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Barley Landrace
Environmental
Association
Plant & Animal Genome 2018
Questions
• Can we identify genetic
variants that contribute
to environmental
adaptation in barley?

• In particular, are their
detectable variants that
could contribute to
cold temperature
adaptation?

• Cold adaptation per se
poorly explored relative
to flowering time
Poets et al. 2015
Study Design
• 784 geo-referenced landraces
from USDA NSGC Core

• Across historic range of
cultivation

• Genotyping on the barley
iSelect 9K - 6,152 SNPs

• Exome capture resequencing
of 137 landraces from our lab

• 137 more from IPK &
collaborators
Barley Landrace
Distribution
• Barley was cultivated from
Europe to East Asia

• By 3 - 5K YBP

• Ultimately from equator to the
Arctic Circle

• 0 - 66˚ N latitude
Barley Adaptation to
Eurasia
Pinhasi et al. 2005
ATGCAGTT
TTGCAGTT
CTGCAGTT
ATGCGGTT
TTGCGGTT
CTGCGGTT
Genetic varaints Phenotypes
Genetic positions of Chromosomes (cM)
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.bio11
−log10(p)
1 2 3 4 5 6 7 8
123456
Mixed model
P-value
Temperature
(C°)
# of plant accessions
Mixed Model Association to bioclimatic variables
Allele Frequency Differentiation - FST
Coop et al. 2009
Fang et al. 2014
Primary Results
• 12 associations to “cloned”
flowering time or cold-
tolerance genes

• 5 associations to loci relate
to cold tolerance

• 14 Fst outliers, primarily
involved flowering time and
cold tolerance
T A T V P Y STOP
A C T G C C A C G G T G C C C T A C T G A
T A T V P Y STOP
A C T G C C A C G G T A C C C T A C T G A
Reference
Alternative
…
…
SNP: 11_20784
Known cold tolerance gene: MLOC_15214.4
0 50 100 150
0
20
40
60
BIO17 Precipitation of Driest Quarter
X11_20784, Chro: 6H, 75.16cM, P−value: 2.563e−04
0
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BIO17 Precipitation of Driest Quarter
100150
tionofDriestQuarter
75.16cM,P−value:2.563e−04
0
200
400
600
800
1000
1200
1400
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100150
tionofDriestQuarter
tionunavailable,P−value:2.597e−04
0
200
400
600
800
1000
1200
1400
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Bio17 Precipitation of Driest Quarter
Li et al. - in prep.
LD around association
“hits”
●
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LD decay for SNPs around SNP: 11_10085
( chr2H : 758851026 bp )
Physical Distance (Kb)
LDestimate(r2
)
−100 −80 −60 −40 −20 0 20 40 60 80 100
0.00.20.40.60.81.0
●
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LD decay for SNPs around SNP: 11_20742
( chr3H : 17409756 bp )
Physical Distance (Kb)
LDestimate(r2
)
−100 −80 −60 −40 −20 0 20 40 60 80 100
0.00.20.40.60.81.0
Li et al. - in prep.
LD around association
“hits”
●
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LD decay for SNPs around SNP: 11_10143
( chr7H : 537951457 bp )
Physical Distance (Kb)
LDestimate(r2
)
−100 −80 −60 −40 −20 0 20 40 60 80 100
0.00.20.40.60.81.0
Li et al. - in prep.
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LD decay for SNPs around SNP: 11_20784
( chr6H : 507569812 bp )
Physical Distance (Kb)
LDestimate(r2
)
−100 −80 −60 −40 −20 0 20 40 60 80 100
0.00.20.40.60.81.0
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LD decay for SNPs around SNP: SCRI_RS_137464
( chr6H : 374867096 bp )
Physical Distance (Kb)
LDestimate(r2
)
−100 −80 −60 −40 −20 0 20 40 60 80 1000.00.20.40.60.81.0
LD decay for SNPs around SNP: SCRI_RS_137464
( chr6H : 374867096 bp )
Physical Distance (Kb)
LDestimate(r2
)
HvPRR1/HvTOC1 HORVU6Hr1G057640
11_10513
556
A/C
535
T/C
580
T/C
592
C/T
608
T/C
609
G/T
618
T/C
621
C/T
632
A/T
644
T/G
645
A/C
647
T/C
658
A/C
669
C/T
670
G/A
673
G/A
710
T/C
714
G/T
716
A/C
717
T/C
726
C/T
728
G/C
734
G/C
745
A/G
754
T/A
856
C/A
1003
G/A
1056
C/G
1169
G/A
1172
G/A
1180
T/C
1188
A/G
1336
C/T
2003
C/T
2011
C/T
2081
T/C
2117
G/T
2318
G/A
2332
C/T
2421
T/C
2579
G/A
2664
G/A
2724
C/T
2737
C/T
2739
T/G
2751
C/G
2767
C/T
2865
C/T
0 2995
Red: Nonsynonymous SNP
Blue: Significant associated SNP
LD with significant associated SNP (r2>0.4)
HvPRR1/HvTOC1
556
A/C
535
T/C
580
T/C
592
C/T
608
T/C
609
G/T
618
T/C
621
C/T
632
A/T
644
T/G
645
A/C
647
T/C
658
A/C
669
C/T
670
G/A
673
G/A
710
T/C
714
G/T
716
A/C
717
T/C
726
C/T
728
G/C
734
G/C
745
A/G
754
T/A
856
C/A
1003
G/A
1056
C/G
1169
G/A
1172
G/A
1180
T/C
1188
A/G
1336
C/T
2003
C/T
2011
C/T
2081
T/C
2117
G/T
2318
G/A
2332
C/T
2421
T/C
2579
G/A
2664
G/A
2724
C/T
2737
C/T
2739
T/G
2751
C/G
2767
C/T
2865
C/T
0 2995
HvPrr1/HvTOC1
SCRI_RS_137464 - chr6H:374867096
Summary
• Recover previously
characterized loci: Cbf3,
PhyC, Ppd-H1, Vrn1

• At least one locus directly
associated with freezing
tolerance

• Environmental association in
barley landraces will benefit
greatly from resequencing,
increased SNP density
Li Lei Chaochih Liu Corey Carter
Skylar Wyant Paul Hoffman
Fumi Katagiri
Code and Data Sharing
• https://github.com/MorrellLab/
Env_Assoc

• https://github.com/MorrellLab/
sequence_handling

• Sequences - NCBI Sequence
Read Archive

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Barley environmental association - Plant & Animal Genome 2018

  • 2. Questions • Can we identify genetic variants that contribute to environmental adaptation in barley? • In particular, are their detectable variants that could contribute to cold temperature adaptation? • Cold adaptation per se poorly explored relative to flowering time Poets et al. 2015
  • 3. Study Design • 784 geo-referenced landraces from USDA NSGC Core • Across historic range of cultivation • Genotyping on the barley iSelect 9K - 6,152 SNPs • Exome capture resequencing of 137 landraces from our lab • 137 more from IPK & collaborators
  • 4. Barley Landrace Distribution • Barley was cultivated from Europe to East Asia • By 3 - 5K YBP • Ultimately from equator to the Arctic Circle • 0 - 66˚ N latitude
  • 6. ATGCAGTT TTGCAGTT CTGCAGTT ATGCGGTT TTGCGGTT CTGCGGTT Genetic varaints Phenotypes Genetic positions of Chromosomes (cM) + ● ● ● ● ● ● ●● ●● ●● ● ●●●● ● ●● ●● ●●● ●● ●●● ● ● ●● ●● ●● ● ●●●● ●●● ● ●● ● ●●● ● ●● ●●● ●● ● ●● ● ●●●●●●● ● ●● ●● ●●● ●●●● ●● ●●● ●● ●● ●● ●● ●●● ●● ●● ●● ●●● ● ● ●●● ● ● ●●● ●● ●● ●● ● ● ●●● ● ●●●●● ●● ●● ● ●● ●●● ●●●● ●● ●●● ●● ●● ●●● ●●● ● ●● ●●● ●●● ● ●●● ●● ●● ● ●● ●●●● ●● ●●● ●● ● ●●● ● ●● ●● ● ●●● ● ● ● ●●●●● ●●● ●● ●● ●●● ● ●● ●● ●●●● ●●● ●●● ●●● ●● ●●● ● ●● ●● ●● ●●● ● ●● ●● ● ●● ●● ● ●● ● ●●● ●● ●●● ●● ● ●● ●●● ● ●●● ●● ●● ●● ●●●● ●● ●●● ● ●● ● ●● ●● ●● ● ●●● ●●● ●●●● ●●● ●● ●●● ●●●● ●●●● ●●● ●● ●●●● ● ● ●● ● ●●● ●● ● ●● ●●● ● ● ●● ●●● ●● ●●●●●● ●● ● ●●● ● ●●● ●● ●● ●● ●●●● ● ● ●● ●● ●● ●● ●●● ●● ●● ●● ● ● ●● ● ● ●●● ●●● ● ●● ● ●● ● ●●●●● ● ● ● ●● ●●● ●● ● ● ● ●● ● ●● ●● ● ●● ●● ●● ●● ● ●● ●● ●● ●●● ●● ●● ●●●● ● ●● ●●● ●●● ● ●● ● ●●●● ●● ●● ●● ●●●● ●● ●●● ●● ●●● ●●●●●● ●● ●● ●● ● ●● ●●● ● ●●● ●● ● ● ●●● ● ● ●● ●●●● ●● ●● ● ●●● ●● ● ●●● ● ● ●● ● ● ●● ● 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  • 7. Fang et al. 2014
  • 8. Primary Results • 12 associations to “cloned” flowering time or cold- tolerance genes • 5 associations to loci relate to cold tolerance • 14 Fst outliers, primarily involved flowering time and cold tolerance T A T V P Y STOP A C T G C C A C G G T G C C C T A C T G A T A T V P Y STOP A C T G C C A C G G T A C C C T A C T G A Reference Alternative … … SNP: 11_20784 Known cold tolerance gene: MLOC_15214.4 0 50 100 150 0 20 40 60 BIO17 Precipitation of Driest Quarter X11_20784, Chro: 6H, 75.16cM, P−value: 2.563e−04 0 200 400 600 800 1000 1200 1400 ● ●●● ● ● ●● ● ● ● ●● ● ● ●● ●●●● ●● ●●●●●● ● ●●●● ● ● ● ● ● ●● ● ●● ● ● ● ●●● ● ● ●●● ● ●● ●● ● ● ●●●●●●●● ● ● ● ● ● ● ● ●●●●●●●●●●●● ● ●●● ● ●● ●●●● ●●●●●●●●●●●● ● ●● ● ● ● ● ● ● ● ●● ● ● ● ●● ● ●●● ● ● ● ●●●●● ● ● ● ● ● ● ● ● ● ●● ●● ● ● ●●●●●●● ● ● ● ● ●●●●●●● ● ●●●●●●●●● ● ● ●● ● ● ●● ● ●●●●●●●● ●● ●●● ● ●●●●● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ●● ● ●●● ● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ● ● ●●●●●● ● ●●●●●● ● ● ● ● ● ●● ● ●● ● ● ● ●●●● ●● ● ●● ● ●● ●● ●●● ● ● ● ●●● ● ● ● ●● ●●●● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ●● ● ●●● ● ● ●● ● ● ● ● ● ● ● ●● ● ● ●● ●● ●●● ●● ●● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ●●● ● ●● ● ●● ● ● ●●●●● ● ● ● ●●● ● ● ● ● ● ●●●● ●● ● ●● ● ●●●● ● ●●● ●●●●● ● ●●●●●● ● ● ● ● ● ● ●●● ● ● ● ●● ● ● ●● ● ● ● ●● ●● ●● ● ● ● ● ● ● ●●●●●●●●● ● ● ●● ●● ● ● ●●●●●●●●●●●●● ● ● ●●●● ●●●●● ●●●●●●●●●●●●●●●●● ● ●● ● ● ●● ● ● ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●● ● ● ●●●●●● ● ●● ●● ● ● ● ●● ● ● ●● ● ● ● ● ● ● ●● ● ● ● ● ●●●●●●●●● ● ● ●●●●●●●●●●●●●●● ●●●●●●●●●●●●● ● ● ● ● ●●● ●● ●● ● ●●●●●●● ● ● ● ● ●● ● ● ● ● ● ●● ●● ● ● ●●● ●● ● ● ● ●● ●●● ● ● ●● ●●●● ●● ● ●●●● ●●●●●●●●● ● ● ● ●●●●●●● ●●●● ●●●● ● ● ●● ●●●●●● ●●● ● ●●●●● ●●● ● ● ●● ●● ● ● ● ● ● ● ●● ●● ●●●● BIO17 Precipitation of Driest Quarter 100150 tionofDriestQuarter 75.16cM,P−value:2.563e−04 0 200 400 600 800 1000 1200 1400 ● ●●●● ●●●●●●●● ●●●● ●●●● ●●●●●●●● ● ● ● ● ● ● ● ●● ● ● ● ● ● ●● ●● ● ● ● ● ● ● ●● ● ● ●● ● ●●●●●●●● ●● ●●● ● ●●●●● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ●● ●●●●●●● ● ●●●●●● ● ●● ●●●● ● ● ●●● ● ● ●●●● ● ● ● ● ● ● ●● ●● ● ● ●● ● ● ● ● ● ● ●● ● ●● ● ●●● ● ●●● ●●● ●● ● ● ● ●●●●●●●●●●●●●●● ●●●●●●● ● ● ● ●●● ●● ● ● ● ●● ●●●● ● ●●●●●●●●● ●●●● ●● ●● ●●●●●● ●●● ● ●●●●●● ● ●● ●● ● ● ● ●● ●●●● 100150 tionofDriestQuarter tionunavailable,P−value:2.597e−04 0 200 400 600 800 1000 1200 1400 ● ●●●● ●●●●●●●● ●●●● ●●●● ●●●●●●●● ● ● ● ● ● ● ● ●● ● ● ● ● ● ●● ●● ● ● ● ● ● ● ●● ● ● ●● ● ●●●●●●●● ●● ●●● ● ●●●●● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ●● ●●●●●● ● ●●●●●● ● ●● ●●●● ● ● ●●● ● ● ●●●● ● ● ● ● ● ● ●● ●● ● ● ●● ● ● ● ● ● ● ●● ● ●● ● ●●● ● ●●● ●●● ●● ● ● ● ●●●●●●●●●●●●●●● ●●●●●●● ● ● ● ●●● ●● ● ● ● ●● ●●●● ● ●●●●●●●●● ●●●● ●● ●● ●●●●●● ●●● ● ●●●●●● ● ●● ●● ● ● ● ●● ●●●● Bio17 Precipitation of Driest Quarter Li et al. - in prep.
  • 9. LD around association “hits” ● ● ● ● ●●●●●●●●●● ● ● ● ● ● ● ● ●●● ● ● ● ● ● ●●● ● ● ● ● ● ●●● ● ●●● ● ● ●● ● ● ● ●● ● ●●●●●●● ● ●● ● ●●●●● ● ●●●● ● ● ● ●●● ● ●● ●●●● ● ● ●● ● ● LD decay for SNPs around SNP: 11_10085 ( chr2H : 758851026 bp ) Physical Distance (Kb) LDestimate(r2 ) −100 −80 −60 −40 −20 0 20 40 60 80 100 0.00.20.40.60.81.0 ● ● ●● ● ●●●●●●●●●● ● ●● ●● ● ● ● ● ● ●●●●●●●● ● ●●● ● ● ●●● ● ● ● ● ● ● ● ● ●●●●●●●●● ● ● ● ●●●● ●●● ● ●●● ●●●●● ● ●● ● ● ● ● ● ● ● ● ● ● ● ●●●●●● ●● ●●● ● ●●●●●● ●●●●●●●●●● ●●●●●●● ● ●●●●●●●●●●●●● ● ● ●●● ● ●●●●●● ● ● ●● ●●● ● ●●●● ● ●●● ● ●● ● ● ● ●●● ● ●●● ● ● ● ●● ● ● ● ●●● ●● ● ● ● ● ●● ● ● ●● ●● ● ●●●●●●●● ●●●●●●●●●●●●●●●●●● ●●●● ● ● ●● ● LD decay for SNPs around SNP: 11_20742 ( chr3H : 17409756 bp ) Physical Distance (Kb) LDestimate(r2 ) −100 −80 −60 −40 −20 0 20 40 60 80 100 0.00.20.40.60.81.0 Li et al. - in prep.
  • 10. LD around association “hits” ● ●●●● ● ● ● ● ● ● ● ● ● ● ●●● ●●● ● ●●●● ● ● ● ● ● ● ●●●● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ● ●●●●● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ●●● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ●● ● ● ●● ● ● ● ● ● ● ●●● LD decay for SNPs around SNP: 11_10143 ( chr7H : 537951457 bp ) Physical Distance (Kb) LDestimate(r2 ) −100 −80 −60 −40 −20 0 20 40 60 80 100 0.00.20.40.60.81.0 Li et al. - in prep. ● ●● ● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ●● ● ●●● ● ●● ● ● ● ● ● ●● ● ● ● ●●● ● ● ● ●● ● LD decay for SNPs around SNP: 11_20784 ( chr6H : 507569812 bp ) Physical Distance (Kb) LDestimate(r2 ) −100 −80 −60 −40 −20 0 20 40 60 80 100 0.00.20.40.60.81.0
  • 11. ●●●● ● ●●●●●●●●●●●●●●●●●●●● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ●● ●● ●● ● ● ● ● ● ● ●● ● LD decay for SNPs around SNP: SCRI_RS_137464 ( chr6H : 374867096 bp ) Physical Distance (Kb) LDestimate(r2 ) −100 −80 −60 −40 −20 0 20 40 60 80 1000.00.20.40.60.81.0 LD decay for SNPs around SNP: SCRI_RS_137464 ( chr6H : 374867096 bp ) Physical Distance (Kb) LDestimate(r2 ) HvPRR1/HvTOC1 HORVU6Hr1G057640 11_10513 556 A/C 535 T/C 580 T/C 592 C/T 608 T/C 609 G/T 618 T/C 621 C/T 632 A/T 644 T/G 645 A/C 647 T/C 658 A/C 669 C/T 670 G/A 673 G/A 710 T/C 714 G/T 716 A/C 717 T/C 726 C/T 728 G/C 734 G/C 745 A/G 754 T/A 856 C/A 1003 G/A 1056 C/G 1169 G/A 1172 G/A 1180 T/C 1188 A/G 1336 C/T 2003 C/T 2011 C/T 2081 T/C 2117 G/T 2318 G/A 2332 C/T 2421 T/C 2579 G/A 2664 G/A 2724 C/T 2737 C/T 2739 T/G 2751 C/G 2767 C/T 2865 C/T 0 2995 Red: Nonsynonymous SNP Blue: Significant associated SNP LD with significant associated SNP (r2>0.4) HvPRR1/HvTOC1 556 A/C 535 T/C 580 T/C 592 C/T 608 T/C 609 G/T 618 T/C 621 C/T 632 A/T 644 T/G 645 A/C 647 T/C 658 A/C 669 C/T 670 G/A 673 G/A 710 T/C 714 G/T 716 A/C 717 T/C 726 C/T 728 G/C 734 G/C 745 A/G 754 T/A 856 C/A 1003 G/A 1056 C/G 1169 G/A 1172 G/A 1180 T/C 1188 A/G 1336 C/T 2003 C/T 2011 C/T 2081 T/C 2117 G/T 2318 G/A 2332 C/T 2421 T/C 2579 G/A 2664 G/A 2724 C/T 2737 C/T 2739 T/G 2751 C/G 2767 C/T 2865 C/T 0 2995
  • 13. Summary • Recover previously characterized loci: Cbf3, PhyC, Ppd-H1, Vrn1 • At least one locus directly associated with freezing tolerance • Environmental association in barley landraces will benefit greatly from resequencing, increased SNP density
  • 14. Li Lei Chaochih Liu Corey Carter Skylar Wyant Paul Hoffman Fumi Katagiri
  • 15. Code and Data Sharing • https://github.com/MorrellLab/ Env_Assoc • https://github.com/MorrellLab/ sequence_handling • Sequences - NCBI Sequence Read Archive