This document summarizes a study on the genetic variation of Aquilegia taxa found within Sardinia. Researchers collected samples from 72 individuals across 15 locations and used AFLP (Amplified Fragment Length Polymorphism) analysis to characterize 371 genetic loci. A neighbor-joining tree grouped samples into two main clusters corresponding to the two Sardinian species A. nuragica and A. barbaricina. Spatial genetic structure analysis also supported delimitation of the two taxa and showed genetic differentiation between populations.

1. FLORA AND EVOLUTION IN THE MEDITERRANEAN AREA
Structure of the genetic variation of
the Sardinian taxa of Aquilegia
J.L. Garrido1, G. Bacchetta2, G. Fenu2, E. Mattana2, and C.M. Herrera1
1
Estación Biológica de Doñana, Consejo Superior de Investigaciones Científicas
(Sevilla, Andalucía, Spain).
2
Centro Conservazione Biodiversità, Dpto. Scienze Botaniche, Università degli
Studi di Cagliari (Cagliari, Sardinia, Italy)

2. INTRODUCTION: METHODS RESULTS CONCLUSIONS
North American Columbines

3. INTRODUCTION: METHODS RESULTS CONCLUSIONS
European Columbines

4. INTRODUCTION METHODS: RESULTS CONCLUSIONS
Study species: Sardinian Columbines (Arrigoni 2006, Flora dell’Isolla di Sardegna)
Delimitation of A. nuragica
Sardinian Aquilegia
taxa
A. barbaricina
A. nugorensis

5. INTRODUCTION METHODS: RESULTS CONCLUSIONS
Study area
NA1
A. nugorensis
A. barbaricina
A. nuragica

6. INTRODUCTION METHODS: RESULTS CONCLUSIONS
Genetic characterization: AFLP Analyses (Vos et al. 1995)
72 individuals on 15 locations
8 (48) primers combinations (PstI +2b / MseI +3b)
NA1
371 loci AFLP (189 – 500 bp)
Contamination, reproducibility (97%), homoplasy controlled
A. nugorensis
A. barbaricina
A. nuragica

7. INTRODUCTION METHODS RESULTS: CONCLUSIONS
NJ Method // Spatial Genetic Structure
Spatial Genetic Structure
Neighbour-Joining
tree distances
Nei-Li
B1 01
B1 03
B1 04
B2 04
45 B1 02
B2 02
2000 bootstrap replicates 34
B2 03
B2 01
B6 01
B6 02
Outgroups: 70
B6 03
B6 04
B6 05
Isopyrum fumarioides 25
B3 04
B4 05
Thalictrum speciosissimun B3 01
B3 03
B3 02
79 B3 12
B4 02
B4 03
58 B4 04
B5 02
B5 03
47 B5 01
B5 07
B5 05
27 B7 01
28 B7 09
B7 07
B7 02
95 B7 03
N5 02
N5 05
12 N5 03
47
N5 01
N5 04
N4 01
N4 04
N4 02
19 52 N4 03
N4 05
N8 01
N8 04
N8 03
45 N8 02
17 N8 05
N2 01
N2 02
N2 03
N1 02
66 N1 03
N3 01
N1 01
N1 05
N1 16
N3 03
38 N3 02
N3 04
N3 09
N7 01
50 N7 07
N7 02
57 N7 03
N7 04
24 N6 01
N6 02
N6 05
100 47 N6 03
IFU 0
TS 03

8. INTRODUCTION METHODS RESULTS: CONCLUSIONS
NJ Method // Spatial Genetic Structure
K = 2 (Evanno et al. 2005)
N7
N6
N1
N3
B6
B1
B2
B7
B5
B3
B4
N4
K=3
N8
N5
N2
Individual multilocus profiles allow us to establish 2 groups (+ 1 hybrid
subgroup) of individuals with homogeneous genetic characters.
(STRUCTURE v.2.3.1, Pritchard et al. 2000, 2007, Falush et al. 2007).

9. INTRODUCTION METHODS RESULTS CONCLUSIONS
Neighbour-Joining tree NJ K=2 K=3
B1 01
B1 03
Nei-Li distances B1 04
B2 04
Aquilegia barbaricina
45 B1 02
B2 02
2000 bootstrap replicates 34
B2 03
B2 01
B6 01
B6 02
Outgroups: 70
B6 03
B6 04
B6 05
Isopyrum fumarioides 25
B3 04
B4 05
Thalictrum speciosissimun B3 01
B3 03
B3 02
79 B3 12
B4 02
B4 03
58 B4 04
B5 02
B5 03
47 B5 01
B5 07
B5 05
27 B7 01
28 B7 09
B7 07
B7 02
95 B7 03
N5 02
N5 05
12 N5 03
47
N5 01
N5 04
N4 01
Aquilegia nugorensis
N4 04
N4 02
19 52 N4 03
N4 05
N8 01
N8 04
N8 03
45 N8 02
17 N8 05
N2 01
N2 02
N2 03
N1 02
66 N1 03
N3 01
N1 01
N1 05
N1 16
N3 03
38 N3 02
N3 04
N3 09
N7 01
50 N7 07
N7 02
57 N7 03
N7 04
24 N6 01
N6 02
N6 05
100 47 N6 03
IFU 0
TS 03

10. INTRODUCTION METHODS RESULTS CONCLUSIONS
Individual genotypes of Sardinian Aquilegia are structured in two main, genetically
homogeneous groups. This conclusion has a high consistency since results from
very different analytical bodies do agree.
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11. INTRODUCTION METHODS RESULTS CONCLUSIONS:
Perspectives
PHYLOGEOGRAPHY
Possible explanations to this genotypes distribution
EVOLUTIONARY ECOLOGY
Adaptive radiation of Sardinian Columbines:
European-like or North American-like differentiation?
MACROEVOLUTION
Relative position in Aquilegia European clade

12. Thanks soooooooo much to:
Cristina Pérez Sample collectors
Laboratorio de Sistemas de Información Geográfica y
Teledetección de la Estación Biológica de Doñana (LAST-EBD-CSIC)
Laboratorio de Ecología Molecular de la Estación Biológica
de Doñana (LEM-EBD-CSIC)