Phylogenetic relationships of the Papuan Swiftlet Aerodramus papuensis and implications for the evolution of avian echolocation

The swiftlets (genera Aerodramus , Collocalia and Hydrochous ) are unusual among birds in that many species can orientate in complete darkness using echolocation. The position of the Papuan Swiftlet Aerodramus papuensis in this group has been uncertain historically, in part due to morphological differences between it and other swiftlets (it has three toes instead of the usual four) and a lack of data on its behaviour (there is uncertainty about whether it echolocates). Here we investigate the phylogenetic affinities of the Papuan Swiftlet using DNA sequence data from two mitochondrial genes, cytochrome b and ND2. We present evidence that it is able to echolocate but, unlike previously studied species that use echolocation primarily while flying in caves, A. papuensis uses this ability while active outside caves at night. We also provide new evidence for placement of the monotypic Waterfall Swiftlet Hydrochous gigas , a species that does not echolocate. Our data provide strong support for a basal relationship between A. papuensis and other Aerodramus taxa and suggest that this species and H. gigas are sister taxa, a relationship that would indicate paraphyly of the genus Aerodramus . Our phylogeny provides new insights into how echolocation has evolved in the swiftlets, in particular by indicating higher levels of homoplasy in this trait than was previously thought.     

Phylogenetic relationships amongst swifts and swiftlets: a multi locus approach

We recently reconstructed the troublesome swiftlet phylogeny using cytochrome-b mitochondrial DNA sequences. The relationship of the giant swiftlet (Hydrochous gigas) with swiftlets of the genus Aerodramus was, however, unresolved. In an attempt to clarify this issue, we now incorporated mitochondrial 12S rRNA and nuclear beta-fibrinogen intron 7 nuclear DNA sequences with the cyt-b sequences of six swiftlet, two swift, and one hummingbird outgroup species. A partition homogeneity (PH) test, used to determine the congruence of phylogenetic signal between two sets of sequences, suggested that cyt-b and Fib7 sequences were incongruent and therefore should not be combined. However, further analyses revealed that the apparent incongruence was probably due to the high amount of variation in cyt-b sequences. Separate and combined analyses of the three sequences unambiguously placed H. gigas as the sister-group of Aerodramus and supported monophyly of the swiftlets. These results were supported by analyses of combined NADH dehydrogenase subunit-2 (ND2) and cyt-b sequences of H. gigas in combination with sequences previously published by other workers. Recently, it was shown that the pygmy swiftlet (C. troglodytes)--in our phylogenetic analyses consistently placed with other, non-echolocating, Collocalia species--is in fact able to echolocate. Echolocation thereby lost its value to distinguish between different swiftlet genera. Furthermore, the phylogenetic distribution of echolocation can be explained either by its single evolution at the base of the swiftlets, with subsequent loss, or by independent evolution in Aerodramus and C. troglodytes. Because yet unpublished data suggest that only the auditory nuclei in swiftlet brains show adaptations to echolocation, the latter explanation seems the more likely one.     

Occurrence of a nonsulfated chondroitin proteoglycan in the dried saliva of Collocalia swiftlets (edible bird's-nest)

Despite their wide occurrence, proteoglycans (PGs) have never been isolated from the saliva of higher animals. We found that the Collocalia glycoproteins isolated from edible birds'-nests (the dried forms of regurgitated saliva of male Collocalia swiftlets) were rich in a PG containing nonsulfated chondroitin glycosaminoglycans (GAGs). We have devised a method to isolate a PG from the water extract of the white nest built by Aerodramus fuciphagus (white nest swiftlets) with a yield of 2-mg PG per gram nest. This PG contained 83% of carbohydrates, of which 79% were GalNAc and GlcUA (D-glucuronic acid) in an equimolar ratio. By using chondroitin AC lyase, the structure of GAGs in this PG was established to be chondroitin ( --> 4GlcUAbeta1 --> 3GalNAcbeta1 --> )(n) chains. The average molecular mass of the chondroitin chain was estimated to be 49 kDa by gel filtration. We have isolated a linkage region hexasaccharide, DeltaHexUAalpha1 --> 3GalNAcbeta1 --> 4GlcUAbeta1 --> 3Galbeta1 --> 3Galbeta1 --> 4Xyl, from this PG by chondroitinase ABC digestion to show that the GAGs in this PG are also linked to the core protein through the common tetrasaccharide linker, GlcUAbeta1 --> 3Galbeta1 --> 3Galbeta1 --> 4Xyl, found in various PGs. As water was not effective in extracting uronic acid-containing glycoconjugates from the black nest built by black nest swiftlets (A. maximus), we used 4 M guanidium chloride and anion-exchange chromatography in the presence of urea to extract and isolate about 30 mg of a chondroitin PG preparation from 10 g of the desialylated black nest. As the biological significance of chondroitin is still not well understood, bird's nest should become a convenient source for preparing this unique GAG to study its biological functions.     

短嘴金丝燕回声定位叫声特征

2012年6月,对湖南省石门县壶瓶山国家级自然保护区神景洞短嘴金丝燕的回声定位叫声进行研究,在黑暗山洞内使用录音仪器录制其自由飞行状态的声音后使用声音软件进行分析.短嘴金丝燕捕食归巢时,快速飞入洞口,在洞内有光区域不发声,到达洞内黑暗区域后开始发出回声定位叫声,且飞行速度减慢.声音分析结果表明其回声定位叫声为双脉冲组的噪声脉冲串型(noise burst),组内脉冲间隔很短[(6.6±0.42)ms],组间脉冲间隔较长[(99.3±3.86) ms],两者差异显著(P＜0.01).对比第一、第二脉冲声音参数发现,主频和脉冲时程差异不显著,第一、第二脉冲主频分别为(6.2±0.08) kHz和(6.2±0.10) kHz (P＞0.05);脉冲时程分别为(2.9±0.12) ms和(3.2±0.17) ms (P＞0.05);最高和最低频率差异显著,第一、第二脉冲最高频率分别为(20.1±1.10) kHz和(15.4±0.98) kHz (P＜0.01),最低频率分别为(3.7±0.12) kHz和(4.0±0.09)kHz (P＜0.05);第一脉冲频宽((16.5±1.17) kHz)宽于第二脉冲((11.4±1.01) kHz) (P＜0.01);且第一脉冲能量[(-32.5±0.60) dB]高于第二脉冲[(-35.2±0.94) dB] (P＜0.05).另外,短嘴金丝燕在黑暗山洞内的回声定位叫声还包含了部分超声波,最高频率可达33.2 kHz.     

Revision of Dennyus (Collodennyus) lice (Phthiraptera: Menoponidae) from swiftlets, with descriptions of new taxa and a comparison of host–parasite relationships

Lice of the subgenus Dennyus ( Collodennyus ) are host specific, permanent parasites of swiftlets (Aves: Apodidae). As a prelude to a test of the hypothesis that these lice have cospeciated with their hosts, we revise the taxonomy of the subgenus, redescribing the seven previously recognized species, and adding thirteen new species and three new subspecies. All twenty-three of these louse taxa are found on swiftlets (Apodiformes: Apodidae), with four from hosts of the genus Collocalia , eighteen from Aerodramus , and one from Hydrochous . Successful identification is associated in most cases with females; males are only tenuously separable. A complete host–parasite list for the subgenus Collodennyus is provided, as well as a key for the identification of these taxa. Limited morphological variation within the subgenus has prevented ready extraction of discrete characters for cladistic analysis. In the absence of such characters, a cluster analysis of female and male lice is presented. Comparison of a dendrogram for Dennyus ( Collodennyus ) with a molecular phylogeny for the swiftlet hosts suggests that the history of the swiftlet–louse association has been complex, including episodes of host switching and independent speciation by the lice.     

A new phylogeny of swiftlets (Aves: Apodidae) based on cytochrome-b DNA

Due to a lack of distinctive morphological characters, swift taxonomy and phylogeny has always been an area of disagreement. To shed more light on this subject, we reconstructed swift(let) phylogeny based on 1143 bp of mitochondrial cytochrome-b DNA sequence. Although this is not the first attempt to reconstruct swift phylogeny using molecular data, our results show higher support for many of the branches due to our much longer sequences. However, placement of Hydrochous is still unexpected. Implementation of more conservative genetic regions and sampling of more taxa could solve this problem. Most importantly, the Collocaliini resolve as a monophyletic group. The internal structure of the group shows that non-echolocating Collocalia and echolocating Aerodramus form two distinct clades. This is in congruence with earlier classifications based on morphological characters, but in contrast with more recent classifications.     

The diets of Malaysian swiftlets

Four species of swiftlet (Aves: Apodidae), two of which produce 'edible nests', occur sympatrically in Malaysia. Food boluses were analysed to investigate (1) diet differences and resource partitioning among the four species and (2) the influence of habitat on the diet of the Glossy Swiftlet Collocalia esculenta. Hymenoptera and diptera were the most abundant prey in all diets. Black-nest Swiftlets Aerodramus maximus were extremely selective; 89% of the items taken were hymenoptera, of which 97% were flying ants. White-nest Swiftlets A. fuciphagus and Mossy-nest Swiftlets A. salanganus took more than twice as many prey items as either of the other species. Their diets were taxonomically diverse and differed only in the average prey size taken, Mossy-nest Swiftlets taking the larger prey. Glossy Swiftlets took a higher proportion of coleoptera than did the other species. For the habitat comparisons, diptera were the main constituents of the Glossy Swiftlet diet in an urban habitat (71% of total prey items), whereas hymenoptera predominated in forest habitat (42% of total prey items). The results suggest that diet separation occurs among the four species of Malaysian swiftlet, in relation to both species and habitat. The implications of potentially habitat-driven dietary changes are discussed for the edible-nest swiftlets.     

Prestin Shows Divergent Evolution Between Constant Frequency Echolocating Bats

The gene Prestin encodes a motor protein that is thought to confer the high-frequency sensitivity and selectivity that characterizes the mammalian auditory system. Recent research shows that the Prestin gene has undergone a burst of positive selection on the ancestral branch of the Old World horseshoe and leaf-nosed bats (Rhinolophidae and Hipposideridae, respectively), and also on the branch leading to echolocating cetaceans. Moreover, these two groups share a large number of convergent amino acid sequence replacements. Horseshoe and leaf-nosed bats exhibit narrowband echolocation, in which the emitted calls are based on the second harmonic of a predominantly constant frequency (CF) component, the frequency of which is also over-represented in the cochlea. This highly specialized form of echolocation has also evolved independently in the neotropical Parnell's mustached bat (Pteronotus parnellii). To test whether the convergent evolution of CF echolocation between lineages has arisen from common changes in the Prestin gene, we sequenced the Prestin coding region (~2,212?bp, >99% coverage) in P. parnellii and several related species that use broadband echolocation calls. Our reconstructed Prestin gene tree and amino acid tree showed that P. parnellii did not group together with Old World horseshoe and leaf-nosed bats, but rather clustered within its true sister species. Comparisons of sequences confirmed that P. parnellii shared most amino acid changes with its congeners, and we found no evidence of positive selection in the branch leading to the genus of Pteronotus. Our result suggests that the adaptive changes seen in Prestin in horseshoe and leaf-nosed bats are not necessary for CF echolocation in P. parnellii.     

Problems in the harvest of edible birds' nests in Sarawak and Sabah, Malaysian Borneo

Due to the value of their nests, there is great pressure on the populations of black-nest swiftlets (Collocalia maximus) and white-nest swiftlets (Collocalia fuciphagus) in the Malaysian provinces of Sarawak and Sabah. The problems are particularly acute at Gunung Mulu National Park, in spite of a complete ban on collection there, and at Niah National Park, where every participant in a complex collection and trading system has an incentive to take more nests than permitted. More successful harvest systems function in Sabah's Gomantong and Madai Caves. Recommendations for improved management of the nest harvest include addressing corruption, ensuring that local people with traditional rights to collect nests do not lose income to illegal immigrant labor and to traders, improving research and education about the swiftlets' behavior and ecology, and moving value-added processing of the nests closer to the caves where they originate and to the people who collect them.     

Things that go bump in the night: evolutionary interactions between cephalopods and cetaceans in the tertiary

Echolocation has evolved independently in several vertebrate groups, and hypotheses about the origin of echolocation in these groups often invoke abiotic mechanisms driving morphological evolution. In bats, for example, the ecological setting associated with the origin of echolocation has been linked to global warming during the Palaeocene–Eocene; similarly, the origin of toothed whales (odontocetes) has been broadly correlated with the establishment of the circum-Antarctic current. These scenarios, and the adaptational hypotheses for the evolution of echolocation with which they are associated, neglect a consideration of possible biotic mechanisms. Here we propose that the origin of echolocation in odontocetes was initially an adaptation for nocturnal epipelagic feeding – primarily on diel migrating cephalopods. We test this hypothesis using data on the temporal, geographical, and water column distributions of odontocetes and cephalopods, and other global events from their respective tertiary histories. From this analysis, we suggest that echolocation in early odontocetes aided nocturnal feeding on cephalopods and other prey items, and that this early system was exapted for deep diving and hunting at depths below the photic zone where abundant cephalopod resources were available 24 h a day. This scenario extends to the evolution of other cephalopod feeding (teuthophagous) marine vertebrates such as pinnipeds and Mesozoic marine reptiles.     

The sensitivity of echolocation by the Grey Swiftlet Aerodramus spodiopygius

Experiments were carried out to study the sensitivity of the echolocation system of the Grey Swiftlet Aerodramus spodiopygius to cylindrical aluminium rods of 1, 5, 10 and 20 mm diameters. Using a method of statistical analysis which enabled the data to be compared with the chance performance of a randomly moving (non-echolocating) missile, the results indicated that this species could echolocate the 20 mm diameter rods but those of smaller diameter were hit in an approximately random fashion. It is concluded that the sensitivity threshold of A. spodiopygius lies between 10 and 20 mm diameter for cylindrical metal rods under the prevailing experimental conditions. Comparisons with other data on the sensitivity of this and other swiftlet species are discussed.     

短嘴金丝燕的回声定位机制与其归巢行为的探究

通过对短嘴金丝燕(Aerodramus brevirostris)的传统遮挡放飞实验和飞行行为的观察,研究短嘴金丝燕回声定位的机制,同时利用SONY MD录音并用Cool Edit2.1软件进行声波分析,证明了该物种是能利用回声定位的鸟类。该物种定位的声波主频较低,所以其定位能力没有蝙蝠(Chiroptera)那样精确,个体主要还是依靠视觉定位,只有在全黑的情况下才利用声音定位的。     

Identification of cryptic species of Miniopterus bats (Chiroptera: Miniopteridae) from Madagascar and the Comoros using bioacoustics overlaid on molecular genetic and morphological characters

The number of Miniopterus bat species on Madagascar and the nearby Comoros islands (Malagasy region) has risen from four to 11. These recently described cryptic taxa have been differentiated primarily based on molecular markers and associated a posteriori morphological characters that corroborate the different clades. Members of this Old World genus are notably conservative in morphology across their range. Several sites on Madagascar hold up to four small‐bodied taxa of this genus that are morphologically similar to one another, although they can be distinguished based on the tragus, an ear structure associated with echolocation. Miniopterus often emit species‐specific calls. In the present study, we analyze the bioacoustics of the 11 species of Miniopterus currently recognized from the Malagasy region, with an initial identification of the 87 recorded and collected individuals based on molecular markers and certain morphological characters. In most cases, bioacoustic parameters differentiate species and have taxonomic utility. Miniopterus griveaudi populations, which occur on three islands (Madagascar, Anjouan, and Grande Comore), showed no significant differences in peak echolocation frequencies. After running a discriminant function analysis based on five bioacoustic parameters, some mismatched assignments of Malagasy species were found, which include allopatric sister‐taxa and sympatric, phylogenetically not closely‐related species of similar body size. Because the peak echolocation frequencies of two species (Miniopterus sororculus and Miniopterus aelleni) were independent of body size, they were acoustically distinguishable from cryptic sympatric congeners. The small variation around the allometric relationship between body size and peak echolocation frequency of Malagasy Miniopterus species suggests that intraspecific communication rather than competition or prey detection may be the driver for the acoustic divergence of these two species. Our well‐defined echolocation data allow detailed ecological work to commence aiming to test predictions about the relative roles of competition, prey availability, and social communication on the evolution of echolocation in Malagasy Miniopterus species. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 104 , 284–302.     

Driving Factors for the Evolution of Species-Specific Echolocation Call Design in New World Free-Tailed Bats (Molossidae)

Phylogeny, ecology, and sensorial constraints are thought to be the most important factors influencing echolocation call design in bats. The Molossidae is a diverse bat family with a majority of species restricted to tropical and subtropical regions. Most molossids are specialized to forage for insects in open space, and thus share similar navigational challenges. We use an unprecedented dataset on the echolocation calls of 8 genera and 18 species of New World molossids to explore how habitat, phylogenetic relatedness, body mass, and prey perception contribute to echolocation call design. Our results confirm that, with the exception of the genus Molossops, echolocation calls of these bats show a typical design for open space foraging. Two lines of evidence point to echolocation call structure of molossids reflecting phylogenetic relatedness. First, such structure is significantly more similar within than among genera. Second, except for allometric scaling, such structure is nearly the same in congeneric species. Despite contrasting body masses, 12 of 18 species call within a relatively narrow frequency range of 20 to 35 kHz, a finding that we explain by using a modeling approach whose results suggest this frequency range to be an adaptation optimizing prey perception in open space. To conclude, we argue that the high variability in echolocation call design of molossids is an advanced evolutionary trait allowing the flexible adjustment of echolocation systems to various sensorial challenges, while conserving sender identity for social communication. Unraveling evolutionary drivers for echolocation call design in bats has so far been hampered by the lack of adequate model organisms sharing a phylogenetic origin and facing similar sensorial challenges. We thus believe that knowledge of the echolocation call diversity of New World molossid bats may prove to be landmark to understand the evolution and functionality of species-specific signal design in bats.     

The evolution of flight and echolocation in bats: another leap in the dark

The earliest known complete bats, from the Eocene (49–53 Mya), were already capable of flapping flight and echolocation. In the absence of direct fossil evidence there have been many speculative scenarios advanced to explain the evolution of these behaviours and their distributions in extant bats. Theories assuming chiropteran monophyly have generally presumed the ancestral pre‐bat was nocturnal, arboreal and insectivorous. Following this assumption hypotheses can be divided into the echolocation first, flight first and tandem development hypotheses, all of which assume that flight evolved only once in the lineage. In contrast, the chiropteran diphyly hypothesis suggests that flight evolved twice. Evidence supporting and refuting the different hypotheses are reviewed. It is concluded that there are significant problems attached to all the current models. A novel hypothesis is advanced, which starts from the assumption that bats are monophyletic and the ancestral pre‐bat was arboreal, but diurnal and frugivorous. After the evolution of flight it is suggested that these animals were driven into the nocturnal niche by the evolution of raptorial birds, and different groups evolved either specialised nocturnal vision (megachiropterans) or echolocation (microchiropterans). A block on sensory modality transfer has retained this distribution of perceptual capabilities ever since, despite some Megachiroptera evolving rudimentary echolocation, and the dietary convergence of some Microchiroptera with the Megachiroptera. The new hypothesis overcomes many of the problems identified in previous treatments.     

Genetic homogeneity among colonies of the white-nest swiftlet (Aerodramus fuciphagus) in Thailand

The white-nest swiftlet, Aerodramus fuciphagus, originally lived in large colonies in natural caves, but now it also occurs in man-made buildings. We investigated the patterns of genetic differentiation in two mitochondrial DNA genes (cyt-b and ND2) and eight microsatellite loci among and within colonies of A. fuciphagus from across recently established man-made colonies in Thailand. Ten white-nest swiftlet colonies were sampled along the coast of the Gulf of Thailand and the Andaman Sea in Thailand during 2003-2006. The genetic diversity of mtDNA was very low, and few significant PhiST values were found between pairs of colonies. Analyses of haplotype relationships did not show genetic structure across the sampled distribution. The level of genetic diversity for microsatellite loci was high, but FST values were not significant. However, due to small sample sizes for some colonies that could limit conclusions on genetic differentiation from PhiST and FST, we also analyzed the microsatellite data using STRUCTURE and found that number of subpopulations of white-nest swiftlets in sampled colonies was one. The lack of genetic differentiation among swiftlet house colonies could be a result of high gene flow between colonies and large population sizes. Our results suggest that A. fuciphagus living in recently established man-made colonies in Thailand should be considered members of a single panmictic population. Future work will be necessary to determine whether this panmixia is stable or a temporary result of the recent explosive expansion of the number of colonies, and comparisons to natural colonies may provide an understanding of mechanisms producing the lack of genetic structure in swiftlet house colonies.     

Phylogeny of the Saprininae reveals interesting ecological shifts in the history of the subfamily (Coleoptera: Histeridae)

A morphological data set for the histerid beetle subfamily Saprininae comprising 95 adult morphological characters scored (multistate coding) from 72 terminal taxa and four outgroups was developed in order to analyse and determine the relationships amongst the genera and subgenera of the Saprininae subfamily. Cladograms were rooted with exemplars of Dendrophilinae (genus Dendrophilus), Bacaniini (genus Bacanius), Abraeinae (genus Chaetabraeus), and Anapleini (genus Anapleus). Parsimony‐based phylogenetic analyses were performed based on the type species of each genus and subgenus of the Saprininae occurring around the world, with the exception of three taxa: Paramyrmetes foveipennis (type species of the genus Paramyrmetes), Satrapister nitens (type species of the genus Satrapister) and Xerosaprinus (Auchmosaprinus) laciniatus (type species of the subgenus Auchmosaprinus) that were not available. In addition, in order to test the monophyly of several questionable genera, multiple exemplars were added in a few cases. The analysis also included an exemplar of an apparently undescribed genus. The results of the analysis confirm the monophyly of the subfamily supported by two unique synapomorphies: (1) presence of sensory structures of the antenna; and (2) presence of the antennal cavity, as well as several other weaker synapomorphies. However, the phylogeny inferred here shows mostly low support for the deeper branches and consequently no major changes in the Saprininae classification are proposed. The presented cladogram is discussed together with its implications for the evolution of the subfamily. The most informative characters and their respective states are outlined. Multiple shifts in lifestyles have evolved during the evolutionary history of the group. Taxa found near the root of the cladogram are mostly nidicolous or myrmecophilous, and inquiliny is presumed to be the plesiomorphic lifestyle of the subfamily. The nidicolous lifestyle has undergone several transformations to other lifestyles and myrmecophily has evolved three times independently during the evolution of the subfamily. Termitoxeny has evolved two times independently in the group whereas ecological adaptation for life in caves has likewise evolved two times independently. The analyses yielded a large clade of predominantly psammophilous taxa; psammophily is thought to have evolved once and has been subsequently lost several times. © 2014 The Linnean Society of London     

The voltage-gated potassium channel subfamily KQT member 4 (KCNQ4) displays parallel evolution in echolocating bats

Bats are the only mammals that use highly developed laryngeal echolocation, a sensory mechanism based on the ability to emit laryngeal sounds and interpret the returning echoes to identify objects. Although this capability allows bats to orientate and hunt in complete darkness, endowing them with great survival advantages, the genetic bases underlying the evolution of bat echolocation are still largely unknown. Echolocation requires high-frequency hearing that in mammals is largely dependent on somatic electromotility of outer hair cells. Then, understanding the molecular evolution of outer hair cell genes might help to unravel the evolutionary history of echolocation. In this work, we analyzed the molecular evolution of two key outer hair cell genes: the voltage-gated potassium channel gene KCNQ4 and CHRNA10, the gene encoding the α10 nicotinic acetylcholine receptor subunit. We reconstructed the phylogeny of bats based on KCNQ4 and CHRNA10 protein and nucleotide sequences. A phylogenetic tree built using KCNQ4 amino acid sequences showed that two paraphyletic clades of laryngeal echolocating bats grouped together, with eight shared substitutions among particular lineages. In addition, our analyses indicated that two of these parallel substitutions, M388I and P406S, were probably fixed under positive selection and could have had a strong functional impact on KCNQ4. Moreover, our results indicated that KCNQ4 evolved under positive selection in the ancestral lineage leading to mammals, suggesting that this gene might have been important for the evolution of mammalian hearing. On the other hand, we found that CHRNA10, a gene that evolved adaptively in the mammalian lineage, was under strong purifying selection in bats. Thus, the CHRNA10 amino acid tree did not show echolocating bat monophyly and reproduced the bat species tree. These results suggest that only a subset of hearing genes could underlie the evolution of echolocation. The present work continues to delineate the genetic bases of echolocation and ultrasonic hearing in bats.     

The spermatozoon of a 'living fossil': Tubiluchus troglodytes (Priapulida)

The spermatozoon of Tubiluchus troglodytes, the first priapulid formally described from the Mediterranean Sea has a head composed of an acrosome and a nucleus. The acrosome is divided in two branches coiled around the nucleus. The nucleus is basally columnar, but apically generates two rods helically coiled one around the other. The midpiece is formed by an axoneme with 27 accessory microtubules, surrounded by three mitochondria. An annulus separates the midpiece from the tail that contains a 9 + 2 axoneme surrounded by nine accessory microtubules. The spermatozoon of T. troglodytes is similar to that of the other two species known from the genus, and completely different from the 'primitive' one of the other priapulids. Since Tubiluchus is considered the most basal of the extant priapulids, and the only genus with an internal fertilization, it may be that in priapulids the external fertilization is a derived character.     

Phylogenetic analyses in cornus substantiate ancestry of xylem supercooling freezing behavior and reveal lineage of desiccation related proteins

The response of woody plant tissues to freezing temperature has evolved into two distinct behaviors: an avoidance strategy, in which intracellular water supercools, and a freeze-tolerance strategy, where cells tolerate the loss of water to extracellular ice. Although both strategies involve extracellular ice formation, supercooling cells are thought to resist freeze-induced dehydration. Dehydrin proteins, which accumulate during cold acclimation in numerous herbaceous and woody plants, have been speculated to provide, among other things, protection from desiccative extracellular ice formation. Here we use Cornus as a model system to provide the first phylogenetic characterization of xylem freezing behavior and dehydrin-like proteins. Our data suggest that both freezing behavior and the accumulation of dehydrin-like proteins in Cornus are lineage related; supercooling and nonaccumulation of dehydrin-like proteins are ancestral within the genus. The nonsupercooling strategy evolved within the blue- or white-fruited subgroup where representative species exhibit high levels of freeze tolerance. Within the blue- or white-fruited lineage, a single origin of dehydrin-like proteins was documented and displayed a trend for size increase in molecular mass. Phylogenetic analyses revealed that an early divergent group of red-fruited supercooling dogwoods lack a similar protein. Dehydrin-like proteins were limited to neither nonsupercooling species nor to those that possess extreme freeze tolerance.