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Focus on the details: morphological evidence supports new cryptic land flatworm (Platyhelminthes) species revealed with molecules

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Abstract

We found numerous dull brown land planarians of the subfamily Geoplaninae within native rainforests and in nearby man-disturbed habitats in Southeastern and South Brazil. Their external and internal morphology are similar to those of Obama ladislavii (Von Graff, 1899), with the exception of the luminous green dorsum of this species, from which a brownish-green variation is also known. Since morphological features commonly used to delimit geoplaninid species failed to distinguish them, we used coalescent and Bayesian-based molecular methods (GMYC, BPP). These methods indicated that we were actually dealing with three species, the greenish O. ladislavii, and two brownish, cryptic species. A meticulous morphological study of the specimens, including type material of O. ladislavii, allowed us to ascertain diagnostic features for each of the species, for which we also propose a molecular diagnosis. This integrative taxonomic study demonstrates the utility of molecular tools to weigh minor morphological features and thus to reveal otherwise cryptic species.

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Abbreviations

cg:

cyanophil glands

cmc:

common muscle coat

cov:

common glandular ovovitelline duct

de:

dorsal epidermis

di:

dorsal insertion

dm:

dorsal cutaneous musculature

e:

eyes

eg:

erythrophil glands

ej:

ejaculatory duct

es:

esophagus

fa:

female atrium

fgd:

female genital duct

gm:

glandular margin

go:

gonopore

i:

intestine

im:

internal pharyngeal musculature

lu:

pharyngeal lumen

m:

mouth

ma:

male atrium

mc:

cutaneous musculature

mg:

glands with mixed secretion

mm:

parenchymal musculature

n:

nerve plate

o:

ovary

om:

outer pharyngeal musculature

ov:

ovovitelline ducts

p:

penis papilla

pp:

pharyngeal pouch

pv:

prostatic vesicle

rg:

rhabditogen glands

sc:

secretory cells

sd:

sperm duct

sg:

shell glands

sp:

sensory pits

sv:

spermiducal vesicle

t:

testes

v:

vitellaria

ve:

ventral epidermis

vi:

ventral insertion

vm:

ventral cutaneous musculature

xg:

xanthophil glands

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Acknowledgements

We thank the Instituto Chico Mendes de Conservação da Biodiversidade, Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renováveis (IBAMA), FATMA (Fundação de Amparo e Tecnologia do Meio Ambiente de Santa Catarina), Departamento de Águas e Energia Elétrica do Governo do Estado de São Paulo (DAEE), for licensing the fieldwork. We are grateful to Ana Lúcia Ramos Seitenfus, Délio Endres Júnior, Lisandro Negrete, Luciméri Teixeira, Márcio Hisayuki Sasamori, Piter Kehoma Boll, Vanessa dos Anjos Baptista, Vanderli Dias and Welcy Santos (UNISINOS), Débora Redivo, Marília Jucá, Leonardo Zerbone, Welton Araújo, Claudia Olivares (USP) for their help in sampling flatworms. Tiago Francoy, Júlio Pedroni (USP) and Guilherme Pinto Cauduro (UNISINOS) are thanked for DNA sequencing part of the specimens. Letícia Ayres Guterres and Rafaela Canello (UNISINOS), Ana Cristina Machado Vasconcelos, Amanda Cseh, Yasmin P. Oliveira, Diego Amorim and Adriano Maximiano (EACH, USP) helped with histological processing. Helma Roggenbuck (Zoological Museum Hamburg, MH) is gratefully thanked for kindest support in loan and permission to section the type material of O. ladislavii. Helmut Sattmann and Stefan Szeiler (NHM) are acknowledged for friendly assistance to FC during stay in the museum. FC has financial support from Fundación de Amparo à Pesquisa do Estado de São Paulo (FAPESP Proc, 2014/13661-8). AMLZ had grants from the Brazilian Research Council (CNPq) and the Fundação de Amparo à Pesquisa do Rio Grande do Sul (FAPERGS) in support of this work. Molecular work done at the Universitat de Barcelona (UB, Spain) has been funded by the Ministerio de Ciencia e Innovación, España (CGL2011-23466). SVA has a scholarship from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). MAP is grateful to the Sociedad Española de Genética (SEG) for the financial support to travel to Brazil where examined and made histological sections of some specimens used in this work. Two anonymous referees are acknowledged for their very valuable suggestions on an early version of the manuscript.

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Authors and Affiliations

  1. Departament de Genètica and Institut de Recerca de la Biodiversitat (IRBio), Universitat de Barcelona, Avinguda Diagonal 643, 08028, Barcelona, Spain

    Marta Álvarez-Presas & Marta Riutort

  2. Instituto de Pesquisas de Planárias and Programa de Pós-Graduacão em Biologia, Universidade do Vale do Rio dos Sinos (UNISINOS), 93022-000, São Leopoldo, Rio Grande do Sul, Brazil

    Silvana V. Amaral & Ana M. Leal-Zanchet

  3. Laboratório de Ecologia e Evolucão (EACH-USP) and Programa de Pós-Graduação em Zoologia (IB-USP), Universidade de São Paulo (USP), Av. Arlindo Bettio, 1000, São Paulo, SP, 03828-000, Brazil

    Fernando Carbayo

Authors
  1. Marta Álvarez-Presas
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  2. Silvana V. Amaral
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  3. Fernando Carbayo
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  4. Ana M. Leal-Zanchet
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  5. Marta Riutort
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Corresponding author

Correspondence to Fernando Carbayo.

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Álvarez-Presas, M., Amaral, S.V., Carbayo, F. et al. Focus on the details: morphological evidence supports new cryptic land flatworm (Platyhelminthes) species revealed with molecules. Org Divers Evol 15, 379–403 (2015). https://doi.org/10.1007/s13127-014-0197-z

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