Species richness in the genus Microplana (Platyhelminthes ... · B. Naturalis Biodiversity Center, PO Box 9517, 2300 RA Leiden, The Netherlands. C. Departament de Genètica, Microbiologia
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Table S1. List of specimens used in the molecular and histological analyses, locality of origin and codes used in the trees, Museum code for histological sections, and accession numbers for the gene sequences
Species Collecting locality Molecular code Museum code GenBank
753C256 KU867224* a North Yorkshire, United Kingdom MT3 JN380018a KR906681 KU872655* KU872706* La Châtelaine, France 713X301 KU867225* a KU872612* KU872656* KU872707* Auliac Dessús, France 737X306 KU867226* a KU872613* KU872657* KU872708*
Unassigned specimens Cagnes sur Mer, France JL1551 KU867227 a 700X299 KU867228* a
Cádiz, Spain 179D054 KU867229* a Asturias, Spain 885A379 KU867230* a Abruzzo, Italy 916I445 KU867231* a Bulgaria 1225U472 KU867232* a
Outgroup Bipalium adventitium Kingston, USA B. adventitium AF178306 DQ666000 DQ665956 KJ599681 B. multilineatum Naka, Nagaoka, Niigata, Japan B. multilineatum HM346600 DQ666012 DQ665957 KU872709
1 Sequence obtained by Jean-Lou Justine a Cox1GMYC Dataset b For this individual there is no molecular information and hence in fact it can not be assigned to either Microplana cf. aixandrei 1 or 2 * Sequences from this study
S1 Fig. S1. Maximum likelihood tree inferred with the Cox1 barcoding dataset. Values at nodes correspond to bootstrap and posterior probability values.
Fig. S2. Maximum likelihood trees inferred with the information of individual nuclear genes and with concatenation of sequences belonging to the three genes: (A) 28S rDNA, (B) elongation factor 1-α, (C) 18S rDNA and (D) nuclear concatenated dataset. Asterisks at nodes correspond to bootstrap and posterior probability values greater than 75% and 0.85, respectively. In D, red asterisks correspond to nodes supported only in the Bayesian inference analysis.
Figs S3–S6. Microplana astricta. Holotype, ZMA V.Pl. 7228.1. S3, Ventral subepidermal and parenchymal musculature, sagittal section; S4, sagittal section through the copulatory apparatus; S5, sagittal section through male atrium and penis papilla; S6, sagittal section of female copulatory apparatus, showing muscular constriction in bursal canal.
S3 S4 200 µm
lpm
cb
pbvnc
ppbc
lpm vd
50 µm go
S5 cb
S6cb
pp bc pp
100 µm go go 100 µm
S7 S8
ph pp
te
200 µm 100 µm
Figs S7–S11. Microplana cephalofusca and M. cingulata. S7, Microplana cephalofusca, holotype ZMA V.Pl. 7229.1, sagittal section, showing testes follicles. S8, Microplana cephalofusca, holotype ZMA V.Pl. 7229.1, sagittal section through penis papilla. S9, Microplana cephalofusca, holotype ZMA V.Pl. 7229.1, sagittal section through copulatory bursa. S10, Microplana cingulata, holotype ZMA V.Pl. 7230.1, sagittal section through pharynx, showing the various muscle layers. S11, Microplana cingulata, holotype ZMA V.Pl. 7230.1, sagittal section of copulatory apparatus, showing the penis papilla, tube-shaped fold and copulatory bursa.
S9 inS10
cb
pp
bc
100 µm lu
S11
ppcb
50 µm
tsf pp 200 µm
S13
S12 50 µm
te vep 100 µm
S14 pb
lu
fgd
100 µm pp go
S15 in
S16
pe gs
fgd
od
50 µm 100 µm vep
Figs S12–S16. Microplana fuscomaculosa. S12, Paratype, ZMA V.Pl. 7231.2, sagittal section through pharynx. S13, ZMA V.Pl. 7232.1, sagittal section, showing the testes. S14, Paratype, ZMA V.Pl. 7231.2, sagittal section through the penis,showing the broad zone of circular muscle around bulbar and penial lumen. S15, Holotype, ZMA V.Pl. 7231.1, sagittalsection of communication between female genital duct and gut via an intestinal appendix. S16, ZMA V.Pl. 7232.1, sagittalsection showing opening of female genital duct into the globular sac.
S17 S18
gid in bc pp
cb pp
100 µm
vep 100 µm
Figs S17–S21. Microplana lutulenta, M. nervosa and M. sparsa. S17, Microplana lutulenta, holotype, ZMA V.Pl. 7234.1, sagittal section through penis and female genital duct. S18, Paratype, ZMA V.Pl. 7236.2, sagittal section of copulatory apparatus. S19, Microplana nervosa, holotype, ZMA V.Pl. 7236.1, sagittal section, showing communication between copulatory bursa and gut, as well as muscularised intestinal extension. S20, Microplana nervosa, holotype, ZMA V.Pl. 7236.1, sagittal section of penis and muscularised intestinal extension. S21, Microplana sparsa, holotype, ZMA V.Pl. 7238.1, sagittal section of the copulatory apparatus.
S19 S20 100 µm
inin
ie pp
cbie
vd
100 µm
S21
sg
fgd pp
100 µm
S22 S23
rh spt cb
50 µm
ch vep
50 µm
Figs S22–S26. Microplana cf. aixandrei and M. henrici. S22, Microplana cf. aixandrei, ZMA V.Pl. 7239.2, sagittal section of bursa, containing remnants of spermatophore. S23, Microplana cf. aixandrei, ZMA V.Pl. 7239.2, sagittal section of anterior ventral epidermis, showing rhamnites and chondrocysts. S24, Microplana henrici, ZMA V.Pl. 7240.1, sagittal section, showing the testes. S25, Microplana henrici, ZMA V.Pl. 7240.1, sagittal section of copulatory apparatus, showing the annular fold. S26, Microplana henrici, ZMA V.Pl. 7240.1, sagittal section of male copulatory apparatus.
S24 S25bl
in
fgd
af
te 100 µm
S26
te
bl af pp
vep 100 µm af
100 µm
S28 S27
ov pp
am af od
af af
200 µm 50 µm vep
Figs S27–S29. Microplana peneckei and M. hyalina. S27, Microplana peneckei, holotype, NHMW, Inv. 2851, sagittal section of penis papilla and atrium, showing three annular folds. S28, Microplana hyalina, ZMA V.Pl. 7241.1, sagittal section, showing ovary, ampulla and oviduct. S29, Microplana hyalina, ZMA V.Pl. 7242.1, sagittal section of copulatory bursa, showing coloured balls in vacuoles of lining epithelium.