Barcoding Anurans from Brazilian Atlantic Forest · AnR1 Primers design 1- Previous works in...

Barcoding Anurans from Brazilian Atlantic Forest

Mariana L. Lyra, Célio F. B. Haddad & Ana Maria L. de Azeredo-Espin

[email protected]

Capes/CNPq Prodoc nº 563.975/05-9

International Symposium on DNA Barcoding –São Paulo 03-04 Dezember 2009

mailto:[email protected]

Laboratório de Herpetologia (UNESP-Rio Claro)

Prof. Célio F. B. Haddad

Biggest tissue andspecimens collection

of Anurans (Brazil)

Laboratório Genética Animal (UNICAMP)

Profª Ana Maria L. Azeredo-Espin

Molecular MarkersmtDNA

+

Pilot Project: Testing COI DNA Barcode in Anurans fromBrazilian Atlantic Forest



[email protected]




Amphibia- 6433 sp.

Anura- 5679 sp. Caudata- 580 sp. Gymnophiona- 174 sp.

Frost, Darrel R. 2009. Amphibian Species of the World: an Online Reference. Version 5.3 (12 February, 2009). Electronic Database accessible at

http://research.amnh.org/herpetology/amphibia/ - American Museum of Natural History, New York, USA.

Siphonops paulensisBolitoglossa paraensisPhyllomedusa sp.

Amphibia- 6433 sp.


Frost, Darrel R. 2009. Amphibian Species of the World: an Online Reference. Version 5.3 (12 February, 2009). Electronic Database accessible at

http://research.amnh.org/herpetology/amphibia/ - American Museum of Natural History, New York, USA.

Amphibia- 857 sp.


Siphonops paulensisBolitoglossa paraensisPhyllomedusa sp.

e.g. Wake 1991, Alford & Richards 1999,Stuart et al. 2004, Vietes et al . 2009….

National Geographic Brazil – April 2009

Decline in amphibian populations worldwide

National Geographic Brazil – April 2009

1- Habitat Loss 2- Climate changes 3- Spread of microbial pathogens

e.g. Wake 1991, Alford & Richards 1999,Stuart et al. 2004, Vietes et al . 2009….

Decline in amphibian populations worldwide

Original area

16% of the Brazilian territory

Today:

~7-8%

~1,2-0,8% intact

Brazilian Atlantic Forest

Data:MMA; INPE/ Fundação SOS mata atlântica, 2009

today

original area

Legend

Biodiversity hotspots



today

original area

Legend

460 amphibian species

~90 endemic species-Bioma

Original area


Today:

~7-8%

~1,2-0,8% intact


today

original area

Legend

New species are described

every year

How many are

disappearing?

Biodiversity hotspots


460 amphibian species

~90 endemic species-Bioma

Original area


Today:

~7-8%

~1,2-0,8% intact

We need to know the species to conservate

DNA barcoding

has emerged at the forefront of efforts to accelerate the rates of taxonomic discovery

DNA barcoding

has emerged at the forefront of efforts to accelerate the rates of taxonomic discovery

Amphibians COI DNA barcoding main problems:

1- the high variability of priming sites that hinder the application of universal primers

2- the observed distinct overlap of intraspecific and interspecific divergence values,

which implies difficulties in the definition of threshold values to identify candidate species

Vences et al 2005; Smith et al. 2007

We need to know the species to conservate

Objectives

1- Design new primers for standard COI amplifications in Anurans

2- Preliminary analysis of inter and intraspecific diversity in anurans

from Brazilian atlantic forest: COI and 16S

3- Case study in Leptodactylus ocellatus (Anura: Leptodactylidae)

4- Case study in Scinax fuscovarius (Anura: Hylidae)

Objectives


2- Preliminary analysis of inter and intraspecfiic diversity in anurans




CFBH tissue collection

(UNESP-Rio Claro)

1994-2009

58 anuran species

14 different families

Images: Célio F B Haddad

Aparasphenodon brunoi

Aplastodiscus callipygius

Xenohyla truncataPhysalaemus atlanticus

Phylomedusa sp.Ceratophrys aurita

Arcovomer passarellii

Flectonotus sp.

Brachycephalus epphipiumIschnocnema sp.

Hypsiboas prasinus

Proceratophrys appendiculata


(UNESP-Rio Claro)

1994-2009

58 anuran species

1 Caudata


Bolitoglossa paraensis



Xenohyla truncata

Phylomedusa sp.


Flectonotus sp.


Hypsiboas prasinus

3 Gymnophiona


(UNESP-Rio Claro)

1994-2009

58 anuran species

1 Caudata


Siphonops paulensis



Xenohyla truncata

Phylomedusa sp.


Flectonotus sp.


Hypsiboas prasinus

Primers design

1- Previous works in Amphibian Barcode

Vences et al. 2005 – (Amphibians from Madagascar)

Smith et al. 2008 (Holoartic amphibians)

2 – 33 complete COI sequences (GeneBank)

COI

W A N C Y S D

Barcode region

COI

AnF1 AnCOI-R

AnR1

Primers design





AnW AnD

Nested or Semi-nested PCR- Museum specimens

COI

W A N C Y S D

Barcode region

COI

AnF1 AnCOI-R

AnR1

Primers design





16S

LV16S

An-16SF An-16SR

COI

W A N C Y S D

Barcode region

COI

AnF1 AnCOI-R

AnR1

Primers design





AnW AnD

COI and 16S - All species amplified

Primers design: Results

0

15

30

45

60

AnF1+AnCOIR AnF1+AnR1 AW-AD 16S

Works Fail

Barcode Region Nested

Objectives


2- Preliminary analysis of inter and intraspecific diversity in anurans




Number species Anura:

58

Number individuals:

170

~3 ind/ sp

Intra and interspecific diversity – COI x 16S

16S

COI

Barcode region

COI: 0.00-0.135 (K2P) 16S: 0.00-0.18 (K2P)

Intraspecific diversity – COI x 16S

0

0,04

0,08

0,12

0,16

COI 16S

K2

P d

ista

nc

e

COI: 0.00-0.135 (K2P) 16S: 0.00-0.18 (K2P)

0

0,04

0,08

0,12

0,16

COI 16S

Intraspecific diversity – COI x 16SK

2P

dis

tan

ce

Highly correlated with geographical distance

0

0,04

0,08

0,12

0,16

COI 16S


2P

dis

tan

ce

Highly correlated with geographical distance

Potential new species? – look for other evidences

0

0,04

0,08

0,12

0,16

COI 16S


2P

dis

tan

ce

COI

Interspecific variation

15-29%

0

2

4

6

8

10

12

14

16

18

20

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

Intra Inter

0

2

4

6

8

10

12

14

16

0 0-1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

intra inter

16S


6-30%

Nº

specie

sN

º specie

s

Interspecific diversity – COI x 16S

K2P distance

K2P distance

Candidate new species?

Sc. catharinae Pseudopadicola cf. falcipes

COI


15-29%

16S


6-30%

0

2

4

6

8

10

12

14

16

18

20

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

Intra Inter

0

2

4

6

8

10

12

14

16

0 0-1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

intra inter

Nº

specie

sN

º specie

s


COI

16S

BUT

Data came from very distinct

anuran families

Interspecific variation are

probably over-estimated

We need to include more

congeneric species to

understand intra and

interspecific variation in

anurans

Can not suggest a threshold

values to identify candidate

species based on this data

0

2

4

6

8

10

12

14

16

18

20

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

Intra Inter

0

2

4

6

8

10

12

14

16

0 0-1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

intra inter

% variation

% variation

Nº

specie

sN

º specie

s


Objectives


2- Preliminary analysis of inter and intraspecic diversity in anurans




Leptodactylus ocellatus (Linnaeus 1758)

21 specimens

http://www.iucnredlist.org/details/57151/0

Célio F B Haddad

A181 L.ocellatus_BA

A243 L.ocellatus_ES

A242 L.ocellatus_BA

A271 L.ocellatus_RJ

A37 L.ocellatus_SP

A237 L.ocellatus_RJ

A238 L.ocellatus_ES

A112 L.ocellatus_ES

A264 L.ocellatus_BA

A235 L.ocellatus_SC

A272 L.ocellatus_SP

A228 L.ocellatus_SP

A253 L.ocellatus_SP

A205 L.ocellatus_MG

A219 L.ocellatus_SP

A222 L.ocellatus_MG

A232 L.ocellatus_PR

A221 L.ocellatus_PR

A260 L.ocellatus_PR

A267 L.ocellatus_RS

A236 L.ocellatus_RS100

90

99

100

100

73

7898

100

96

0.02

Neighbor-joining/ K2P

AND

Gen. Mix.Yule-Coalescent model – GMYC

(Bayesian analysis)

Leptodactylus ocellatus: tree

A181 L.ocellatus_BA

A243 L.ocellatus_ES

A242 L.ocellatus_BA

A271 L.ocellatus_RJ

A37 L.ocellatus_SP

A237 L.ocellatus_RJ

A238 L.ocellatus_ES

A112 L.ocellatus_ES

A264 L.ocellatus_BA

A235 L.ocellatus_SC

A272 L.ocellatus_SP

A228 L.ocellatus_SP

A253 L.ocellatus_SP

A205 L.ocellatus_MG

A219 L.ocellatus_SP

A222 L.ocellatus_MG

A232 L.ocellatus_PR

A221 L.ocellatus_PR

A260 L.ocellatus_PR

A267 L.ocellatus_RS


90

99

100

100

73

7898

100

96

0.02

COI – NJ K2P

Leptodactylus ocellatus: Clades

A181 L.ocellatus_BA

A243 L.ocellatus_ES

A242 L.ocellatus_BA

A271 L.ocellatus_RJ

A37 L.ocellatus_SP

A237 L.ocellatus_RJ

A238 L.ocellatus_ES

A112 L.ocellatus_ES

A264 L.ocellatus_BA

A235 L.ocellatus_SC

A272 L.ocellatus_SP

A228 L.ocellatus_SP

A253 L.ocellatus_SP

A205 L.ocellatus_MG

A219 L.ocellatus_SP

A222 L.ocellatus_MG

A232 L.ocellatus_PR

A221 L.ocellatus_PR

A260 L.ocellatus_PR

A267 L.ocellatus_RS


90

99

100

100

73

7898

100

96

0.02

A181 16S BA

A037 16S SP

A243 16S ES

A242 16S BA

A238 16S ES

A271 16S RJ

A237 16S RJ

A112 16S ES

A264 16S BA

A272 16S SP

A228 16S SP

A235 16S SC

A253 16S SP

A221 16S PR

A219 16S SP

A222 16S MG

A232 16S PR

A211 16S SC

A205 16S MG

A260 16S PR

A267 16S RS

A236 16S RS

100

89

68

88

99

0.02

COI – NJ K2P 16S – NJ K2P

Leptodactylus ocellatus: clades COI x 16S

clade 1

clade 2: D=0.016

clade 3: D=0.004

clade 4: D=0.006

clade 5: D=0.011

A181 L.ocellatus_BA

A243 L.ocellatus_ES

A242 L.ocellatus_BA

A271 L.ocellatus_RJ

A37 L.ocellatus_SP

A237 L.ocellatus_RJ

A238 L.ocellatus_ES

A112 L.ocellatus_ES

A264 L.ocellatus_BA

A235 L.ocellatus_SC

A272 L.ocellatus_SP

A228 L.ocellatus_SP

A253 L.ocellatus_SP

A205 L.ocellatus_MG

A219 L.ocellatus_SP

A222 L.ocellatus_MG

A232 L.ocellatus_PR

A221 L.ocellatus_PR

A260 L.ocellatus_PR

A267 L.ocellatus_RS


90

99

100

100

73

7898

100

96

0.02

Leptodactylus ocellatus: intra-clade diversity

K2P

D=0.038

D=0.147

A181 L.ocellatus_BA

A243 L.ocellatus_ES

A242 L.ocellatus_BA

A271 L.ocellatus_RJ

A37 L.ocellatus_SP

A237 L.ocellatus_RJ

A238 L.ocellatus_ES

A112 L.ocellatus_ES

A264 L.ocellatus_BA

A235 L.ocellatus_SC

A272 L.ocellatus_SP

A228 L.ocellatus_SP

A253 L.ocellatus_SP

A205 L.ocellatus_MG

A219 L.ocellatus_SP

A222 L.ocellatus_MG

A232 L.ocellatus_PR

A221 L.ocellatus_PR

A260 L.ocellatus_PR

A267 L.ocellatus_RS


90

99

100

100

73

7898

100

96

0.02

Leptodactylus ocellatus: inter-clade diversity

K2P

Strong correlated with geographical distribution

A181 L.ocellatus_BA

A243 L.ocellatus_ES

A242 L.ocellatus_BA

A271 L.ocellatus_RJ

A37 L.ocellatus_SP

A237 L.ocellatus_RJ

A238 L.ocellatus_ES

A112 L.ocellatus_ES

A264 L.ocellatus_BA

A235 L.ocellatus_SC

A272 L.ocellatus_SP

A228 L.ocellatus_SP

A253 L.ocellatus_SP

A205 L.ocellatus_MG

A219 L.ocellatus_SP

A222 L.ocellatus_MG

A232 L.ocellatus_PR

A221 L.ocellatus_PR

A260 L.ocellatus_PR

A267 L.ocellatus_RS


90

99

100

100

73

7898

100

96

0.02

Leptodactylus ocellatus: geographic distribution

Bioacoustics and chromosomal differences

– coastal plain x plateau areas

(Silva et al. 2000)

A181 L.ocellatus_BA

A243 L.ocellatus_ES

A242 L.ocellatus_BA

A271 L.ocellatus_RJ

A37 L.ocellatus_SP

A237 L.ocellatus_RJ

A238 L.ocellatus_ES

A112 L.ocellatus_ES

A264 L.ocellatus_BA

A235 L.ocellatus_SC

A272 L.ocellatus_SP

A228 L.ocellatus_SP

A253 L.ocellatus_SP

A205 L.ocellatus_MG

A219 L.ocellatus_SP

A222 L.ocellatus_MG

A232 L.ocellatus_PR

A221 L.ocellatus_PR

A260 L.ocellatus_PR

A267 L.ocellatus_RS


90

99

100

100

73

7898

100

96

0.02


Diversity is underestimated

Bioacoustics and chromosomal differences

– coastal plain x plateau areas

(Silva et al. 2000)


A181 L.ocellatus_BA

A243 L.ocellatus_ES

A242 L.ocellatus_BA

A271 L.ocellatus_RJ

A37 L.ocellatus_SP

A237 L.ocellatus_RJ

A238 L.ocellatus_ES

A112 L.ocellatus_ES

A264 L.ocellatus_BA

A235 L.ocellatus_SC

A272 L.ocellatus_SP

A228 L.ocellatus_SP

A253 L.ocellatus_SP

A205 L.ocellatus_MG

A219 L.ocellatus_SP

A222 L.ocellatus_MG

A232 L.ocellatus_PR

A221 L.ocellatus_PR

A260 L.ocellatus_PR

A267 L.ocellatus_RS


90

99

100

100

73

7898

100

96

0.02

Objectives


2- Preliminary analysis of inter and intraspecic diversity in anurans




Scinax fuscovarius (Lutz, 1925)

Foto: Raúl Maneyoro

Foto: Lucas Grandinetti

http://www.iucnredlist.org/apps/redlist/details/55958/0

A258 S.fuscov SP

A200 S.fuscov SP

A234 S.fuscov SC

A220 S.fuscov SP

A202 S.fuscov SP

A257 S.fuscov SP

A254 S.fuscov SP

A146 S.fuscov SP

A227 S.fuscov SP

A233 S.fuscov MG

A51 S.fuscov SP

A248 S.fuscov SP

A109 S.fuscov SP

A183 S.fuscov SP

A201 S.fuscov SP

A250 S.fuscov SP

A214 S.fuscov MS

A230 S.fuscov SC

A245 S.fuscov MS

A212 S.fuscov SC

A179 S.fuscov Es

A204 S.fuscov GO

A203 S.fuscov GO

A247 S.fuscov MT

A226 S.fuscov MG

A266 S.fuscov RS

A241 S.fuscov SP

A197 S.fuscov SP

67

80

89

100

100

92

9691

96

63 49

47

38

54

15

11

34

33

38

2

1

3

6

13

0.02

COI: NJ-K2P

Scinax fuscovarius: tree

Diversity

0 – 24%

A258 S.fuscov SP

A200 S.fuscov SP

A234 S.fuscov SC

A220 S.fuscov SP

A202 S.fuscov SP

A257 S.fuscov SP

A254 S.fuscov SP

A146 S.fuscov SP

A227 S.fuscov SP

A233 S.fuscov MG

A51 S.fuscov SP

A248 S.fuscov SP

A109 S.fuscov SP

A183 S.fuscov SP

A201 S.fuscov SP

A250 S.fuscov SP

A214 S.fuscov MS

A230 S.fuscov SC

A245 S.fuscov MS

A212 S.fuscov SC

A179 S.fuscov Es

A204 S.fuscov GO

A203 S.fuscov GO

A247 S.fuscov MT

A226 S.fuscov MG

A266 S.fuscov RS

A241 S.fuscov SP

A197 S.fuscov SP

67

80

89

100

100

92

9691

96

63 49

47

38

54

15

11

34

33

38

2

1

3

6

13

0.02

COI: NJ-K2P

Scinax fuscovarius: geographic distribution

Not correlated with geographic distance

Specimens collection:

Correction

A197 Scinax sp.

A226 Sc. eurydice

A241 Sc. cf. hayii

A266 Scinax sp.

A258 S.fuscov SP

A200 S.fuscov SP

A234 S.fuscov SC

A220 S.fuscov SP

A202 S.fuscov SP

A257 S.fuscov SP

A254 S.fuscov SP

A146 S.fuscov SP

A227 S.fuscov SP

A233 S.fuscov MG

A51 S.fuscov SP

A248 S.fuscov SP

A109 S.fuscov SP

A183 S.fuscov SP

A201 S.fuscov SP

A250 S.fuscov SP

A214 S.fuscov MS

A230 S.fuscov SC

A245 S.fuscov MS

A212 S.fuscov SC

A179 S.fuscov Es

A204 S.fuscov GO

A203 S.fuscov GO

A247 S.fuscov MT

A226 S.fuscov MG

A266 S.fuscov RS

A241 S.fuscov SP

A197 S.fuscov SP

67

80

89

100

100

92

9691

96

63 49

47

38

54

15

11

34

33

38

2

1

3

6

13

0.02

COI: NJ-K2P

Scinax fuscovarius: collection

COI: NJ-K2P

Scinax fuscovarius: synonymy

A258 S.fuscov SP

A200 S.fuscov SP

A234 S.fuscov SC

A220 S.fuscov SP

A202 S.fuscov SP

A257 S.fuscov SP

A254 S.fuscov SP

A146 S.fuscov SP

A227 S.fuscov SP

A233 S.fuscov MG

A51 S.fuscov SP

A248 S.fuscov SP

A109 S.fuscov SP

A183 S.fuscov SP

A201 S.fuscov SP

A250 S.fuscov SP

A214 S.fuscov MS

A230 S.fuscov SC

A245 S.fuscov MS

A212 S.fuscov SC

A179 S.fuscov Es

A204 S.fuscov GO

A203 S.fuscov GO

A247 S.fuscov MT

89

100

100

92

9691

96

63 49

47

38

54

15

11

34

33

38

2

1

3

6

13

0.02

Diversity:

3.6 - 4.4%

Scinax megapodia x Scinax fuscovarius

A247 - Sc. megapodia(?) distribution

Synonym revalidation?

Look for other evidences

COI: NJ-K2P

Scinax fuscovarius: synonymy

A258 S.fuscov SP

A200 S.fuscov SP

A234 S.fuscov SC

A220 S.fuscov SP

A202 S.fuscov SP

A257 S.fuscov SP

A254 S.fuscov SP

A146 S.fuscov SP

A227 S.fuscov SP

A233 S.fuscov MG

A51 S.fuscov SP

A248 S.fuscov SP

A109 S.fuscov SP

A183 S.fuscov SP

A201 S.fuscov SP

A250 S.fuscov SP

A214 S.fuscov MS

A230 S.fuscov SC

A245 S.fuscov MS

A212 S.fuscov SC

A179 S.fuscov Es

A204 S.fuscov GO

A203 S.fuscov GO

A247 S.fuscov MT

89

100

100

92

9691

96

63 49

47

38

54

15

11

34

33

38

2

1

3

6

13

0.02

Diversity:

3.6 - 4.4%

COI: NJ-K2P

Scinax fuscovarius: criptic diversity?

A258 S.fuscov SP

A200 S.fuscov SP

A234 S.fuscov SC

A220 S.fuscov SP

A202 S.fuscov SP

A257 S.fuscov SP

A254 S.fuscov SP

A146 S.fuscov SP

A227 S.fuscov SP

A233 S.fuscov MG

A51 S.fuscov SP

A248 S.fuscov SP

A109 S.fuscov SP

A183 S.fuscov SP

A201 S.fuscov SP

A250 S.fuscov SP

A214 S.fuscov MS

A230 S.fuscov SC

A245 S.fuscov MS

A212 S.fuscov SC

A179 S.fuscov Es

A204 S.fuscov GO

A203 S.fuscov GO89

100

92

9691

96

63 49

47

38

54

15

11

34

33

38

2

1

3

6

13

0.02

Diversity

~2.6%

COI: NJ-K2P


A258 S.fuscov SP

A200 S.fuscov SP

A234 S.fuscov SC

A220 S.fuscov SP

A202 S.fuscov SP

A257 S.fuscov SP

A254 S.fuscov SP

A146 S.fuscov SP

A227 S.fuscov SP

A233 S.fuscov MG

A51 S.fuscov SP

A248 S.fuscov SP

A109 S.fuscov SP

A183 S.fuscov SP

A201 S.fuscov SP

A250 S.fuscov SP

A214 S.fuscov MS

A230 S.fuscov SC

A245 S.fuscov MS

A212 S.fuscov SC

A179 S.fuscov Es

A204 S.fuscov GO

A203 S.fuscov GO89

100

92

9691

96

63 49

47

38

54

15

11

34

33

38

2

1

3

6

13

0.02

COI: NJ-K2P


A258 S.fuscov SP

A200 S.fuscov SP

A234 S.fuscov SC

A220 S.fuscov SP

A202 S.fuscov SP

A257 S.fuscov SP

A254 S.fuscov SP

A146 S.fuscov SP

A227 S.fuscov SP

A233 S.fuscov MG

A51 S.fuscov SP

A248 S.fuscov SP

A109 S.fuscov SP

A183 S.fuscov SP

A201 S.fuscov SP

A250 S.fuscov SP

A214 S.fuscov MS

A230 S.fuscov SC

A245 S.fuscov MS

A212 S.fuscov SC

A179 S.fuscov Es

A204 S.fuscov GO

A203 S.fuscov GO89

100

92

9691

96

63 49

47

38

54

15

11

34

33

38

2

1

3

6

13

0.02

Geographic distribution of diversity?

Look for other evidences

1- Our primers for COI DNA Barcoding Anurans amplified all

species sampled

SUMARY AND CONCLUSIONS


species sampled

2- Although both COI and 16S rRNA have a great potential for

DNA barcoding amphibians, we strongly suggest a large-scale

effort to barcode the amphibians using COI



species sampled




3- Anuran diversity in Brazil is underestimated, and COI DNA

barcodes may help flag potential new species



species sampled




3- Anuran diversity in Brazil is underestimated, and COI DNA

barcodes may help flag potential new species

4- DNA barcodes may help to accelerate taxonomic revision

(bioacustics and morpho-anatomy)


5 - DNA barcode may help identify misclassification in collection

(Biological collections certification)




6- DNA barcode may help resolve problems with synonymy




6- DNA barcode may help resolve problems with synonymy

7- The use of DNA barcode will help make a better estimative of

amphibian species richness and may yield important findings for

the knowledge and conservation of these species.


Obrigada

Thank you

CNPq

Conselho Nacional de Desen volvimentoCi ent ífico e Te cno lógico

MLL Fellowship

Prodoc nº 563.975/05-9

Laboratory projects UNICAMP

Phylomedusa lemur –

Photo: ANAM

CFBH collection- UNESP



[email protected]




Ultrametric tree - MrBayes 3.2

Analise no R (split, ape, gee)

GMYC

Pons et al 2006 – Syst.Biology 55: 595-609 – “single”

Fontanero et al 2007 – Plos Biology 5: 914-921 – “single”

Monaghan et al 2009 - Syst.Biology – advance access publ jul 2009 – “multiple”

“General Mixed Yule-Coalescent model – GMYC”

Identificar limites de “espécies/grupos” baseado nos ramos de uma árvore que

contém espécies e populações.

Processos coalescentes neutros (pops) X especiação

“Single thresholds”: 6 unidades coalescentes

Outgroup

“Multiple thresholds”: 7 unidades coalescentes

Outgroup

A m a z o n i a

C e r r a d o

Coastal Ecosystems

Coastal Ecosystems

Southern Grasslands

Pantanal

A t

l a

n t i c

F

o r

e s

t

t r a

n s

i t i

o n

z o

n e s

Created a new account

Order: Anura

15 families

76 species

Order: Gymnophiona

1 family

3 species

Order: Caudata

1 family

1 species

Total: 80 sp.

“ DNA Barcoding Anuran from Brazilian of Atlantic forest” Fellowship: Capes/CNPqProdoc nº 563.975/05-9

Barcoding Anurans from Brazilian Atlantic Forest · AnR1 Primers design 1- Previous works in...

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