2013 11-25-sheffield

A social chromosome in the red fire ant.

[email protected] Sheffield 2013-11-25

mailto:[email protected]

© National Geographic

Atta leaf-cutter ants

Oecophylla Weaver ants

© ameisenforum.de

© ameisenforum.de

Fourmis tisserandes

© ameisenforum.de

Oecophylla Weaver ants

Tofilski et al 2008

Forelius pusillus

Tofilski et al 2008

Forelius pusillus hides the nest entrance at night

Avant

Workers staying outside die« preventive self-sacrifice »

Tofilski et al 2008

Forelius pusillus hides the nest entrance at night

Dorylus driver ants: ants with no home

© BBC

Animal biomass (Brazilian rainforest)

from Fittkau & Klinge 1973

Other insects 49.6

Amphibians 2.8

Reptiles 3.7

Birds 5.3

Mammals 14.5

!Earthworms

17.3

!!

Spiders 4.7

Soil fauna excluding earthworms,

ants & termites 148

Ants & termites 114

www.sciencemag.org SCIENCE VOL 331 25 FEBRUARY 2011 1067

REPORTS

on

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Solenopsis invicta fire ants are a big problem!very well studied!

Ascunce et al 2011

Solenopsis invicta fire ant: two social forms

!

•1 large queen"•Independent founding"•Highly territorial"•Many sizes of workers

!

•2-100 smaller queens"•Dependent founding"•No inter-colony aggression"•All workers similar size

Single-queen form: Multiple-queen form:

Fire ants+

Population genetics: Allozyme screen

Ken Ross L. Keller

“starch gel”+

1 2 3

Allozyme screen Social form associated to Gp-9 locus

Frequency of the most

common allele!

Locus!

0.3!0.4!0.5!0.6!0.7!0.8!0.9!1.0!

Single queen!Multiple queen!

Est-6!Est-4!G3pdh-1!Ca-4!Pgm-4!Ddh-1!Pro-5!

Pgm-3!

Acoh-5!

acoh-1!

Acy-1!

Pgm-1!

Aat-2!

Gp-9!

Ken Ross and colleagues"Laurent Keller and colleagues

Single queen form Multiple queen form


Social form completely associated to Gp-9 locus

bbbbBB BB Bb bb




bbBB BB Bb

x

Gp-9 bb females rareKen Ross and colleagues"

Laurent Keller and colleagues



BB BB Bb




BB BB Bb

xKen Ross and colleagues"




BB BB Bb

x xKen Ross and colleagues"




BB BB Bb

x x xKen Ross and colleagues"


Single queen form Multiple queen form(>15% ) (< 5% )


•Is this gene the single überregulator?


Hypothesis: influences queen odor & how workers « smell » queens"

Gp-9 is an odorant binding protein

Krieger & Ross

dN/dS = infinityKrieger & Ross

•Is this gene the single überregulator?


maybe 1/14th of the genome?•Only 14 allozyme markers were used

Locus!

0.3!0.4!0.5!0.6!0.7!0.8!0.9!1.0!

Single queen!Multiple queen!

Est-6!Est-4!G3pdh-1!Ca-4!Pgm-4!Ddh-1!Pro-5!

Pgm-3!

Acoh-5!

acoh-1!

Acy-1!

Pgm-1!

Aat-2!

Gp-9!

This changes everything.454"

Illumina"Solid...

Any lab can sequence anything!

Are other genes linked to Gp-9?

Sequenced:"•a Gp-9 B � genome"

!

!

Single ♂:His brothers:

11×4×

(330bp-insert paired reads) (normal single-end reads)45× +Sequencing from haploid males (for easier assembly):

(8,000 & 20,000bp-insert paired reads)

B 20x

The genome of a Gp-9 B � fire ant

Single ♂:His brothers:

11×4×

(330bp-insert paired reads) (normal single-end reads)45× +Sequencing from haploid males (for easier assembly):

(8,000 & 20,000bp-insert paired reads)

B 20x

The genome of a Gp-9 B � fire ant

Assembly approach:1. Assemble short Illumina reads with SOAPdenovo→N50: 3600 bp"2. Chop assembly into “fake 454 reads” (300bp)"3. Assemble fake + real 454 reads with Newbler→N50: 720,000 bp

10,000 scaffolds (100 biggest scaffolds: 50% of genome)

Total: 350,000,000 bp assembled. The rest: repeats→Wurm et al 2011

★ Expansion of lipid-processing gene families (for Cuticular Hydrocarbons?)"

★ 420 putative olfactory receptors (more than any other insect!)

The genome of the fire antSome findings:

0.05

SiOR

04648+10

SiOR01968+4

SiOR0

0899+7

SiOR02814+3

SiOR04171+6

SiO

R04609+4

SiOR00330+28

SiO

R02

694+25

SiO

R04609+20

SiOR05285+6

SiOR00330+25

SiOR04510+15

SiOR00330+18

SiOR04609+23

SiOR01

968+23

SiO

R03

952+4

SiOR0

4648+16

SiO

R05

901+2

SiO

R02

944+4

SiOR01968+5

SiOR04171+19 SiOR

04648+5

SiOR10535+3

SiOR0

1224+3

SiOR06723+2

SiOR01968+9

SiOR02883+1

SiOR0

0899+3

SiOR01629+1

SiOR04171+1

SiOR01629+11

SiOR04171+10

SiOR04171+13

SiOR02694+3

SiOR04171+20

SiO

R02694+35

SiOR04171+15

SiOR04609+7

SiOR05118+2

SiOR07

837+2

SiO

R02

694+27

SiOR01968+10

SiOR04648+17

SiOR0196

8+19

SiO

R02

694+17

SiOR04648+13

SiOR01968+6

SiOR

00899+1

2

SiOR0

5901+1

SiOR00330+20

SiO

R02

648+2

SiO

R02

659+2

SiOR01968+1

6

SiOR0

0899+11

SiO

R02974

SiOR04171+2

SiOR03952+2

SiOR06792+2

SiOR04510+4

SiOR04171+28

SiOR05285+5

SiOR05285+9

SiOR

00899+1

5

SiOR

04648+3

SiO

R02694+36

SiOR10535+1

SiO

R02

694+19

SiO

R02

694+23

SiOR02694+1

SiO

R04609+14

SiOR01122

SiOR

04648+9

SiO

R02694+34

SiOR01629+8

SiOR

04648+8

SiOR04510+8

SiOR06573

SiO

R02

944+1

SiOR00330+26

SiO

R00330+1S

iOR02

694+15

SiO

R03038

SiOR05285+7

SiOR0

0899+5

SiOR04609+10

SiO

R04609+3

SiOR04339

SiOR0

8068

SiOR

04648+6

SiOR04510+2

SiOR05285+8

SiOR01

573+4

SiOR0366

3

SiOR04171+8

SiOR01858+2

SiOR01968+2SiOR01968+1

SiOR02694+5

SiOR01968+3

SiOR06723+3

SiOR01968+15

SiOR05285+1

SiOR0

0899+4

SiOR04609+22

SiOR04171+9

SiOR02694+9

SiO

R02

648+1

SiOR06792+3

SiOR0

1573+2

SiO

R02

694+20

SiOR1

0542

SiO

R04609+15

SiOR02694+8

SiOR00330+16

SiOR0

0899+2

SiOR02694+10

SiOR04510+9

SiOR05285+3

SiOR04171+24

SiO

R04609+2

SiOR05285+11

SiOR02

694+14

SiOR01

573+1

SiOR05285+2

SiOR00613

SiOR0196

8+22

SiOR

00899+9

SiO

R06

843+2

SiO

R02694+37

SiOR0

0899+1

SiOR04609+9

SiOR05431+2

SiOR10535+2

SiOR00330+15

SiO

R02

694+18

SiOR0

1224+2

SiOR04510+11

SiOR00330+23

SiO

R02

694+29

SiOR05416

SiOR05285+10

SiOR02694+2

SiOR01629+9

SiOR05431+1

SiOR02883+2

SiOR04171+14

SiO

R08

341

SiO

R02

694+22

SiOR01

224+1

SiOR01968+12

SiOR05431+3

SiOR04510+6

SiOR02694+7

SiOR04609+5

SiO

R02

944+2

SiOR03

952+3

SiOR01968+8

SiOR04609+24

SiO

R02

694+30

SiOR01629+10

SiOR04510+14

SiOR00565

SiOR05118+3

SiO

R01321

SiO

R04609+19

SiOR00330+14

SiO

R02694+38

SiOR04609+8

SiOR04171+16

SiOR10455

SiO

R04609+16

SiO

R04609+21

SiO

R02

694+28

SiO

R02

659+1

SiOR04171+5

SiOR00330+29

SiOR01968+14

SiOR03983

SiOR00330+27

SiOR05285+4

SiOR04510+1

SiOR

00899+8

SiOR

04648+7

SiO

R04609+17

SiO

R00330+5

SiO

R02

694+21

SiOR02814+4

SiOR00330+7

SiOR01

629+3

SiOR01

968+26

SiO

R02694+31

SiOR0

4648+2

SiO

R02694+39

SiOR04171+25

SiOR06577

SiOR01

968+25

SiOR04171+21

SiOR06792+6

SiO

R04609+11

SiOR02694+11

SiOR06792+1

SiOR04171+4

SiOR01629+5

SiOR00330+21

SiOR0

4648+15

SiO

R00330+6

SiO

R02

694+16

SiOR

04648+11

SiOR

04648+4

SiO

R00330+3

SiO

R06

535

SiOR04171+7

SiOR10493

SiO

R02694+32

SiOR06792+4

SiOR04510+3

SiO

R06

890

SiOR0196

8+20

SiO

R04609+12

SiOR04171+3

SiOR01968

+18

SiOR01968+11

SiO

R04609+13

SiOR01629+12

SiOR00330+22

SiO

R02694+33

SiOR00330+13

SiOR01

573+3

SiOR04510+16

SiOR04171+12

SiOR04510+13

SiOR

05118+1

SiO

R02

944+3

SiOR04171+26

SiOR04171+17

SiOR

00899+14

SiOR04171+29

SiOR

00899+1

3

SiOR02694+13

SiOR00330+24

SiOR00330+19

SiOR04171+27

SiO

R02

694+24

SiOR04510+5

SiOR07090

SiOR03952+1

SiOR04510+10

SiOR00330+17

SiO

R02

694+26

SiO

R04609+1

SiOR0

0899+6

SiOR04648+12

SiOR02814+2

SiOR00330+11

SiOR04171+18

SiOR01968+1

7

SiOR00330+10

SiOR00330+9

SiOR0162

9+2

SiOR04171+11

SiOR04510+12

SiOR0196

8+21

SiOR00330+8

SiOR01858+1

SiOR04510+7

SiOR05431+4

SiOR02694+6

SiOR01968+13

SiO

R00330+4

SiO

R04609+18

SiOR0

0899+10

SiOR00330+12

SiOR00330+31

SiO

R06

843+1

SiOR07

837+1

SiO

R00330+2

SiOR01629+4

SiOR0

4648+1

SiOR01

968+24

SiOR04171+23

SiOR0162

9+7

SiOR0

4648+14

SiO

R06

843+3

SiOR06792+5

SiOR01968+7

SiOR01629

+6

SiOR02883+3

SiOR02694+12

SiOR

05118+4

SiOR04171+22

SiO

R01080

SiOR04609+6

SiOR02814+1

SiOR00330+30

SiOR05285+12

SiOR06723+1

SiOR02694+4

Wurm et al 2011

> 400Harvester ant 399 344Nasonia wasp 225honey bee 165

6510

★ Expansion of lipid-processing gene families (for Cuticular Hydrocarbons)"

★ 420 putative olfactory receptors (more than any other insect!)"

★ Functional DNA-methylation system


Wurm et al 2011

0.05

SiOR

04648+10

SiOR01968+4

SiOR0

0899+7

SiOR02814+3

SiOR04171+6

SiO

R04609+4

SiOR00330+28

SiO

R02

694+25

SiO

R04609+20

SiOR05285+6

SiOR00330+25

SiOR04510+15

SiOR00330+18

SiOR04609+23

SiOR01

968+23

SiO

R03

952+4

SiOR0

4648+16

SiO

R05

901+2

SiO

R02

944+4

SiOR01968+5

SiOR04171+19 SiOR

04648+5

SiOR10535+3

SiOR0

1224+3

SiOR06723+2

SiOR01968+9

SiOR02883+1

SiOR0

0899+3

SiOR01629+1

SiOR04171+1

SiOR01629+11

SiOR04171+10

SiOR04171+13

SiOR02694+3

SiOR04171+20

SiO

R02694+35

SiOR04171+15

SiOR04609+7

SiOR05118+2

SiOR07

837+2

SiO

R02

694+27

SiOR01968+10

SiOR04648+17

SiOR0196

8+19

SiO

R02

694+17

SiOR04648+13

SiOR01968+6

SiOR

00899+1

2

SiOR0

5901+1

SiOR00330+20

SiO

R02

648+2

SiO

R02

659+2

SiOR01968+1

6

SiOR0

0899+11

SiO

R02974

SiOR04171+2

SiOR03952+2

SiOR06792+2

SiOR04510+4

SiOR04171+28

SiOR05285+5

SiOR05285+9

SiOR

00899+1

5

SiOR

04648+3

SiO

R02694+36

SiOR10535+1

SiO

R02

694+19

SiO

R02

694+23

SiOR02694+1

SiO

R04609+14

SiOR01122

SiOR

04648+9

SiO

R02694+34

SiOR01629+8

SiOR

04648+8

SiOR04510+8

SiOR06573

SiO

R02

944+1

SiOR00330+26

SiO

R00330+1S

iOR02

694+15

SiO

R03038

SiOR05285+7

SiOR0

0899+5

SiOR04609+10

SiO

R04609+3

SiOR04339

SiOR0

8068

SiOR

04648+6

SiOR04510+2

SiOR05285+8

SiOR01

573+4

SiOR0366

3

SiOR04171+8

SiOR01858+2

SiOR01968+2SiOR01968+1

SiOR02694+5

SiOR01968+3

SiOR06723+3

SiOR01968+15

SiOR05285+1

SiOR0

0899+4

SiOR04609+22

SiOR04171+9

SiOR02694+9

SiO

R02

648+1

SiOR06792+3

SiOR0

1573+2

SiO

R02

694+20

SiOR1

0542

SiO

R04609+15

SiOR02694+8

SiOR00330+16

SiOR0

0899+2

SiOR02694+10

SiOR04510+9

SiOR05285+3

SiOR04171+24

SiO

R04609+2

SiOR05285+11

SiOR02

694+14

SiOR01

573+1

SiOR05285+2

SiOR00613

SiOR0196

8+22

SiOR

00899+9

SiO

R06

843+2

SiO

R02694+37

SiOR0

0899+1

SiOR04609+9

SiOR05431+2

SiOR10535+2

SiOR00330+15

SiO

R02

694+18

SiOR0

1224+2

SiOR04510+11

SiOR00330+23

SiO

R02

694+29

SiOR05416

SiOR05285+10

SiOR02694+2

SiOR01629+9

SiOR05431+1

SiOR02883+2

SiOR04171+14

SiO

R08

341

SiO

R02

694+22

SiOR01

224+1

SiOR01968+12

SiOR05431+3

SiOR04510+6

SiOR02694+7

SiOR04609+5

SiO

R02

944+2

SiOR03

952+3

SiOR01968+8

SiOR04609+24

SiO

R02

694+30

SiOR01629+10

SiOR04510+14

SiOR00565

SiOR05118+3

SiO

R01321

SiO

R04609+19

SiOR00330+14

SiO

R02694+38

SiOR04609+8

SiOR04171+16

SiOR10455

SiO

R04609+16

SiO

R04609+21

SiO

R02

694+28

SiO

R02

659+1

SiOR04171+5

SiOR00330+29

SiOR01968+14

SiOR03983

SiOR00330+27

SiOR05285+4

SiOR04510+1

SiOR

00899+8

SiOR

04648+7

SiO

R04609+17

SiO

R00330+5

SiO

R02

694+21

SiOR02814+4

SiOR00330+7

SiOR01

629+3

SiOR01

968+26

SiO

R02694+31

SiOR0

4648+2

SiO

R02694+39

SiOR04171+25

SiOR06577

SiOR01

968+25

SiOR04171+21

SiOR06792+6

SiO

R04609+11

SiOR02694+11

SiOR06792+1

SiOR04171+4

SiOR01629+5

SiOR00330+21

SiOR0

4648+15

SiO

R00330+6

SiO

R02

694+16

SiOR

04648+11

SiOR

04648+4

SiO

R00330+3

SiO

R06

535

SiOR04171+7

SiOR10493

SiO

R02694+32

SiOR06792+4

SiOR04510+3

SiO

R06

890

SiOR0196

8+20

SiO

R04609+12

SiOR04171+3

SiOR01968

+18

SiOR01968+11

SiO

R04609+13

SiOR01629+12

SiOR00330+22

SiO

R02694+33

SiOR00330+13

SiOR01

573+3

SiOR04510+16

SiOR04171+12

SiOR04510+13

SiOR

05118+1

SiO

R02

944+3

SiOR04171+26

SiOR04171+17

SiOR

00899+14

SiOR04171+29

SiOR

00899+1

3

SiOR02694+13

SiOR00330+24

SiOR00330+19

SiOR04171+27

SiO

R02

694+24

SiOR04510+5

SiOR07090

SiOR03952+1

SiOR04510+10

SiOR00330+17

SiO

R02

694+26

SiO

R04609+1

SiOR0

0899+6

SiOR04648+12

SiOR02814+2

SiOR00330+11

SiOR04171+18

SiOR01968+1

7

SiOR00330+10

SiOR00330+9

SiOR0162

9+2

SiOR04171+11

SiOR04510+12

SiOR0196

8+21

SiOR00330+8

SiOR01858+1

SiOR04510+7

SiOR05431+4

SiOR02694+6

SiOR01968+13

SiO

R00330+4

SiO

R04609+18

SiOR0

0899+10

SiOR00330+12

SiOR00330+31

SiO

R06

843+1

SiOR07

837+1

SiO

R00330+2

SiOR01629+4

SiOR0

4648+1

SiOR01

968+24

SiOR04171+23

SiOR0162

9+7

SiOR0

4648+14

SiO

R06

843+3

SiOR06792+5

SiOR01968+7

SiOR01629

+6

SiOR02883+3

SiOR02694+12

SiOR

05118+4

SiOR04171+22

SiO

R01080

SiOR04609+6

SiOR02814+1

SiOR00330+30

SiOR05285+12

SiOR06723+1

SiOR02694+4

★ Expansion of lipid-processing gene families (for Cuticular Hydrocarbons)"

★ 420 putative olfactory receptors (more than any other insect!)"

★ Functional DNA-methylation system "★Ant-specific duplication and subfunctionalization

of vitellogenin (in bees: involved in reproduction & division of labor)


Wurm et al 2011

significance of these duplication events in vitellogenins, odorperception genes, and a family of lipid-processing genes. We alsodiscuss additional features of interest in the fire ant genome rel-evant to the complex social biology of this species, including sexdetermination genes, DNA methylation genes, telomerase, andthe insulin and juvenile hormone pathways.

Vitellogenins. In contrast to other insects that mainly have only oneor two vitellogenins, the fire ant genome harbors four adjacent

copies of vitellogenin (Vg1-4) (Fig. 3A), all of which are at leastpartially supported by EST reads. A phylogenetic analysis revealsthat an ancestral vitellogenin gene first underwent duplication,followed by possibly independent duplications of each of thedaughter vitellogenins, thus giving rise to Vg1 and Vg4 and to Vg2andVg3. All of these duplications occurred after the ancestor offireants split from wasps and bees (Fig. 3B). The single vitellogeninfound inA.mellifera is amultifunctional protein (26) involved in theregulation of life span (27, 28) and division of labor (29). Quanti-tative RT-PCR shows thatVg1 andVg4 are preferentially expressedin workers and Vg2 and Vg3 in queens (Fig. 3C, SI Materials andMethods, and Table S1G). Vitellogenin expression in S. invictaworkers is surprising because they lack ovaries. Given the super-organism properties of ant societies, the expression patterns sug-gest that vitellogenins underwent neo- or subfunctionalizationafter duplication to acquire caste-specific functions.

Odor Perception. Consistent with studies in other insects, we finda single S. invicta ortholog to DmOr83b, a broadly expressed ol-factory receptor (OR) required to interact with other ORs forDrosophila and Tribolium castaneum olfaction (30–32). BeyondOR83b, OR number varies greatly between insect species. Blastsearches and GeneWise searches using an HMM profile con-structed with aligned ORs from N. vitripennis (33) and Pogono-myrmex barbatus identified more than 400 loci in the S. invictagenome with significant sequence similarity to ORs. Preliminarywork on gene model reconstruction identified 297 intact full-length proteins. Many S. invicta ORs are in tandem arrays (Fig.S2A) and derive from recent expansions. S. invicta may thus har-bor the largest identified insect OR repertoire because there are10 ORs in Pediculus humanus (34), 60 in Drosophila, 165 inA. mellifera, 225 in N. vitripennis (33), and 259 in T. castaneum(32). The large numbers of N. vitripennis and T. castaneum ORsare thought to be due to current or past difficulties in host andfood finding. As has been suggested for A. mellifera (35), the largenumber of S. invicta ORs may result from the importance ofchemical communication in ants. The odorant-binding proteins(OBPs) are another family of genes also known to play roles inchemosensation in Drosophila (36). Intriguingly, the social orga-nization of S. invicta colonies is completely associated with se-

Eum

etaz

oa

No hits 3424

Not assigned 274

Cnidaria 100

Art

hro

po

da

Ne

op

tera

Coe

lom

ata

Bila

teri

a

Nematoda 25

Deuterostomia 173

Arachnida 50

Paraneoptera 577

End

opte

rygo

ta

Hym

en

op

tera

Apoidea 7398

Chalcidoidea 3271

Ichneumonoidea 94

Diptera 404

Lepidoptera 29

Coleoptera 713

Fig. 2. Taxonomic distribution of best blastp hits of S. invicta proteins to thenonredundant (nr) protein database (E < 10−5). Results were first plottedusing MEGAN software (22) and then branches with fewer than 20 hits wereremoved, branch lengths were reduced for compactness, and tree topologywas adjusted to reflect consensus phylogenies (23, 24).

Vg1Vg4 Vg3 Vg2

2,330,000 bp 2,360,000 bpA

B CSolenopsis Vg1Solenopsis Vg4Solenopsis Vg2Solenopsis Vg3

Apis VgBombus Vg

Nasonia Vg1

Nasonia Vg2Pteromalus Vg

Encarsia VgPimpla Vg

Athalia Vg

Apocrita

Tenthedinoidea

Vespoidea

Apoidea

Aculeata

Chalcidoidea

Ichneumonoidea 0

5000

10000

15000

20000

25000 Vg3Vg2

142 389 17820 1.4 9269 0.61 40WQ WQW Q WQ

*** ***Vg1 Vg4* ***

0

100

200

300

400

500

600

Fig. 3. S. invicta vitellogenins. (A) Four vitellogenins are located within a single 40,000-bp region of the S. invicta genome. (B) Parsimony tree of known hy-menopteran vitellogenin protein sequences suggests that two rounds of vitellogenin duplication occurred after the split between ants and other hyme-nopterans including bees and wasps. (C) Quantitative RT-PCR of the four putative S. invicta vitellogenins on whole bodies of major workers (W) and matedqueens (Q) (n = 10). The y axis indicates mRNA concentrations for the different vitellogenins. Values depicted by each bar are shown below the x-axis labels.Error bars represent SEs. Expression differences between queens and workers were significant (Bonferroni-corrected two-tailed t tests: *P < 0.05, ***P < 10−10).

Wurm et al. PNAS Early Edition | 3 of 6

EVOLU

TION

0.05

SiOR

04648+10

SiOR01968+4

SiOR0

0899+7

SiOR02814+3

SiOR04171+6

SiO

R04609+4

SiOR00330+28

SiO

R02

694+25

SiO

R04609+20

SiOR05285+6

SiOR00330+25

SiOR04510+15

SiOR00330+18

SiOR04609+23

SiOR01

968+23

SiO

R03

952+4

SiOR0

4648+16

SiO

R05

901+2

SiO

R02

944+4

SiOR01968+5

SiOR04171+19 SiOR

04648+5

SiOR10535+3

SiOR0

1224+3

SiOR06723+2

SiOR01968+9

SiOR02883+1

SiOR0

0899+3

SiOR01629+1

SiOR04171+1

SiOR01629+11

SiOR04171+10

SiOR04171+13

SiOR02694+3

SiOR04171+20

SiO

R02694+35

SiOR04171+15

SiOR04609+7

SiOR05118+2

SiOR07

837+2

SiO

R02

694+27

SiOR01968+10

SiOR04648+17

SiOR0196

8+19

SiO

R02

694+17

SiOR04648+13

SiOR01968+6

SiOR

00899+1

2

SiOR0

5901+1

SiOR00330+20

SiO

R02

648+2

SiO

R02

659+2

SiOR01968+1

6

SiOR0

0899+11

SiO

R02974

SiOR04171+2

SiOR03952+2

SiOR06792+2

SiOR04510+4

SiOR04171+28

SiOR05285+5

SiOR05285+9

SiOR

00899+1

5

SiOR

04648+3

SiO

R02694+36

SiOR10535+1

SiO

R02

694+19

SiO

R02

694+23

SiOR02694+1

SiO

R04609+14

SiOR01122

SiOR

04648+9

SiO

R02694+34

SiOR01629+8

SiOR

04648+8

SiOR04510+8

SiOR06573

SiO

R02

944+1

SiOR00330+26

SiO

R00330+1S

iOR02

694+15

SiO

R03038

SiOR05285+7

SiOR0

0899+5

SiOR04609+10

SiO

R04609+3

SiOR04339

SiOR0

8068

SiOR

04648+6

SiOR04510+2

SiOR05285+8

SiOR01

573+4

SiOR0366

3

SiOR04171+8

SiOR01858+2

SiOR01968+2SiOR01968+1

SiOR02694+5

SiOR01968+3

SiOR06723+3

SiOR01968+15

SiOR05285+1

SiOR0

0899+4

SiOR04609+22

SiOR04171+9

SiOR02694+9

SiO

R02

648+1

SiOR06792+3

SiOR0

1573+2

SiO

R02

694+20

SiOR1

0542

SiO

R04609+15

SiOR02694+8

SiOR00330+16

SiOR0

0899+2

SiOR02694+10

SiOR04510+9

SiOR05285+3

SiOR04171+24

SiO

R04609+2

SiOR05285+11

SiOR02

694+14

SiOR01

573+1

SiOR05285+2

SiOR00613

SiOR0196

8+22

SiOR

00899+9

SiO

R06

843+2

SiO

R02694+37

SiOR0

0899+1

SiOR04609+9

SiOR05431+2

SiOR10535+2

SiOR00330+15

SiO

R02

694+18

SiOR0

1224+2

SiOR04510+11

SiOR00330+23

SiO

R02

694+29

SiOR05416

SiOR05285+10

SiOR02694+2

SiOR01629+9

SiOR05431+1

SiOR02883+2

SiOR04171+14

SiO

R08

341

SiO

R02

694+22

SiOR01

224+1

SiOR01968+12

SiOR05431+3

SiOR04510+6

SiOR02694+7

SiOR04609+5

SiO

R02

944+2

SiOR03

952+3

SiOR01968+8

SiOR04609+24

SiO

R02

694+30

SiOR01629+10

SiOR04510+14

SiOR00565

SiOR05118+3

SiO

R01321

SiO

R04609+19

SiOR00330+14

SiO

R02694+38

SiOR04609+8

SiOR04171+16

SiOR10455

SiO

R04609+16

SiO

R04609+21

SiO

R02

694+28

SiO

R02

659+1

SiOR04171+5

SiOR00330+29

SiOR01968+14

SiOR03983

SiOR00330+27

SiOR05285+4

SiOR04510+1

SiOR

00899+8

SiOR

04648+7

SiO

R04609+17

SiO

R00330+5

SiO

R02

694+21

SiOR02814+4

SiOR00330+7

SiOR01

629+3

SiOR01

968+26

SiO

R02694+31

SiOR0

4648+2

SiO

R02694+39

SiOR04171+25

SiOR06577

SiOR01

968+25

SiOR04171+21

SiOR06792+6

SiO

R04609+11

SiOR02694+11

SiOR06792+1

SiOR04171+4

SiOR01629+5

SiOR00330+21

SiOR0

4648+15

SiO

R00330+6

SiO

R02

694+16

SiOR

04648+11

SiOR

04648+4

SiO

R00330+3

SiO

R06

535

SiOR04171+7

SiOR10493

SiO

R02694+32

SiOR06792+4

SiOR04510+3

SiO

R06

890

SiOR0196

8+20

SiO

R04609+12

SiOR04171+3

SiOR01968

+18

SiOR01968+11

SiO

R04609+13

SiOR01629+12

SiOR00330+22

SiO

R02694+33

SiOR00330+13

SiOR01

573+3

SiOR04510+16

SiOR04171+12

SiOR04510+13

SiOR

05118+1

SiO

R02

944+3

SiOR04171+26

SiOR04171+17

SiOR

00899+14

SiOR04171+29

SiOR

00899+1

3

SiOR02694+13

SiOR00330+24

SiOR00330+19

SiOR04171+27

SiO

R02

694+24

SiOR04510+5

SiOR07090

SiOR03952+1

SiOR04510+10

SiOR00330+17

SiO

R02

694+26

SiO

R04609+1

SiOR0

0899+6

SiOR04648+12

SiOR02814+2

SiOR00330+11

SiOR04171+18

SiOR01968+1

7

SiOR00330+10

SiOR00330+9

SiOR0162

9+2

SiOR04171+11

SiOR04510+12

SiOR0196

8+21

SiOR00330+8

SiOR01858+1

SiOR04510+7

SiOR05431+4

SiOR02694+6

SiOR01968+13

SiO

R00330+4

SiO

R04609+18

SiOR0

0899+10

SiOR00330+12

SiOR00330+31

SiO

R06

843+1

SiOR07

837+1

SiO

R00330+2

SiOR01629+4

SiOR0

4648+1

SiOR01

968+24

SiOR04171+23

SiOR0162

9+7

SiOR0

4648+14

SiO

R06

843+3

SiOR06792+5

SiOR01968+7

SiOR01629

+6

SiOR02883+3

SiOR02694+12

SiOR

05118+4

SiOR04171+22

SiO

R01080

SiOR04609+6

SiOR02814+1

SiOR00330+30

SiOR05285+12

SiOR06723+1

SiOR02694+4


BB BB Bb

x x x

Single queen form Multiple queen form(>15% ) (< 5% )



Sequenced:"•a Gp-9 B � genome"

!

!•a Gp-9 b � genome

“Next Generation Genotyping.”RAD sequencing

“Next Generation Genotyping.”

Bb

unfertilised eggs

haploid �

Gp-9 B Gp-9 b Gp-9 B Gp-9 b Gp-9 b Gp-9 B

RAD sequencing

38 B� & 38 b�

EcoR1 EcoR1 EcoR1

RAD sequencing of haploid � for SNP discovery & genotypingGp-9 B

EcoR1 EcoR1 EcoR1

AACTG

AACTG

AACTG

AACTG

Gp-9 B

RAD sequencing of haploid � for SNP discovery & genotypingGp-9 B

AACTGGp-9 B

Gp-9 B

GGCCT

Gp-9 B

Gp-9 B

AAGGTGp-9 B

Gp-9 B

CCAGTGp-9 b

Gp-9 b

TAAATGp-9 b

Gp-9 b

GGAATGp-9 b

Gp-9 b

38 Gp-9 B "males

38 Gp-9 b "males

RAD sequencing of haploid � for SNP discovery & genotyping

Identify polymorphismindividual x locus

genotype table

RADseq: sequencing the same 0.01% of the genome in many individuals

A B C D E FL1 A C A A C CL2 G G T - T GL3 - A G A - GL4 C - - G G CL5 T T C T C -L6 G A A - - G

2419

loci

38 B� & 38 b�

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20+

Amount of variance explained per principal component

Principal Component

% V

aria

nce

Exp

lain

ed

05

1015

2025

30

12.7%

6.1% 5.4% 4.8% 4.7% 3.9% 3.5% 3.2% 3.1% 2.9% 2.8% 2.6% 2.4% 2.3% 2.2% 2.0% 1.9% 1.7% 1.6%

30.2%

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20+

Amount of variance explained per principal component

Principal Component

% V

aria

nce

Exp

lain

ed

05

1015

2025

30

PCA: Principal Component Analysis

pc: 2 % variance: 6.073

pc: 3

%

var

ianc

e: 5

.441

-0.2

-0.1

0.0

0.1

0.2

-0.2 -0.1 0.0 0.1 0.2pc: 2 % variance: 6.073

pc: 3

%

var

ianc

e: 5

.441

-0.2

-0.1

0.0

0.1

0.2

-0.2 -0.1 0.0 0.1 0.2

Principal Components: PC2 vs PC3

Gp-9 B �Gp-9 b �


pc: 2

%

var

ianc

e: 6

.073

-0.2

-0.1

0.0

0.1

0.2

-0.10 -0.05 0.00 0.05 0.10 0.15

Principal Components: PC1 vs PC2


pc: 2

%

var

ianc

e: 6

.073

-0.2

-0.1

0.0

0.1

0.2

-0.10 -0.05 0.00 0.05 0.10 0.15

Gp-9 B �Gp-9 b �

Amount of sequence linked to Gp-9

Confirmed in 8 additional families (sons of Bb queens) with RAD (3 families) and PCR (5 families). BBqueens: normal recombination throughout (RAD 3 families) ⟹ Gp-9 is not a “recombination cold-spot”

brc_m013_0001..brc_m013_0005brc_m013_0006..brc_m013_0014brc_m013_0015..brc_m013_0017brc_m013_0018brc_m013_0019..brc_m013_0020brc_m013_0021..brc_m013_0029brc_m013_0030..brc_m013_0031brc_m013_0032..brc_m013_0034brc_m013_0035..brc_m013_0036brc_m013_0037..brc_m013_0038brc_m013_0039..brc_m013_0043brc_m013_0044brc_m013_0045brc_m013_0046..brc_m013_0048brc_m013_0049brc_m013_0050..brc_m013_0051brc_m013_0052..brc_m013_0056brc_m013_0057..brc_m013_0061

brc_m013_0062..brc_m013_0075brc_m013_0076..brc_m013_0078brc_m013_0079..brc_m013_0081

brc_m013_0082..brc_m013_0088

brc_m013_0089..brc_m013_0092

brc_m013_0093..brc_m013_0096brc_m013_0097brc_m013_0098..brc_m013_0113brc_m013_0114..brc_m013_0119brc_m013_0120..brc_m013_0130brc_m013_0131brc_m013_0132..brc_m013_0134brc_m013_0135..brc_m013_0136brc_m013_0137..brc_m013_0139brc_m013_0140..brc_m013_0142brc_m013_0143brc_m013_0144..brc_m013_0146brc_m013_0147..brc_m013_0154brc_m013_0155

brc_m013_0156brc_m013_0157..brc_m013_0180brc_m013_0181brc_m013_0182brc_m013_0183..brc_m013_0188brc_m013_0189..brc_m013_0208brc_m013_0209brc_m013_0210..brc_m013_0211brc_m013_0212..brc_m013_0215brc_m013_0216brc_m013_0217brc_m013_0218..brc_m013_0224brc_m013_0225..brc_m013_0228brc_m013_0229..brc_m013_0237brc_m013_0238..brc_m013_0245

brc_m013_0246..brc_m013_0270

brc_m013_0271..brc_m013_0274brc_m013_0275brc_m013_0276..brc_m013_0278

brc_m013_0279

brc_m013_0280

brc_m013_0281..brc_m013_0294

brc_m013_0295..brc_m013_0303brc_m013_0304brc_m013_0305..brc_m013_0307brc_m013_0308brc_m013_0309..brc_m013_0314brc_m013_0315..brc_m013_0317brc_m013_0318..brc_m013_0320brc_m013_0321..brc_m013_0326brc_m013_0327brc_m013_0328brc_m013_0329..brc_m013_0330brc_m013_0331..brc_m013_0333brc_m013_0334..brc_m013_0339brc_m013_0340brc_m013_0341..brc_m013_0343brc_m013_0344brc_m013_0345..brc_m013_0349brc_m013_0350

brc_m013_0351..brc_m013_0354brc_m013_0355brc_m013_0356brc_m013_0357..brc_m013_0361brc_m013_0362..brc_m013_0376brc_m013_0377..brc_m013_0390brc_m013_0391..brc_m013_0393brc_m013_0394..brc_m013_0400brc_m013_0401..brc_m013_0439brc_m013_0440..brc_m013_0478brc_m013_0479..brc_m013_0480

0

20

40

60

80

100

120

140

160

180

LG1brc_m013_0481brc_m013_0482..brc_m013_0484brc_m013_0485..brc_m013_0488brc_m013_0489..brc_m013_0502brc_m013_0503brc_m013_0504..brc_m013_0519brc_m013_0520..brc_m013_0532brc_m013_0533..brc_m013_0534brc_m013_0535..brc_m013_0537brc_m013_0538..brc_m013_0540brc_m013_0541..brc_m013_0543brc_m013_0544..brc_m013_0545brc_m013_0546..brc_m013_0549

brc_m013_0550..brc_m013_0554

brc_m013_0555..brc_m013_0560

brc_m013_0561..brc_m013_0562brc_m013_0563..brc_m013_0565

brc_m013_0566brc_m013_0567brc_m013_0568..brc_m013_0570brc_m013_0571..brc_m013_0573

brc_m013_0574..brc_m013_0575brc_m013_0576..brc_m013_0578brc_m013_0579..brc_m013_0580brc_m013_0581brc_m013_0582..brc_m013_0584brc_m013_0585brc_m013_0586brc_m013_0587brc_m013_0588..brc_m013_0591brc_m013_0592brc_m013_0593brc_m013_0594..brc_m013_0612brc_m013_0613..brc_m013_0614brc_m013_0615..brc_m013_0632brc_m013_0633brc_m013_0634..brc_m013_0648brc_m013_0649..brc_m013_0655brc_m013_0656brc_m013_0657..brc_m013_0694brc_m013_0695..brc_m013_0703brc_m013_0704brc_m013_0705brc_m013_0706..brc_m013_0707brc_m013_0708brc_m013_0709..brc_m013_0711brc_m013_0712..brc_m013_0715brc_m013_0716..brc_m013_0721brc_m013_0722brc_m013_0723brc_m013_0724..brc_m013_0728brc_m013_0729brc_m013_0730brc_m013_0731brc_m013_0732..brc_m013_0735brc_m013_0736..brc_m013_0769brc_m013_0770..brc_m013_0771brc_m013_0772..brc_m013_0773brc_m013_0774..brc_m013_0775brc_m013_0776..brc_m013_0782brc_m013_0783brc_m013_0784..brc_m013_0795brc_m013_0796..brc_m013_0798brc_m013_0799..brc_m013_0801brc_m013_0802..brc_m013_0805brc_m013_0806..brc_m013_0809brc_m013_0810..brc_m013_0811brc_m013_0812..brc_m013_0824brc_m013_0825..brc_m013_0826brc_m013_0827..brc_m013_0829brc_m013_0830..brc_m013_0831brc_m013_0832..brc_m013_0842brc_m013_0843..brc_m013_0854brc_m013_0855..brc_m013_0861brc_m013_0862brc_m013_0863..brc_m013_0864brc_m013_0865..brc_m013_0867brc_m013_0868..brc_m013_0883brc_m013_0884..brc_m013_0893brc_m013_0894brc_m013_0895..brc_m013_0897brc_m013_0898..brc_m013_0906brc_m013_0907..brc_m013_0910brc_m013_0911..brc_m013_0925brc_m013_0926..brc_m013_0928brc_m013_0929..brc_m013_0931

0

20

40

60

80

100

120

140

LG2brc_m013_0932..brc_m013_0941brc_m013_0942..brc_m013_0943brc_m013_0944..brc_m013_0945brc_m013_0946..brc_m013_0949brc_m013_0950..brc_m013_0952

brc_m013_0953..brc_m013_0975

brc_m013_0976..brc_m013_1019brc_m013_1020brc_m013_1021brc_m013_1022..brc_m013_1061brc_m013_1062brc_m013_1063

brc_m013_1064..brc_m013_1065

brc_m013_1066..brc_m013_1068brc_m013_1069brc_m013_1070

brc_m013_1071..brc_m013_1074

brc_m013_1075

brc_m013_1076..brc_m013_1081

brc_m013_1082..brc_m013_1086brc_m013_1087..brc_m013_1088brc_m013_1089..brc_m013_1098brc_m013_1099..brc_m013_1106brc_m013_1107..brc_m013_1116brc_m013_1117brc_m013_1118..brc_m013_1121brc_m013_1122..brc_m013_1127brc_m013_1128brc_m013_1129..brc_m013_1136brc_m013_1137..brc_m013_1138brc_m013_1139..brc_m013_1141brc_m013_1142..brc_m013_1144brc_m013_1145..brc_m013_1156brc_m013_1157brc_m013_1158..brc_m013_1170brc_m013_1171..brc_m013_1181brc_m013_1182..brc_m013_1185brc_m013_1186brc_m013_1187..brc_m013_1205brc_m013_1206..brc_m013_1218brc_m013_1219..brc_m013_1220brc_m013_1221..brc_m013_1224brc_m013_1225..brc_m013_1228brc_m013_1229brc_m013_1230..brc_m013_1236brc_m013_1237brc_m013_1238..brc_m013_1247brc_m013_1248..brc_m013_1251

brc_m013_1252brc_m013_1253..brc_m013_1268brc_m013_1269..brc_m013_1270brc_m013_1271..brc_m013_1273brc_m013_1274brc_m013_1275..brc_m013_1280brc_m013_1281

brc_m013_1282..brc_m013_1286brc_m013_1287..brc_m013_1298brc_m013_1299..brc_m013_1307brc_m013_1308brc_m013_1309..brc_m013_1313brc_m013_1314..brc_m013_1317brc_m013_1318..brc_m013_1319brc_m013_1320..brc_m013_1326brc_m013_1327..brc_m013_1340brc_m013_1341..brc_m013_1362brc_m013_1363..brc_m013_1385

0

20

40

60

80

100

120

140

LG3brc_m013_1386..brc_m013_1388brc_m013_1389..brc_m013_1398brc_m013_1399..brc_m013_1406brc_m013_1407..brc_m013_1411brc_m013_1412..brc_m013_1413brc_m013_1414..brc_m013_1416

brc_m013_1417brc_m013_1418..brc_m013_1420brc_m013_1421..brc_m013_1424brc_m013_1425..brc_m013_1432brc_m013_1433..brc_m013_1442brc_m013_1443brc_m013_1444..brc_m013_1450brc_m013_1451brc_m013_1452brc_m013_1453..brc_m013_1455brc_m013_1456..brc_m013_1467brc_m013_1468..brc_m013_1469brc_m013_1470brc_m013_1471..brc_m013_1474brc_m013_1475brc_m013_1476brc_m013_1477brc_m013_1478..brc_m013_1482brc_m013_1483

brc_m013_1484brc_m013_1485..brc_m013_1487brc_m013_1488..brc_m013_1490


brc_m013_1497..brc_m013_1500brc_m013_1501brc_m013_1502..brc_m013_1513brc_m013_1514..brc_m013_1562brc_m013_1563..brc_m013_1565brc_m013_1566..brc_m013_1567

brc_m013_1568..brc_m013_1580brc_m013_1581..brc_m013_1587brc_m013_1588..brc_m013_1591brc_m013_1592..brc_m013_1593brc_m013_1594..brc_m013_1604brc_m013_1605..brc_m013_1607brc_m013_1608..brc_m013_1609brc_m013_1610..brc_m013_1611brc_m013_1612..brc_m013_1616brc_m013_1617..brc_m013_1618brc_m013_1619..brc_m013_1620brc_m013_1621..brc_m013_1629brc_m013_1630..brc_m013_1633brc_m013_1634..brc_m013_1638brc_m013_1639..brc_m013_1647brc_m013_1648..brc_m013_1649brc_m013_1650..brc_m013_1656brc_m013_1657..brc_m013_1665brc_m013_1666..brc_m013_1672brc_m013_1673..brc_m013_1674brc_m013_1675..brc_m013_1678brc_m013_1679..brc_m013_1682brc_m013_1683brc_m013_1684brc_m013_1685..brc_m013_1686brc_m013_1687..brc_m013_1700brc_m013_1701..brc_m013_1702brc_m013_1703brc_m013_1704..brc_m013_1707brc_m013_1708..brc_m013_1709brc_m013_1710..brc_m013_1714brc_m013_1715..brc_m013_1728brc_m013_1729..brc_m013_1742

0

20

40

60

80

100

120

140

LG4brc_m013_1743..brc_m013_1750


brc_m013_1769..brc_m013_1772

brc_m013_1773..brc_m013_1779brc_m013_1780brc_m013_1781brc_m013_1782..brc_m013_1783brc_m013_1784brc_m013_1785..brc_m013_1786

brc_m013_1787

brc_m013_1788brc_m013_1789..brc_m013_1790brc_m013_1791..brc_m013_1793brc_m013_1794..brc_m013_1797brc_m013_1798..brc_m013_1800brc_m013_1801..brc_m013_1804brc_m013_1805brc_m013_1806..brc_m013_1808brc_m013_1809brc_m013_1810..brc_m013_1813brc_m013_1814..brc_m013_1818brc_m013_1819..brc_m013_1820brc_m013_1821..brc_m013_1822brc_m013_1823..brc_m013_1824brc_m013_1825brc_m013_1826..brc_m013_1840brc_m013_1841..brc_m013_1842brc_m013_1843brc_m013_1844..brc_m013_1848brc_m013_1849brc_m013_1850brc_m013_1851..brc_m013_1853brc_m013_1854..brc_m013_1858brc_m013_1859..brc_m013_1866brc_m013_1867..brc_m013_1868brc_m013_1869brc_m013_1870..brc_m013_1874brc_m013_1875..brc_m013_1876brc_m013_1877..brc_m013_1878brc_m013_1879..brc_m013_1883brc_m013_1884brc_m013_1885brc_m013_1886..brc_m013_1888brc_m013_1889..brc_m013_1895brc_m013_1896..brc_m013_1899brc_m013_1900..brc_m013_1913brc_m013_1914brc_m013_1915..brc_m013_1922brc_m013_1923brc_m013_1924..brc_m013_1928brc_m013_1929..brc_m013_1942brc_m013_1943..brc_m013_1946brc_m013_1947..brc_m013_1948brc_m013_1949brc_m013_1950..brc_m013_1953brc_m013_1954..brc_m013_1955brc_m013_1956brc_m013_1957..brc_m013_1963brc_m013_1964..brc_m013_1966brc_m013_1967..brc_m013_1968brc_m013_1969..brc_m013_1970brc_m013_1971brc_m013_1972brc_m013_1973..brc_m013_1980brc_m013_1981..brc_m013_1983brc_m013_1984..brc_m013_1990brc_m013_1991..brc_m013_1993brc_m013_1994..brc_m013_1996brc_m013_1997..brc_m013_2009

0

20

40

60

80

100

120

LG5brc_m013_2010..brc_m013_2028

brc_m013_2029..brc_m013_2038brc_m013_2039

brc_m013_2040..brc_m013_2041brc_m013_2042..brc_m013_2047brc_m013_2048..brc_m013_2050brc_m013_2051..brc_m013_2053brc_m013_2054..brc_m013_2062brc_m013_2063brc_m013_2064..brc_m013_2065brc_m013_2066..brc_m013_2067brc_m013_2068brc_m013_2069..brc_m013_2071brc_m013_2072..brc_m013_2081brc_m013_2082

brc_m013_2083brc_m013_2084brc_m013_2085..brc_m013_2099brc_m013_2100brc_m013_2101..brc_m013_2102brc_m013_2103..brc_m013_2107brc_m013_2108brc_m013_2109..brc_m013_2112

brc_m013_2113..brc_m013_2114brc_m013_2115..brc_m013_2123brc_m013_2124..brc_m013_2131brc_m013_2132brc_m013_2133brc_m013_2134..brc_m013_2136

brc_m013_2137

brc_m013_2138..brc_m013_2139

brc_m013_2140..brc_m013_2142brc_m013_2143..brc_m013_2150brc_m013_2151..brc_m013_2152brc_m013_2153..brc_m013_2161brc_m013_2162..brc_m013_2163

brc_m013_2164..brc_m013_2165brc_m013_2166..brc_m013_2170brc_m013_2171..brc_m013_2172brc_m013_2173brc_m013_2174..brc_m013_2182brc_m013_2183..brc_m013_2186brc_m013_2187..brc_m013_2190brc_m013_2191..brc_m013_2193brc_m013_2194brc_m013_2195..brc_m013_2201brc_m013_2202..brc_m013_2203brc_m013_2204..brc_m013_2220brc_m013_2221..brc_m013_2232brc_m013_2233..brc_m013_2239brc_m013_2240..brc_m013_2261brc_m013_2262..brc_m013_2267brc_m013_2268..brc_m013_2269brc_m013_2270..brc_m013_2271brc_m013_2272..brc_m013_2282brc_m013_2283..brc_m013_2284brc_m013_2285..brc_m013_2299brc_m013_2300..brc_m013_2301brc_m013_2302..brc_m013_2305brc_m013_2306..brc_m013_2307brc_m013_2308..brc_m013_2330brc_m013_2331..brc_m013_2337brc_m013_2338..brc_m013_2352

0

20

40

60

80

100

120

LG6brc_m013_2353..brc_m013_2365brc_m013_2366..brc_m013_2369

brc_m013_2370..brc_m013_2372brc_m013_2373..brc_m013_2378brc_m013_2379..brc_m013_2386brc_m013_2387brc_m013_2388..brc_m013_2394brc_m013_2395..brc_m013_2397brc_m013_2398brc_m013_2399brc_m013_2400brc_m013_2401brc_m013_2402..brc_m013_2407brc_m013_2408..brc_m013_2411brc_m013_2412..brc_m013_2416brc_m013_2417brc_m013_2418brc_m013_2419..brc_m013_2436brc_m013_2437..brc_m013_2441brc_m013_2442brc_m013_2443..brc_m013_2444brc_m013_2445brc_m013_2446..brc_m013_2453brc_m013_2454brc_m013_2455..brc_m013_2460brc_m013_2461brc_m013_2462..brc_m013_2470brc_m013_2471..brc_m013_2474brc_m013_2475..brc_m013_2482brc_m013_2483brc_m013_2484..brc_m013_2487brc_m013_2488brc_m013_2489..brc_m013_2492brc_m013_2493..brc_m013_2496brc_m013_2497brc_m013_2498..brc_m013_2504brc_m013_2505brc_m013_2506..brc_m013_2510brc_m013_2511..brc_m013_2523brc_m013_2524..brc_m013_2531brc_m013_2532..brc_m013_2536brc_m013_2537..brc_m013_2555brc_m013_2556..brc_m013_2571brc_m013_2572..brc_m013_2573brc_m013_2574..brc_m013_2579brc_m013_2580..brc_m013_2581brc_m013_2582..brc_m013_2587brc_m013_2588brc_m013_2589..brc_m013_2594

brc_m013_2595

brc_m013_2596..brc_m013_2597brc_m013_2598..brc_m013_2604brc_m013_2605brc_m013_2606..brc_m013_2616brc_m013_2617..brc_m013_2619brc_m013_2620..brc_m013_2623


brc_m013_2629..brc_m013_2630

0

20

40

60

80

100

LG7brc_m013_2631..brc_m013_2632brc_m013_2633brc_m013_2634..brc_m013_2635brc_m013_2636..brc_m013_2642brc_m013_2643..brc_m013_2657brc_m013_2658..brc_m013_2659brc_m013_2660..brc_m013_2661brc_m013_2662

brc_m013_2663

brc_m013_2664brc_m013_2665..brc_m013_2666

brc_m013_2667..brc_m013_2668brc_m013_2669..brc_m013_2670brc_m013_2671..brc_m013_2680brc_m013_2681..brc_m013_2682brc_m013_2683brc_m013_2684..brc_m013_2685brc_m013_2686..brc_m013_2694


brc_m013_2714..brc_m013_2725brc_m013_2726..brc_m013_2727brc_m013_2728..brc_m013_2731brc_m013_2732brc_m013_2733..brc_m013_2753brc_m013_2754..brc_m013_2758brc_m013_2759..brc_m013_2763brc_m013_2764..brc_m013_2779brc_m013_2780brc_m013_2781brc_m013_2782..brc_m013_2784brc_m013_2785..brc_m013_2787brc_m013_2788..brc_m013_2791brc_m013_2792..brc_m013_2797brc_m013_2798..brc_m013_2799brc_m013_2800brc_m013_2801..brc_m013_2804brc_m013_2805..brc_m013_2809

brc_m013_2810..brc_m013_2811

brc_m013_2812..brc_m013_2813brc_m013_2814..brc_m013_2817brc_m013_2818..brc_m013_2827brc_m013_2828brc_m013_2829..brc_m013_2832brc_m013_2833brc_m013_2834..brc_m013_2840brc_m013_2841..brc_m013_2846brc_m013_2847brc_m013_2848..brc_m013_2852brc_m013_2853brc_m013_2854..brc_m013_2856brc_m013_2857..brc_m013_2862brc_m013_2863..brc_m013_2868brc_m013_2869..brc_m013_2874brc_m013_2875..brc_m013_2896

0

20

40

60

80

100

LG8

brc_m013_2897..brc_m013_2920brc_m013_2921..brc_m013_2928brc_m013_2929..brc_m013_2931brc_m013_2932

brc_m013_2933

brc_m013_2934..brc_m013_2935brc_m013_2936brc_m013_2937..brc_m013_2943brc_m013_2944brc_m013_2945..brc_m013_2946brc_m013_2947brc_m013_2948brc_m013_2949..brc_m013_2950brc_m013_2951..brc_m013_2957brc_m013_2958..brc_m013_2961brc_m013_2962..brc_m013_2970brc_m013_2971..brc_m013_2980brc_m013_2981..brc_m013_2992brc_m013_2993..brc_m013_2996brc_m013_2997..brc_m013_2998brc_m013_2999..brc_m013_3000brc_m013_3001brc_m013_3002..brc_m013_3003brc_m013_3004brc_m013_3005brc_m013_3006..brc_m013_3010brc_m013_3011..brc_m013_3014brc_m013_3015brc_m013_3016..brc_m013_3019brc_m013_3020brc_m013_3021..brc_m013_3030brc_m013_3031..brc_m013_3032brc_m013_3033..brc_m013_3034brc_m013_3035..brc_m013_3036brc_m013_3037..brc_m013_3045brc_m013_3046..brc_m013_3052brc_m013_3053brc_m013_3054..brc_m013_3061brc_m013_3062..brc_m013_3066brc_m013_3067..brc_m013_3068brc_m013_3069..brc_m013_3076brc_m013_3077..brc_m013_3084brc_m013_3085..brc_m013_3087brc_m013_3088..brc_m013_3089brc_m013_3090..brc_m013_3096brc_m013_3097..brc_m013_3100brc_m013_3101..brc_m013_3104brc_m013_3105brc_m013_3106..brc_m013_3112brc_m013_3113..brc_m013_3122brc_m013_3123..brc_m013_3124brc_m013_3125..brc_m013_3127brc_m013_3128..brc_m013_3145brc_m013_3146..brc_m013_3159brc_m013_3160..brc_m013_3172

0

20

40

60

80

100

LG9brc_m013_3173..brc_m013_3175brc_m013_3176..brc_m013_3180brc_m013_3181..brc_m013_3189brc_m013_3190..brc_m013_3198brc_m013_3199brc_m013_3200..brc_m013_3201brc_m013_3202..brc_m013_3203brc_m013_3204brc_m013_3205..brc_m013_3206brc_m013_3207..brc_m013_3211brc_m013_3212..brc_m013_3214brc_m013_3215..brc_m013_3227brc_m013_3228..brc_m013_3230brc_m013_3231..brc_m013_3235brc_m013_3236..brc_m013_3238brc_m013_3239..brc_m013_3242brc_m013_3243..brc_m013_3244brc_m013_3245brc_m013_3246..brc_m013_3247brc_m013_3248..brc_m013_3249brc_m013_3250..brc_m013_3252brc_m013_3253..brc_m013_3257brc_m013_3258brc_m013_3259brc_m013_3260..brc_m013_3261brc_m013_3262..brc_m013_3263brc_m013_3264brc_m013_3265..brc_m013_3269brc_m013_3270..brc_m013_3274brc_m013_3275..brc_m013_3276brc_m013_3277..brc_m013_3281brc_m013_3282..brc_m013_3284brc_m013_3285brc_m013_3286..brc_m013_3289brc_m013_3290..brc_m013_3296brc_m013_3297brc_m013_3298..brc_m013_3300brc_m013_3301..brc_m013_3302brc_m013_3303..brc_m013_3305brc_m013_3306..brc_m013_3308brc_m013_3309..brc_m013_3314brc_m013_3315..brc_m013_3317brc_m013_3318..brc_m013_3329brc_m013_3330..brc_m013_3331brc_m013_3332..brc_m013_3338brc_m013_3339..brc_m013_3340brc_m013_3341..brc_m013_3344brc_m013_3345..brc_m013_3349brc_m013_3350..brc_m013_3357brc_m013_3358..brc_m013_3359brc_m013_3360brc_m013_3361..brc_m013_3368brc_m013_3369..brc_m013_3372brc_m013_3373..brc_m013_3376brc_m013_3377brc_m013_3378..brc_m013_3386brc_m013_3387..brc_m013_3388brc_m013_3389..brc_m013_3395brc_m013_3396..brc_m013_3399

0

20

40

60

80

LG10brc_m013_3400..brc_m013_3411brc_m013_3412brc_m013_3413..brc_m013_3424brc_m013_3425brc_m013_3426brc_m013_3427..brc_m013_3429

brc_m013_3430

brc_m013_3431..brc_m013_3432brc_m013_3433..brc_m013_3435brc_m013_3436brc_m013_3437..brc_m013_3439

brc_m013_3440..brc_m013_3441brc_m013_3442

brc_m013_3443..brc_m013_3445brc_m013_3446..brc_m013_3447brc_m013_3448..brc_m013_3449brc_m013_3450..brc_m013_3454brc_m013_3455brc_m013_3456..brc_m013_3462brc_m013_3463..brc_m013_3464brc_m013_3465brc_m013_3466..brc_m013_3467brc_m013_3468..brc_m013_3472brc_m013_3473brc_m013_3474..brc_m013_3476brc_m013_3477..brc_m013_3487brc_m013_3488brc_m013_3489..brc_m013_3491brc_m013_3492..brc_m013_3500brc_m013_3501..brc_m013_3512brc_m013_3513..brc_m013_3514brc_m013_3515..brc_m013_3524brc_m013_3525..brc_m013_3527brc_m013_3528..brc_m013_3531brc_m013_3532..brc_m013_3547brc_m013_3548..brc_m013_3557brc_m013_3558..brc_m013_3566brc_m013_3567..brc_m013_3568brc_m013_3569..brc_m013_3570brc_m013_3571..brc_m013_3574brc_m013_3575..brc_m013_3582brc_m013_3583..brc_m013_3592brc_m013_3593..brc_m013_3605brc_m013_3606..brc_m013_3616brc_m013_3617..brc_m013_3618brc_m013_3619..brc_m013_3622brc_m013_3623..brc_m013_3624brc_m013_3625..brc_m013_3628brc_m013_3629..brc_m013_3635

0

20

40

60

80

LG11



brc_m013_3672


brc_m013_3683..brc_m013_3685

brc_m013_3686..brc_m013_3688

brc_m013_3689..brc_m013_3693

brc_m013_3694..brc_m013_3698brc_m013_3699

brc_m013_3700brc_m013_3701..brc_m013_3702brc_m013_3703..brc_m013_3704brc_m013_3705..brc_m013_3712brc_m013_3713brc_m013_3714..brc_m013_3716brc_m013_3717..brc_m013_3724brc_m013_3725..brc_m013_3730brc_m013_3731..brc_m013_3752brc_m013_3753..brc_m013_3758brc_m013_3759..brc_m013_3789brc_m013_3790..brc_m013_3801brc_m013_3802..brc_m013_3814brc_m013_3815..brc_m013_3818brc_m013_3819..brc_m013_3822brc_m013_3823..brc_m013_3826brc_m013_3827..brc_m013_3832brc_m013_3833..brc_m013_3837brc_m013_3838brc_m013_3839..brc_m013_3841brc_m013_3842..brc_m013_3847

brc_m013_3848..brc_m013_3853brc_m013_3854..brc_m013_3858brc_m013_3859..brc_m013_3868brc_m013_3869..brc_m013_3871brc_m013_3872..brc_m013_3901brc_m013_3902brc_m013_3903..brc_m013_3909brc_m013_3910brc_m013_3911..brc_m013_3926brc_m013_3927..brc_m013_3931brc_m013_3932..brc_m013_3948

0

20

40

60

80

LG12


brc_m013_3972..brc_m013_3975brc_m013_3976

brc_m013_3977..brc_m013_3985

brc_m013_3986brc_m013_3987..brc_m013_3994brc_m013_3995..brc_m013_3997brc_m013_3998..brc_m013_4004

brc_m013_4005..brc_m013_4006brc_m013_4007..brc_m013_4008brc_m013_4009..brc_m013_4010brc_m013_4011..brc_m013_4013brc_m013_4014brc_m013_4015brc_m013_4016..brc_m013_4019brc_m013_4020..brc_m013_4021brc_m013_4022..brc_m013_4025brc_m013_4026..brc_m013_4032brc_m013_4033..brc_m013_4036brc_m013_4037..brc_m013_4041brc_m013_4042..brc_m013_4043

brc_m013_4044..brc_m013_4046brc_m013_4047..brc_m013_4056brc_m013_4057brc_m013_4058..brc_m013_4063brc_m013_4064..brc_m013_4071brc_m013_4072..brc_m013_4075brc_m013_4076brc_m013_4077brc_m013_4078..brc_m013_4085brc_m013_4086..brc_m013_4089brc_m013_4090..brc_m013_4091brc_m013_4092..brc_m013_4093brc_m013_4094..brc_m013_4095brc_m013_4096..brc_m013_4114brc_m013_4115..brc_m013_4117brc_m013_4118..brc_m013_4131brc_m013_4132..brc_m013_4133brc_m013_4134..brc_m013_4146

0

20

40

60

80

LG13


brc_m013_4168brc_m013_4169

brc_m013_4170brc_m013_4171..brc_m013_4172brc_m013_4173..brc_m013_4175brc_m013_4176brc_m013_4177..brc_m013_4178brc_m013_4179..brc_m013_4183brc_m013_4184..brc_m013_4185brc_m013_4186..brc_m013_4187brc_m013_4188..brc_m013_4191brc_m013_4192brc_m013_4193..brc_m013_4194brc_m013_4195..brc_m013_4206brc_m013_4207..brc_m013_4210brc_m013_4211..brc_m013_4213brc_m013_4214brc_m013_4215..brc_m013_4217brc_m013_4218..brc_m013_4221brc_m013_4222..brc_m013_4223brc_m013_4224..brc_m013_4231brc_m013_4232..brc_m013_4234brc_m013_4235..brc_m013_4239brc_m013_4240..brc_m013_4246brc_m013_4247..brc_m013_4248brc_m013_4249..brc_m013_4258brc_m013_4259..brc_m013_4260brc_m013_4261..brc_m013_4269brc_m013_4270..brc_m013_4271brc_m013_4272..brc_m013_4278brc_m013_4279..brc_m013_4280brc_m013_4281..brc_m013_4284brc_m013_4285brc_m013_4286..brc_m013_4288brc_m013_4289..brc_m013_4294brc_m013_4295..brc_m013_4296brc_m013_4297..brc_m013_4301brc_m013_4302..brc_m013_4313brc_m013_4314..brc_m013_4320brc_m013_4321..brc_m013_4322brc_m013_4323..brc_m013_4345brc_m013_4346..brc_m013_4351

0

20

40

60

LG14brc_m013_4352..brc_m013_4366brc_m013_4367brc_m013_4368brc_m013_4369..brc_m013_4373brc_m013_4374..brc_m013_4381brc_m013_4382..brc_m013_4383brc_m013_4384..brc_m013_4385brc_m013_4386brc_m013_4387..brc_m013_4388brc_m013_4389brc_m013_4390..brc_m013_4404brc_m013_4405brc_m013_4406..brc_m013_4409brc_m013_4410..brc_m013_4411brc_m013_4412..brc_m013_4418brc_m013_4419..brc_m013_4434brc_m013_4435..brc_m013_4442brc_m013_4443..brc_m013_4448brc_m013_4449..brc_m013_4451brc_m013_4452..brc_m013_4461brc_m013_4462..brc_m013_4471brc_m013_4472..brc_m013_4475brc_m013_4476..brc_m013_4477brc_m013_4478brc_m013_4479brc_m013_4480..brc_m013_4485brc_m013_4486brc_m013_4487..brc_m013_4491brc_m013_4492brc_m013_4493brc_m013_4494..brc_m013_4495brc_m013_4496..brc_m013_4501brc_m013_4502..brc_m013_4510brc_m013_4511..brc_m013_4531brc_m013_4532brc_m013_4533..brc_m013_4534brc_m013_4535..brc_m013_4541brc_m013_4542..brc_m013_4543brc_m013_4544..brc_m013_4545brc_m013_4546..brc_m013_4548brc_m013_4549..brc_m013_4551brc_m013_4552..brc_m013_4555brc_m013_4556..brc_m013_4561

0

20

40

60

LG15

brc_m013_4562..brc_m013_4577brc_m013_4578..brc_m013_4594brc_m013_4595..brc_m013_4599brc_m013_4600..brc_m013_4625brc_m013_4626..brc_m013_4638brc_m013_4639..brc_m013_4642brc_m013_4643..brc_m013_4644brc_m013_4645..brc_m013_4650brc_m013_4651..brc_m013_4663brc_m013_4664..brc_m013_4668brc_m013_4669..brc_m013_4670brc_m013_4671..brc_m013_4674brc_m013_4675..brc_m013_4679brc_m013_4680..brc_m013_4681brc_m013_4682brc_m013_4683..brc_m013_4688brc_m013_4689..brc_m013_4692brc_m013_4693..brc_m013_4695brc_m013_4696..brc_m013_4701brc_m013_4702brc_m013_4703..brc_m013_4712brc_m013_4713..brc_m013_4717brc_m013_4718..brc_m013_4720brc_m013_4721brc_m013_4722..brc_m013_4726brc_m013_4727..brc_m013_4728brc_m013_4729..brc_m013_4742brc_m013_4743brc_m013_4744..brc_m013_4746brc_m013_4747brc_m013_4748..brc_m013_4749brc_m013_4750..brc_m013_4752

brc_m013_4753..brc_m013_4756brc_m013_4757..brc_m013_4759brc_m013_4760..brc_m013_4762brc_m013_4763brc_m013_4764..brc_m013_4766brc_m013_4767..brc_m013_4769brc_m013_4770brc_m013_4771..brc_m013_4774brc_m013_4775..brc_m013_4776brc_m013_4777..brc_m013_4778brc_m013_4779..brc_m013_4780brc_m013_4781..brc_m013_4793brc_m013_4794brc_m013_4795..brc_m013_4798brc_m013_4799..brc_m013_4802brc_m013_4803..brc_m013_4806brc_m013_4807brc_m013_4808..brc_m013_4814brc_m013_4815..brc_m013_4819brc_m013_4820brc_m013_4821brc_m013_4822..brc_m013_4823brc_m013_4824brc_m013_4825..brc_m013_4855brc_m013_4856..brc_m013_4858brc_m013_4859..brc_m013_4863brc_m013_4864..brc_m013_4865brc_m013_4866..brc_m013_4875brc_m013_4876..brc_m013_4881brc_m013_4882..brc_m013_4891brc_m013_4892..brc_m013_4895brc_m013_4896..brc_m013_4911brc_m013_4912..brc_m013_4938brc_m013_4939brc_m013_4940..brc_m013_4957brc_m013_4958..brc_m013_4959brc_m013_4960..brc_m013_4972brc_m013_4973..brc_m013_4981brc_m013_4982..brc_m013_4983

0

20

40

60

80

100

120

LGSB

Gp-9

Figure 1a b

Si_gnF.scaffold00779_nt2778431.7

Si_gnF.scaffold00779_nt1255229Si_gnF.scaffold02684_nt10884.2


Si_gnF.scaffold00779_nt3746833Si_gnF.scaffold00779_nt374687927.3Si_gnF.scaffold00779_nt382158728.5




Si_gnF.scaffold05266_nt63430678.4Si_gnF.scaffold05266_nt65952779.7





Si_gnF.scaffold03404_nt128925Si_gnF.scaffold03404_nt228606Si_gnF.scaffold03404_nt241461

104.4


Si_gnF.scaffold00413_nt1035856109.6Si_gnF.scaffold01573_nt108462110.8Si_gnF.scaffold01573_nt447618112.1Si_gnF.scaffold00899_nt377419Si_gnF.scaffold00899_nt686574114.4Si_gnF.scaffold00899_nt236146Si_gnF.scaffold00899_nt332715Si_gnF.scaffold00899_nt335756

115.8

Si_gnF.scaffold00469_nt794118.0Si_gnF.scaffold00690_nt229012Si_gnF.scaffold00690_nt415290119.3



LGS Bfrom M013




22.2



Si_gnF.scaffold01573_nt10846280.7Si_gnF.scaffold00255_nt314067Si_gnF.scaffold00255_nt407778Si_gnF.scaffold00413_nt1035856Si_gnF.scaffold00413_nt676115Si_gnF.scaffold00469_nt794Si_gnF.scaffold00690_nt229012Si_gnF.scaffold00690_nt415290Si_gnF.scaffold00899_nt236146Si_gnF.scaffold00899_nt332715Si_gnF.scaffold00899_nt335756Si_gnF.scaffold00899_nt377419Si_gnF.scaffold00899_nt686574Si_gnF.scaffold01573_nt447618Si_gnF.scaffold01957_nt412242Si_gnF.scaffold03404_nt128925Si_gnF.scaffold03404_nt228606Si_gnF.scaffold03404_nt241461Si_gnF.scaffold05266_nt634306Si_gnF.scaffold05266_nt659527Si_gnF.scaffold05266_nt733643Si_gnF.scaffold05266_nt753644Si_gnF.scaffold06914_nt297673Si_gnF.scaffold07090_nt1051771Si_gnF.scaffold07090_nt710010Si_gnF.scaffold09758_nt222732

81.8


LGS B/bfrom P034

Gp-9

Total285 non-recombiningmarkersbrc_m013_0001..brc_m013_0005

brc_m013_0006..brc_m013_0014brc_m013_0015..brc_m013_0017brc_m013_0018brc_m013_0019..brc_m013_0020brc_m013_0021..brc_m013_0029brc_m013_0030..brc_m013_0031brc_m013_0032..brc_m013_0034brc_m013_0035..brc_m013_0036brc_m013_0037..brc_m013_0038brc_m013_0039..brc_m013_0043brc_m013_0044brc_m013_0045brc_m013_0046..brc_m013_0048brc_m013_0049brc_m013_0050..brc_m013_0051brc_m013_0052..brc_m013_0056brc_m013_0057..brc_m013_0061

brc_m013_0062..brc_m013_0075brc_m013_0076..brc_m013_0078brc_m013_0079..brc_m013_0081

brc_m013_0082..brc_m013_0088

brc_m013_0089..brc_m013_0092

brc_m013_0093..brc_m013_0096brc_m013_0097brc_m013_0098..brc_m013_0113brc_m013_0114..brc_m013_0119brc_m013_0120..brc_m013_0130brc_m013_0131brc_m013_0132..brc_m013_0134brc_m013_0135..brc_m013_0136brc_m013_0137..brc_m013_0139brc_m013_0140..brc_m013_0142brc_m013_0143brc_m013_0144..brc_m013_0146brc_m013_0147..brc_m013_0154brc_m013_0155

brc_m013_0156brc_m013_0157..brc_m013_0180brc_m013_0181brc_m013_0182brc_m013_0183..brc_m013_0188brc_m013_0189..brc_m013_0208brc_m013_0209brc_m013_0210..brc_m013_0211brc_m013_0212..brc_m013_0215brc_m013_0216brc_m013_0217brc_m013_0218..brc_m013_0224brc_m013_0225..brc_m013_0228brc_m013_0229..brc_m013_0237brc_m013_0238..brc_m013_0245

brc_m013_0246..brc_m013_0270


brc_m013_0279

brc_m013_0280

brc_m013_0281..brc_m013_0294

brc_m013_0295..brc_m013_0303brc_m013_0304brc_m013_0305..brc_m013_0307brc_m013_0308brc_m013_0309..brc_m013_0314brc_m013_0315..brc_m013_0317brc_m013_0318..brc_m013_0320brc_m013_0321..brc_m013_0326brc_m013_0327brc_m013_0328brc_m013_0329..brc_m013_0330brc_m013_0331..brc_m013_0333brc_m013_0334..brc_m013_0339brc_m013_0340brc_m013_0341..brc_m013_0343brc_m013_0344brc_m013_0345..brc_m013_0349brc_m013_0350

brc_m013_0351..brc_m013_0354brc_m013_0355brc_m013_0356brc_m013_0357..brc_m013_0361brc_m013_0362..brc_m013_0376brc_m013_0377..brc_m013_0390brc_m013_0391..brc_m013_0393brc_m013_0394..brc_m013_0400brc_m013_0401..brc_m013_0439brc_m013_0440..brc_m013_0478brc_m013_0479..brc_m013_0480

0

20

40

60

80

100

120

140

160

180

LG1brc_m013_0481brc_m013_0482..brc_m013_0484brc_m013_0485..brc_m013_0488brc_m013_0489..brc_m013_0502brc_m013_0503brc_m013_0504..brc_m013_0519brc_m013_0520..brc_m013_0532brc_m013_0533..brc_m013_0534brc_m013_0535..brc_m013_0537brc_m013_0538..brc_m013_0540brc_m013_0541..brc_m013_0543brc_m013_0544..brc_m013_0545brc_m013_0546..brc_m013_0549

brc_m013_0550..brc_m013_0554

brc_m013_0555..brc_m013_0560


brc_m013_0566brc_m013_0567brc_m013_0568..brc_m013_0570brc_m013_0571..brc_m013_0573

brc_m013_0574..brc_m013_0575brc_m013_0576..brc_m013_0578brc_m013_0579..brc_m013_0580brc_m013_0581brc_m013_0582..brc_m013_0584brc_m013_0585brc_m013_0586brc_m013_0587brc_m013_0588..brc_m013_0591brc_m013_0592brc_m013_0593brc_m013_0594..brc_m013_0612brc_m013_0613..brc_m013_0614brc_m013_0615..brc_m013_0632brc_m013_0633brc_m013_0634..brc_m013_0648brc_m013_0649..brc_m013_0655brc_m013_0656brc_m013_0657..brc_m013_0694brc_m013_0695..brc_m013_0703brc_m013_0704brc_m013_0705brc_m013_0706..brc_m013_0707brc_m013_0708brc_m013_0709..brc_m013_0711brc_m013_0712..brc_m013_0715brc_m013_0716..brc_m013_0721brc_m013_0722brc_m013_0723brc_m013_0724..brc_m013_0728brc_m013_0729brc_m013_0730brc_m013_0731brc_m013_0732..brc_m013_0735brc_m013_0736..brc_m013_0769brc_m013_0770..brc_m013_0771brc_m013_0772..brc_m013_0773brc_m013_0774..brc_m013_0775brc_m013_0776..brc_m013_0782brc_m013_0783brc_m013_0784..brc_m013_0795brc_m013_0796..brc_m013_0798brc_m013_0799..brc_m013_0801brc_m013_0802..brc_m013_0805brc_m013_0806..brc_m013_0809brc_m013_0810..brc_m013_0811brc_m013_0812..brc_m013_0824brc_m013_0825..brc_m013_0826brc_m013_0827..brc_m013_0829brc_m013_0830..brc_m013_0831brc_m013_0832..brc_m013_0842brc_m013_0843..brc_m013_0854brc_m013_0855..brc_m013_0861brc_m013_0862brc_m013_0863..brc_m013_0864brc_m013_0865..brc_m013_0867brc_m013_0868..brc_m013_0883brc_m013_0884..brc_m013_0893brc_m013_0894brc_m013_0895..brc_m013_0897brc_m013_0898..brc_m013_0906brc_m013_0907..brc_m013_0910brc_m013_0911..brc_m013_0925brc_m013_0926..brc_m013_0928brc_m013_0929..brc_m013_0931

0

20

40

60

80

100

120

140

LG2brc_m013_0932..brc_m013_0941brc_m013_0942..brc_m013_0943brc_m013_0944..brc_m013_0945brc_m013_0946..brc_m013_0949brc_m013_0950..brc_m013_0952

brc_m013_0953..brc_m013_0975

brc_m013_0976..brc_m013_1019brc_m013_1020brc_m013_1021brc_m013_1022..brc_m013_1061brc_m013_1062brc_m013_1063

brc_m013_1064..brc_m013_1065


brc_m013_1071..brc_m013_1074

brc_m013_1075

brc_m013_1076..brc_m013_1081

brc_m013_1082..brc_m013_1086brc_m013_1087..brc_m013_1088brc_m013_1089..brc_m013_1098brc_m013_1099..brc_m013_1106brc_m013_1107..brc_m013_1116brc_m013_1117brc_m013_1118..brc_m013_1121brc_m013_1122..brc_m013_1127brc_m013_1128brc_m013_1129..brc_m013_1136brc_m013_1137..brc_m013_1138brc_m013_1139..brc_m013_1141brc_m013_1142..brc_m013_1144brc_m013_1145..brc_m013_1156brc_m013_1157brc_m013_1158..brc_m013_1170brc_m013_1171..brc_m013_1181brc_m013_1182..brc_m013_1185brc_m013_1186brc_m013_1187..brc_m013_1205brc_m013_1206..brc_m013_1218brc_m013_1219..brc_m013_1220brc_m013_1221..brc_m013_1224brc_m013_1225..brc_m013_1228brc_m013_1229brc_m013_1230..brc_m013_1236brc_m013_1237brc_m013_1238..brc_m013_1247brc_m013_1248..brc_m013_1251

brc_m013_1252brc_m013_1253..brc_m013_1268brc_m013_1269..brc_m013_1270brc_m013_1271..brc_m013_1273brc_m013_1274brc_m013_1275..brc_m013_1280brc_m013_1281

brc_m013_1282..brc_m013_1286brc_m013_1287..brc_m013_1298brc_m013_1299..brc_m013_1307brc_m013_1308brc_m013_1309..brc_m013_1313brc_m013_1314..brc_m013_1317brc_m013_1318..brc_m013_1319brc_m013_1320..brc_m013_1326brc_m013_1327..brc_m013_1340brc_m013_1341..brc_m013_1362brc_m013_1363..brc_m013_1385

0

20

40

60

80

100

120

140

LG3brc_m013_1386..brc_m013_1388brc_m013_1389..brc_m013_1398brc_m013_1399..brc_m013_1406brc_m013_1407..brc_m013_1411brc_m013_1412..brc_m013_1413brc_m013_1414..brc_m013_1416

brc_m013_1417brc_m013_1418..brc_m013_1420brc_m013_1421..brc_m013_1424brc_m013_1425..brc_m013_1432brc_m013_1433..brc_m013_1442brc_m013_1443brc_m013_1444..brc_m013_1450brc_m013_1451brc_m013_1452brc_m013_1453..brc_m013_1455brc_m013_1456..brc_m013_1467brc_m013_1468..brc_m013_1469brc_m013_1470brc_m013_1471..brc_m013_1474brc_m013_1475brc_m013_1476brc_m013_1477brc_m013_1478..brc_m013_1482brc_m013_1483



brc_m013_1497..brc_m013_1500brc_m013_1501brc_m013_1502..brc_m013_1513brc_m013_1514..brc_m013_1562brc_m013_1563..brc_m013_1565brc_m013_1566..brc_m013_1567

brc_m013_1568..brc_m013_1580brc_m013_1581..brc_m013_1587brc_m013_1588..brc_m013_1591brc_m013_1592..brc_m013_1593brc_m013_1594..brc_m013_1604brc_m013_1605..brc_m013_1607brc_m013_1608..brc_m013_1609brc_m013_1610..brc_m013_1611brc_m013_1612..brc_m013_1616brc_m013_1617..brc_m013_1618brc_m013_1619..brc_m013_1620brc_m013_1621..brc_m013_1629brc_m013_1630..brc_m013_1633brc_m013_1634..brc_m013_1638brc_m013_1639..brc_m013_1647brc_m013_1648..brc_m013_1649brc_m013_1650..brc_m013_1656brc_m013_1657..brc_m013_1665brc_m013_1666..brc_m013_1672brc_m013_1673..brc_m013_1674brc_m013_1675..brc_m013_1678brc_m013_1679..brc_m013_1682brc_m013_1683brc_m013_1684brc_m013_1685..brc_m013_1686brc_m013_1687..brc_m013_1700brc_m013_1701..brc_m013_1702brc_m013_1703brc_m013_1704..brc_m013_1707brc_m013_1708..brc_m013_1709brc_m013_1710..brc_m013_1714brc_m013_1715..brc_m013_1728brc_m013_1729..brc_m013_1742

0

20

40

60

80

100

120

140

LG4brc_m013_1743..brc_m013_1750


brc_m013_1769..brc_m013_1772

brc_m013_1773..brc_m013_1779brc_m013_1780brc_m013_1781brc_m013_1782..brc_m013_1783brc_m013_1784brc_m013_1785..brc_m013_1786

brc_m013_1787

brc_m013_1788brc_m013_1789..brc_m013_1790brc_m013_1791..brc_m013_1793brc_m013_1794..brc_m013_1797brc_m013_1798..brc_m013_1800brc_m013_1801..brc_m013_1804brc_m013_1805brc_m013_1806..brc_m013_1808brc_m013_1809brc_m013_1810..brc_m013_1813brc_m013_1814..brc_m013_1818brc_m013_1819..brc_m013_1820brc_m013_1821..brc_m013_1822brc_m013_1823..brc_m013_1824brc_m013_1825brc_m013_1826..brc_m013_1840brc_m013_1841..brc_m013_1842brc_m013_1843brc_m013_1844..brc_m013_1848brc_m013_1849brc_m013_1850brc_m013_1851..brc_m013_1853brc_m013_1854..brc_m013_1858brc_m013_1859..brc_m013_1866brc_m013_1867..brc_m013_1868brc_m013_1869brc_m013_1870..brc_m013_1874brc_m013_1875..brc_m013_1876brc_m013_1877..brc_m013_1878brc_m013_1879..brc_m013_1883brc_m013_1884brc_m013_1885brc_m013_1886..brc_m013_1888brc_m013_1889..brc_m013_1895brc_m013_1896..brc_m013_1899brc_m013_1900..brc_m013_1913brc_m013_1914brc_m013_1915..brc_m013_1922brc_m013_1923brc_m013_1924..brc_m013_1928brc_m013_1929..brc_m013_1942brc_m013_1943..brc_m013_1946brc_m013_1947..brc_m013_1948brc_m013_1949brc_m013_1950..brc_m013_1953brc_m013_1954..brc_m013_1955brc_m013_1956brc_m013_1957..brc_m013_1963brc_m013_1964..brc_m013_1966brc_m013_1967..brc_m013_1968brc_m013_1969..brc_m013_1970brc_m013_1971brc_m013_1972brc_m013_1973..brc_m013_1980brc_m013_1981..brc_m013_1983brc_m013_1984..brc_m013_1990brc_m013_1991..brc_m013_1993brc_m013_1994..brc_m013_1996brc_m013_1997..brc_m013_2009

0

20

40

60

80

100

120

LG5brc_m013_2010..brc_m013_2028

brc_m013_2029..brc_m013_2038brc_m013_2039

brc_m013_2040..brc_m013_2041brc_m013_2042..brc_m013_2047brc_m013_2048..brc_m013_2050brc_m013_2051..brc_m013_2053brc_m013_2054..brc_m013_2062brc_m013_2063brc_m013_2064..brc_m013_2065brc_m013_2066..brc_m013_2067brc_m013_2068brc_m013_2069..brc_m013_2071brc_m013_2072..brc_m013_2081brc_m013_2082

brc_m013_2083brc_m013_2084brc_m013_2085..brc_m013_2099brc_m013_2100brc_m013_2101..brc_m013_2102brc_m013_2103..brc_m013_2107brc_m013_2108brc_m013_2109..brc_m013_2112

brc_m013_2113..brc_m013_2114brc_m013_2115..brc_m013_2123brc_m013_2124..brc_m013_2131brc_m013_2132brc_m013_2133brc_m013_2134..brc_m013_2136

brc_m013_2137

brc_m013_2138..brc_m013_2139

brc_m013_2140..brc_m013_2142brc_m013_2143..brc_m013_2150brc_m013_2151..brc_m013_2152brc_m013_2153..brc_m013_2161brc_m013_2162..brc_m013_2163

brc_m013_2164..brc_m013_2165brc_m013_2166..brc_m013_2170brc_m013_2171..brc_m013_2172brc_m013_2173brc_m013_2174..brc_m013_2182brc_m013_2183..brc_m013_2186brc_m013_2187..brc_m013_2190brc_m013_2191..brc_m013_2193brc_m013_2194brc_m013_2195..brc_m013_2201brc_m013_2202..brc_m013_2203brc_m013_2204..brc_m013_2220brc_m013_2221..brc_m013_2232brc_m013_2233..brc_m013_2239brc_m013_2240..brc_m013_2261brc_m013_2262..brc_m013_2267brc_m013_2268..brc_m013_2269brc_m013_2270..brc_m013_2271brc_m013_2272..brc_m013_2282brc_m013_2283..brc_m013_2284brc_m013_2285..brc_m013_2299brc_m013_2300..brc_m013_2301brc_m013_2302..brc_m013_2305brc_m013_2306..brc_m013_2307brc_m013_2308..brc_m013_2330brc_m013_2331..brc_m013_2337brc_m013_2338..brc_m013_2352

0

20

40

60

80

100

120

LG6brc_m013_2353..brc_m013_2365brc_m013_2366..brc_m013_2369

brc_m013_2370..brc_m013_2372brc_m013_2373..brc_m013_2378brc_m013_2379..brc_m013_2386brc_m013_2387brc_m013_2388..brc_m013_2394brc_m013_2395..brc_m013_2397brc_m013_2398brc_m013_2399brc_m013_2400brc_m013_2401brc_m013_2402..brc_m013_2407brc_m013_2408..brc_m013_2411brc_m013_2412..brc_m013_2416brc_m013_2417brc_m013_2418brc_m013_2419..brc_m013_2436brc_m013_2437..brc_m013_2441brc_m013_2442brc_m013_2443..brc_m013_2444brc_m013_2445brc_m013_2446..brc_m013_2453brc_m013_2454brc_m013_2455..brc_m013_2460brc_m013_2461brc_m013_2462..brc_m013_2470brc_m013_2471..brc_m013_2474brc_m013_2475..brc_m013_2482brc_m013_2483brc_m013_2484..brc_m013_2487brc_m013_2488brc_m013_2489..brc_m013_2492brc_m013_2493..brc_m013_2496brc_m013_2497brc_m013_2498..brc_m013_2504brc_m013_2505brc_m013_2506..brc_m013_2510brc_m013_2511..brc_m013_2523brc_m013_2524..brc_m013_2531brc_m013_2532..brc_m013_2536brc_m013_2537..brc_m013_2555brc_m013_2556..brc_m013_2571brc_m013_2572..brc_m013_2573brc_m013_2574..brc_m013_2579brc_m013_2580..brc_m013_2581brc_m013_2582..brc_m013_2587brc_m013_2588brc_m013_2589..brc_m013_2594

brc_m013_2595

brc_m013_2596..brc_m013_2597brc_m013_2598..brc_m013_2604brc_m013_2605brc_m013_2606..brc_m013_2616brc_m013_2617..brc_m013_2619brc_m013_2620..brc_m013_2623


brc_m013_2629..brc_m013_2630

0

20

40

60

80

100

LG7brc_m013_2631..brc_m013_2632brc_m013_2633brc_m013_2634..brc_m013_2635brc_m013_2636..brc_m013_2642brc_m013_2643..brc_m013_2657brc_m013_2658..brc_m013_2659brc_m013_2660..brc_m013_2661brc_m013_2662

brc_m013_2663

brc_m013_2664brc_m013_2665..brc_m013_2666

brc_m013_2667..brc_m013_2668brc_m013_2669..brc_m013_2670brc_m013_2671..brc_m013_2680brc_m013_2681..brc_m013_2682brc_m013_2683brc_m013_2684..brc_m013_2685brc_m013_2686..brc_m013_2694


brc_m013_2714..brc_m013_2725brc_m013_2726..brc_m013_2727brc_m013_2728..brc_m013_2731brc_m013_2732brc_m013_2733..brc_m013_2753brc_m013_2754..brc_m013_2758brc_m013_2759..brc_m013_2763brc_m013_2764..brc_m013_2779brc_m013_2780brc_m013_2781brc_m013_2782..brc_m013_2784brc_m013_2785..brc_m013_2787brc_m013_2788..brc_m013_2791brc_m013_2792..brc_m013_2797brc_m013_2798..brc_m013_2799brc_m013_2800brc_m013_2801..brc_m013_2804brc_m013_2805..brc_m013_2809

brc_m013_2810..brc_m013_2811

brc_m013_2812..brc_m013_2813brc_m013_2814..brc_m013_2817brc_m013_2818..brc_m013_2827brc_m013_2828brc_m013_2829..brc_m013_2832brc_m013_2833brc_m013_2834..brc_m013_2840brc_m013_2841..brc_m013_2846brc_m013_2847brc_m013_2848..brc_m013_2852brc_m013_2853brc_m013_2854..brc_m013_2856brc_m013_2857..brc_m013_2862brc_m013_2863..brc_m013_2868brc_m013_2869..brc_m013_2874brc_m013_2875..brc_m013_2896

0

20

40

60

80

100

LG8


brc_m013_2933

brc_m013_2934..brc_m013_2935brc_m013_2936brc_m013_2937..brc_m013_2943brc_m013_2944brc_m013_2945..brc_m013_2946brc_m013_2947brc_m013_2948brc_m013_2949..brc_m013_2950brc_m013_2951..brc_m013_2957brc_m013_2958..brc_m013_2961brc_m013_2962..brc_m013_2970brc_m013_2971..brc_m013_2980brc_m013_2981..brc_m013_2992brc_m013_2993..brc_m013_2996brc_m013_2997..brc_m013_2998brc_m013_2999..brc_m013_3000brc_m013_3001brc_m013_3002..brc_m013_3003brc_m013_3004brc_m013_3005brc_m013_3006..brc_m013_3010brc_m013_3011..brc_m013_3014brc_m013_3015brc_m013_3016..brc_m013_3019brc_m013_3020brc_m013_3021..brc_m013_3030brc_m013_3031..brc_m013_3032brc_m013_3033..brc_m013_3034brc_m013_3035..brc_m013_3036brc_m013_3037..brc_m013_3045brc_m013_3046..brc_m013_3052brc_m013_3053brc_m013_3054..brc_m013_3061brc_m013_3062..brc_m013_3066brc_m013_3067..brc_m013_3068brc_m013_3069..brc_m013_3076brc_m013_3077..brc_m013_3084brc_m013_3085..brc_m013_3087brc_m013_3088..brc_m013_3089brc_m013_3090..brc_m013_3096brc_m013_3097..brc_m013_3100brc_m013_3101..brc_m013_3104brc_m013_3105brc_m013_3106..brc_m013_3112brc_m013_3113..brc_m013_3122brc_m013_3123..brc_m013_3124brc_m013_3125..brc_m013_3127brc_m013_3128..brc_m013_3145brc_m013_3146..brc_m013_3159brc_m013_3160..brc_m013_3172

0

20

40

60

80

100

LG9brc_m013_3173..brc_m013_3175brc_m013_3176..brc_m013_3180brc_m013_3181..brc_m013_3189brc_m013_3190..brc_m013_3198brc_m013_3199brc_m013_3200..brc_m013_3201brc_m013_3202..brc_m013_3203brc_m013_3204brc_m013_3205..brc_m013_3206brc_m013_3207..brc_m013_3211brc_m013_3212..brc_m013_3214brc_m013_3215..brc_m013_3227brc_m013_3228..brc_m013_3230brc_m013_3231..brc_m013_3235brc_m013_3236..brc_m013_3238brc_m013_3239..brc_m013_3242brc_m013_3243..brc_m013_3244brc_m013_3245brc_m013_3246..brc_m013_3247brc_m013_3248..brc_m013_3249brc_m013_3250..brc_m013_3252brc_m013_3253..brc_m013_3257brc_m013_3258brc_m013_3259brc_m013_3260..brc_m013_3261brc_m013_3262..brc_m013_3263brc_m013_3264brc_m013_3265..brc_m013_3269brc_m013_3270..brc_m013_3274brc_m013_3275..brc_m013_3276brc_m013_3277..brc_m013_3281brc_m013_3282..brc_m013_3284brc_m013_3285brc_m013_3286..brc_m013_3289brc_m013_3290..brc_m013_3296brc_m013_3297brc_m013_3298..brc_m013_3300brc_m013_3301..brc_m013_3302brc_m013_3303..brc_m013_3305brc_m013_3306..brc_m013_3308brc_m013_3309..brc_m013_3314brc_m013_3315..brc_m013_3317brc_m013_3318..brc_m013_3329brc_m013_3330..brc_m013_3331brc_m013_3332..brc_m013_3338brc_m013_3339..brc_m013_3340brc_m013_3341..brc_m013_3344brc_m013_3345..brc_m013_3349brc_m013_3350..brc_m013_3357brc_m013_3358..brc_m013_3359brc_m013_3360brc_m013_3361..brc_m013_3368brc_m013_3369..brc_m013_3372brc_m013_3373..brc_m013_3376brc_m013_3377brc_m013_3378..brc_m013_3386brc_m013_3387..brc_m013_3388brc_m013_3389..brc_m013_3395brc_m013_3396..brc_m013_3399

0

20

40

60

80

LG10brc_m013_3400..brc_m013_3411brc_m013_3412brc_m013_3413..brc_m013_3424brc_m013_3425brc_m013_3426brc_m013_3427..brc_m013_3429

brc_m013_3430

brc_m013_3431..brc_m013_3432brc_m013_3433..brc_m013_3435brc_m013_3436brc_m013_3437..brc_m013_3439

brc_m013_3440..brc_m013_3441brc_m013_3442

brc_m013_3443..brc_m013_3445brc_m013_3446..brc_m013_3447brc_m013_3448..brc_m013_3449brc_m013_3450..brc_m013_3454brc_m013_3455brc_m013_3456..brc_m013_3462brc_m013_3463..brc_m013_3464brc_m013_3465brc_m013_3466..brc_m013_3467brc_m013_3468..brc_m013_3472brc_m013_3473brc_m013_3474..brc_m013_3476brc_m013_3477..brc_m013_3487brc_m013_3488brc_m013_3489..brc_m013_3491brc_m013_3492..brc_m013_3500brc_m013_3501..brc_m013_3512brc_m013_3513..brc_m013_3514brc_m013_3515..brc_m013_3524brc_m013_3525..brc_m013_3527brc_m013_3528..brc_m013_3531brc_m013_3532..brc_m013_3547brc_m013_3548..brc_m013_3557brc_m013_3558..brc_m013_3566brc_m013_3567..brc_m013_3568brc_m013_3569..brc_m013_3570brc_m013_3571..brc_m013_3574brc_m013_3575..brc_m013_3582brc_m013_3583..brc_m013_3592brc_m013_3593..brc_m013_3605brc_m013_3606..brc_m013_3616brc_m013_3617..brc_m013_3618brc_m013_3619..brc_m013_3622brc_m013_3623..brc_m013_3624brc_m013_3625..brc_m013_3628brc_m013_3629..brc_m013_3635

0

20

40

60

80

LG11



brc_m013_3672


brc_m013_3683..brc_m013_3685

brc_m013_3686..brc_m013_3688

brc_m013_3689..brc_m013_3693

brc_m013_3694..brc_m013_3698brc_m013_3699

brc_m013_3700brc_m013_3701..brc_m013_3702brc_m013_3703..brc_m013_3704brc_m013_3705..brc_m013_3712brc_m013_3713brc_m013_3714..brc_m013_3716brc_m013_3717..brc_m013_3724brc_m013_3725..brc_m013_3730brc_m013_3731..brc_m013_3752brc_m013_3753..brc_m013_3758brc_m013_3759..brc_m013_3789brc_m013_3790..brc_m013_3801brc_m013_3802..brc_m013_3814brc_m013_3815..brc_m013_3818brc_m013_3819..brc_m013_3822brc_m013_3823..brc_m013_3826brc_m013_3827..brc_m013_3832brc_m013_3833..brc_m013_3837brc_m013_3838brc_m013_3839..brc_m013_3841brc_m013_3842..brc_m013_3847

brc_m013_3848..brc_m013_3853brc_m013_3854..brc_m013_3858brc_m013_3859..brc_m013_3868brc_m013_3869..brc_m013_3871brc_m013_3872..brc_m013_3901brc_m013_3902brc_m013_3903..brc_m013_3909brc_m013_3910brc_m013_3911..brc_m013_3926brc_m013_3927..brc_m013_3931brc_m013_3932..brc_m013_3948

0

20

40

60

80

LG12


brc_m013_3972..brc_m013_3975brc_m013_3976

brc_m013_3977..brc_m013_3985

brc_m013_3986brc_m013_3987..brc_m013_3994brc_m013_3995..brc_m013_3997brc_m013_3998..brc_m013_4004

brc_m013_4005..brc_m013_4006brc_m013_4007..brc_m013_4008brc_m013_4009..brc_m013_4010brc_m013_4011..brc_m013_4013brc_m013_4014brc_m013_4015brc_m013_4016..brc_m013_4019brc_m013_4020..brc_m013_4021brc_m013_4022..brc_m013_4025brc_m013_4026..brc_m013_4032brc_m013_4033..brc_m013_4036brc_m013_4037..brc_m013_4041brc_m013_4042..brc_m013_4043

brc_m013_4044..brc_m013_4046brc_m013_4047..brc_m013_4056brc_m013_4057brc_m013_4058..brc_m013_4063brc_m013_4064..brc_m013_4071brc_m013_4072..brc_m013_4075brc_m013_4076brc_m013_4077brc_m013_4078..brc_m013_4085brc_m013_4086..brc_m013_4089brc_m013_4090..brc_m013_4091brc_m013_4092..brc_m013_4093brc_m013_4094..brc_m013_4095brc_m013_4096..brc_m013_4114brc_m013_4115..brc_m013_4117brc_m013_4118..brc_m013_4131brc_m013_4132..brc_m013_4133brc_m013_4134..brc_m013_4146

0

20

40

60

80

LG13


brc_m013_4168brc_m013_4169

brc_m013_4170brc_m013_4171..brc_m013_4172brc_m013_4173..brc_m013_4175brc_m013_4176brc_m013_4177..brc_m013_4178brc_m013_4179..brc_m013_4183brc_m013_4184..brc_m013_4185brc_m013_4186..brc_m013_4187brc_m013_4188..brc_m013_4191brc_m013_4192brc_m013_4193..brc_m013_4194brc_m013_4195..brc_m013_4206brc_m013_4207..brc_m013_4210brc_m013_4211..brc_m013_4213brc_m013_4214brc_m013_4215..brc_m013_4217brc_m013_4218..brc_m013_4221brc_m013_4222..brc_m013_4223brc_m013_4224..brc_m013_4231brc_m013_4232..brc_m013_4234brc_m013_4235..brc_m013_4239brc_m013_4240..brc_m013_4246brc_m013_4247..brc_m013_4248brc_m013_4249..brc_m013_4258brc_m013_4259..brc_m013_4260brc_m013_4261..brc_m013_4269brc_m013_4270..brc_m013_4271brc_m013_4272..brc_m013_4278brc_m013_4279..brc_m013_4280brc_m013_4281..brc_m013_4284brc_m013_4285brc_m013_4286..brc_m013_4288brc_m013_4289..brc_m013_4294brc_m013_4295..brc_m013_4296brc_m013_4297..brc_m013_4301brc_m013_4302..brc_m013_4313brc_m013_4314..brc_m013_4320brc_m013_4321..brc_m013_4322brc_m013_4323..brc_m013_4345brc_m013_4346..brc_m013_4351

0

20

40

60

LG14brc_m013_4352..brc_m013_4366brc_m013_4367brc_m013_4368brc_m013_4369..brc_m013_4373brc_m013_4374..brc_m013_4381brc_m013_4382..brc_m013_4383brc_m013_4384..brc_m013_4385brc_m013_4386brc_m013_4387..brc_m013_4388brc_m013_4389brc_m013_4390..brc_m013_4404brc_m013_4405brc_m013_4406..brc_m013_4409brc_m013_4410..brc_m013_4411brc_m013_4412..brc_m013_4418brc_m013_4419..brc_m013_4434brc_m013_4435..brc_m013_4442brc_m013_4443..brc_m013_4448brc_m013_4449..brc_m013_4451brc_m013_4452..brc_m013_4461brc_m013_4462..brc_m013_4471brc_m013_4472..brc_m013_4475brc_m013_4476..brc_m013_4477brc_m013_4478brc_m013_4479brc_m013_4480..brc_m013_4485brc_m013_4486brc_m013_4487..brc_m013_4491brc_m013_4492brc_m013_4493brc_m013_4494..brc_m013_4495brc_m013_4496..brc_m013_4501brc_m013_4502..brc_m013_4510brc_m013_4511..brc_m013_4531brc_m013_4532brc_m013_4533..brc_m013_4534brc_m013_4535..brc_m013_4541brc_m013_4542..brc_m013_4543brc_m013_4544..brc_m013_4545brc_m013_4546..brc_m013_4548brc_m013_4549..brc_m013_4551brc_m013_4552..brc_m013_4555brc_m013_4556..brc_m013_4561

0

20

40

60

LG15

brc_m013_4562..brc_m013_4577brc_m013_4578..brc_m013_4594brc_m013_4595..brc_m013_4599brc_m013_4600..brc_m013_4625brc_m013_4626..brc_m013_4638brc_m013_4639..brc_m013_4642brc_m013_4643..brc_m013_4644brc_m013_4645..brc_m013_4650brc_m013_4651..brc_m013_4663brc_m013_4664..brc_m013_4668brc_m013_4669..brc_m013_4670brc_m013_4671..brc_m013_4674brc_m013_4675..brc_m013_4679brc_m013_4680..brc_m013_4681brc_m013_4682brc_m013_4683..brc_m013_4688brc_m013_4689..brc_m013_4692brc_m013_4693..brc_m013_4695brc_m013_4696..brc_m013_4701brc_m013_4702brc_m013_4703..brc_m013_4712brc_m013_4713..brc_m013_4717brc_m013_4718..brc_m013_4720brc_m013_4721brc_m013_4722..brc_m013_4726brc_m013_4727..brc_m013_4728brc_m013_4729..brc_m013_4742brc_m013_4743brc_m013_4744..brc_m013_4746brc_m013_4747brc_m013_4748..brc_m013_4749brc_m013_4750..brc_m013_4752

brc_m013_4753..brc_m013_4756brc_m013_4757..brc_m013_4759brc_m013_4760..brc_m013_4762brc_m013_4763brc_m013_4764..brc_m013_4766brc_m013_4767..brc_m013_4769brc_m013_4770brc_m013_4771..brc_m013_4774brc_m013_4775..brc_m013_4776brc_m013_4777..brc_m013_4778brc_m013_4779..brc_m013_4780brc_m013_4781..brc_m013_4793brc_m013_4794brc_m013_4795..brc_m013_4798brc_m013_4799..brc_m013_4802brc_m013_4803..brc_m013_4806brc_m013_4807brc_m013_4808..brc_m013_4814brc_m013_4815..brc_m013_4819brc_m013_4820brc_m013_4821brc_m013_4822..brc_m013_4823brc_m013_4824brc_m013_4825..brc_m013_4855brc_m013_4856..brc_m013_4858brc_m013_4859..brc_m013_4863brc_m013_4864..brc_m013_4865brc_m013_4866..brc_m013_4875brc_m013_4876..brc_m013_4881brc_m013_4882..brc_m013_4891brc_m013_4892..brc_m013_4895brc_m013_4896..brc_m013_4911brc_m013_4912..brc_m013_4938brc_m013_4939brc_m013_4940..brc_m013_4957brc_m013_4958..brc_m013_4959brc_m013_4960..brc_m013_4972brc_m013_4973..brc_m013_4981brc_m013_4982..brc_m013_4983

0

20

40

60

80

100

120

LGSB

Gp-9

Figure 1a b




Si_gnF.scaffold00779_nt3746833Si_gnF.scaffold00779_nt374687927.3Si_gnF.scaffold00779_nt382158728.5




Si_gnF.scaffold05266_nt63430678.4Si_gnF.scaffold05266_nt65952779.7






104.4


Si_gnF.scaffold00413_nt1035856109.6Si_gnF.scaffold01573_nt108462110.8Si_gnF.scaffold01573_nt447618112.1Si_gnF.scaffold00899_nt377419Si_gnF.scaffold00899_nt686574114.4Si_gnF.scaffold00899_nt236146Si_gnF.scaffold00899_nt332715Si_gnF.scaffold00899_nt335756

115.8




LGS Bfrom M013




22.2



Si_gnF.scaffold01573_nt10846280.7Si_gnF.scaffold00255_nt314067Si_gnF.scaffold00255_nt407778Si_gnF.scaffold00413_nt1035856Si_gnF.scaffold00413_nt676115Si_gnF.scaffold00469_nt794Si_gnF.scaffold00690_nt229012Si_gnF.scaffold00690_nt415290Si_gnF.scaffold00899_nt236146Si_gnF.scaffold00899_nt332715Si_gnF.scaffold00899_nt335756Si_gnF.scaffold00899_nt377419Si_gnF.scaffold00899_nt686574Si_gnF.scaffold01573_nt447618Si_gnF.scaffold01957_nt412242Si_gnF.scaffold03404_nt128925Si_gnF.scaffold03404_nt228606Si_gnF.scaffold03404_nt241461Si_gnF.scaffold05266_nt634306Si_gnF.scaffold05266_nt659527Si_gnF.scaffold05266_nt733643Si_gnF.scaffold05266_nt753644Si_gnF.scaffold06914_nt297673Si_gnF.scaffold07090_nt1051771Si_gnF.scaffold07090_nt710010Si_gnF.scaffold09758_nt222732

81.8


LGS B/bfrom P034

Gp-9

Total285 non-recombiningmarkers

No recombination between B and b over 4% of genome: ⅔ of a chromosme!

Sex chromosomes

X Y

Gp-9 B

Gp-9 b

SB Sb

“Social chromosomes”

?

Why non-recombining?

SB

Sb

difficult to detect in sequence alignments

Why non-recombining? Structural differences

Sb SB Gp-9B

genetic map

E3

E17

A22

Gp-9 B male Gp-9 b male Figure 2 a

b Diploid Gp-9 Bb

Sb SB Gp-9B

genetic map

E3

E17

A22


b Diploid Gp-9 Bb

Sb SB Gp-9B

genetic map

E3

E17

A22


b Diploid Gp-9 Bb

Sb SB Gp-9B

genetic map

E3

E17

A22


b Diploid Gp-9 Bb

Why non-recombining? Structural differencesusing Flourescence in situ Hybridization

John Wang @ Taipei

X

ʁ

X X Y

ʂ

Maybe several rearrangements

Predictions: •genes in S are responsible for phenotype?

SBSB SB Sb

Single queen colony Multiple queen colony

Most BB vs Bb gene expression differences map to S

Non-recombing region of S contains 800 genes

polygyne colonies (Table 2 and Table S2; all P,0.05). The 11genes encoding mitochondrial and prefoldin complex (molecularchaperone) proteins were all up-regulated in polygyne comparedto monogyne workers. Increased mitochondrial gene expressionmay reflect increased oxidative metabolism, while increasedprefoldin expression may indicate higher protein synthesis rates,possibly in relation to the relatively smaller size and highermetabolic rates of polygyne workers [42].

The pattern of expression of the six genes in the viral genecategory is consistent with the expectation that differences in socialorganization affect susceptibility to pathogens and parasites. In themonogyne form, there is intense selection against susceptibleinfected individuals during independent colony founding, a stagethat colonies of the polygyne form never display [26]. Accordingly,we found that workers in the polygyne form express moresequences corresponding to viral genes than their counterparts inthe monogyne form, presumably because of relaxed selection andgenerally greater susceptibility in the former (see also [43]). Basedon sequence similarity and correlated expression across ourexperiments, we identified six gene products that likely representthree different viruses, a ssRNA negative-strand (2) virus and twossRNA positive-strand (+) viruses, one of which is the SINV-2 virus[44]. All 20 polygyne study colonies showed evidence of infectionwith at least one virus (mean number of viral types per colony,2.560.67), whereas only three of the 20 monogyne coloniesshowed evidence of infection, in all cases by a single viral type.Finally, the pattern of expression of another socially-regulated

gene, this one encoding a defensin (a class of small proteinantibiotics active against viruses, bacteria, and fungi [45]), also isconsistent with greater selection for resistance in the monogyneform, as this gene was more highly expressed in monogyne thanpolygyne workers.

The numbers of genes differentially expressed in the genotype (39)and social form (91) comparisons are relatively low compared to thenumbers expected based on other published microarray experi-ments. There are several possible explanations for this. First, the useof whole worker bodies as the source of RNA may decrease theprobability of detecting genes whose level of expression varies amongcells or tissues. Second, our comparisons were performed on groupsof workers originating from different colonies, thus adding a colony-level effect to, and thus increasing the total variance in, geneexpression. Finally, workers of alternate genotype or social formapparently exhibit fewer phenotypic differences than queens[20,21,46,47], possibly reflecting the involvement of fewer differen-tially expressed genes in the former caste.

Remarkably, there was almost no overlap between genes whoselevel of expression was influenced by the focal workers’ Gp-9genotypes and genes whose expression was influenced by the socialenvironment, with only one of the 129 differentially expressedgenes appearing in both categories. This demonstrates an almostcomplete decoupling of the direct effects of the genomic regionmarked by Gp-9 and the indirect effects mediated by socialinteractions within colonies. Moreover, there is little indication ofan interaction between these direct and indirect effects; genes

Figure 1. Expression profiles between S. invicta adult workers with the BB and Bb genotypes of Gp-9. Expression profiles for 39differentially expressed genes are depicted (ANOVA, 10% false discovery rate [FDR]). Each row represents data for one gene, and each columnrepresents data for a pool of 7–10 nestmates with the same Gp-9 genotype sampled from each of twenty colonies of each social form. Colonies werecollected from Georgia (2004, checkered peach bar; 2006, solid peach bar) and Louisiana (2006, solid red bar), USA (data from each polygyne colonyare presented in the identical order for the alternate genotype groupings). Expression levels for each gene are depicted relative to the average levelacross all experimental samples (blue, low levels; yellow, high levels). Genes are arranged by hierarchical clustering. See Text S1 and Table S1 forconfirmation of selected gene expression results with quantitative RT-PCR (qRT-PCR).doi:10.1371/journal.pgen.1000127.g001

Gene Expression Patterns for a Social Trait

PLoS Genetics | www.plosgenetics.org 3 July 2008 | Volume 4 | Issue 7 | e1000127

Gene expression: Gp-9 Bb vs BB workers in multiple queen colonies

29 significant genes "are in the SB/Sb region

(p<10-10)

20 of

Similar for BB vs Bb queens; & for B vs b males. Wang et al 2008

Predictions: •genes in S are responsible for phenotype?

•Sb is degenerating?

probably!⟹ directional (antagonistic?) selection?

X

ʁ

X X Y

ʂ

Maybe several rearrangements

SBSB SB Sb

Single queen colony Multiple queen colony

Is Sb degenerating?

99.8% of non-gap sequences are identical

SB

Sb

(Almost) no SB or Sb-specific sequenceActually quite similar to SB:

But clearly: relaxation of purifying selectionSb contains more small repeats

genes seem to be intact in Sb

Introns bigger in Sb than SB

Sb is degenerating: repeats cause bad assembly

[b] vs. [c]: p < 10-4

[a] vs. [c]: p < 10-7

Gp-9B male Gp-9b male

Region:

Genome assembly:

Normally recombiningregions from all 16

linkage groups

Normally recombiningregions from all 16

linkage groups

Sb region without recombination

in Gp-9 Bb queens

SB region without recombination

in Gp-9 Bb queens

Sca

ffold

leng

th (b

p)

0

1,000,000

2,000,000

3,000,000

4,000,000

5,000,000

6,000,000

[a] [a], [b] [a] [c]

SB Sb

Is Sb degenerating?

Sb contains more big repeats ⟹ bad assemblydN/dS bigger in S than rest of the genome

Probably � haploidy = strong purifying selection ⟹ slower degeneration

99.8% of non-gap sequences are identical(Almost) no SB or Sb-specific sequence

Actually quite similar to SB:

But clearly: relaxation of purifying selectionSb contains more small repeats

genes seem to be intact in Sb

Introns bigger in Sb than SB

Age of the region based on dS

0

50

100

150

200

250

0.00 0.05 0.10 0.15 0.20 0.25leafcutterDndsSubset$dS

coun

tleafcutterdS

0

50

100

150

0.00 0.05 0.10 0.15 0.20 0.25subset(dndsdata, gp9linked == TRUE)$dS

coun

t

gp9linkedSolenopsisdS

## Min. 1st Qu. Median Mean 3rd Qu. Max.

## 0.00000 0.08160 0.10300 0.10559 0.12600 0.32190

Richard thinks the mega-dS values are likely not orthologs. WARNING: We still have some leafcutterD-ndsSubset dS > 1 here!!!!

1.3.4 Calculating age using mean based on dS

10000000 * median(subset(dndsdata, gp9linked == TRUE)$dS)/median(leafcutterDndsSubset$dS)

## [1] 330097.09


## [1] 264077.67

Richard Nichols says I can use Maximum Likelihood to compare such distributions. See section below.

1.3.5 Calculating age based on dN

What do my dN distributions look like?

multiplot(qplot(x = subset(leafcutterDndsSubset, dN < 0.1)$dN, binwidth = 0.002, main = "leafcutterdNs"), qplot(x = subset(dndsdata,

gp9linked == TRUE & dN < 0.1)$dN, binwidth = 0.0005, main = "gp9linkedSolenopsisdNs"), cols = 1)

3

0

50

100

150

200

250

0.00 0.05 0.10 0.15 0.20 0.25leafcutterDndsSubset$dS

coun

tleafcutterdS

0

50

100

150

0.00 0.05 0.10 0.15 0.20 0.25subset(dndsdata, gp9linked == TRUE)$dS

coun

t

gp9linkedSolenopsisdS

## Min. 1st Qu. Median Mean 3rd Qu. Max.

## 0.00000 0.08160 0.10300 0.10559 0.12600 0.32190

Richard thinks the mega-dS values are likely not orthologs. WARNING: We still have some leafcutterD-ndsSubset dS > 1 here!!!!

1.3.4 Calculating age using mean based on dS


## [1] 330097.09


## [1] 264077.67

Richard Nichols says I can use Maximum Likelihood to compare such distributions. See section below.

1.3.5 Calculating age based on dN

What do my dN distributions look like?

multiplot(qplot(x = subset(leafcutterDndsSubset, dN < 0.1)$dN, binwidth = 0.002, main = "leafcutterdNs"), qplot(x = subset(dndsdata,

gp9linked == TRUE & dN < 0.1)$dN, binwidth = 0.0005, main = "gp9linkedSolenopsisdNs"), cols = 1)

3

Leafcutter common ancestor: 8,000,000-10,000,000 years ago

Maximum Likelihood Estimation of SB/Sb age:280,000-425,000⟹ little time for degeneration

Summary

Solenopsis invicta queen number determined by Gp-9 genotypes: !•only BB workers ➔ single BB queen"•with Bb workers ➔ multiple Bb queens

Genome sequencing + RAD Genotyping•Gp-9 marks ~4% of genome"•social like sex chromosomes: SB is like X; Sb is like Y

some relaxation of purifying selection"but haploid males ➔ strong purifying selection

Structural differences between SB and Sb ➔ no recombination

Ants are cool.

SB and Sb stopped recombining ~400,000 years ago.

genome evolution social evolutionGenerally

Single- vs. Multiple queennessin fire antsin similar independent species

•one or many loci?"•one or many genes?"•convergence?

Social parasitism

Strengths of selection in social evolution

concepts & mechanisms

Medically relevant questionsQueen longevity? Candidate gene studies

VitellogeninSex determination genes

functional testing....

Tools for genomics work on emerging model organisms

Molecular response to social upheaval

AGATCCCCGCTGGGCGAATTCGGCTTAGAGAGGTGGAGTGCAGCCTGGTGCCACGGTGTCGTGTTCGCATTGTTAATTGGCCGGCCGTTtCATTTtCCGTTGCGGCGGCGTAACGTGAGAGACAGCGTAACGGGAAGCGAAACGACACGGCGCGTCGAGCGTGCACAGAGAAGTTGAATTATCGAGGGaGAAAAAGGGCGCGAGGGAGGGGCAGGACGGTCAAACACTGAGAAAGCAGGCAACATCCGCGAACCCGTAAATTAATTAGGGCCAACATAATAGCTGGGAATGGCTGGTAGTGGTGGTTCGTCTTCGTGGTCCAGCTTGCGGTTCCCGAGTGCCCTCGCAGTTCGCGCGGTCATGCCTCGGAAACTACACGATACACCTTGCGTCATCCTTACACGAGGATACCTGTTTGCGATCCCCTGAGGTATTAACGAGCGTATAAGCAGACTCAAAGTACGAACACACTGCTAGTTTCGCGTGCATGTTGCTCGTCGCCGTGCAGACGAGTTAGTACTGTCGACAAAGTAGTCGTAAAGTACGTAAGAGTCTTCCTCTAGTAGCGAGGACTAATCTCCCGGTACATTAATCTATGTATATTTTATGATATACACACCTGTATTTATCGAAAGTTTATGTTTATCTCGAAAATTAACGTTAATTTTCGAGGACGCAAAAGCTGTCGCAAGTTTaTTTAATTTTGTTTGtATTTTTTTTTCTTGCTCATTTTTATTTCGAAGTATGCAAATTGAATCTCTTGGTGGCATAACGAGAATTTTCGAAGCTTAAAGGGATCTGCGTTGGCGCGAAGAGAATGGGACTTGTTTTAAACTTTTCTTTCCAACGAACACCGTTTCCTTCATTGTAACGTAAAAATGGTGAGCTTCTGCGGCGCGATTGGTCTCATCTTTCCTCTCGTATCGTTCGTCATTTTGTCCGAGAGCGTGTAGATATTGTACCCGATAGAACGGAGACGGAATCACGCTATACGGTTCCCTCCCAAATGTTCGTTCCTCACGGGCAAAAGTACAAGTAAAAGTAGGAGGGTCCGACTTTATGACCCTACGAGGCACAATAAAGGTTGATGACTACTAAACGTAGAAGAACACGTATACAGTCCGACGTAAAGTTATTTTTCAATTCCGCGGTCCTCCGCCGCAGTCGTTTTGCCTTGACGTGGAAAAGGAAATTTCCCGGGTTCTAGTCGTCCGGTCTTCTTTtCGTTCTACTGACAATACCATAtTTTCGACATAGATCGATCTCCTTTtCTCTTtCTCTCTCTCTCTCTCTCTCTCTCTTGAAAATGAAAGCGCGGAAGAGCCGGATCGTGCAACGCACAATGCGAGGCCGCTTTGTACGAGGTACGAGGAGTCCGTCATGCGCCGTAAAACGCCGGAATATGCAATACTACTTTCGCAGCGACGGGGCTTGCACGAAGTAATATCGTGAAACGTAGAACCGTTCTTTTCATACGGATATGCGGGAGAAGTTGCTCGTCCGCCCTCCGGCGCATACACGTCGCGCGAGAGTATCGTGCATCCGAAACTCTGAGATGAAACTCTTCTGTAGTCGATATTCGTCGCGATAAATAGATAAGTCTCGGATAAGGTAGAGACATCGATAATTCCGTTACGAGAATACTCGAGAATAAGATCCAAGTGAAGTGATCACGCTCCATATGGTTTCAGATTAATCTCCAATCGGCTGACGAAGGAGGATCATCCTTTTACCGGTGGAAAATAGGGTGGATTGGCGGAGCAAGAAGGCTAAACAGAGAATAAACAGGAGCTATAGCCGAACGAGGGAGAAGGTAAGTAGATGCCTCGGAGAACGTGAGCGAAAAAAGGAAACGGGTACGAGAGAAAAAGAAACGGACGGAATAGCGGCTCGGGATTCGCATCCCAAGGAAAACCAAGGCTATACCGGGGTCTTGGATTATTCGAGGCACGACGACGATCTTCGAGTCAGCCGACTGTCTGTACCGTGAAAGTGGCACGTATCGATCGCACGGCTGGATTATCTTCCACTTCGATCTACGACGATTACTTCCGCCATCGTATATCCGGGCTTTGCACTAGCGAGGCTATTTAAAAATCTGCGCTCAGTAACTACTTATGATTTTTCCATCAGAAACGATTGTGGAGAGAAAAGAGGGAAAAAAAGAGATAGACAGCCTCTGGCTCGAAATGCTAATTTCGCAATCGAGAATTAAAATGCACTTCTTTGTATCTAAATTTTCGTAGAATTAAAATAAAATTGAATAAAGCAATGAATAAATTGAATAAACTAAAATATAGCTAAATATTTTTTCCTCTATACAAGGTGAATATAATTATCAAATATTTAAGTATGTAGATTGAATTTAAACAGCCTCGGAAGAGAAAAGAATCGGATAAACGAAATGCTTTTGCCTCTATTTTCAAGCACGTGACGAATAAAATCTAGCAAAGCTTTTCGACACAATATGTCGACGCAAATGTGGTCTATTTTGGCTAATGATTATTACCGGGAGTCCCGGGCACGGTGTGTCCGCGGCGCGATAAATTAGAGCGCGAATCGACTTCCACGGCCGTTGTAGAAGGTACTTTGGCAAACGTTATTTCTTCCTGTCTCGAAGGAAGCGCCACTCGAAAACTTGGAAAAGTTCGGCCAGCTGCACGCACCGCGATCTCGGGTCCGTTCTGGCTCGGTGGCGTGCGCAGGACGGTTGTGAGACGAGAGAAAGAGAGAAAGGAGGATAGAGCGCGAGAGGGAAAGAAGAGGGAACATCCGCGTGCGTGGTTGTATATGGCGTTAATGGCGGCGAGCATAAAGCATCCCCGcGCCTGCACGCTCGACCACGGTCGTTTACAGTGCCACTATTTATGTTGATAACTTCGGAATGGAGTCAGATACACGGTACCGAGTGCCGGCGGTTCGGCGGTGGTCCGGCAGCGGTCTGGTAGCACTTGCAAAGTATGGGAGAAGAAGGGGGATGGTGGATTCGCGGATCTTTTTGGTCGTGCAAGGAAGGCGGGTCGGTTAGGGTAGGTAGAGAGGAGACGAGCCGAGTCGAGAACAAGATTCAAGCGGAAAGTTTTGCGAGTTAATGGTGCGGAGGTAATGCCGCGGCCGCAACAGCAACAGCCATCCCCCTGCTTGTGTGTTCGCTCGCTCGCTCACTtCTTCCGTTCTCTCCTCTTCGCGCTAGTTCTCTCTCTCTCTCTCTCTCTCTCTCTGtCTTTCTTGGAATAGCCGTAGAAAAAaGGAAAAGAAAGAGAGAGAAAGAACGAGACTCCTTTCTCCCCACGAATTCTCTCCTCTCTTTAAGCACACTCTCTCTTCCTGCaccCCCCCCCcTCcTCTCTCTCTCTTTCTCTCTCTATTTTCTTGTCTTCCTCACTTGCATCATCCCTCGTCCATTTCCcTCTGGAGAACGTGCCACGTTCTCACTTCCTCGTTCGTCGCTACTTTTTTTCTTCTTTAGTTGTCTCCTCTCACACCCTCGAGACGGCCCGATCTTTTCCTCGTACAGCTCCTATCACGAACAGTCGCTAACAAGTGCATCGAATGCAAGTTGCCGACAAACTTCTTCCACCGATTTGTGCTTGTTCTCTGTGCATGCGCGGGCATGTATATCTCTATTAGGCGACATCTGCTCTCAGCTTTTTCATAACGAGGTAGCGCGGATTCGCGCTCCGAGAGACTTCACGAAGCACTTCCGATCTCGCTACAGTAGAATGCGTATTGTATTTTCTTGTCTCTCTCATTTACTCTTTTCTATCTTTCGTATCTAGCGTGAATACTCCCATGAGGAATGTAGAAACCAGATTTTGAAACGGCTTCTTCGTATCTAAATTTTCTTAACTATTTGTTCCTCAAAGTCCATTTTGACGTTATAAATTTTTATTTTTTAATGAGAATGTTTTTATGTGGAGAAGAAAAGTACAACTTTTTTCAAAATGCAATTAAATTTATACAAGACTACTAAGATAAAAAAAGATGCAAATAATAATGTTCAACTTACTAACTGCTATATTATTAAGGCCAAGTTAAATTACAGATTTATGATGTTGCAGAATATAATAGAAAAGTTTCAAGAAAAAAaCATTTTAAAGCTTAAAGAGTTTGTTTTCAAACCCATCAAATTTTTTtCTCTGGCAGTGCTGCTGCTCTGCGACGTCATTTTCAAGTCGATCAATGCAAAGTTAACAAAAGTATTTCAACTTTAAAATCATGCAGAAAGTTGGGAAAAATTTGTTTTCAATTTTACTTAAATTTTAGTTACATTTTTTCTAAAAGCTTAAAATCTCCTCTTTAAAATATGTCTTTAAAAATTGAGCTATGATTTTTCTTCACCGAGTTATCGTAATTTGAAATGGCCAATCGACGTTTTCTTTTCATCACTAGGAATAATGCCGGCGGATTTAAGCTTTCAGAAGATTCCAAGCAAAGTTGAAAACAATTTGTTTGAATTCCGCATGATCTCAAAGTTGAAGCTAAAAAAATTTCACAAGACTTTAAAAGACAAAAAGTCAATGTTGCACAATCGACTTGGAAATAACGCCACTTGGAGCAGAGAAGTATTGCCGAGAAAAAGCTTCGACCGGGTTTGAAAGCAGGATCTATAGGCTTCCAAACTTTTTTTTtGAAAAATGGAGCTTTAAGGTTACTTTTCACGAAATTGTATAAATTGTGTTTTGTTGtTTTTATCATCGATTCGAAATGATCCTTTCATTTTGTTCAAGAATGTATCATTAACTTAATGCAATGATATCTTTTAATAATTCAATACTTCTTACTAATTAGATTTAGAAAAGTTTCATGAACAATTTTAGAACGGTTATTGCTAATTATTCGCCAAAAACAGACGCCTTTCTTCGAGGAGTAACAACTCAAGCTGCATTCGCGGTTGACGGCTTTCCGCGGCGCGTCGCGGTAATATGGCTCCTATTTAATTACCCGGTCCTTTGGGAGCTTAAACCAGCCAGAGCTGGAACGGCTGCGCCGTTATTCTATCTAGACTCCTTTGCTTTCTCAAGCAGGCGCGCGATCAAACCTTCTCGCATAAAAGACAATCGCAGCTGGCAGTCGACGACGCGcGGGACAGTCGAATCATGACCCGCTCCTCTCTAGTTACCGCCGTCAGTCTGCTCTACTTCCAGACGCCGCGCGTAATCTATTCGACATTAGTTGCTTGATTGCACCGTAAAATGCGACGGCGACGTGAACGAGAACGACGACGACGACGACGACGACGACAACGACGACGACAATGACAACGACGACGACGACAAGAGTGGGTTCGTCGGTGCTCGATGGCGCCTCCGATTTCAACCGCAGCACGATGCAAGCCCATTACTATCGCCCGGAGCTAAACGGCACCCGGAGCTCGTGCCATTAAGGGAATCTAGGGTCCGATCCACCTCATTGAATTCCGTTcAATTGCGATTATGATAATGCGTGAACGATCGCCGTGGACGTGAGCTACGGaCAACgAGGGTGTTCGTTCTCGGCTCAGAGAAACGCAGCGATAAAATTATCAGTGACAGCTTCATTTTTGTTACATTTGACGTTGAAAAATTTGCGAAAGAAATGTGCATTATTCAACTACATTGACAATTGTAAATCTTACATGACCTTTTATTAATATACATATATGTAAGATATATGTTCCATCTCTAGTCTCGTTCGATAATGAAGAAATTATAGCTGCAACATAGACGCCGGATTATTCGCGGGGGAAACCGTTGATTATTTGGACTCCGTGGGCTCGGGGTCTGGTTTACTTCTTCCCTTATGTCCGGAGATAATGGACACTATTACCTTAACGAGGCCCCAGTCTCTTAGCCGGTAATTGCTTCGAGATATCCGAGAGAGCTCCGGCGTACGTTGCCGCCTGGTGTTGCAGGCAGAGAACCCaGACGGTATTATTGCCGCCGAGGCTACTCGCCCGTTTCATGGTGGTTATTGTTATGGGCCTGCGGCATTAAGATGTACACCGCACTCTCATACGGGACCCACCACCCCATATACGAGCTGCATATATACTTATGCGGGGAGGATTTCATTACGCCGCTTCATAACTGGCGGTCTTCAAAATCGCGATAAATCGGGATTGTCTTCCTTGTTAAATCAGCTCCCGTTCCCCTTCCTCATATCGCTGACGAAGCCAACGAGACGGATTAATCTGCGCGATTGAATGGCCTCTATGACGTAAGCGCACCGTTTACTGGCACGGCTCCTCGTGTTCACGTGAAACGATTTGCGTCGTAAATATTTTATTTTAACATCGCAACATCAAGACAGACGAGGATCGGCTATTGCCTCGTGATCCTAAAAGGGAGATTCTCAAGGCGGAATCGGGGTAACGCGTTCGTTGATCTCGCCAAACTACGGCATCTTGAGGACTAGTCTTAGaGGAAAAAAAGACGACGAGGAGACACGGTGAGCATTAGATGAGAAAGAGACGGCGCGGCGCGGTGCGGCGGAGCGAGACGGAAAGAGATCAAATCTGGATATCAGGATTAGGGTGGGTACGTAATCCGCAGGACGACGGGTGGTAGGAACGGTGGATCCGTCGGCAGATCTCATCGCGCGGAGGAACTCTGCGGGTACTTCGCCCGGCCAAATACGACAAGAGCAGCAGCCTAACTCGAGTAGAGCCGCCGAGATGGTTTACGGCTCGTCGCAAGTTGGTGAAATTTAAGAGCGCATTTAAAACTGGTTTGGCGCGATGCCCTGCGCCTCCCTGCAGCAGGTACTGCCGGCGAGAGATACCGCTGGGCTCCGACAGGAGTTTATCGCCCTAACATTCTTGGAAAGCTTCGGATGGATTTCCCTCCTCAACCCATTTCTTCCGGCAGCACCTCAAATCGTCGCTTCTTTCGAGCTCCCTGTGTCGTCCGTTCTTTCCCCTTGCCACGAAAATTCTTCCAACAGACTCGACACCAAATCGTCACGACGATCATCGGTCAAGATACCCGGAGAAACGTCGCGGACGATGTTACCGTCAGATCGGACGCTTTTAGCTCCGCATTGGAGCTTTTTCCTCGACCGTCTCGGAGGAGATTTAGCGGACCGACAACAAATGAAGGTCATCTTTCGCGCGAGAAAGCGTCTCACCTTGATTTCCGTCTCGTTTCTCCGGAAATTGGATATCAACCGGACGAACTGTAATGTGTACGTTAAATTGCCAATTATATATATAATGTAACAGCTGATTATCTCAAGTGTCCTAAAAACCATTATACTCTTAATTTCTGTGAAAAATGGCGAAAATAAAAAAGAAACCGATCTTAATAAAGATATTCTTCCTGATAGATGCCATGACCCACGTGGAAAACTTTTTAGTTTTGTACAGTGGTATTATACGTTATCTTCCGCTGAACGTAAGACGTGCCTATCGCGCAATTTCATCGCGACGTCGTCGTATAGCGATTATGGCTACTCCATTAAAAATGAATTTTATAAAGGCAATCTTTCCAAGCGATCGTTGTAGGAGAAAAAGGCGAAAGCCGGAGCCAAAGGGGATGAGGCCACTACCTTTGGCTGATCCACTTCGAATGATAATCACCTCTAGGAGACTCAATTTCGCCCTGCTCCGCGTCCTTACCCGTTCCTATCTTCGGAAGGTTCAACGCCGCAGCGGACTGCATCTTTCACTCCCTTCGTCACCACCGCCCTATTCCTATCGCCCTCCGCGCGCCTACCGCCCCTATATCCTTCCCTTCCTTCACtCCTAGACTATTCTGAACGACCTCTTCCCCCATTCGCCAACGCTCACTCCTAACTGATTGGAGTACCAATCAATGCGGCATTCAGGCGGCCGTGCTGAAaCTTTAGGAAATTAACTATTCACTCTCTGGAAATGGTTATTTGGAAGGCCGGAAAGGCAGTCGGGACTACGTTACGGAGAATATGAGCGAAAGGATATGGGCAGACGATGACGATAGATAAGACAGTTCAATTTTCAATTAGGCGTGGACGTAGCGCGCGTAGCTCGGCTCCTACCACGATGCTTACCACGCGTCCCATACGTTAGGAGGAATCCGA

But making it useable is hard.

Sequencing a genome is now cheap & easy.

Tools for genomics work on emerging model organisms

Antgenomes.org Sequenceserver.com: "BLAST made easy

1: identifying sequences.

Gene predictionDozens of software algorithms: dozens of predictions

20% failure rate: "•missing pieces"•extra pieces"•incorrect merging"•incorrect splitting

Visual inspection... and manual fixing required. 1 gene = 20 minutes to 3 days"15,000 genes * 20 species = impossible. Ya

ndell

& E

nce

2013

NRG

GTCTACAATGCGATTGTAAAATAGCACGAgAGGTGCATATGATGAACGACTATGTTCCACAACCACAGCTCATATATAACATGATTTtGTTTGCCGAATTCATACACGCATTACAACACACATTGAATTCAATAATAATATCAAATTCACATTCAAAGCTTTCAAGTTAGACAAAAGTTTTAATGCCGTTTTtACCTGTTTTtGAAAAGGTAATTTTCTTTAGATATATACAGTTTGTAATaTTAGGTATTTTATAAACAGTGTGTATATTTCTTACAATATAAAAGACACAATTGCAAACTAGCATGATTGTAAACAATTGCTAAACGGATCAATATAAATTAAAATTGTAATATTAAGTATCAAACCGATAATTTTTATTTATTGTTCATTGTTTGTTCTTTATTTTGTTATTTGTAAATAATGAAA

Evidence

Evidence

Consensus:

Monica Dragan https://github.com/monicadragan/GeneValidator

2.

https://github.com/monicadragan/GeneValidator

Gene predictionDozens of software algorithms: dozens of predictions

20% failure rate: "•missing pieces"•extra pieces"•incorrect merging"•incorrect splitting

Visual inspection... and manual fixing required. 1 gene = 20 minutes to 3 days"15,000 genes * 20 species = impossible. Ya

ndell

& E

nce

2013

NRG

GTCTACAATGCGATTGTAAAATAGCACGAgAGGTGCATATGATGAACGACTATGTTCCACAACCACAGCTCATATATAACATGATTTtGTTTGCCGAATTCATACACGCATTACAACACACATTGAATTCAATAATAATATCAAATTCACATTCAAAGCTTTCAAGTTAGACAAAAGTTTTAATGCCGTTTTtACCTGTTTTtGAAAAGGTAATTTTCTTTAGATATATACAGTTTGTAATaTTAGGTATTTTATAAACAGTGTGTATATTTCTTACAATATAAAAGACACAATTGCAAACTAGCATGATTGTAAACAATTGCTAAACGGATCAATATAAATTAAAATTGTAATATTAAGTATCAAACCGATAATTTTTATTTATTGTTCATTGTTTGTTCTTTATTTTGTTATTTGTAAATAATGAAA

Evidence

Consensus:

Begin

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Being curated

Curate

Being curated

Curate

Being curated

Curate

Submit Submit Submit

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http://afra.sbcs.qmul.ac.ukAnurag Priyam

3. Crowd-sourcing inspection & correction of gene predictions Scientific software + Facebook + Points + Badges + Redundancy

http://afra.sbcs.qmul.ac.uk

Ecology & Evolution & Vital-IT @ LausanneEvolve & Psych @ Queen Mary

LAURENT KELLER JOHN WANG DEWAYNE SHOEMAKER OKSANA RIBA-GROGNUZ MINGKWAN NIPITWATTANAPHON !

[email protected]

M Corona, S Nygaard, BG Hunt, KK Ingram, L Falquet, M Nipitwattanaphon, D Gotzek, MB Dijkstra, J Oettler, F Comtesse, CJ Shih, WJ Wu, CC Yang, J Thomas, E Beaudoing, S Pradervand, V Flegel, ED Cook, R Fabbretti, H Stockinger, L Long, WG Farmerie, J Oakey, JJ Boomsma, P Pamilo, SV Yi, J Heinze, MAD Goodisman, L Farinelli, K Harshman, N

Hulo, L Cerutti, Ioannis Xenarios

Anurag Priyam & Monica Dragan

http://www.unil.ch/dee/page32298_en.html

mailto:[email protected]

2013 11-25-sheffield

Science

multiplequeen form

colleagues social form

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colleagues laurent keller