Two tightly-linked genes controlling grain length underlie a major grain weight QTL in polyploid wheat

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1 Two tightly-linked genes controlling grain length underlie a major grain weight QTL in polyploid wheat Jemima

2 Final grain yield is an important, but complex trait Number of spikes Number of grains Grain weight

3 Dissecting Grain Size: Grain Length and Grain Width Grain weight Grain Size Grain Filling Grain Length Grain Width

4 A QTL Reducing for Thousand complexity Grain using Weight Near (TGW) Isogenic on chromosome Lines (NILs) 5A Doubled Haploid population Charger x Badger Number of SNPs Inner tracks SNP variation (iselect 90k chip) 1. Parental SNPs 2. BC 4 NILs SNPs Outer track Combined LOD score (12 environments) Brinton et al. biorxiv 2017

5 TGW (g) 5A NILs have increased TGW 5A- NILs 5A NILs *** * *** 9% *** 7% 7% *** 8% 4% BC 2 BC 4 Brinton et al. biorxiv 2017

6 Grain length (mm) Grain width (mm) TGW (g) Grain length 5A is NILs the have main increased driver of increased TGW TGW 5A- NILs 5A NILs 5A - *** * 5A *** 9% *** 7% 7% *** *** 8% 4% *** BC 2 BC 4 *** 4% *** 4% *** 4% *** 5% *** 4% ** *** 1% ** 1% *** 0% 2% 2% BC 2 BC 4 BC 2 BC 4 Brinton et al. biorxiv 2017

7 5A QTL increases grain length uniformly 5A- NILs 5A NILs 5A - 5A BC 2 BC 4 Brinton et al. biorxiv 2017

8 Longer cells or more cells? 5A - 5A Cell expansion Cell division Brinton et al. biorxiv 2017

9 5A grains Longer have cells increased or more cells? length 5A - 5A Per genotype group: Grains measured = 24 Cells measured = >1500 5A- average 5A average 8.33% P = µm Brinton et al. biorxiv 2017

10 5A grains have increased cell length 5A - 5A 5A- average 5A average 8.33% P = Brinton et al. biorxiv 2017

11 5A grains have increased cell length independent of absolute grain size 5A - 5A 5A- large 5A average 9.53% P = Brinton et al. biorxiv 2017

12 Brinton et al. biorxiv 2017 Using RNAseq: 5A QTL When acts differential does during the grain expression 5A QTL development act? between NILs 5A- NILs 5A NILs 50 transcripts differentially expressed

13 20% of DE genes relate to ubiquitin & protein turnover E1 E2 E3 Proteasome

14 20% of DE genes relate to ubiquitin & protein turnover GW2: RING-type E3 ligase DA2 da2 E1 E2 E3 GW2 gw2 GW2 gw2 Song et al Nature 2007 Xia et al Plant Cell 2013 GW2-A gw2-a GW2-A gw2-a Proteasome Simmonds et al TAG 2016 GWAS: Su et al TAG 2011, Zhang et al J Integr Plant Biol 2013

15 20% of DE genes relate to ubiquitin & protein turnover E1 E2 E3 Proteasome

16 Rec group Fine mapping of the 5A QTL for grain length 5A 3/4 365 Mb 75 Mb 1/ A - 5A n=1 n=4 n=4 n=1 n=8 n=2 n=5 n=4 n=6 n=5 n=10 n=4 n= A - 5A 5A- 5A Grain length (mm) Brinton et al. biorxiv 2017

17 New Rec group Fine mapping of the 5A QTL for grain length Mean 5A- 5A n=3 n=3 n=33 n=3 n=13 n=21 n=4 n=21 5A- 5A A- CCCC 5A BBBB BBBC BBCC BCCC CBBB CCBB CCCB Grain length (mm)

18 New Rec group New Rec group Fine mapping of the 5A QTL for grain length Mean 5A- 5A 5A- 5A- 5A n=3 n=3 n=33 n=3 n=13 n=21 n=4 n=21 CCCC 5A BBBB BBBC BBCC BCCC CBBB CCBB CCCB Grain length (mm) mean length n=27bbbc-c 1.1 n=6 BBBC-B 1.2 CCCB-C CCCB-b n=10bccc-c 3.1 n=3 BCCC-b Grain length (mm)

19 New Rec group New Rec group Fine mapping of the 5A QTL for grain length Mean 5A- 5A 5A- 5A- 5A n=3 n=3 n=33 n=3 n=13 n=21 n=4 n=21 CCCC 5A BBBB BBBC BBCC BCCC CBBB CCBB CCCB Grain length (mm) mean length n=27bbbc-c 1.1 n=6 BBBC-B 1.2 CCCB-C CCCB-b n=10bccc-c 3.1 n=3 BCCC-b Grain length (mm)

20 New Rec group New Rec group Fine mapping of the 5A QTL for grain length Mean 5A- 5A 5A- 5A- 5A n=3 n=3 n=33 n=3 n=13 n=21 n=4 n=21 CCCC 5A BBBB BBBC BBCC BCCC CBBB CCBB CCCB Grain length (mm) mean length 1 3 n=27bbbc-c 1.1 n=6 BBBC-B 1.2 CCCB-C CCCB-b n=10bccc-c 3.1 n=3 BCCC-b Grain length (mm)

21 New Rec group New Rec group Fine mapping of the 5A QTL for grain length Mean 5A- 5A 5A- 5A- 5A n=3 n=3 n=33 n=3 n=13 n=21 n=4 n=21 CCCC 5A BBBB BBBC BBCC BCCC CBBB CCBB CCCB Grain length (mm) mean length 1 3 n=27bbbc-c 1.1 n=6 BBBC-B 1.2 CCCB-C CCCB-b n=10bccc-c 3.1 n=3 BCCC-b Grain length (mm)

22 Two tightly linked genes underlie the 5A QTL GL-A1 5.5 Mb 76 genes GL-A2 9.6 Mb 152 genes

23 GL-A1 Two tightly and GL-A2 linked are genes not fixed underlie in UK the the germplasm 5A 5A QTL 5A- Charger Short grain Dickens Panorama Solstice Relay Santiago Claire 5A Badger Long grain GL-A1 GL-A2 Cadenza Gallant GL-A1 GL-A2

24 GL-A1 Two tightly and GL-A2 linked are genes not fixed underlie in UK the germplasm 5A QTL 5A- Charger Short grain Dickens Panorama Solstice Relay Santiago Claire Cadenza Charger Malacca Xi19 5A Badger Long grain GL-A1 Gallant GL-A2 Cadenza Gallant GL-A1 GL-A2

25 GL-A1 Two tightly and GL-A2 linked are genes not fixed underlie in UK the germplasm 5A QTL 5A- Charger Short grain Dickens Panorama Claire Relay Santiago Solstice Avalon GL-A1 5A Badger Long grain GL-A1 GL-A2 Cadenza Gallant GL-A1 GL-A2

26 Testing candidates using exome-sequenced TILLING mutants Gene position Gene expression Gene function 15 candidate genes x??? A genome mutant B genome mutant Kronos TILLING mutants (AB) Krasileva et al. PNAS 2017

27 Can we combine different grain size genes? Cell expansion 5% Cell expansion Cell division Cell division 10%?? 5%

28 Can we combine different grain size genes? Cell expansion Grain length (5A QTL) Cell expansion Cell division Cell division 10%?? 5%

29 Can we combine different grain size genes? Cell expansion Grain length (5A QTL) Cell expansion Cell division Cell division 10%?? Grain width (6A QTL) Simmonds et al BMC Plant Biol 2014

30 Thousand grain weight (g) Can we combine different grain size genes? Cell expansion Grain length (5A QTL) Cell division Grain width (6A QTL) Simmonds et al BMC Plant Biol 2014 BC 4 Paragon NILs 2016

31 Understanding the genes and mechanisms controlling grain size will allow us to increase final yield in wheat. x??

32 James Simmonds Francesca Minter Abdul Kader Alabdullah Tobin Florio Cristobal

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