Contrasting patterns in the genetic diversity and population structure of land snails from Peru

Pedro Romero, M. Sc
Martin Fellow Museo de Historial Natural Universidad Nacional Mayor de San Marcos Lima - Per

Cerro de Agua

Lachay

Bostryx scalariformis (Mollusca, Orthalicidae)

Ancn

Pasamayo

Morphometrics: Shell design make the difference in B. scalariformis populations

Shape

Shell design

DNA extraction: CTAB+Proteinase K+-ME, OH 96

PCR: 16S rRNA mitochondrial gene.

H12 Pasamayo

16S rRNA could be use for intraspecific analyses in land snails.

H13 Pasamayo 81 H14 Pasamayo H15 Pasamayo

Clades correspond to B. scalariformis morphotypes.

97

73 99

H16 Pasamayo H11 Ancn

89 H10 Ancn H9 Ancn H8 Lachay H7 Lachay 87 H5 Cerro de Agua H6 Cerro de Agua 77 H4 Lachay H1 Cerro de Agua H3 Lachay / Cerro de Agua

Haplotype 17 is related to B. sordidus.

H2 Cerro de Agua B. sordidus Ancn - t1 100 H17 Pasamayo B. sordidus Ancn - t2 B. sordidus Iguanil - Ig B. sordidus Ancn - t3 99 B. sordidus Ancn - Y

91 B. sordidus Carabayllo - Y B. modestus B. conspersus

Intraspecific phylogeny

0.02

Haplotypes (different sequences=17) in B. scalariformis populations. Two groups: Cerro de Agua Lachay and Pasamayo Ancn. H17 show a high number of mutations .

Hap 4 8 H_1 T H_2 . H_3 . H_4 . H_5 . H_6 . H_7 . H_8 . H_9 A H_10 . H_11 . H_12 . H_13 . H_14 . H_15 . H_16 . H_17 .

5 9 T . . . . . C . . . . . . . . . .

6 0 A . . . . . . . . . T . . . . . .

6 8 G . . . . . . . . . . . . . . . A

6 9 T . . . . C A A A A A A A A A A A

7 0 A . . . . . . . . . . G G G G G G

8 1 T . . . . . . . . . . C C C . . .

8 2 G . . . . . . . . . . . . . . . A

8 4 C . . . . . . . . . . . . . . . T

8 8 A . . . . . . . . . . . . . . T .

8 9 T . . . . . . . A A A A A A A A .

9 1 G . . . . . . . . . . . . . . . A

1 0 2 A . . . . . . . . . . . T . . . T

1 2 3 G . . . . . . . A A A A A A A A .

1 2 4 T . . . C . . . A A A A A A A A A

1 2 5 C . . . . . T T T T T T T T T T T

1 2 6 G . . . . . . . . . . A A A A A .

1 2 8 A . . . . . . . . . . . . . . . T

1 6 0 T . . . . . . . . . . . . . . . A

1 6 4 T . . . . . . . . . . . . . . . A

1 7 5 T C . . . . . . . . . . . . . . .

1 7 0 A . . . . . . . . . . . . . . . T

1 7 1 T . . . . . . . . . . . . . . . C

1 8 3 A . . . . . . . G G G G G G G G .

1 8 0 A . . . . . . . . . . . . . . . G

2 0 1 T . . . . . . . . . . . . . . . A

2 1 7 A . . . . . . . G G G G G G G G .

2 1 7 T . . . . . . . . . . . . . . . A

2 3 8 T . . . . . . . . . . . . . . . G

2 3 1 T . . . . . . . . . . . . . . . A

2 3 3 T . . . A . . . . . . . . . . . A

2 3 4 T . . . . . . . . . . . . . . . A

2 4 8 A . . . . . . . . G G G G G G G .

2 4 0 T . . . . . . . G G G G G G G G C

2 4 2 T . . . . . . . C C C C C G G G .

2 4 5 G . . A . . . . . . . . . . . . A

2 4 6 A . . . . . . . T T T T T T T T .

2 5 3 C T T T T T T T T T T T T T T T T

2 6 4 A . . . . . . . T T T T T T T T .

2 6 7 A . . . . . . . . . . . . . G . .

2 6 8 A . . G . . . . . . . . . . . . .

2 7 9 G . . . . . . . . . . . . . . . A

3 0 1 T . . . . . . . C C C C C C C C .

3 0 4 A . . . . . . . . . . . . . . . C

N Pob. 1 C 1 C 8 L/C 1 L 1 C 3 C 1 L 1 L 2 A 11 A 1 P 3 P 1 P 7 P 1 P 1 P 7 P

There is a high geographical structure of the genetic diversity

Distribucin geogrfica de los haplotipos de B. scalariformis
Localidad Ancn Pasamayo Lachay Cerro de Agua N Individuos Haplotipo H1 H2 H3 H4 H5 H6 H7 H8 H9 2 H10 H11 H12 H13 H14 H15 H16 H17 11 1 3 1 7 1 1 7 Total 14 20 7 10 51

1 1

1 1

4 4 8

1 1 1 1 3 3

1 1

1 1 2 11 1 3 1 7 1 1 7

Haplotipos H1 H8

Haplotipos H9 H17

Fst
Ancn Pasamayo Lachay Cerro de Agua Ancn 0.0000 0.7943 0.9149 0.9362 Pasamayo 0.0000 0.9226 0.9424 Lachay Cerro de Agua

0.0000 0.0560

0.0000

Fst values vary between 0 and 1. 0 = No differentiation 1 = High differentiation

Haplotype network

Cerro de Agua
-

Lachay

11

Pasamayo

Ancn

Pasamayo

Ancn

Lineage expansion may be related to historic rise of superficial sea temperatures (SST), and El Nio/Southern Oscillation (ENSO). Coastal region could have had cycles of vegetation expansion and hiperarid periods. This could have influenced the genetic structure of land snail populations, and also the divergence among populations

Lachay

Cerro de Agua

400 0.020

300 0.015

200 0.010

100 0.005

0.0 0.000

Kya

100 km

1. CICRA y CM1 (Rio Madre de Dios / Rio Los Amigos = AMI)

2. Concepcin, Palmereto y Gamitana (Rio Madre de Dios = MD).

Systrophia helicyloides

Systrophia helicycloides
(dOrbigny, 1835)

(a) CICRA, (b) CM1, (c) Concepcin, (d) Gamitana, (e) Palmereto.

Haplotype distribution
Regiones Los Amigos Localidades S1 2 3 1 1 7 S2 1 S3 1 S4 Haplotipos S5 S6 S7 3 3 1 8 7 1 18 4 1 S8 2 S9 1 1 1 2 5 6 S10 1 S11 Total 11 7 10 10 8 46

CICRA CM1 Concepcin Bajo Madre Gamitana de Dios Palmereto Total

1 2 1 1 3

More frecuent haplotypes are in both river basins

Hap 4 6 S1 G S2 . S3 . S4 . S5 . S6 A S7 A S8 A S9 A S10 A S11 A 5 4 T . . . . A A A A . . 5 8 A . . . . . . . . T T 6 2 C . . . . . . . . T T 6 4 T . . . . . . . . A A 6 7 C . . . . . . . . T T 7 0 A . . . . . . . . C C 8 1 T . . . . . . . . A A 8 3 A . . . . . . . . G G 8 6 T . . . . . . . . . A 8 9 T . . . . . . . . A A 9 2 C . . . . . . . . T T 9 9 A . . . . . . . . G G 1 2 0 C . . . . . . . . T T 1 2 5 A . . . . . . . . G G 1 2 6 A . . . . . . . . T T 1 2 7 T . . . . . . . . A A 1 2 9 T . . . . . . . . A A 1 3 2 A . . . . . . . . T T 1 5 3 T . . . . . . . . C C 1 5 6 A . . G . . . . . . . 0 6 1 T . . . . . . . . A A 1 6 4 A . . . . . . . . T T 1 8 0 A . . . . . . . G . . 1 8 1 C . . . . . . . . T T 1 8 9 A . . . . G G G G G G 1 9 0 A . . . . C C C C . . 1 9 1 A . . . . . . . T . . 1 9 7 A T . . . . . . . . . 2 0 7 T . . . . . . . A A A 2 0 8 A . . . . . . . . T . 2 1 1 T . . . . . . . . A A 2 1 4 T . . . . A A . . G G 2 1 9 A . . . . . T . . . . 2 2 0 A . . . . . . . . T T 2 2 1 G . . . . . . . . A A 2 2 7 A . . . . . . . . T T 2 3 0 A . . . . . . . . T T 2 3 1 T C C C C . . . . . . 2 3 2 C . . . . T T T T . . 2 3 4 A G G . . . . . . T T 2 3 5 T . . . . . . . . A A 2 3 6 G . . . . . . . . A A 2 3 8 A . . . . . . . . T T 2 4 1 G . . . . . . . . A A 2 4 2 A . . . . G G G G . . 2 4 6 A . . . . . . . . G G 2 7 2 T . . . . . . . . A A 2 7 3 A . . . . T T T T T T N Pob

7 CM1/Cic/Gam/Pal 1 Cic 1 Cic 3 Gam/Con 18 Cic/Gam/Con 4 CM1/Cic 1 Pal 2 CM1 1 CM1 2 Cic/Pal 6 Con/Pal

S1-S5: Both S6-S9: Los Amigos S10 y S11 with high differences

S4 S5 S2
96

S3 S1 S8

100

S9
93 99

S6 S7

79

S10
100

S11

Systrophia stenostrepta Systrophia eatoni

Wayampia sp.

Concepcin
CICRA Gamitana Palmereto

0.05

CM1

Intraspecific phylogeny

CICRA CM1 Concepcin

Gamitana Palmereto

Fst values
CM1 CICRA Concepcin Gamitana Palmereto CM1 0,00000 0,05701 0,25515 0,55046 -0,31250 CICRA 0,00000 0,06648 0,20062 -0,15864 Concepcin Gamitana Palmereto 0,00000 -0,00891 0,07640 0,00000 0,65686

0,00000

(a) (b) CM1-CICRA Con-Gam-Pal CM1-CICRA Con-Gam-Pal CM1-CICRA 0,00000 0,00000 Con-Gam-Pal 0,09735 0,00000 0,16699 0,00000

(tau) /=2ut/

Tau (0,752)
u (3,66 x 10-5), Tiempo desde la expansin 10 15 mil aos atrs. Rise of temperatures, vegetation expansion.

LGM

River influence

Last period of Andean orogenesis

Amazon Basin formation

Marine incursions in the Amazon
Millions of years

What we know until know

Land snails show a high mutation rate. Intraspecific analysis showed different lineages, sometimes resemble differences in morphological data.

Land snails could allow us to understand past events related with changes in the landscape.
Their low vagility does not permit them to escape to changing enviromental conditions. Therefore it is possible to see how the habitat could influence the population genetic structure and how these changes may be observed studiyng their genomes.