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Fig. 7 in Comparative spermatozoa ultrastructure of neotropical grass frogs (genus Leptodactylus) with comments on anuran reproductive modes and phylogeny

Fig. 7. Optimization of ultrastructural characters (characters 6 to 8, and 11 to 13) of the spermatozoa on a pruned phylogenetic hypothesis for Leptodactylidae. See list of characters and bibliographic sources in Table 3. Colors correspond to: gray, ambiguity; blue, state 0; red, state 1.

Fig. 8 in Comparative spermatozoa ultrastructure of neotropical grass frogs (genus Leptodactylus) with comments on anuran reproductive modes and phylogeny

Fig. 8. Taxonomic distribution and optimization of 11 selected ultrastructural characters of the spermatozoa on the phylogenetic hypothesis of Leptodactylus proposed by de S´a et al. (2014) and subsequent modifications (see materials and methods section). Numbers refer to characters described in Results section.

Fig. 6 in Comparative spermatozoa ultrastructure of neotropical grass frogs (genus Leptodactylus) with comments on anuran reproductive modes and phylogeny

Fig. 6. Optimization of reproductive modes and ultrastructural characters (characters 1 to 5) of the spermatozoa on a pruned phylogenetic hypothesis for Leptodactylidae (see section "Optimization" in Materials and Methods). See list of characters and bibliographic sources in Tables 1 and 3 Colors correspond to: gray, ambiguity; blue, state 0; red, state 1; green, state 2; orange, state 3; black, state 4.

Fig. 4 in Comparative spermatozoa ultrastructure of neotropical grass frogs (genus Leptodactylus) with comments on anuran reproductive modes and phylogeny

Fig. 4. Transmission electron microscopy of spermatozoa of species of the Leptodactylus melanonotus group. A: L. melanonotus. LS of the acrosomal vesicle and nuclei (n). Note the long and thick acrosomal vesicle (av) and the reduced sub-acrosomal space (ss). B: L. validus. TS showing acrosomal vesicle (ac) and nuclei (n) at different levels. Note the thickness of the acrosomal vesicle and of the acrosomal space. The conical shape of the nucleus is observed in the different cross-sections. C: L. podicipinus. LS of the acrosomal complex-nuclei (n), with the organelle-free cytoplasmic region (black arrowhead) present. D. L. wagneri. LS of the posterior region of the nucleus (n), showing the asymmetrical nuclear fossa (nf) and the transverse striations (ts). E: L.validus. LS of the posterior nuclear region. Note the symmetrical fossa with the proximal (pc) and distal (dc) centrioles. F: L. wagneri. TS of the axoneme (a) and paraxonemal rod (pr) separated from the mitochondrial collar (mc) by the cytoplasmic canal (cc). G-H: L. wagneri, L. melanonotus. TSs of the tail showing the axoneme (a) associated to the juxtaxonemal fiber (jf), and the axial fiber (af) attached to the other components of the tail by the undulating membrane (o). I: L. leptodactyloides. Terminal region of the tail where the axoneme (a) is the only element of the tail. Scales bars: C, D, H, I = 0.5 µm; A, E-G = 0.2 µm; B = 1 µm.

Fig. 1. A-B in Comparative spermatozoa ultrastructure of neotropical grass frogs (genus Leptodactylus) with comments on anuran reproductive modes and phylogeny

Fig. 1. A-B: Schematic reconstruction of the morphology of the two types of spermatozoa observed in the genus Leptodactylus. Note that the main difference between the two morpho-types is the presence of the organelle-free cytoplasmic region between the acrosomal vesicle and the other end of the nucleus. Dashed lines indicate the cross-section represented on the right-hand side of the image.

FIGURE 7 in A new grass frog with rupicolous tadpoles from northern Zambia (Anura: Ptychadenidae)

FIGURE 7. Two habitat photos of the Mutinondo site showing the steep slope with a film of water seeping out of the higher vegetation, and the associated granite inselbergs.

FIGURE 4. A in A new grass frog with rupicolous tadpoles from northern Zambia (Anura: Ptychadenidae)

FIGURE 4. A—Part of the advertisement call of Ptychadena mutinondoensis, showing the clicks and brief trills. B—The advertisement call of Ptychadena broadleyi showing a typical trill of 10 pulses. Both recorded at 24°C.

FIGURE 1 in A new grass frog with rupicolous tadpoles from northern Zambia (Anura: Ptychadenidae)

FIGURE 1. Map showing breeding sites of Ptychadena mutinondoensis in the Mutinondo Wilderness locality in northern Zambia: A—Lodge Rock, B—Camper's Rock, C—Klipspringer Rock, D—Kankonde Rock, E—Between Charlies Rock and Big Chipundu Stream, F—Downstream of Mulinso Falls, and G—Top end of Mafonie Mushitu.

FIGURE 3. A in A new grass frog with rupicolous tadpoles from northern Zambia (Anura: Ptychadenidae)

FIGURE 3. A—Lateral view of holotype of Ptychadena mutinondoensis (ZMB 88379). B—Sharp snout of Ptychadena mutinondoensis (ZMB 88380) compared to C—snout of Ptychadena broadleyi (ZMB 88389). Scale bars = 10 mm.

FIGURE 6 in A new grass frog with rupicolous tadpoles from northern Zambia (Anura: Ptychadenidae)

FIGURE 6. Top—Eggs of Ptychadena mutinondoensis developing in a film of water under vegetation. Below—A stage 40 tadpole ZMB 88378 from Mutinondo Wilderness. Tadpole scale bar = 10 mm.

FIGURE 5 in A new grass frog with rupicolous tadpoles from northern Zambia (Anura: Ptychadenidae)

FIGURE 5. Rainfall and timing of breeding of Ptychadena mutinondoensis at Lodge Rock, Mutinondo Wilderness, during the 2016/2017 season. Vertical arrows: initial two egg-laying events with % of known oviposition sites active; horizontal arrow: period eggs and/or tadpoles were present. Rainfall data collected at Lodge Rock, 200 m north of type locality.

FIGURE 2 in A new grass frog with rupicolous tadpoles from northern Zambia (Anura: Ptychadenidae)

FIGURE 2. Maximum Likelihood phylogeny of representatives of all available Ptychadena species. Branch support is given as SH-aLRT/UFBoot, both as percentages. Scale bar is substitutions/site.