glu-B1al, Glu-A3d, Glu-B3g etc) Tightening glutenin polymer (Glu-D1d) Decreasing glutenin polymer (Glu-A1c, Glu-A3e, Glu-B3j etc) 8

9 Effect of subunit 5 (Glu-D1d) on glutenin polymer Without subunit 5 With subunit mm 0.04mm

10 Subunit 5 (Glu-D1d) improves bread volume Without subunit 5 With subunit 5

11 Glutenin subunits and gluten strength Strong Glu-D1d Glu-B1al Strong ExtraStrong Glu-1 (HMW-GS) Medium Medium Glu-B1e Glu-A1a Glu-A1c Weak Weak Glu-A3e/f Glu-B3j Glu-B3ae Glu-3 (LMW-GS) Strong Glu-B3g Glu-A3d Glu-B3b Ikeda et al, Cereal Chem 2014

12 Glutenin subunit composition Cultivar Country Glu-A1 Glu-B1 Glu-D1 Glu-A3 Glu-B3 Glu-D3 Abbot UK a d a e g d Hereward UK c u b f g l Shango UK a b a c ab a Apache France b b b d g c Arezzo France c c d f g c Soissons France b b d d b c Altos Germany c i d c g c Aspirant Germany a a d d g l Bussard Germany a c d c g l Cardos Germany c c d c g c Dekan Germany c d d c g c Flair Germany c d d e g c Ritmo Germany a d b f g l Zentos Germany c c d f g b Karl USA (HRW) a u d c b c Palo Duro USA (HRW) b u d f g a Jagalene USA (HRW) b i d c g l Glenlea Canada (ES) b al d g g c Katepwa Canada (HRS) b c d e h c AC Barrie Canada (HRS) b al d e h c Alsen USA (HRS) b c d d h a Thatcher USA (HRS) b c d e h e Parshall USA (HRS) b c d d h a

13 Glu-D1d Glu-B1al Glu-1 (HMW-GS) Glutenin subunits and gluten strength Glu-B1e Glu-A1a Glu-A1c Strong Weak US soft Club Glu-A3e/f Glu-B3j Glu-B3ae US/Canadian hard red spring Glu-3 (LMW-GS) Australian noodle/bread Glu-A3d US hard red winter European hard red winter Japanese noodle Glu-B3g Glu-B3b Strong Ikeda et al, Cereal Chem 2014

14 Grain hardness and puroindoline Soft wheat Hard wheat A-starch B-starch 6μm Protein matrix Starch Starch binding : PIN-a : PIN-b Wild Pina null Hardness: Pina null > Pinb point mutation > wild type

15 Damaged starch generated during milling of hard wheat Cracked starch Worn starch Damaged starch amylase maltose maltase sucrose fermentation CO2 ethanol Yeast

16 Amylose content and usages More Amylose content Less 30% Normal 27% Partial waxy 23% Partial waxy 0% Waxy Wild Wx-A1null Wx-B1null Wx-A1/Wx-B1 double null Wx-A1/Wx-B1/ Wx-D1 all null Less Springiness More Pullman-type bread White salted noodle Rice European bread Sponge cake

17 Classification based on starch quality More damaged starch Pina-D1b Hard Hard/normal Hard/partial waxy Hard/partial waxy Hard/waxy Pinb-D1b Pinb-D1d Soft Soft/normal Soft/partial waxy Soft/partial waxy Soft/waxy Wx-B1b Wx-A1b/Wx-B1b All null Springi -ness

18 More damaged starch Classification of wheat class based on starch properties Pina-D1b Hard Pinb-D1b Pinb-D1d US/Canadian hard red spring European & US/Canadian hard red winter US soft Japanese hard red spring/winter Australian noodle Japanese waxy Soft Japanese noodle Wx-B1b Wx-A1b/Wx-B1b All null Springi -ness

19 Expert Working Group Improving Wheat Quality for Processing and Health

20 7 subgroups works together Genetic resources & gene nomenclature Resources Seed components Nutrients Seed proteins Carbohydrates Allergy Functionality Processing Food safety Improving Wheat Quality for Processing and Health

21 EWG for Improving wheat quality for processing and health Subgroups Organizers Memeber Seed proteins Tatsuya M. Ikeda (Japan) 27 Allergy Angela Juhasz (Hungary) 9 Carbohydrates Ravindra Chibbar (Canada) 5 Nutrition Peter Shewry (UK) 13 Processing Valérie Lullien-Pellerin (France) 4 Food safety Sofia Chulze (Argentina) 3 Genetic resources and gene nomenclature 7 subgroups, total 63 members. John W. Rogers (Argentina) & Carlos Guzmán (CIMMYT) 7

22 Seed protein analysis Other proteins Dough viscoelasticity Glutenins & Gliadins Puroindolins Polyphenol oxidases Noodle color Grain hardness Unifying nomenclature Selecting standard methods Sharing valuable cultivars

23 Unify nomenclature with durum Glu/Gli alleles kda Glu-A3 Glu-B3 Glu-B2 Gli-B1 Alaga (m) new (y) ad (b) b (44) Andalucia 344 (-) new2 (-) new (b) b (null) Ardente d* (w) d (a) a (45) Blancal de Nules (-)? (aa) h (-) b (-) Clarofino (o) d d (a) a (45) Cocorit (l)? d (a) a (45) Fanfarrón (q) d c? (-) b n.a. Granja Badajoz d* (v) i (a) a (45) Jiloca (p) e (t) d (b) b (45) Langdon (k) new2 (s) c? (b) b (42) Mexicali (j)? (r) d (a) a (45) Mourisco Fino (i) d* i (-) b (-) Mundial (n) d* (u) d (a) a (45) Mexicali, 2. Langdon, 3. Jiloca, 4. Mundial, 5. Granja Badajoz 6. Ardente, 7. Fanfarron, 8. Clarofino, 9. Alga, 10. Andalcia Blancal de Nules, 12. Cocorit, 13. Mourisco Fino Ikeda et al. unpublished

24 Wheat allergy analysis Genetic effects Transcriptome Environmental effects Human factors Proteome Effect of processing Seed allergome Understanding wheat allergens and human health Breeding for special needs

25 Not only the Gluten allergy Non-Celiac Wheat Sensitivity (NCWS, NCGS) FODMAPs

26 Starch % Polymeric protein % Providing rigorous information to consumers NO FRANKE-WHEAT! Chibber et al. Old cultivars 1860~ Modren cultivars

27 Franken-wheat Modern wheat has new gluten, citing a wrong paper! 1~5.Progenies of somatic hybrid, 6.Wheat-related wild species (Agropyron elongatum),7. Chinese bread wheat Xin, Gao et al. (2009). High frequency of HMW-GS sequence variation through somatic hybridization between Agropyron elongatum and common wheat. Planta, 231, doi: /s

28 Addressing wheat/gluten avoidance and consumers wellbeing Collaboration with Global Research and Communication Project is now Ready to start!

29 Fusarium toxins analysis for food safety Biodiversity of toxigenic species Impact of climate change Risk Maps at a global level Effects of processing on toxin distribution MYCOTOXINS Strategies to reduce their impact Masked mycotoxins Monitoring human and animal exposure BIOMARKERS BREEDING for resistance against fungal infection and toxin accumulation- Transgenic cultivars? Plant-pathogen interaction METABOLOME BIOCONTROL as part of a IPM (Integrated Pest Management)

30 Improving wheat nutrients Minerals Mg/Se/Zn/Fe/Cd Dietary fiber Arabinoxylan, β-glucan Resistant starch Analysis of mapping populations to identify QTLs Determine variation in content and composition Multisite trials to select Master sets of varieties exhibiting wide diversity Establish molecular marker systems for major QTLs Deposit the Master sets to public germplasm banks Determine heritability of major groups of bioactive components

31 Collaboration with Genomicists for connecting gene annotation and the gene catalog Whole genome sequence data Genomics Gene annotation CDS, predicted CDS, non-cds Genetics Gene Catalog Gene class Gene symbol Allele Breeding New cultivars

32 Collaboration with Genomicists for unifying gene nomenclature among Triticeae Triticum & Aegilops Hordeum Hordeum vulgare Triticum urartu Gxx-A1 Aegilops tauchii Gxx-D1 Gxx-H1 Gxx-A1 Gxx-B1 Gxx-A1 Gxx-B1 Gxx-D1 Triticum durum Triticum aestivum

33 Development of master sets for quality analyses Identification of diversity described for quality traits (alleles, genes, accessions). Collection of accessions representing the variability Diversity evaluated and confirmed Update the Catalogue of Gene Symbols. multiplication of the master sets of seed Sharing seeds in public Germplasm Banks

34 Selection of standard core cultivar sets for Glu-3

35 Master set of glutenins Go to CIMMYT Germplasm Bank web: Search for: %GluStd Advantages of Master Sets: Different groups working with the same materials/accessions Promote the use of more genetic diversity

36 Sharing the master sets among germplasm banks International Genebank e.g. CIMMYT Public National Germplasm bank Public Natinal Germplasm bank Public National Germplasm bank Sharing identical materials Classified master sets based on breeding purposes (glutenin, gliadins, puroindolines, Waxy, PPOs, etc)

37 Genetic resources & gene nomenclature Selecting and distributing Master sets. Refining the Gene Catalog. Supporting gene annotation. Nutrients Seed proteins Carbohydrates Allergy Processing Food safety Sustainability context Higher yield Withstanding climate change Providing rigorous information Sharing methods, materials & databases. Developing innovative strategies with other groups. Good for human health Meeting demands of food industries and consumers

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