Molecular taxonomy in

Neotropical Passeriformes

Fabrício R. Santos

Associate Professor

Dep. de Biologia Geral, ICB

Universidade Federal de Minas Gerais

Belo Horizonte, MG, Brazil

Megadiversity

Mexico450

Colombia456

China499

Indonesia515

Brazil524

Mammals

Brazil468

Indonesia511

Colombia520

Mexico717

Australia755

Reptiles

China274

Mexico284

Ecuador402

Brazil517

Colombia 583

Amphibians

Australia25 000

Mexico26 000

China30 000

Colombia45 000

Brazil55 000

Vascular plants

Country and species numbersGroup

Venezuela1308

Ecuador1435

Brazil1573

Peru1705

Colombia1721

Birds

10 % of 1.4 million known species

Brazilian Amazon

Biodiversity hotspots

Atlantic Rain Forest

Cerrado

(Brazilian savanna)

Taxonomic difficulties

• Many species are largely distributed and fragmented;

• Phylogeographic studies indicate existence of many intraspecific divergent clades, even for species with samples limited geographically;

• Majority of species are still to be formally described;

• Collections are poor sources of vouchers, with few exceptions;

• A large investment is needed for new and representative samplings.

Molecular identification of known spp

All 260 known species of North-American birds

Bird Genetics in the lab

• Population Genetics

• Phylogeography

• Systematics and

phylogeny

• Reproductive behavior

• Molecular taxonomy

Molecular Methodology

Extraction,

storage, catalog

Tissue/specimen

collection

PCR

Sequencing data

MegaBACE

Data Analysis

mtDNA

D-Loop

ND5

H-strand

ND4

ND4L

ND3COIII

L-strand

ND6

ND2

ND1

COII

Small ribosomal RNA

Large ribosomal RNA

ATPase subunit 8

ATPase subunit 6

Cytochrome b

COICOI

COI sequences

• Divergent individuals chosen for COI

sequencing after phylogeographic inspection.

• High quality sequence data (510-800 bp):

large PCR amplicons as template,

two independent PCR products,

double strand sequencing,

careful edition of consensus sequences

based on phred/phrap/consed.

Molecular taxonomy in Thamnophilidae

Passeriformes; Suboscines

Inter x intraspecific diversity of COI sequences in Thamnophilidae

II

III IV

IT. caerulescens

T. ambiguus / T. pelzelni

0

1

2

3

4

5

6

7

8

9

10

11

12

0 1 2 3 4 5 6 7 8 9 10 11 12

Maximum intraspecific divergence(%)

Min

imum

inte

rspe

cific

dive

rge

nce

(%)

II

III IV

I

0

1

2

3

4

5

6

7

8

9

10

11

12

0 1 2 3 4 5 6 7 8 9 10 11 12

Maximum intraspecific divergence(%)

Min

imum

inte

rspe

cific

dive

rge

nce

(%)

B0665B0663

B0655B0653

B0654B0656B0369

B0367B0368

B0609B0647

B0317B0329B0326

B0658B0644B0657

B0677B0678

B0646B0291B0292

B0290B0680

B0643B0660

B0281B0285B1189

B1301B1188

B1296B1186B1303

B0525B0524

B0515B0511

B0513B0528

B0535B0562

B1148B1147B1150

99

99

99

99

90

99

99

99

99

99

99

99

99

99

99

91

99

99

99

82

90

D. ochropyga

D. ferruginea

R. ardesiaca

D. squamata

B0335

P. leucoptera

M. loricata

T. major

D. plumbeus

F. serrana

D. mentalis

H. atricapillus

S. cristatus

T. doliatus

T. caerulescens

T. ambiguus

T. pelzelni

Phylogeneticanalysis of COI sequences in Thamnophilidae

Thamnophilus caerulescens Thamnophilus ambiguusThamnophilus pelzelniThamnophilus doliatusHerpsilochmus atricapillusSakesphorus cristatusDsythamnus_mentalisDsythamnus plumbeusDrymophila ochropygaDrymophila ferrugineaDrymophila squamataMyrmeciza loricataRhopornis ardesiacaPyriglena leucopteraFormicivora serranaTaraba major

11111122222222222333333444 001123500344801345566779001569011

2451769253681928436

CACAAAACAACAAACCCAACAAAGCCCATAAAA .................G.T...A......... ....G..............T...A....C.... ............G.....GT...A.T.C..... .TT.............T..A.C.A.A...C... T..................AG..ATA....... .....GG............A...A.A......G .G.........G..T....A..GA.A....... ..............A....A...A.A....... ..............T..C.A...A.AA...... ........G..........G...A.AT...... ............C......A...A.A....... ..............T....A...T.AT...T.. .........G.........A...T.A.....G. .......T.....GT....A...A.AT...... ...G......T....T...A...A.A.......

Autapomorphic sites in ThamnophilidaeSynapomorphic in each species

Character basedbarcodes 16 spp

COI variable positions

0000000

58149564710650

Molecular taxonomy in Neotropical Passeriformes

• A broad geographic sampling should be priority;

• Details of population structure and phylogeography should be used, particularly for species with large geographic distribution, or likely susceptible to sympatric substructuring due to sexual selection etc;

• High quality sequences should be used to provide a more discriminating power;

• These studies should be accomplished closely by other taxonomists, as many more taxa are expected to be found.

Colaborators

• Miguel A. Marini and team - UnB;• Marcos Rodrigues and team - UFMG;• Cristina Miyaki and team - USP;• Marcelo Ferreira Vasconcelos;• Lucas Carrara and team – UFMG.

Team

DanielaEloísa

Sibelle Anderson, Juliana e Camila

Support

PG Ciência Animal - UFMG