Spectral sensitivity of cone photoreceptors and opsin expression in two colour-divergent lineages of the lizard Ctenophorus decresii

Intraspecific differences in sensory perception are rarely reported but may occur when a species range extends across varying sensory environments, or there is coevolution between the sensory system and a varying signal. Examples in colour vision and colour signals are rare in terrestrial systems. The tawny dragon lizard Ctenophorus decresii is a promising candidate for such intraspecific variation, because the species comprises two geographically and genetically distinct lineages in which throat colour (a social signal used in intra- and inter-specific interactions) is locally adapted to the habitat and differs between lineages. Male lizards from the southern lineage have UV-blue throats, whereas males from the northern lineage are polymorphic with four discrete throat colours that all show minimal UV reflectance. Here, we determine the cone photoreceptor spectral sensitivities and opsin expression of the two lineages, to test whether they differ, particularly in the UV wavelengths. Using microspectrophotometry on retinal cone photoreceptors, we identified a long-wavelength-sensitive (LWS) visual pigment, a 'short' and 'long' medium-wavelength-sensitive (MWS) pigment and a short-wavelength-sensitive (SWS) pigment, all of which did not differ in λmax between lineages. Through transcriptome analysis of opsin genes we found that both lineages express four cone opsin genes, including the SWS1 opsin with peak sensitivity in the UV range, and that amino acid sequences did not differ between lineages with the exception of a single leucine to valine substitution in the RH2 opsin. Counts of yellow and transparent oil droplets associated with LWS+MWS and SWS+UVS cones, respectively, showed no difference in relative cone proportions between lineages. Therefore, contrary to predictions, we find no evidence of differences between lineages in single cone photoreceptor spectral sensitivity or opsin expression. However, we confirm the presence of four single cone classes, suggesting tetrachromacy in C. decresii, and we also provide the first evidence of UV sensitivity in agamid lizards.

Sexual selection predicts brain structure in dragon lizards

Phenotypic traits such as ornaments and armaments are generally shaped by sexual selection, which often favours larger and more elaborate males compared to females. But can sexual selection also influence the brain? Previous studies in vertebrates report contradictory results with no consistent pattern between variation in brain structure and the strength of sexual selection. We hypothesize that sexual selection will act in a consistent way on two vertebrate brain regions that directly regulate sexual behaviour: the medial preoptic nucleus (MPON) and the ventromedial hypothalamic nucleus (VMN). The MPON regulates male reproductive behaviour whereas the VMN regulates female reproductive behaviour and is also involved in male aggression. To test our hypothesis, we used high-resolution magnetic resonance imaging combined with traditional histology of brains in 14 dragon lizard species of the genus Ctenophorus that vary in the strength of precopulatory sexual selection. Males belonging to species that experience greater sexual selection had a larger MPON and a smaller VMN. Conversely, females did not show any patterns of variation in these brain regions. As the volumes of both these regions also correlated with brain volume (BV) in our models, we tested whether they show the same pattern of evolution in response to changes in BV and found that the do. Therefore, we show that the primary brain nuclei underlying reproductive behaviour in vertebrates can evolve in a mosaic fashion, differently between males and females, likely in response to sexual selection, and that these same regions are simultaneously evolving in concert in relation to overall brain size.

Afternoon T : Testosterone level is higher in red than yellow male polychromatic lizards

Recent work on within-species polymorphism across a broad range of taxa has renewed and considerably increased the attention to this classic evolutionary area, notably in lizard species where colors covary with reproductive strategies. We demonstrate elsewhere that red-headed males beat yellow-headed males in staged contests for females in the Australian painted dragon lizard Ctenophorus pictus. This morph difference in behaviour is linked to what appears to be a convention of red dominance in male–male interactions set very early in ontogeny, long before coloration has developed. In the current note, we investigate the relationship between time of day, which is directly linked to vigilance time in territorial males, and plasma levels of testosterone and corticosterone. We show that red males have higher testosterone levels in late afternoon following a day of territory patrolling and a non-significant trend in plasma corticosterone levels that decline with time of day. In conclusion, there are significant differences in testosterone profile between the two color morphs, providing a potential proximate link to the behavioural differences between them.

Sexual selection and the evolution of complex color patterns in dragon lizards

Many species have elaborate and complex coloration and patterning, which often differ between the sexes. Sexual selection may increase the size or intensity of color patches (elaboration) in one sex or drive the evolution of novel signal elements (innovation). The latter potentially increases color pattern complexity. Color pattern complexity may also be influenced by ecological factors related to predation and environment; however, very few studies have investigated the effects of both sexual and natural selection on color pattern complexity across species. We used a phylogenetic comparative approach to examine these effects in 85 species and subspecies of Australian dragon lizards (family Agamidae). We quantified color pattern complexity by adapting the Shannon–Wiener diversity index. There were clear sex differences in color pattern complexity, which were positively correlated with both sexual dichromatism and sexual size dimorphism, consistent with the idea that sexual selection plays a significant role in the evolution of color pattern complexity. By contrast, we found little evidence of a link between environmental factors and color pattern complexity on body regions exposed to predators. Our results suggest that sexual selection rather than natural selection has led to increased color pattern complexity in males.

Isolation breeds naivety : island living robs Australian varanid lizards of toad-toxin immunity via four-base-pair mutation

Since their introduction to the toad-free Australian continent cane toads (Bufo marinus) have caused a dramatic increase in naïve varanid mortality when these large lizards attempt to feed on this toxic amphibian. In contrast Asian–African varanids, which have coevolved with toads, are resistant to toad toxin. Toad toxins, such as Bufalin target the H1-H2 domain of the α1 subunit of the sodium-potassium-ATPase enzyme. Sequencing of this domain revealed identical nucleotide sequences in four Asian as well as in three African varanids, and identical sequences in all 11 Australian varanids. However, compared to the Asian–African varanids, the Australian varanids showed four-base-pair substitutions, resulting in the alteration in three of the 12 amino acids representing the H1-H2 domain. The phenotypic effect of the substitutions was investigated in human embryonic kidney (HEK) 293 cells stably transfected with the Australian and the Asian–African H1-H2 domains. The transfections resulted in an approximate 3000-fold reduction in resistance to Bufalin in the Australian HEK293 cells compared to the Asian–African HEK293 cells, demonstrating the critical role of this minor mutation in providing Bufalin resistance. Our study hence presents a clear link between genotype and phenotype, a critical step in understanding the evolution of phenotypic diversity.

Climate change, multiple paternity and offspring survival in lizards

Recent work suggests that rising spring temperatures over recent decades have eliminated many lizard populations, and threaten many more worldwide. However, because ambient temperatures constrain activity times in ectotherms, warming conditions (as expected under global climate change scenarios) can increase the duration of seasonal opportunities for courtship and mating. Thus, in species where polyandry results in enhanced offspring viability, a warming climate may not necessarily impair long-term survival. Our nine-year study of a sand lizard (Lacerta agilis) population near the northern range limit in Sweden revealed consistently higher incidence of multiple paternity of clutches in warmer years, and higher viability of offspring from multiply-sired clutches (presumably reflecting the advantages of more intense sperm competition). Any trend to warmer spring temperatures likely will benefit offspring viability in this system, by increasing a female's opportunities to mate with additional males.

Detecting the impact of invasive species on native fauna: Cane toads (Bufo marinus), frillneck lizards (Chlamydosaurus kingii) and the importance of spatial replication

An understanding of which native species are severely impacted by an anthropogenic change (such as the arrival of an invasive species) and which are not is critical to prioritizing conservation efforts. However, it is difficult to detect such impacts if the native taxa exhibit strong stochastic variations in abundance; a ‘natural’ population decline might be wrongly interpreted as an impact of the invader. Frillneck lizards (Chlamydosaurus kingii) are large iconic Australian agamids, and have been reported to decline following the invasion of toxic cane toads. We monitored three populations of the species in the savanna woodland of tropical Australia over a 7-year period bracketing toad arrival. One population crashed, one remained stable and one increased. Hence, studies on any single population might have inferred that cane toads have negative, negligible or positive effects on frillneck lizards. With the benefit of spatial replication, and in combination with observations of prey choice by captive lizards, our data suggest that invasive cane toads have had little or no effect on frillneck abundance.

In hot pursuit : fluctuating mating system and sexual selection in sand lizards

A changing climate is expected to have profound effects on many aspects of ectotherm biology. We report on a decade-long study of free-ranging sand lizards (Lacerta agilis), exposed to an increasing mean mating season temperature and with known operational sex ratios. We assessed year-to-year variation in sexual selection on body size and postcopulatory sperm competition and cryptic female choice. Higher temperature was not linked to strength of sexual selection on body mass, but operational sex ratio (more males) did increase the strength of sexual selection on body size. Elevated temperature increased mating rate and number of sires per clutch with positive effects on offspring fitness. In years when the “quality” of a female's partners was more variable (in standard errors of a male sexual ornament), clutches showed less multiple paternity. This agrees with prior laboratory trials in which females exercised stronger cryptic female choice when male quality varied more. An increased number of sires contributing to within-clutch paternity decreased the risk of having malformed offspring. Ultimately, such variation may contribute to highly dynamic and shifting selection mosaics in the wild, with potential implications for the evolutionary ecology of mating systems and population responses to rapidly changing environmental conditions.

Increased mortality of naive varanid lizards after the invasion of non-native can toads (bufo marinus)

Exotic animal and plant species introduced into the Australian continent often imparted catastrophic effects on the indigenous fauna and flora. Proponents of biological control introduced the South American Cane Toad (Bufo marinus) into the sugar cane fields of Queensland in 1935. The Cane Toad is one of the most toxic bufonids and when seized by naive Australian predators, the toxin usually kills the attacker. One group of Australian squamate reptiles that are very susceptible to Cane Toad toxins is varanid lizards. Prior to Cane Toad invasion of our study area, the Adelaide River floodplain of the Northern Territory of Australia, annual mortality of adult male radio-tagged yellow-spotted Goannas (Varanus panoptes) was very low. After the arrival of toads in October 2005, all radio-tracked goannas were found dead in August 2006, most likely attempting to feed on the toads. Our results suggest that invasive Cane Toads place naive adult male Yellow-spotted Goannas at risk of possibly >90% mortality. This increase in mortality could reduce the genetic diversity and hamper long-term survival of these large carnivorous lizards.

Population genetic structure, gene flow and sex-biased dispersal in frillneck lizards (Chlamydosaurus kingii)

By using both mitochondrial and nuclear multiloci markers, we explored population genetic structure, gene flow and sex-specific dispersal of frillneck lizards (Chlamydosaurus kingii) sampled at three locations, separated by 10 to 50 km, in a homogenous savannah woodland in tropical Australia. Apart from a recombinant lizard, the mitochondrial analyses revealed two nonoverlapping haplotypes/populations, while the nuclear markers showed that the frillneck lizards represented three separate clusters/populations. Due to the small population size of the mtDNA, fixation may occur via founder effects and/or drift. We therefore suggest that either of these two processes, or a combination of the two, are the most likely causes of the discordant results obtained from the mitochondrial and the nuclear markers. In contrast to the nonoverlapping mitochondrial haplotypes, in 12 out of 74 lizards, mixed nuclear genotypes were observed, hence revealing a limited nuclear gene flow. Although gene flow should ultimately result in a blending of the populations, we propose that the distinct nuclear population structure is maintained by frequent fires resulting in local bottlenecks, and concomitant spatial separation of the frillneck lizard populations. Limited mark-recapture data and the difference in distribution of the mitochondrial and nuclear markers suggest that the mixed nuclear genotypes were caused by juvenile male-biased dispersal.