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Fig. 5 in Morphological versus molecular delimitation of ciliate species: a case study of the family Clevelandellidae (Protista, Ciliophora, Armophorea)

Fig. 5. Clevelandella hastula (Kidder, 1937). Vietnamese specimens isolated from Panesthia angustipennis cognata Bey-Bienko, 1969 from life (A, F–H) and after protargol impregnation (B–E). A–E. Ventral view of specimens with well-preserved body shape. Arrows mark the proximal end of the peristomial opening, black arrowheads mark the proximal end of the adoral zone of membranelles. F. Ventral view, showing general organization of body. G–H. Ciliary pattern of ventral and dorsal sides. Conspicuous cilia of adoral membranelles emerge out of the peristomial opening in (G). Asterisks indicate the position of the ciliary whorl (posterior suture), arrow marks the proximal end of the peristomial opening. Scale bars = 30 μm.

Fig. 10 in Morphological versus molecular delimitation of ciliate species: a case study of the family Clevelandellidae (Protista, Ciliophora, Armophorea)

Fig. 10. Clevelandella parapanesthiae (Kidder, 1937). Vietnamese specimens isolated from Panesthia angustipennis cognata Bey-Bienko, 1969 after protargol impregnation. A–J. Variability of body shape and size as well as of the nuclear (shaded grey) and oral (shaded yellow) apparatus. Scale bar = 30 μm.

Fig. 1 in Morphological versus molecular delimitation of ciliate species: a case study of the family Clevelandellidae (Protista, Ciliophora, Armophorea)

Fig. 1. Clevelandella constricta (Kidder, 1937). Vietnamese specimens isolated from Panesthia angustipennis cognata Bey-Bienko, 1969 from life (A) and after protargol impregnation (B–N). A. Ventral view of a representative specimen, length 120 μm. B–K. Variability of body shape and size as well as of the nuclear (shaded grey) and oral (shaded yellow) apparatus. L. Semi-schematic diagram, showing the general body organization. Black double arrowhead marks densely packed, oval, refractile bodies (probably paraglycogen platelets). M–N. Ciliary pattern of ventral and dorsal sides. Arrow marks the right suture, black arrowheads indicate the position of the ciliary whorl (posterior suture). O. Prokaryotes freely scattered throughout the cytoplasm posterior to the macronucleus. P. Detail of oval, refractile bodies (probably paraglycogen platelets) anterior to the macronucleus. Scale bars = 50 μm.

Fig. 3 in Morphological versus molecular delimitation of ciliate species: a case study of the family Clevelandellidae (Protista, Ciliophora, Armophorea)

Fig. 3. Clevelandella constricta (Kidder, 1937). Vietnamese (A, E–G) and Cambodian (D) specimens isolated from Panesthia angustipennis cognata Bey-Bienko, 1969, as well as Thai I specimens (B– C) isolated from Panesthia angustipennis angustipennis (Illiger, 1801) from life (A, D–G) and after protargol impregnation (B–C). A–C. Ventral view of specimens with well-preserved body shape. D–E. Ventral view, showing the general body organization. Arrows mark oval, refractile bodies anterior to the macronucleus, black arrowheads mark the proximal end of the adoral zone of membranelles, white arrowheads denote the karyophore attached to the right and left body margins and black double arrowhead marks the canal leading from the contractile vacuole to the cytopyge. F–G. Ciliary pattern of ventral and dorsal sides. Asterisks mark the position of the ciliary whorl (posterior suture), white double arrowhead denotes the right suture. Scale bars: A–C, E–G = 50 μm; D = 20 μm.

Fig. 11 in Morphological versus molecular delimitation of ciliate species: a case study of the family Clevelandellidae (Protista, Ciliophora, Armophorea)

Fig. 11. Clevelandella parapanesthiae (Kidder, 1937). Vietnamese specimens (A–B, E–G) isolated from Panesthia angustipennis cognata Bey-Bienko, 1969 and Thai I specimens (C–D) isolated from Panesthia angustipennis angustipennis (Illiger, 1801) from life (A, E–G) and after protargol impregnation (B–D). A–D. Ventral views of specimens with well-preserved body shape. Black arrowheads mark the proximal end of the adoral zone of membranelles. E–G. A strongly squeezed specimen by pressure of the cover slip, causing the body to become markedly wider and the notch at the base of the peristomial projection to be lost. The general body organization is shown in (E), the ciliary pattern of ventral and dorsal sides is shown in (F) and (G). Asterisks mark the position of the ciliary whorl (posterior suture), white arrowhead denotes the karyophore attaching to right body margin, white double arrowhead denotes the right suture. Scale bars = 30 μm.

Figure 1 in DNA barcoding and coalescent-based delimitation of endosymbiotic clevelandellid ciliates (Ciliophora: Clevelandellida): a shift to molecular taxonomy in the inventory of ciliate diversity in panesthiine cockroaches

Figure 1. Phylogenetic tree based on the 18S rRNA gene, ITS1-5.8S-ITS2 region and 28S rRNA gene, showing relationships among ciliates isolated from the digestive tract of panesthiine cockroaches. All tree-building methods resulted in very similar topologies. The single exception is the Anteclevelandella constricta cluster, where the IQTree topology differs from that of both Bayesian trees (shown in the box). Bootstrap values for the maximum likelihood conducted in IQTrees and posterior probabilities for Bayesian inferences conducted in MrBayes and Phycas are listed at corresponding nodes of the best scoring IQTree. Specimen codes and further details are listed in Table 1. The scale bar denotes two substitutions per one hundred nucleotide positions.

Figure 4 in DNA barcoding and coalescent-based delimitation of endosymbiotic clevelandellid ciliates (Ciliophora: Clevelandellida): a shift to molecular taxonomy in the inventory of ciliate diversity in panesthiine cockroaches

Figure 4. Consensus secondary structure of the ITS2 molecule of 54 members of the order Clevelandellida isolated from the digestive tract of cockroaches. Note that the central loop radiates four highly conserved helices. The structure logo of helices is shown on the right side. The height of a base is proportional to its frequency in the multiple sequence alignment.

Figure 3 in DNA barcoding and coalescent-based delimitation of endosymbiotic clevelandellid ciliates (Ciliophora: Clevelandellida): a shift to molecular taxonomy in the inventory of ciliate diversity in panesthiine cockroaches

Figure 3. MDS diagrams (A, C, E) and TCS networks (B, D, F) based on 18S rRNA gene (A, B), ITS1-5.8S-ITS2 region (C, D) and 28S rRNA gene (E, F) sequences of the family Clevelandellidae. The MDS diagrams show the distribution of specimens in the genotype space. Species belonging to the same genus form a cluster distinctly isolated from other such clusters. The TCS networks reflect the most parsimonious relationships among species given the individual markers. Numbers along edges indicate mutational steps between adjacent nodes. Species belonging to the same genus are marked by the same colour code.

Figure 7 in DNA barcoding and coalescent-based delimitation of endosymbiotic clevelandellid ciliates (Ciliophora: Clevelandellida): a shift to molecular taxonomy in the inventory of ciliate diversity in panesthiine cockroaches

Figure 7. Putative secondary structure models of the highly divergent helix c3-1 in the D2 domain of the 28S rRNA molecule of 17 species of the order Clevelandellida isolated from the digestive tract of cockroaches. Arrowheads denote the molecular diagnostic characters.

Figure 2. Coalescent species trees with 13 in DNA barcoding and coalescent-based delimitation of endosymbiotic clevelandellid ciliates (Ciliophora: Clevelandellida): a shift to molecular taxonomy in the inventory of ciliate diversity in panesthiine cockroaches

Figure 2. Coalescent species trees with 13 (A) and 14 (B) assumed species based on 18S rRNA gene, ITS1-5.8S-ITS2 region and 28S rRNA gene sequences. Posterior probabilities of clades are provided along internal branches and posterior probabilities for the presence of individual species are provided behind the terminal branches. Scale bars denote the fraction of substitutions per site.

Figure 9 in DNA barcoding and coalescent-based delimitation of endosymbiotic clevelandellid ciliates (Ciliophora: Clevelandellida): a shift to molecular taxonomy in the inventory of ciliate diversity in panesthiine cockroaches

Figure 9. MDS diagram (A) and TCS network (B) based on concatenated 18S rRNA gene, ITS1-5.8S-ITS2 region and 28S rRNA gene sequences of the family Clevelandellidae. The MDS diagram shows the distribution of specimens in the genotype space. Species belonging to the same genus form a cluster distinctly isolated from other such clusters. The TCS network reflects the most parsimonious relationships among species given the concatenated dataset. Numbers along edges indicate mutational steps between adjacent nodes. Species belonging to the same genus are marked by the same colour code.

Figure 6 in DNA barcoding and coalescent-based delimitation of endosymbiotic clevelandellid ciliates (Ciliophora: Clevelandellida): a shift to molecular taxonomy in the inventory of ciliate diversity in panesthiine cockroaches

Figure 6. Putative secondary structure models of helices 39‒44 in the V7 region of the 18S rRNA molecule of four Nyctotherus species isolated from the digestive tract of cockroaches. Arrowheads denote the molecular diagnostic characters. CBC, compensatory base change; hemi-CBC, hemi-compensatory base change.

Figure 5 in DNA barcoding and coalescent-based delimitation of endosymbiotic clevelandellid ciliates (Ciliophora: Clevelandellida): a shift to molecular taxonomy in the inventory of ciliate diversity in panesthiine cockroaches

Figure 5. Putative secondary structure models of the highly divergent helix 23e1 in the V4 region of the 18S rRNA molecule of 17 species of the order Clevelandellida isolated from the digestive tract of cockroaches. Arrowheads denote the molecular diagnostic characters.

Fig. 18 in Morphological versus molecular delimitation of ciliate species: a case study of the family Clevelandellidae (Protista, Ciliophora, Armophorea)

Fig. 18. Phylogeny of the family Clevelandellidae Kidder, 1938 based on 18S rRNA gene sequences. Bootstrap values for maximum likelihood analyses performed in IQTrees and PhyML as well as posterior probabilities for Bayesian interference performed in MrBayes and Phycas were mapped onto the best scoring IQTree. Dashes indicate statistical support below 50%. Sequences obtained in this study are in bold face. Members of each morphospecies are marked by the same color, as shown in Fig. 17. For specimen codes and further details, see Table 6. The scale bar denotes six substitutions per one thousand nucleotide positions.

Fig. 14. Paraclevelandia brevis Kidder, 1937. Thai I in Morphological versus molecular delimitation of ciliate species: a case study of the family Clevelandellidae (Protista, Ciliophora, Armophorea)

Fig. 14. Paraclevelandia brevis Kidder, 1937. Thai I specimens (A–C, F–G) isolated from Panesthia angustipennis angustipennis (Illiger, 1801) and Vietnamese specimens (D–E) isolated from Panesthia angustipennis cognata Bey-Bienko, 1969 from life (A–C) and after protargol impregnation (D–G). A. Ventral overview, showing the general body organization. Arrows mark the proximal end of the peristomial opening, black arrowheads denote the proximal end of the adoral zone of membranelles, white arrowheads denote the karyophore attached to the anterior body pole. B–C. Ciliary pattern of ventral and dorsal sides. Asterisks mark the position of the ciliary whorl (posterior suture). D–G. Ventral views of specimens with well-preserved body shape. Scale bars = 20 μm.

Fig. 13. Paraclevelandia brevis Kidder, 1937. Thai I in Morphological versus molecular delimitation of ciliate species: a case study of the family Clevelandellidae (Protista, Ciliophora, Armophorea)

Fig. 13. Paraclevelandia brevis Kidder, 1937. Thai I specimens isolated from Panesthia angustipennis angustipennis (Illiger, 1801) after protargol impregnation. A–J. Variability of body shape and size as well as of the nuclear (shaded grey) and oral (shaded yellow) apparatus. Scale bar = 20 μm.

Fig. 4 in Morphological versus molecular delimitation of ciliate species: a case study of the family Clevelandellidae (Protista, Ciliophora, Armophorea)

Fig. 4. Clevelandella hastula (Kidder, 1937). Vietnamese specimens isolated from Panesthia angustipennis cognata Bey-Bienko, 1969 from life (A) and after protargol impregnation (B–N). A. Ventral view of a representative specimen, length 90 μm. B–K. Variability of body shape and size as well as of the nuclear (shaded grey) and oral (shaded yellow) apparatus. L. Semi-schematic diagram, showing the general body organization. M–N. Ciliary pattern of ventral and dorsal sides. Arrow marks the right suture and black arrowheads denote the position of the ciliary whorl (posterior suture). Scale bars = 30 μm.

Fig. 2 in Morphological versus molecular delimitation of ciliate species: a case study of the family Clevelandellidae (Protista, Ciliophora, Armophorea)

Fig. 2. Clevelandella constricta (Kidder, 1937). Thai I specimens isolated from Panesthia angustipennis angustipennis (Illiger, 1801) after protargol impregnation. A–J. Variability of body shape and size as well as of the nuclear (shaded grey) and oral (shaded yellow) apparatus. Scale bar = 50 μm.

Fig. 7 in Morphological versus molecular delimitation of ciliate species: a case study of the family Clevelandellidae (Protista, Ciliophora, Armophorea)

Fig. 7. Clevelandella panesthiae (Kidder, 1937). Vietnamese specimens isolated from Panesthia angustipennis cognata Bey-Bienko, 1969 after protargol impregnation. A–D. Variability of body shape and size as well as of the nuclear (shaded grey) and oral (shaded yellow) apparatus. Scale bar = 50 μm.

Fig. 17 in Morphological versus molecular delimitation of ciliate species: a case study of the family Clevelandellidae (Protista, Ciliophora, Armophorea)

Fig. 17. Phylogeny of the order Clevelandellida de Puytorac & Grain, 1976 based on 18S rRNA gene sequences. Some metopids were used as outgroup. Bootstrap values for maximum likelihood analyses performed in IQTrees and PhyML as well as posterior probabilities for Bayesian interference performed in MrBayes and Phycas were mapped onto the best scoring IQTree. Dashes indicate statistical support below 50%, asterisks denote mismatch with the topology shown. Sequences obtained in this study are in bold face. Members of each morphospecies are marked by the same color, following the code in the inset. For specimen codes and further details, see Table 6. The scale bar denotes three substitutions per one hundred nucleotide positions.