3.2. Cytogenetic Analyses
Bradypodion thamnobates: the karyotype is composed of 2n = 34 chromosomes as previously described ([
36] reported as unpublished ex [
3]). There are 12 biarmed macrochromosomes and 22 microchromosomes, with difficult to identify morphologies (
Figure 1a). C-banding revealed the presence of heterochromatin in the pericentromeric regions (
Figure 1b) and interstitial telomeric signal (ITS) signals were detected in a pair of macrochromosomes (
Figure 1c). FISH with rRNA probe showed a signal in pair 2 (
Figure 1d).
Brookesia stumpffi: we only had one individual available and due to methodological complications were only able to obtain preliminary data for this species. The karyotype is composed of 2n = 36 with 12 macrochromosomes and 24 microchromosomes as described previously [
33]. As seen in
Figure 1e, the macrochromosomes are biarmed. C-banding revealed heterochromatin in the centromeres of several microchromosomes (
Figure 1f). We also detected ITS in at least two chromosomal pairs (
Figure 1g) and the rRNA probe produced a signal in one pair of microchromosomes (
Figure 1h).
Calumma brevicorne: the karyotype is composed of 2n = 32 as described previously [
34,
37]. There are 18 biarmed macrochromosomes and 14 microchromosomes (
Figure 1i). C-banding revealed heterochromatin in one arm of a middle-sized metacentric pair (
Figure 1j). We detected ITS signals in several pairs of macrochromosomes (
Figure 1k). FISH with rRNA probe showed a signal in pair 1 (
Figure 1l). CGH did not detect any sexual differences (Figure 4a,b).
Calumma cf.
crypticum: the karyotype is composed of 2n = 32 chromosomes. Eighteen chromosomes are biarmed macrochromosomes and 14 are microchromosomes (
Figure 1m). In pair 9 we detected intraspecific polymorphism in the chromosome morphology. In four studied individuals both chromosomes were metacentric, in one male both were acrocentric and in one female one chromosome was metacentric and the other acrocentric (
Figure 1m in box). C-banding showed heterochromatin in the centromeric/pericentromeric regions of several macrochromosomes and in one arm of a pair of middle-sized macrochromosomes (
Figure 1n). On three pairs of macrochromosomes we detected ITS signals (
Figure 1o). FISH with rRNA probe revealed a signal in pair 1 (
Figure 1p). CGH did not uncover any sexual differences (Figure 4c,d).
Calumma globifer: the karyotype consists of 2n = 36 chromosomes as previously described [
38]. Twelve chromosomes are biarmed macrochromosomes, 24 chromosomes can be assigned as microchromosomes (
Figure 1q). C-banding revealed heterochromatin in the pericentromeric regions of four pairs of macrochromosomes (
Figure 1r). No ITS signals were detected (
Figure 1s). FISH with rRNA probe showed a signal in pair 2 (
Figure 1t). CGH did not uncover any sexual differences (Figure 4e,f).
Calumma malthe: the karyotype is composed of 2n = 36 chromosomes. There are 12 macro- and 24 microchromosomes. Pair 2 is acrocentric while all other macrochromosomes are biarmed (
Figure 2a). C-banding revealed heterochromatin in a pair of microchromosomes (
Figure 2b). ITS signals were detected in several macrochromosomes (
Figure 2c) and rRNA probe produced a signal in pair 1 (
Figure 2d).
Calumma parsonii: the karyotype is composed of 2n = 36 as previously described [
37]. There are 12 biarmed macrochromosomes and 24 microchromosomes (
Figure 2e). C-banding revealed heterochromatin in pericentromeric regions in four pairs of macrochromosomes and one microchromosome pair (
Figure 2f). No ITS signals were detected (
Figure 2g). FISH with rRNA probe showed a signal in pair 2 (
Figure 2h). CGH did not uncover any sexual differences (Figure 4g,h).
Chamaeleo calyptratus: the karyotype is composed of 2n = 24 as previously described [
39]. There are 12 biarmed macrochromosomes and 12 microchromosomes (
Figure 2i). C-banding uncovered heterochromatin in the pericentomeric regions centromeres (
Figure 2j). ITSs were present in a pair of macrochromosomes (
Figure 2k). FISH with rRNA probe showed a signal in pair 1 (
Figure 2l) and CGH did not reveal any sexual differences (Figure 4i,j).
Kinyongia boehmei: the karyotype is composed of 2n = 36 chromosomes of which 12 are biarmed macrochromosomes and 24 microchromosomes (
Figure 2m). C-banding uncovered heterochromatin in the centromeres and on a pair of microchromosomes (
Figure 2n). ITSs were present in several chromosomal pairs (
Figure 2o). FISH with rRNA probe produced a signal in pair 2 (
Figure 2p).
Kinyongia xenorhina: the karyotype consists of 2n = 36 of which 12 are biarmed macrochromosomes and 24 microchromosomes (
Figure 2q). Pair 6 can also be classified as acrocentric as the p-arms are rather short. C-banding uncovered heterochromatin in the pericentromeric regions of one pair of macrochromosomes (
Figure 2r). We detected ITS signals in several macrochromosomes (
Figure 2s). FISH with rRNA probe showed a signal in pair 2 (
Figure 2t).
Rhampholeon temporalis: the karyotype consists of 2n = 22 where 10 chromosomes are biarmed macrochromosomes, six are intermediate-sized chromosomes and six are microchromosomes (
Figure 3a). Heterochromatin accumulation was found in pair 8 of the intermediate-sized chromosomes (
Figure 3b). C-banding revealed larger blocks of heterochromatin in one chromosome from this pair in the female and also a different position of its centromere. ITSs were found in the intermediate-sized chromosomes (
Figure 3c). FISH with rRNA probe produced a signal in pair 2 (
Figure 3d).
Rieppeleon brevicaudatus: the karyotype consists of 2n = 62 (note the large number) and subsequently the chromosomes decline in size and are most likely all acrocentric (
Figure 3e). C-banding uncovered heterochromatin in the centromeres/pericentromeric regions (
Figure 3f). No ITS signals were detected, however in several chromosomes we observed a higher accumulation of telomeric sequences in the telomeric regions (
Figure 3g). FISH with rRNA probe showed a signal in one pair, probably pair 6 or 7 (
Figure 3h). CGH did not uncover any sexual differences (
Figure 4k,l).
Trioceros bitaeniatus: the karyotype consists of 2n = 24 of which 20 are macrochromosomes and 4 microchromosomes as described previously [
40]. All macrochromosomes are biarmed (
Figure 3i). C-banding uncovered heterochromatin in the centromeres/pericentromeric regions (
Figure 3j). ITSs were detected in pairs 1 and 2 (
Figure 3k). In pair 2 we observed a secondary constriction at the site where the rRNA probe was bound (
Figure 3l).
Trioceros hoehnelii: the karyotype is composed of 2n = 24 of which 20 are macrochromosomes and 4 microchromosomes as previously described [
41]. All macrochromosomes are biarmed (
Figure 3m). C-banding uncovered heterochromatin in the centromeres/pericentromeric regions and in a pair of microchromosomes (
Figure 3n). We detected ITS accumulations in two pairs of macrochromosomes (
Figure 3o). In pair 2 we observed a secondary constriction with the rRNA probe signal (
Figure 3p). CGH did not uncover any sexual differences (
Figure 4m,n).
Trioceros johnstoni: the karyotype is composed of 2n = 36 as described previously [
33]. There are 14 macrochromosomes out of which 12 are biarmed and 22 microchromosomes (
Figure 3q). In pair 7 we observed possible heteromorphy. In the female, it seems that the chromosomes in this pair differ in size while in the male they may differ in morphology (
Figure 3q). C-banding revealed heterochromatin in the pericentromeric regions and also strongly in the only acrocentric macrochromosomes—pair 7, in both sexes (
Figure 3r). We detected ITS signals in several chromosomal pairs (
Figure 3s). FISH with rRNA probe produced a signal in pair 2 (
Figure 3t). CGH revealed enrichment of female sequences in chromosome pair 7 in both sexes (
Figure 4o,p). Chromosomes from pair 7 were microdissected and hybridised separately to female metaphases and the probes prepared from them displayed a signal in both chromosomes from each pair (
Figure 4q,r). The probe prepared from male pair 7 also hybridized to both chromosomes of pair 7 in female metaphase (
Figure 4s).
Trioceros melleri: the karyotype consists of 2n = 36 with 12 predominantly biarmed macrochromosomes and 24 microchromosomes (
Figure 3u). C-banding revealed heterochromatin in one pair of microchromosome (
Figure 3v). No ITS signals were detected (
Figure 3w). In pair 2 we observed a secondary constriction where the rRNA probe produced a signal (
Figure 3x).