S. Uthicke, J. Doyle, S. Duggan, N. Yasuda, A. D mcKinnon




Дата канвертавання21.04.2016
Памер170.17 Kb.
Supplementary Information

Outbreak of coral-eating Crown-of-Thorns creates continuous cloud of larvae over 320km of the Great Barrier Reef



S. Uthicke, J. Doyle, S. Duggan, N. Yasuda, A.D McKinnon

Supplementary Table 1 Detailed list of sample locations. VT= vertical tows from the research vessel; HT = near or on reef horizontal tows from the tender; NS = no sequence obtained (i.e. not counted as positive sample).



Station Name

Latitude

(S)


Longitude

(E)


Date


Type of tow

Water Depth (m)

Location


Genbank Accession No.

COT001

16.0793

145.7033

17/12/2014

VT

29

S of Undine Rf.

NS

COT002

15.6958

145.7290

18/12/2014

VT

52

S of Osterlund Rf.

KT271503

COT003

15.6638

145.6607

18/12/2014

VT

40

S of Osterlund Rf.

KT271504

COT004

15.6250

145.5950

18/12/2014

VT

35

S of Osterlund Rf.

KT271505

COT005

15.5867

145.5447

18/12/2014

VT

35

S of Osterlund Rf.

KT271506

COT006

15.5455

145.4560

18/12/2014

VT

32

S of Osterlund Rf.

KT271507

COT007

15.5927

145.5748

18/12/2014

HT

2 – 10

Emily/Osterlund Rf.

KT271508

COT008

15.5914

145.5777

18/12/2014

HT

2 – 10

Emily/Osterlund Rf.

NS

COT009

15.5913

145.5799

18/12/2014

HT

2 – 10

Emily/Osterlund Rf.

NS

COT010

15.5810

145.5791

18/12/2014

HT

2 – 10

Emily/Osterlund Rf.

NS

COT011

15.5827

145.5804

18/12/2014

HT

2 – 10

Emily/Osterlund Rf.

NS

COT012

15.6155

145.7700

19/12/2014

VT

48

Irene Rf./Ribbon Rf. 1

KT271509

COT013

15.5862

145.7993

19/12/2014

VT

44

Ribbon Rf. 1/2

KT271510

COT014

15.5712

145.8838

19/12/2014

VT

1400*

Coral Sea

NS

COT015

15.5558

145.9700

19/12/2014

VT

2000*

Coral Sea

NS

COT016

15.6197

145.8067

19/12/2014

HT

2 – 10

Ribbon Rf. 1

NS

COT017

15.6160

145.8095

19/12/2014

HT

2 – 10

Ribbon Rf. 1

NS

COT018

15.6120

145.8091

19/12/2014

HT

2 – 10

Ribbon Rf. 1

NS

COT019

15.6008

145.7983

19/12/2014

HT

2 –- 10

Ribbon Rf. 1

KT271511

COT020

15.6037

145.8001

19/12/2014

HT

2 – 10

Ribbon Rf. 1

NS

COT021

15.6010

145.8024

19/12/2014

HT

2 – 10

Ribbon Rf. 1

NS

COT022

15.6470

145.5763

20/12/2014

VT

38

E of Ribbon Rf. 1

KT271512

COT023

15.7422

145.5710

20/12/2014

VT

31

Cairns Rf./Endeavour Rf.

NS

COT024

15.8035

145.5847

20/12/2014

VT

33

Endeavour Rf./Pickersgill Rf.

KT271513

COT025

15.8496

145.5883

20/12/2014

VT

34

N of Pickersgill Rf.

KT271514

COT026

15.9092

145.5750

20/12/2014

VT

32

S of Pickersgill Rf.

KT271515

COT027

15.9385

145.6527

20/12/2014

VT

39

Evening Rf.

KT271516

COT028

16.0328

145.6317

20/12/2014

VT

33

Mackay Rf.

KT271517

COT029

16.1530

145.6743

20/12/2014

VT

42

N of Rudder Rf.

KT271518

COT030

16.1777

145.6743

20/12/2014

HT

2 – 10

Rudder Rf.

KT271519

COT031

16.1753

145.6790

20/12/2014

HT

2 – 10

Rudder Rf.

KT271520

COT032

16.1848

145.6690

20/12/2014

HT

2 – 10

Rudder Rf.

KT271521

COT033

16.1878

145.6687

20/12/2014

HT

2 – 10

Rudder Rf.

KT271522

COT034

16.2135

145.6661

20/12/2014

HT

2 – 10

Rudder Rf.

KT271523

COT035

16.2159

145.6695

20/12/2014

HT

2 – 10

Rudder Rf.

KT271524

COT036

16.2325

145.6430

21/12/2014

VT

42

Rudder Rf./Tongue Rf.

KT271525

COT037

16.3938

145.6245

21/12/2014

VT

31

E of Batt Rf.

KT271526

COT038

16.4878

145.8273

21/12/2014

VT

39

S of Batt Rf.

KT271527

COT039

16.5973

145.8703

21/12/2014

VT

39

E of Oyster Rf.

KT271528

COT040

16.7303

145.9717

21/12/2014

VT

43

Arlington Rf./Green Isl.

KT271529

COT041

16.9042

146.1355

22/12/2014

VT

39

E of Sudbury Rf.

KT271530

COT042

17.2202

146.2545

22/12/2014

VT

38

S of Flora Rf.

KT271531

COT043

17.4918

146.3413

22/12/2014

VT

38

E of Peart Rf.

KT271532

COT044

17.6320

146.3516

22/12/2014

VT

42

N of Ellison Rf.

NS

COT045

17.9402

146.4468

22/12/2014

VT

36

S of Beaver Rf.

KT271533

COT046

18.0801

146.4744

22/12/2014

VT

32

E of Otter Rf.

KT271534

COT047

18.2140

146.5213

22/12/2014

VT

35

E of Britomart Rf.

NS

COT048

19.1010

146.8630

23/12/2014

VT

15

Magnetic Isl.

NS

COT049

16.7538

145.9760

13/01/2015

HT

2 – 8

Green Isl.

NS

COT050

16.7538

145.9765

13/01/2015

HT

2 – 8

Green Isl.

NS

COT051

17.7591

146.3573

17/02/2015

VT

41

S of Ellison Rf.

NS

COT052

16.7713

145.9540

18/02/2015

VT

39

Green Isl.

NS

COT053

16.9208

145.9967

20/02/2015

VT

18

Fitzroy Isl.

NS

COT054

17.7591

146.3573

16/06/2015

VT

40

S of Ellison Rf.

NS

COT055

16.7716

145.9540

17/06/2015

VT

38

Green Isl.

NS

COT056

16.9207

145.9967

17/06/2015

VT

18

Fitzroy Isl.

NS

 

 

 

 







 




*: Two deep stations in the Coral Sea only towed to 100 m.

Supplementary Table 2 Asteroid and other Echinoderm samples used for specificity testing of the primers developed; pc= pyloric caeca.



Sample number

Date


Class


GenBank No.

Species


Tissue


Location


3

21/2/14

Asteroidea

KT271482

Nardoa novaecaledoniae

skin

Pelorus Is.

5

20/2/14

Asteroidea

KT271483

Echinaster sp.

skin

Picnic Bay

14

24/2/14

Asteroidea

KT271484

Cryptasterina sp.

skin

Orpheus Is.

18

26/2/14

Asteroidea

KT271485

Fromia indica

skin

Rib Rf.

20

26/2/14

Asteroidea

KT271486

Echinaster callosus

skin

Rib Rf.

22

26/2/14

Asteroidea

KT271487

Echinaster sp.

skin

Rib Rf.

28

26/2/14

Asteroidea

KT271488

Linckia multifora

skin

Rib Rf.

31

1/3/14

Asteroidea

KT271489

Choriaster sp.

skin

Davies Rf.

32

16/3/14

Asteroidea

KT271490

Neoferdina cummingii

skin

Lizard Is.

33

16-/3/14

Asteroidea

KT271491

Fromia monilis

skin

Lizard Is.

38

13/2/14

Asteroidea

KT271492

Linckia laevigata

pc

Davies Rf.

41

13/2/14

Holothurioidea

KT271493

Stichopus chloronotus

pc

Davies Rf.

72

8/11/13

Asteroidea

KT271494

Acanthaster planci

gonads

Arlington Rf.

74

8/11/13

Asteroidea

KT271495

Acanthaster planci

gonads

Arlington Rf.

75

8/11/13

Asteroidea

KT271496

Acanthaster planci

gonads

Arlington Rf.

76

8/11/13

Asteroidea

KT271497

Acanthaster planci

gonads

Arlington Rf.

78

8/11/13

Asteroidea

KT271498

Acanthaster planci

gonads

Arlington Rf.

80

6/12/13

Asteroidea

KT271499

Acanthaster planci

gonads

Arlington Rf.

81

8/11/13

Asteroidea

KT271500

Acanthaster planci

gonads

Arlington Reef

82

8/11/13

Asteroidea

KT271501

Acanthaster planci

gonads

Arlington Rf.

84

12/11/13

Asteroidea

KT271502

Acanthaster planci

gonads

Arlington Rf.

Supplementary Table 3 Details of Echinoderm embryos and larvae picked from a subset of plankton samples (‘Sample’), each larva received a separate identification number (‘Larva’). ‘Species’ and ‘Class’ describe the nearest GenBank match, with percentage of identity given. Interspecific divergence of the Asteroidea and Holothuroidea is usually < 2% , thus identity values below that range can be regarded as actual species matches. Even general can diverge by > 20%, thus in matches below 90% even the inferred Class of the sample will be unreliable.




Larval samples




GenBank







Sample

Larva

Accession No.

Species

Class

Identity.

Accession No.

COT007

L001

KT271547

Acanthaster planci

Asteroidea

99%

AB231475.1

COT007

L005

KT271548

Acanthaster planci

Asteroidea

99%

AB231475.1

COT003

L011

KT271535

Thelenota anax

Holothuroidea

99%

FJ971404.1

COT003

L012

KT271536

Acanthaster planci

Asteroidea

99%

AB231475.1

COT003

L021

KT271544

Holothuria cf. cinerascens

Holothuroidea

83%

EU848262.1

COT033

L024

KT271560

Patiriella parvivipara

Asteroidea

82%

U50055.1

COT033

L026

KT271561

Acanthaster planci

Asteroidea

99%

AB231475.1

COT033

L027

KT271562

Acanthaster planci

Asteroidea

99%

AB231475.1

COT033

L028

KT271563

Acanthaster planci

Asteroidea

99%

AB231475.1

COT033

L031

KT271564

Nectria ocellata

Asteroidea

82%

EU869956.1

COT006

L034

KT271546

Linckia laevigata

Asteroidea

100%

KF834635.1

COT005

L039

KT271545

Luidia maculata

Asteroidea

91%

JQ740635.1

COT039

L048

KT271566

Holothuria notabilis

Holothuroidea

85%

KJ801945.1

COT039

L050

KT271567

Patiriella parvivipara

Asteroidea

82%

U50055.1

COT039

L051

KT271568

Holothuria notabilis

Holothuroidea

99%

KJ801945.1

COT026

L052

KT271552

Holothuria fuscopunctata

Holothuroidea

99%

EU848272.1

COT026

L054

KT271553

Actinopyga sp.

Holothuroidea

99%

EU848232.1

COT038

L056

KT271565

Patiriella parvivipara

Asteroidea

82%

U50055.1

COT022

L059

KT271551

Acanthaster planci

Asteroidea

99%

AB231475.1

COT003

L201

KT271537

Oreaster occidentalis

Asteroidea

93%

U50059.1

COT003

L202

KT271538

Holothuria leucospilota

Holothuroidea

84%

FJ589211.1

COT003

L204

KT271539

Tropiometra afra

Crinoidea

80%

GU327867.1

COT003

L206

KT271540

Holothuria impatiens

Holothuroidea

100%

KF142183.1

COT003

L207

KT271541

Holothuria notabilis

Holothuroidea

99%

KJ801945.1

COT003

L208

KT271542

Holothuria coluber

Holothuroidea

99%

EU848297.1

COT003

L209

KT271543

Holothuria arenicola

Holothuroidea

99%

JN207610.1

COT033

L210

KT271554

Patiriella parvivipara

Asteroidea

82%

U50055.1

COT033

L211

KT271555

Patiriella parvivipara

Asteroidea

82%

U50055.1

COT033

L212

KT271556

Oreaster reticulatus

Asteroidea

83%

U50060.1

COT033

L213

KT271557

Patiriella parvivipara

Asteroidea

82%

U50055.1

COT033

L214

KT271558

Patiriella parvivipara

Asteroidea

82%

U50055.1

COT033

L215

KT271559

Oreaster reticulatus

Asteroidea

83%

U50060.1

COT022

L218

KT271549

Luidia maculata

Asteroidea

91%

JQ740635.1

COT022

L219

KT271550

Thelenota ananas

Holothuroidea

82%

EU848258.1

Supplementary References

Uthicke, S., M. Byrne, and C. Conand. 2010. Genetic Barcoding of commercial Bêche-de-mer species (Echinodermata: Holothuroidea). Molecular Ecology Resources 10:634-646.

Ward, R., B. Holmes, and T. O'Hara. 2008. DNA barcoding discriminates echinoderm species. Molecular Ecology Resources 8:1202-1211.

Supplementary Figure 1 Specificity tests of CoTS primers against other Echinoderm samples from the GBR (see Supplementary Table 1 for sample numbers). Note only 72 and 78 from A. planci GBR tissue collection were used as representatives of the 9 CoTS specimens collected at Arlington Reef. Neg = negative control.



Supplementary Figure 2 Sensitivity test of CoTS primers developed. CoTS larvae from cultures were mixed with 100 (1 in100), 1000 (1 in 1000) and 10,000 (1 in 10,000) other organisms from a plankton sample. Pos = positive control, Neg = negative control.











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