重庆市城口县大巴山国家级自然保护区发现宁陕线形蛇与该物种的遗传分化分析

宁陕线形蛇Stichophanes ningshaanensis Yuan,1983是游蛇科Colubridae的单型属单型种,为中国特有种,至今仅被报道分布于陕西省宁陕县火地塘(模式产地)、湖北省神农架国家级自然保护区和重庆市五里坡国家级自然保护区。目前对该物种的研究和野外采集记录甚少。2017年7月8日,在重庆市城口县大巴山国家级自然保护区采到1号疑似宁陕线形蛇的雌性成体标本。经形态比较和分子系统发育分析,鉴定为宁陕线形蛇。基于线粒体细胞色素b(cyt b)基因和核基因片段cmos的系统发育分析表明,城口大巴山标本嵌入宁陕线形蛇支系中,与该物种其他种群在cyt b基因上的遗传距离(0.001~0.009)远小于与其近缘种的(0.174~0.292)。本研究揭示宁陕线形蛇种群间存在形态和遗传上的分化。形态上,城口大巴山标本上唇鳞数量为5/5,而神农架和火地塘种群均为6/6,前者尾下鳞数量为61对,少于后者(64~73对)。在cyt b基因上,3个种群各自拥有1个单倍型。城口大巴山单倍型与湖北神农架单倍型之间有1个突变、与宁陕火地塘单倍型之间有9个突变,而分子系统树支持宁陕火地塘种群与湖北神农架种群关系更近。研究结果为宁陕线形蛇的地理分布和系统学研究提供了重要信息。     

Molecular systematics of New World lampropeltinine snakes (Colubridae): implications for biogeography and evolution of food habits

We used mitochondrial gene sequences to infer phylogenetic relationships among North American snakes of the colubrid tribe Lampropeltini (Arizona, Bogertophis, Cemophora , New World Elaphe, Lampropellis, Pituophis, Rhinocheilus, Senticolis, Stilosoma) , and assessed the implications of our findings for the biogeography and evolution of food habits among these serpents. The maximum likelihood phylogeny identified Rhinocheilus as the sister taxon to all other lampropeltinines, and supported the monophyly of Lampropeltis (including Stilosoma) , New World Elaphe , and Pituophis , but not that of Bogertophis. This phylogeny also suggested a sister group relationship between Cemophora and Lampropeltis , and between New World Elaphe and Pituophis , and strongly supported that Sentkolis belongs within Lampropeltini, thus contradicting previous suggestions that Senticolis is not a lampropeltinine. Using a method for approximating ancestral areas of clades, we determined that western North America was most likely the ancestral area of lampropeltinines. Our survey of published studies, combined with unpublished data, indicated that lampropeltinines as a group feed mainly on mammals, less frequently on lizards, birds, and bird eggs, and only rarely on squamate eggs, snakes, anurans, and insects. Some individual species indeed emphasize mammals in their diets, but others most frequently eat lizards, squamate eggs, bird eggs, or snakes, whereas others take two prey types with similar frequency. Our reconstruction of the evolution of food habits among lampropeltinines suggests that a diet emphasizing lizards is ancestral, and therefore diets that mosdy consist of mammals, squamate and bird eggs, and snakes are derived within the clade. In at least some species, smaller individuals prey mostly on lizards and larger ones add mammals to their diets.     

Heterologous amplification of microsatellite markers from colubroid snakes in European natricines (serpentes: natricinae)

Eighteen microsatellite loci developed for a range of snake species (New World natricines, elapids, crotalids) were tested against European natricines (Natrix natrix, N. maura, and N. tessellata) in cross-species amplification experiments. Five loci were polymorphic (average expected heterozygosity 0.749 for a population of N. natrix in Amsterdam, mean sample size 47.8) and three loci were monomorphic. The remainder could not be consistently scored or failed to amplify. Further tests on single individuals of a diverse set of eight species of colubroid snakes showed that 15 of the 18 loci could be cross-amplified in at least one of these species. We conclude that our results show promise for the utilization of these markers for experimental assessments of genetic variation in the phylogenetically closely related group of European natricine snakes with emphasis on N. natrix. The full suite of microsatellite markers now available for snakes may show additional potential for subsequent investigation across a broader range of colubroid snakes.     

Genetic Connectivity among Populations of an Endangered Snake Species from Southeastern Australia (Hoplocephalus bungaroides, Elapidae)

For endangered species that persist as apparently isolated populations within a previously more extensive range, the degree of genetic exchange between those populations is critical to conservation and management. A lack of gene flow can exacerbate impacts of threatening processes and delay or prevent colonization of sites after local extirpation. The broad-headed snake, Hoplocephalus bungaroides, is a small venomous species restricted to a handful of disjunct reserves near Sydney, Australia. Mark-recapture studies have indicated low vagility for this ambush predator, suggesting that gene flow also may be low. However, our analyses of 11 microsatellite loci from 163 snakes collected in Morton National Park, from six sites within a 10-km diameter, suggest relatively high rates of gene flow among sites. Most populations exchange genes with each other, with one large population serving as a source area and smaller populations apparently acting as sinks. About half of the juvenile snakes, for which we could reliably infer parentage, were collected from populations other than those in which we collected their putative parents. As expected from the snakes' reliance on rocky outcrops during cooler months of the year, most gene flow appears to be along sandstone plateaux rather than across the densely forested valleys that separate plateaux. The unexpectedly high rates of gene flow on a landscape scale are encouraging for future conservation of this endangered taxon. For example, wildlife managers could conserve broad-headed snakes by restoring habitats near extant source populations in areas predicted to be least affected by future climate change.     

Coral snake mimicry: live snakes not avoided by a mammalian predator

The occurrence of coral snake coloration among unrelated venomous and non-venomous New World snake species has often been explained in terms of warning coloration and mimicry. The idea that snake predators would avoid coral snakes in nature seems widely established and is postulated in many discussions on coral snake mimicry. However, the few workers that have tested a potential aposematic function of the conspicuous colour pattern focused exclusively on behaviour of snake predators towards coloured abstract models. Here we report on behaviour of temporarily caged, wild coatis (Nasua narica) when confronted with co-occurring live snakes, among which were two species of venomous coral snakes. Five different types of responses have been observed, ranging from avoidance to predation, yet none of the coatis avoided either of the two coral snake species or other species resembling these. As in earlier studies coatis appeared to avoid coral snake models, our findings show that results from studies with abstract snake models cannot unconditionally serve as evidence for an aposematic function of coral snake coloration.     

Interspecific patterns of species richness, geographic range size, and body size among New World venomous snakes

Many higher taxa exhibit latitudinal gradients in species richness, geographic range size, and body size. However, these variables are often interdependent, such that examinations of univariate or bivariate patterns alone may be misleading. Therefore, I examined latitudinal gradients in, and relationships between, species richness, geographic range size, and body size among 144 species of New World venomous snakes [families Elapidae (coral snakes) and Viperidae (pitvipers)]. Both lineages are monophyletic, collectively span 99° of latitude, and are extremely variable in body size and geographic range sizes. Coral snakes exhibit highest species richness near the equator, while pitviper species richness peaks in Central America. Species – range size distributions were strongly right-skewed for both families. There was little support for Bergmann's rule or Rapoport's rule for snakes of either family, as neither body size nor range size increased significantly with latitude. However, range area and median range latitude were positively correlated above 15° N, indicating a possible "Rapoport effect" at high northern latitudes. Geographic range size was positively associated with body size. Available continental area strongly influenced range size. Comparative (phylogenetically-based) analyses revealed that shared history is a poor predictor of range size variation within clades. Among vipers, trends in geographic range sizes may have been structured more by historical biogeography than by macroecological biotic factors.     

Marine invasions by non-sea snakes, with thoughts on terrestrial-aquatic-marine transitions

Few species of snakes show extensive adaptations to aquatic environments and even fewer exploit the oceans. A survey of morphology, lifestyles, and habitats of 2552 alethenophidian snakes revealed 362 (14%) that use aquatic environments, are semi-aquatic, or aquatic; about 70 (2.7%) of these are sea snakes (Hydrophiinae and Laticaudinae). The ancient and aquatic family Acrochordidae contains three extant species, all of which have populations inhabiting brackish or marine environments, as well as freshwater. The Homalopsidae have the most ecologically diverse representatives in coastal habitats. Other families containing species exploiting saline waters with populations in freshwater environments include: the Dipsadidae of the western hemisphere, the cosmopolitan Natricidae, the African Grayinae, and probably a few Colubridae. Species with aquatic and semi-aquatic lifestyles are compared with more terrestrial (fossorial, cryptozoic, and arboreal) species for morphological traits and life histories that are convergent with those found in sea snakes; this may provide clues to the evolution of marine snakes and increase our understanding of snake diversity.     

Phylogenetic relationships of the dwarf boas and a comparison of Bayesian and bootstrap measures of phylogenetic support

Four New World genera of dwarf boas (Exiliboa, Trachyboa, Tropidophis, and Ungaliophis) have been placed by many systematists in a single group (traditionally called Tropidophiidae). However, the monophyly of this group has been questioned in several studies. Moreover, the overall relationships among basal snake lineages, including the placement of the dwarf boas, are poorly understood. We obtained mtDNA sequence data for 12S, 16S, and intervening tRNA-val genes from 23 species of snakes representing most major snake lineages, including all four genera of New World dwarf boas. We then examined the phylogenetic position of these species by estimating the phylogeny of the basal snakes. Our phylogenetic analysis suggests that New World dwarf boas are not monophyletic. Instead, we find Exiliboa and Ungaliophis to be most closely related to sand boas (Erycinae), boas (Boinae), and advanced snakes (Caenophidea), whereas Tropidophis and Trachyboa form an independent clade that separated relatively early in snake radiation. Our estimate of snake phylogeny differs significantly in other ways from some previous estimates of snake phylogeny. For instance, pythons do not cluster with boas and sand boas, but instead show a strong relationship with Loxocemus and Xenopeltis. Additionally, uropeltids cluster strongly with Cylindrophis, and together are embedded in what has previously been considered the macrostomatan radiation. These relationships are supported by both bootstrapping (parametric and nonparametric approaches) and Bayesian analysis, although Bayesian support values are consistently higher than those obtained from nonparametric bootstrapping. Simulations show that Bayesian support values represent much better estimates of phylogenetic accuracy than do nonparametric bootstrap support values, at least under the conditions of our study.     

Out of Asia: natricine snakes support the Cenozoic Beringian Dispersal Hypothesis

Based on a combination of six mitochondrial gene fragments (12S RNA, cyt b, ND1, ND2, ND4 and CO1) and one nuclear gene (c-mos) from 22 genera we infer phylogenetic relationships among natricine snakes and examine the date and area of origin of these snakes. Our phylogenetic results indicate: (1) the subfamily Natricinae is strongly supported as monophyletic including a majority of extant genera, and a poorly known and previously unassigned species Trachischium monticola, (2) two main clades are inferred within Natricinae, one containing solely taxa from the Old World (OW) and the other comprising taxa from a monophyletic New World (NW) group with a small number of OW relatives. Within the first clade, the genera Xenochrophis and Amphiesma are apparently not monophyletic. Divergence dating and ancestral area estimation indicate that the natricines originated in tropical Asia during the later Eocene or the Oligocene. We recover two major dispersals events out of Asia, the first to Africa in the Oligocene (28 Ma) and the second to the Western Palearctic and the New World at 27 Ma. This date is consistent with the dispersal of numerous other OW groups into the NW.     

DID EGG‐LAYING BOAS BREAK DOLLO'S LAW? PHYLOGENETIC EVIDENCE FOR REVERSAL TO OVIPARITY IN SAND BOAS (ERYX: BOIDAE)

Re-evolution of lost complex morphological characters has been proposed for several characters, including insect wings, limbs, eyes in snakes, and digits in lizards, among others. There has also been much interest in whether the transition from oviparity to viviparity is reversible, particularly in squamate reptiles where the transition to viviparity has occurred more times than in any other lineage. Here, we present a phylogenetic analysis of boid snakes based on a concatenated multigene study of all genera of erycines, New and Old World boines, plus other groups thought to be closely related with boines such as monotypic species Calabaria and Casarea . We reconstruct ancestral parity mode on this phylogeny and present statistical evidence that oviparity reevolved in a species of Old World sand boa in the genus Eryx nearly 60 million years after the initial boid transition to viviparity. Remarkably, like other viviparous boas hatchlings of oviparous Eryx lack an egg-tooth providing independent evidence that oviparity is a derived state in these species.     

What are the closest relatives of the hot-spring snakes (Colubridae, Thermophis), the relict species endemic to the Tibetan Plateau?

We conducted phylogenetic analyses to identify the closest related living relatives of the Xizang and Sichuan hot-spring snakes (T. baileyi and T. zhaoermii) endemic to the Tibetan Plateau, using mitochondrial DNA sequences (cyt b, ND4) from eight specimens, together with sequences from 95 additional caenophidian and five henophidian genera that were downloaded from GenBank. Phylogenetic trees were obtained using Bayesian Inference and Maximum likelihood methods. Results suggest that hot-spring snakes, which are adapted to high and cold environments, were clustered in the monophyletic Xenodontinae. Xenodontinae is one of the largest subfamilies of colubrid snakes, with about 90 genera and more than 500 species known, and are primarily tropical snakes previously thought to be restricted to the New World. Our data failed to provide any evidence that the New World xenodontines diverged from Thermophis and dispersed into the New World, also failed to suggest a colonization of Asia by New World xenodontines by dispersal from the New World. An alternative plausible scenario may be that Thermophis and the New World xenodontines evolved independently in Asia and America, respectively, after the divergence of their common ancestor. The divergence of the two species in Thermophis was caused by the barrier of the Hengduan Mountains, and the speciation had almost occurred when Tibetan Plateau attained present elevation.     

An immunological assessment of the phylogenetic position of New World coral snakes

The New World coral snakes (micrurines), genera Micrurus and Micruroides have recently been seen as derived from a lineage of South American colubrids, rather than from a common lineage with Old World elapids and sea snakes as traditionally accepted. We compared serum albumins of representative coral snakes, Old World elapids, sea snakes, and neotropical colubrids immunologically. Phylogenetic analysis of the biochemical data unambiguously allies the micrurines with the family Elapidae as it is currently understood. Using the albumin molecular clock calibration derived from other terrestrial vertebrates. we suggest a late Oligocene-early Miocene separation between the New and Old World elapid lineages. This requires a movement of elapid stocks from Asia into North America, and supporting evidence for this model is derived from several paleontological sources. We suggest that a number of extant micrurine lineages have had long independent histories.     

Effects of season, sex and body size on the feeding ecology of turtle-headed sea snakes (Emydocephalus annulatus) on IndoPacific inshore coral reefs

In terrestrial snakes, many cases of intraspecific shifts in dietary habits as a function of predator sex and body size are driven by gape limitation and hence are most common in species that feed on relatively large prey and exhibit a wide body-size range. Our data on sea snakes reveal an alternative mechanism for intraspecific niche partitioning, based on sex-specific seasonal anorexia induced by reproductive activities. Turtle-headed sea snakes (Emydocephalus annulatus) on coral reefs in the New Caledonian Lagoon feed entirely on the eggs of demersal-spawning fishes. DNA sequence data (cytochrome b gene) on eggs that we palpated from stomachs of 37 snakes showed that despite this ontogenetic stage specialization, the prey comes from a taxonomically diverse array of species including damselfish (41 % of samples, at least 5 species), blennies (41 %, 4 species) and gobies (19 %, 5 species). The composition of snake diets shifted seasonally (with damselfish dominating in winter but not summer), presumably reflecting seasonality of fish reproduction. That seasonal shift affects male and female snakes differently, because reproduction is incompatible with foraging. Adult female sea snakes ceased feeding when they became heavily distended with developing embryos in late summer, and males ceased feeding while they were mate searching in winter. The sex divergence in foraging habits may be amplified by sexual size dimorphism; females grow larger than males, and larger snakes (of both sexes) feed more on damselfish (which often lay their eggs in exposed sites) than on blennies and gobies (whose eggs are hidden within narrow crevices). Specific features of reproductive biology of coral reef fish (seasonality and nest type) have generated intraspecific niche partitioning in these sea snakes, by mechanisms different from those that apply to terrestrial snakes.     

Phylogeny of snake trypanosomes inferred by SSU rDNA sequences, their possible transmission by phlebotomines, and taxonomic appraisal by molecular, cross-infection and morphological analysis

Blood examination by microhaematocrit and haemoculture of 459 snakes belonging to 37 species revealed 2.4% trypanosome prevalence in species of Viperidae (Crotalus durissus and Bothrops jararaca) and Colubridae (Pseudoboa nigra). Trypanosome cultures from C. durissus and P. nigra were behaviourally and morphologically indistinguishable. In addition, the growth and morphological features of a trypanosome from the sand fly Viannamyia tuberculata were similar to those of snake isolates. Cross-infection experiments revealed a lack of host restriction, as snakes of 3 species were infected with the trypanosome from C. durissus. Phylogeny based on ribosomal sequences revealed that snake trypanosomes clustered together with the sand fly trypanosome, forming a new phylogenetic lineage within Trypanosoma closest to a clade of lizard trypanosomes transmitted by sand flies. The clade of trypanosomes from snakes and lizards suggests an association between the evolutionary histories of these trypanosomes and their squamate hosts. Moreover, data strongly indicated that these trypanosomes are transmitted by sand flies. The flaws of the current taxonomy of snake trypanosomes are discussed, and the need for molecular parameters to be adopted is emphasized. To our knowledge, this is the first molecular phylogenetic study of snake trypanosomes.     

Molecular evolution of the infrared sensory gene TRPA1 in snakes and implications for functional studies

TRPA1 is a calcium ion channel protein recently identified as the infrared receptor in pit organ-containing snakes. Therefore, understanding the molecular evolution of TRPA1 may help to illuminate the origin of "heat vision" in snakes and reveal the molecular mechanism of infrared sensitivity for TRPA1. To this end, we sequenced the infrared sensory gene TRPA1 in 24 snake species, representing nine snake families and multiple non-snake outgroups. We found that TRPA1 is under strong positive selection in the pit-bearing snakes studied, but not in other non-pit snakes and non-snake vertebrates. As a comparison, TRPV1, a gene closely related to TRPA1, was found to be under strong purifying selection in all the species studied, with no difference in the strength of selection between pit-bearing snakes and non-pit snakes. This finding demonstrates that the adaptive evolution of TRPA1 specifically occurred within the pit-bearing snakes and may be related to the functional modification for detecting infrared radiation. In addition, by comparing the TRPA1 protein sequences, we identified 11 amino acid sites that were diverged in pit-bearing snakes but conserved in non-pit snakes and other vertebrates, 21 sites that were diverged only within pit-vipers but conserved in the remaining snakes. These specific amino acid substitutions may be potentially functional important for infrared sensing.     

两种蛇Sox基因的PCR-SSCP分析 *

采用PCR技术,以特异扩增人SRY基因HMG-box保守区的一对引物,扩增了乌梢蛇和赤链蛇的Sox基因。结果 两种蛇雌雄个体与人一样,均能扩增出大小为220bp左右的基因片段。对扩增产物进行的SSCP分析发现两种蛇之间以及种内雌雄个体间单链迁移率略有差异,而与人有较大差异。本文为研究蛇类的性别决定机制及Sox基因的演化提供了分子生物学资料。     

Integrating phylogeny,environment and space to explore variation in macroecological traits of Viperidae and Elapidae (Squamata: Serpentes)

We used eigenvector mapping in space and phylogeny to investigate the relationships among space, phylogeny and environment on body size and range size variation across two groups of venomous snakes – Viperidae and Elapidae – from the New World. Data on species geographic range sizes, maximum body sizes and phylogenetic relationships were compiled from the available literature. The distributional data were also used to calculate the latitudinal and longitudinal midpoint and the environmental centroids for each species. The eigenvectors extracted from the pair wise spatial and phylogenetic distance matrices were integrated with environmental variables into a method of variation partitioning where the variation in each trait was quantitatively attributed to ‘pure’ and/or shared effects of phylogeny, environment and space. Our results showed that variation in body size was predominantly determined by phylogeny in both groups of snakes. For Viperidae, we found that pure ‘effects’ of phylogeny were the strongest, indicating that most of the body size evolution that was phylogenetically determined in this group occurred independently of environment and geographical proximity. Regarding range sizes, pure phylogenetic influences were very low in both groups, whereas the largest single fraction of explained variation corresponded to overlapped influences of the three sets of predictors, especially for Elapidae. Along with this, we found evidence that niche conservatism is an important processes underlying variation in body size and range size in both groups of snakes.     

河北兴隆县雾灵山蝮蛇分类地位的探讨

河北兴隆县的蝮蛇分类问题,以往有国外学者进行过报道.国内文献记载河北省只分布有一种蝮蛇即短尾蝮.2006年实地考察和饲养观察研究发现,雾灵山的蝮蛇不是同一种,而是两种蝮蛇.它们的形态、分布和习性均不同,依照中国动物志爬行纲第三卷鉴定,分别为短尾蝮和中介蝮,从而恢复了河北省中介蝮分布的纪录.