Mitogenomes have been widely used in phylogenetic and taxonomic research,due to their maternal inheritance,accelerated evolutionary rates,compact circular structure,and the absence of introns.Although their utility is...Mitogenomes have been widely used in phylogenetic and taxonomic research,due to their maternal inheritance,accelerated evolutionary rates,compact circular structure,and the absence of introns.Although their utility is well-established,efficient mitogenome sequencing still poses a challenge for evolutionary studies.Here,we presented a novel hybridization capture protocol that enables high-fidelity sequencing of the complete mitogenome(16535 bp)of Odorrana nasuta.This genome exhibits typical anuran characteristics,including an A+T nucleotide bias(56.37%)and conserved gene composition.Notably,we identified a lineage-specific trnH transposition event that reorganizes the typical gene order into a unique trnH-trnL1-trnT-trnP-trnF cluster,which may be a potential synapomorphic trait for the genus Odorrana.Phylogenetic reconstruction using 13 protein-coding genes from 59 anuran species strongly supported the monophyly of Ranidae and its constituent genera(Odorrana,Rana,Lithobates,Amolops,Pelophylax,Glandirana),yet reveals the incongruence in the placement of Hylarana.The conflicting topologies observed for Hylarana and Glandirana across different studies highlight critical methodological limitations,including taxonomic sampling biases and data type dependency,which collectively compromise phylogenetic resolution.By integrating mitogenomic structure signatures with sequence-based phylogenies,this work advances our understanding of the systematics within Odorrana and highlights the need of multi-locus genomic datasets for resolving deep phylogenetic conflicts within Ranidae.展开更多
Taxonomic studies of the Odorrana versabilis group have been consistently disorganized,and they have often been incorrectly associated with an abandoned subgenus or genus Bamburana in existing literature.In this study...Taxonomic studies of the Odorrana versabilis group have been consistently disorganized,and they have often been incorrectly associated with an abandoned subgenus or genus Bamburana in existing literature.In this study,we conducted an integrative taxonomic analysis using molecular phylogenetic analysis of two mitochondrial gene fragments(16S rRNA and CO1)and morphological examination of 41 specimens collected during herpetological surveys in southern China.Our results confirmed the monophyly of O.versabilis group which exhibited distinct morphological traits.We also identified a cryptic lineage sister with O.exiliversabilis.Additionally,our study revealed that O.trankieni was nested within O.nasuta.Consequently,we redefined the morphological characteristics of the O.versabilis group,described a new species in this group,O.confusa sp.nov.,and proposed that O.trankieni be considered a junior synonym of O.nasuta.One of the most significant findings of this study was the observation of distinct morphological changes in adult males across all species within the O.versabilis group between the non-breeding and breeding seasons.This study contributes to research on various aspects of Asian amphibian fauna,including taxonomy,ecology,evolution,and physiology.展开更多
The Odorrana schmackeri sensu la to is a group of cascade frogs that are widely distributed in southern China.However,their taxonomy has long been unresolved and several cryptic species have been reported in the past ...The Odorrana schmackeri sensu la to is a group of cascade frogs that are widely distributed in southern China.However,their taxonomy has long been unresolved and several cryptic species have been reported in the past decade.Previous analyses revealed the existence of three sympatrie species(i.c,O.schmackeri,O.hejiangensis and O.sp2) at the type locality of O.schmackeri(Gaojiayan Town,Yichang City,Hubei Province,China).However,the lack of an original description for the holotype specimen(SMF6241) of O.schmackeri poses a confusing and controversial problem in species nomination within this group.Here,we reevaluate the taxonomic affinities of SMF6241 and examine the distinctness of O.sp2 based on specimens from the type locality.We use integrative approaches that combine morphology with 10 microsatellite loci and bioacoustics.Canonical discriminant analysis of morphological characteristics revealed three distinct groupings(i.e.,O.schmackeri,O.hejiangensis and O.sp2) and attributed SMF6241 to O.schmackeri.Genetic clustering analysis of nuclear DNA also identified three corresponding clusters.In addition,analysis of interspecific variation in morphometrics and acoustics showed divergence between O.sp2 and its sympatric congeners.All lines of evidence from morphological,molecular and bioacoustic approaches suggest the species status of O.sp2,namely Odorrana ichangensis Chen,sp.nov.Overall,this study resolves the taxonomic assignment of the holotype of O.schmackeri and provides an example for species delimitation using integrative approaches.展开更多
We describe Odorrana liboensis sp.nov.,a new species from the Maolan National Nature Reserve,Libo County,Guizhou Province,China.Phylogenetic analyses based on DNA sequences of the mitochondrial 12 S rRNA,16 S rRNA,and...We describe Odorrana liboensis sp.nov.,a new species from the Maolan National Nature Reserve,Libo County,Guizhou Province,China.Phylogenetic analyses based on DNA sequences of the mitochondrial 12 S rRNA,16 S rRNA,and ND2 genes supported the new species as an independent lineage,closely related to O.lipuensis.The uncorrected genetic distances between the 12 S rRNA and16 S rRNA in the new species and its closest congener,O.lipuensis,were 6.06%and 5.19%,respectively.The new species is distinguished from its congeners by a combination of the following morphological characters:(1)having medium body size,with the snout-vent length(SVL)of adult females approximately 1.2 times as long as males at 56.9±1.0(55.8-58.2 mm,n=9)in females and 48.7±1.2(47.1-49.9 mm,n=5)in males;(2)head length greater than width in males and females;(3)tympanum distinctly visible,greater than one-half the diameter of the eye;(4)eyes big and prominent,width of upper eyelid(UEW)approximately3/4 of eye diameter(ED);(5)dorsolateral folds absent;(6)two metacarpal tubercles;(7)relative finger lengths:Ⅱ<Ⅰ<Ⅳ<Ⅲ;(8)subarticular tubercles on fingers prominent:1,1,2,2;(9)one metatarsal tubercle;(10)tibiotarsal articulation reaching to between the eye and nostril when the leg is stretched forward;(11)toes with entire webbing to disks;(12)subarticular tubercles on toes prominent:1,1,2,3,2;(13)dorsal surfaces of limbs with distinct brownish-black bands;(14)smooth,grass-green dorsum with irregular brown mottling;(15)venter smooth,lacking black spots;and(16)lacking pectoral spinules,lacking vocal sacs,and light white nuptial pad present on finger I in males.The new species is currently only known from the type locality.展开更多
Antimicrobial peptides(AMPs) are a group of gene-encoded small peptides that play pivotal roles in the host immune system of multicellular organisms.Cathelicidins are an important family of AMPs that exclusively exist...Antimicrobial peptides(AMPs) are a group of gene-encoded small peptides that play pivotal roles in the host immune system of multicellular organisms.Cathelicidins are an important family of AMPs that exclusively exist in vertebrates. Many cathelicidins have been identified from mammals, birds, reptiles and fish. To date, however, cathelicidins from amphibians are poorly understood. In the present study, two novel cathelicidins(OL-CATH1 and 2) were identified and studied from the odorous frog Odorrana livida.Firstly, the cDNAs encoding the OL-CATHs(780 and735 bp in length, respectively) were successfully cloned from a lung cDNA library constructed for the frog. Multi-sequence alignment was carried out to analyze differences between the precursors of the OL-CATHs and other representative cathelicidins.Mature peptide sequences of OL-CATH1 and 2 were predicted(33 amino acid residues) and their secondary structures were determined(OL-CATH1 showed a random-coil conformation and OL-CATH2 demonstrated α-helical conformation). Furthermore,OL-CATH1 and 2 were chemically synthesized and their in vitro functions were determined. Antimicrobial and bacterial killing kinetic analyses indicated that OL-CATH2 demonstrated relatively moderate and rapid antimicrobial potency and exhibited strong anti-inflammatory activity. At very low concentrations(10 μg/mL), OL-CATH2 significantly inhibited the lipopolysaccharide(LPS)-induced transcription and production of pro-inflammatory cytokines TNF-α, IL-1βand IL-6 in mouse peritoneal macrophages. In contrast, OL-CATH1 did not exhibit any detectableantimicrobial or anti-inflammatory activities. Overall,identification of these OL-CATHs from O. livida enriches our understanding of the functions of cathelicidins in the amphibian immune system. The potent antimicrobial and anti-inflammatory activities of OL-CATH2 highlight its potential as a novel candidate in anti-infective drug development.展开更多
A new species of the genusOdorrana is described from a completely dark karst cave of northeastern Guangxi, southern China. The new species,Odorranalipuensissp. nov., can be distinguished from its congeners by a combin...A new species of the genusOdorrana is described from a completely dark karst cave of northeastern Guangxi, southern China. The new species,Odorranalipuensissp. nov., can be distinguished from its congeners by a combination of the following characters： medium size （SVL： 40.7–47.7 mm in males, 51.1–55.4 mm in females）;tips of all but ifrst ifnger expanded with circummarginal grooves; smooth, grass-green dorsum with irregular brown mottling; pineal body invisible; throat to upper abdomen with gray mottling; dorsal surfaces of limbs with brown bands; dorsolateral fold absent; tiny spinules on lateral body, temporal region, and anterior and posterior edge of tympanum;white nuptial pad present on ifnger I; males lacking vocal sacs; females having creamy yellow eggs, without black poles. Uncorrected sequence divergences betweenO.lipuensissp. nov. and all homologous 16S rRNA sequences ofOdorrana available on GenBank is equal to or greater than 4.9%. Currently, the new species is only known from the type locality.展开更多
We isolated and characterized 17 microsatellite DNA loci for the odorous frog Odorrana margaretae from its transcriptome sequence data. These loci were screened with 24 individuals from Mt. Emei. All loci were polymor...We isolated and characterized 17 microsatellite DNA loci for the odorous frog Odorrana margaretae from its transcriptome sequence data. These loci were screened with 24 individuals from Mt. Emei. All loci were polymorphic, with the number of alleles ranging from 2 to 8. The observed and expected heterozygosity, polymorphism information content, ranged from 0.04 to 1, 0.04 to 0.81, and 0.040 to 0.763, respectively. All loci were in linkage equilibrium and six loci were significantly deviated from Hardy-Weinberg equilibrium after sequential Bonferroni corrections. Cross- species amplification test was conducted for ten odorous frog species, and 12 loci were amplifiable in most species. With the high cross-species amplification rates, thesc markers will provide useful molecular tools for conservation genetic and phylogeographic studies on the genus Odorrana and Bamburana.展开更多
Male-biased sexual dimorphism in hind limb muscles is widespread in anuran species where scramble competition is common among males. Such sexual difference is thought to result from sexual selection. In this view, we ...Male-biased sexual dimorphism in hind limb muscles is widespread in anuran species where scramble competition is common among males. Such sexual difference is thought to result from sexual selection. In this view, we tested the differences in muscle mass between the sexes and between amplectant and non-amplectant males by quantifying the mass of four hindlimb muscles （triceps femoris, sartorius, gracilis and plantaris longus） of females and males of Odorrana schmackeri. The results showed that females significantly exceeded males for muscle triceps femoris, gracilis, plantaris longus and total mass when controlled for body size. There are no significant differences between amplectant and non-amplectant males. It is probable that the maintenance of the amplectant position in O. schmackeri may depend on the strength of hindlimb muscles in females to support the pair.展开更多
Odorrana margaretae （Anura： Ranidae） is widely distributed in the southern provinces of China. Previously, 72 antimicrobial peptides （AMPs） belonging to 21 families were identified from the skin of O. margaretae,...Odorrana margaretae （Anura： Ranidae） is widely distributed in the southern provinces of China. Previously, 72 antimicrobial peptides （AMPs） belonging to 21 families were identified from the skin of O. margaretae, which were captured in the Hunan province. In the present study, five O. margaretae frogs were captured from the Guizhou province and a total of 28 cDNAs encoding 17 host defense peptides （HDPs） belonging to 14 families were cloned from the skin cDNA library of O. margaretae. Among the 17 HDPs, only one （brevinin-1-Omar5） had been characterized. The distinct HDP expression profiles for O. margaretae in the previous and present study may be attributed to the environmental differences between the sampling locations and the genetic divergence among O. margaretae populations. Besides, 11 of the 17 HDPs identified in the present study were novel for ranids. In order to understand their roles in host defense reactions, three HDPs （odorranain-H-OM1, odorranain-M-OM and ranatuerin-2-OM）, which possess low sequence similarity with the known amphibian HDPs, were selected for further chemical synthesis and functional analysis. Odorranain-H-OM1 showed direct antimicrobial activity against bacteria and fungi. Odorranain-M-OM exhibited concentration-dependent anti-oxidant activity. Ranatuerin-2-OM showed lectin-like activity and could strongly hemagglu -tinate human intact erythrocytes with or without the presence of Ca2＋. The diverse activities of HDPs implied that they may play different roles in host defense reactions of O. margaretae.展开更多
Acoustic communication is essential for anuran survival and reproduction, and masking background noise can affect the effective acoustic communication. The larger odorous frog(Odorrana graminea) inhabits noise montane...Acoustic communication is essential for anuran survival and reproduction, and masking background noise can affect the effective acoustic communication. The larger odorous frog(Odorrana graminea) inhabits noise montane streams, and it has shown an ultrasound communication adaptation. However, the molecular mechanism underlying their ultrasonic hearing adaptation remains unknown. To characterize and investigate the molecular characteristics and evolution of the high-frequency hearing-sensitive gene(KCNQ4) in O. graminea, termed as OgKCNQ4, the rapid amplification of cDNA ends(RACE) was performed to amplify the cDNA of OgKCNQ4. Different bioinformatics analyses were used to investigate the molecular characteristics. Multiple nucleotide and amino acid sequence alignment were conducted, and phylogenies were reconstructed under the maximum likelihood and Bayesian approaches. The full-length cDNA of OgKCNQ4 was 2065 bp, and the open reading frame(ORF) was 2046 bp encoding for a putative protein with 681 amino acids. The relative molecular weight of OgKCNQ4 was 76.453 kD and the putative PI was 9.69. Secondary structure prediction analyses suggested 42.29% alpha helixes and 43.76% random coils in OgKCNQ4. Gene homology and Phylogenetic analyses revealed the closest relationship between OgKCNQ4 and KCNQ4 of Nanorana parkeri with 96.9% similarity and 95.0% identity. We first determined the full-length cDNA of OgKCNQ4 and the results here could provide foundations for further study on the evolution of KCNQ4 and its relationship to ultrasonic communication in amphibians.展开更多
Nonlinear phenomena are commonly shown in the vocalization of animals and exerts different adaptive functions.Although some studies have pointed out that nonlinear phenomena can enhance the individual identification o...Nonlinear phenomena are commonly shown in the vocalization of animals and exerts different adaptive functions.Although some studies have pointed out that nonlinear phenomena can enhance the individual identification of male Odorrana tormota,whether the nonlinear phenomena play a specific role in the sexual selection of O.tormota remain unclear.Here we presented evidence that there was a significant negative correlation(Pearson:n=30,r=0.65,P<0.001)between the nonlinear phenomena content and snout-vent length in the male O.tormota,and two-choice amplexus experiments showed that female O.tormota preferred male with smaller body size containing higher nonlinear phenomena content in its calls.Phonotaxis experiments also revealed that females preferred calls with higher nonlinear phenomena content.Additionally,compared to the calls with lower nonlinear phenomena content and higher fundamental frequency,there was shorter response time in phonotactic behaviour of female induced by the calls with higher nonlinear phenomena content and lower fundamental frequency.We argue that the nonlinear phenomena content in the calls of male O.tormota can convey its body size information and may provide important clues for female frogs in darkened surroundings to identify males’body size during mate choice,meanwhile,higher nonlinear phenomena content in males’calls may increase the attractiveness of males to females.The results of this study provide confirmation that,for O.tormota,nonlinear phenomena have specific function in mate choice.展开更多
The green odorous frog(Odorrana margaretae)has an interesting ring-shaped divergence pattern around the Sichuan Basin and documenting its morphological variations is essential in understanding its evolutionary history...The green odorous frog(Odorrana margaretae)has an interesting ring-shaped divergence pattern around the Sichuan Basin and documenting its morphological variations is essential in understanding its evolutionary history.Using curvilinear models,we detected significant geographical clinal variations in morphological traits,particularly sizes,of female O.margaretae.Males had significantly smaller sizes than females,and also had smaller variation ranges than females.One major trend of morphological variations was clinal:populations from the west tended to have a larger size with wider head and longer posterior limbs than populations from the east.Species history,with an early extended isolation and two subsequent secondary contacts,may explain most of the geographical clinal variations of O.margaretae.Bioclimatic factors may also contribute in explaining the variance of morphology.展开更多
Body surface area（BSA）was regarded as a more readily quantifiable parameter relative to body mass in the normalization of comparative biochemistry and physiology.The BSA prediction has attracted unceasing research b...Body surface area（BSA）was regarded as a more readily quantifiable parameter relative to body mass in the normalization of comparative biochemistry and physiology.The BSA prediction has attracted unceasing research back more than a century on animals,especially on humans and rats.Few studies in this area for anurans were reported,and the equation for body surface area（S）and body mass（W）：S=9.9 W 0.56,which was concluded from toads of four species in 1969,was generally adopted to estimate the body surface areas for anurans until recent years.However,this equation was not applicable to Odorrana grahami.The relationship between body surface area and body mass for this species was established as：S=15.4 W 0.579.Our current results suggest estimation equations should be used cautiously across different species and body surface area predictions on more species need to be conducted.展开更多
基金the National Natural Science Foundation of China(NSFC)to Zhuo CHEN(Grant Nos.32270440 and 31601848)and Xiaohong CHEN(Grant No.U21A20192).
文摘Mitogenomes have been widely used in phylogenetic and taxonomic research,due to their maternal inheritance,accelerated evolutionary rates,compact circular structure,and the absence of introns.Although their utility is well-established,efficient mitogenome sequencing still poses a challenge for evolutionary studies.Here,we presented a novel hybridization capture protocol that enables high-fidelity sequencing of the complete mitogenome(16535 bp)of Odorrana nasuta.This genome exhibits typical anuran characteristics,including an A+T nucleotide bias(56.37%)and conserved gene composition.Notably,we identified a lineage-specific trnH transposition event that reorganizes the typical gene order into a unique trnH-trnL1-trnT-trnP-trnF cluster,which may be a potential synapomorphic trait for the genus Odorrana.Phylogenetic reconstruction using 13 protein-coding genes from 59 anuran species strongly supported the monophyly of Ranidae and its constituent genera(Odorrana,Rana,Lithobates,Amolops,Pelophylax,Glandirana),yet reveals the incongruence in the placement of Hylarana.The conflicting topologies observed for Hylarana and Glandirana across different studies highlight critical methodological limitations,including taxonomic sampling biases and data type dependency,which collectively compromise phylogenetic resolution.By integrating mitogenomic structure signatures with sequence-based phylogenies,this work advances our understanding of the systematics within Odorrana and highlights the need of multi-locus genomic datasets for resolving deep phylogenetic conflicts within Ranidae.
基金This work was supported by DFGP Project of Fauna of Guangdong-202115Nanling National Park Construction Project(No.ZD22-ZC016AG)2022 Wildlife Monitoring Projects of the Guangdong Forestry Bureau,and the National Animal Collection Resource Center,China.
文摘Taxonomic studies of the Odorrana versabilis group have been consistently disorganized,and they have often been incorrectly associated with an abandoned subgenus or genus Bamburana in existing literature.In this study,we conducted an integrative taxonomic analysis using molecular phylogenetic analysis of two mitochondrial gene fragments(16S rRNA and CO1)and morphological examination of 41 specimens collected during herpetological surveys in southern China.Our results confirmed the monophyly of O.versabilis group which exhibited distinct morphological traits.We also identified a cryptic lineage sister with O.exiliversabilis.Additionally,our study revealed that O.trankieni was nested within O.nasuta.Consequently,we redefined the morphological characteristics of the O.versabilis group,described a new species in this group,O.confusa sp.nov.,and proposed that O.trankieni be considered a junior synonym of O.nasuta.One of the most significant findings of this study was the observation of distinct morphological changes in adult males across all species within the O.versabilis group between the non-breeding and breeding seasons.This study contributes to research on various aspects of Asian amphibian fauna,including taxonomy,ecology,evolution,and physiology.
基金supported by the National Natural Sciences Foundation of China (NSFC-31872220, 31572245, 31372164)。
文摘The Odorrana schmackeri sensu la to is a group of cascade frogs that are widely distributed in southern China.However,their taxonomy has long been unresolved and several cryptic species have been reported in the past decade.Previous analyses revealed the existence of three sympatrie species(i.c,O.schmackeri,O.hejiangensis and O.sp2) at the type locality of O.schmackeri(Gaojiayan Town,Yichang City,Hubei Province,China).However,the lack of an original description for the holotype specimen(SMF6241) of O.schmackeri poses a confusing and controversial problem in species nomination within this group.Here,we reevaluate the taxonomic affinities of SMF6241 and examine the distinctness of O.sp2 based on specimens from the type locality.We use integrative approaches that combine morphology with 10 microsatellite loci and bioacoustics.Canonical discriminant analysis of morphological characteristics revealed three distinct groupings(i.e.,O.schmackeri,O.hejiangensis and O.sp2) and attributed SMF6241 to O.schmackeri.Genetic clustering analysis of nuclear DNA also identified three corresponding clusters.In addition,analysis of interspecific variation in morphometrics and acoustics showed divergence between O.sp2 and its sympatric congeners.All lines of evidence from morphological,molecular and bioacoustic approaches suggest the species status of O.sp2,namely Odorrana ichangensis Chen,sp.nov.Overall,this study resolves the taxonomic assignment of the holotype of O.schmackeri and provides an example for species delimitation using integrative approaches.
基金supported by programs of the Biodiversity Survey and Assessment Project of the Ministry of Ecology and Environment,China(2019HJ2096001006)the Strategic Priority Research Program B of the Chinese Academy of Sciences(CAS)(No.XDB31000000)+2 种基金the National Natural Science Foundation of China(NSFC 31460091)the National Animal Collection Resource Center,China(No.2005DKA21402)the Application of Amphibian Natural Antioxidant Peptides as Cosmetic Raw Material Antioxidants(No.QKZYD[2020]4002)。
文摘We describe Odorrana liboensis sp.nov.,a new species from the Maolan National Nature Reserve,Libo County,Guizhou Province,China.Phylogenetic analyses based on DNA sequences of the mitochondrial 12 S rRNA,16 S rRNA,and ND2 genes supported the new species as an independent lineage,closely related to O.lipuensis.The uncorrected genetic distances between the 12 S rRNA and16 S rRNA in the new species and its closest congener,O.lipuensis,were 6.06%and 5.19%,respectively.The new species is distinguished from its congeners by a combination of the following morphological characters:(1)having medium body size,with the snout-vent length(SVL)of adult females approximately 1.2 times as long as males at 56.9±1.0(55.8-58.2 mm,n=9)in females and 48.7±1.2(47.1-49.9 mm,n=5)in males;(2)head length greater than width in males and females;(3)tympanum distinctly visible,greater than one-half the diameter of the eye;(4)eyes big and prominent,width of upper eyelid(UEW)approximately3/4 of eye diameter(ED);(5)dorsolateral folds absent;(6)two metacarpal tubercles;(7)relative finger lengths:Ⅱ<Ⅰ<Ⅳ<Ⅲ;(8)subarticular tubercles on fingers prominent:1,1,2,2;(9)one metatarsal tubercle;(10)tibiotarsal articulation reaching to between the eye and nostril when the leg is stretched forward;(11)toes with entire webbing to disks;(12)subarticular tubercles on toes prominent:1,1,2,3,2;(13)dorsal surfaces of limbs with distinct brownish-black bands;(14)smooth,grass-green dorsum with irregular brown mottling;(15)venter smooth,lacking black spots;and(16)lacking pectoral spinules,lacking vocal sacs,and light white nuptial pad present on finger I in males.The new species is currently only known from the type locality.
基金supported by grants from the Jiangsu Students' Innovation and Entrepreneurship Training Program(2017suda098)the National Natural Science Foundation of China(31772455)+2 种基金Natural Science Foundation of Jiangsu Province(BK20160336 and BK20171214)Natural Science Foundation of College in Jiangsu Province(16KJB350004)Suzhou Science and Technology Development Project(SYN201504 and SNG2017045)
文摘Antimicrobial peptides(AMPs) are a group of gene-encoded small peptides that play pivotal roles in the host immune system of multicellular organisms.Cathelicidins are an important family of AMPs that exclusively exist in vertebrates. Many cathelicidins have been identified from mammals, birds, reptiles and fish. To date, however, cathelicidins from amphibians are poorly understood. In the present study, two novel cathelicidins(OL-CATH1 and 2) were identified and studied from the odorous frog Odorrana livida.Firstly, the cDNAs encoding the OL-CATHs(780 and735 bp in length, respectively) were successfully cloned from a lung cDNA library constructed for the frog. Multi-sequence alignment was carried out to analyze differences between the precursors of the OL-CATHs and other representative cathelicidins.Mature peptide sequences of OL-CATH1 and 2 were predicted(33 amino acid residues) and their secondary structures were determined(OL-CATH1 showed a random-coil conformation and OL-CATH2 demonstrated α-helical conformation). Furthermore,OL-CATH1 and 2 were chemically synthesized and their in vitro functions were determined. Antimicrobial and bacterial killing kinetic analyses indicated that OL-CATH2 demonstrated relatively moderate and rapid antimicrobial potency and exhibited strong anti-inflammatory activity. At very low concentrations(10 μg/mL), OL-CATH2 significantly inhibited the lipopolysaccharide(LPS)-induced transcription and production of pro-inflammatory cytokines TNF-α, IL-1βand IL-6 in mouse peritoneal macrophages. In contrast, OL-CATH1 did not exhibit any detectableantimicrobial or anti-inflammatory activities. Overall,identification of these OL-CATHs from O. livida enriches our understanding of the functions of cathelicidins in the amphibian immune system. The potent antimicrobial and anti-inflammatory activities of OL-CATH2 highlight its potential as a novel candidate in anti-infective drug development.
基金supported by the Natural Science Foundation of Guangxi Province, China (Grant Nos: 2012GXNSFAA053057, 0832238)the ‘Xibuzhiguang’ Project of Chinese Academy of Sciences
文摘A new species of the genusOdorrana is described from a completely dark karst cave of northeastern Guangxi, southern China. The new species,Odorranalipuensissp. nov., can be distinguished from its congeners by a combination of the following characters： medium size （SVL： 40.7–47.7 mm in males, 51.1–55.4 mm in females）;tips of all but ifrst ifnger expanded with circummarginal grooves; smooth, grass-green dorsum with irregular brown mottling; pineal body invisible; throat to upper abdomen with gray mottling; dorsal surfaces of limbs with brown bands; dorsolateral fold absent; tiny spinules on lateral body, temporal region, and anterior and posterior edge of tympanum;white nuptial pad present on ifnger I; males lacking vocal sacs; females having creamy yellow eggs, without black poles. Uncorrected sequence divergences betweenO.lipuensissp. nov. and all homologous 16S rRNA sequences ofOdorrana available on GenBank is equal to or greater than 4.9%. Currently, the new species is only known from the type locality.
基金supported by the National Natural Science Foundation of China(Grant No.31172061)
文摘We isolated and characterized 17 microsatellite DNA loci for the odorous frog Odorrana margaretae from its transcriptome sequence data. These loci were screened with 24 individuals from Mt. Emei. All loci were polymorphic, with the number of alleles ranging from 2 to 8. The observed and expected heterozygosity, polymorphism information content, ranged from 0.04 to 1, 0.04 to 0.81, and 0.040 to 0.763, respectively. All loci were in linkage equilibrium and six loci were significantly deviated from Hardy-Weinberg equilibrium after sequential Bonferroni corrections. Cross- species amplification test was conducted for ten odorous frog species, and 12 loci were amplifiable in most species. With the high cross-species amplification rates, thesc markers will provide useful molecular tools for conservation genetic and phylogeographic studies on the genus Odorrana and Bamburana.
基金supported by National Sciences Foundation of China (No. 31372164, 30870277)the Scientific Research Foundation of Henan Normal University (No. 01046500145)+1 种基金Joint Funds for Fostering Talents of NSFC the People’s Government of Henan Province (Grant No. U1304309)
文摘Male-biased sexual dimorphism in hind limb muscles is widespread in anuran species where scramble competition is common among males. Such sexual difference is thought to result from sexual selection. In this view, we tested the differences in muscle mass between the sexes and between amplectant and non-amplectant males by quantifying the mass of four hindlimb muscles （triceps femoris, sartorius, gracilis and plantaris longus） of females and males of Odorrana schmackeri. The results showed that females significantly exceeded males for muscle triceps femoris, gracilis, plantaris longus and total mass when controlled for body size. There are no significant differences between amplectant and non-amplectant males. It is probable that the maintenance of the amplectant position in O. schmackeri may depend on the strength of hindlimb muscles in females to support the pair.
基金supported by the grants from Guiyang Science and Technology Plan Projects (2010-01-Z-24) to Jiang ZHOU
文摘Odorrana margaretae （Anura： Ranidae） is widely distributed in the southern provinces of China. Previously, 72 antimicrobial peptides （AMPs） belonging to 21 families were identified from the skin of O. margaretae, which were captured in the Hunan province. In the present study, five O. margaretae frogs were captured from the Guizhou province and a total of 28 cDNAs encoding 17 host defense peptides （HDPs） belonging to 14 families were cloned from the skin cDNA library of O. margaretae. Among the 17 HDPs, only one （brevinin-1-Omar5） had been characterized. The distinct HDP expression profiles for O. margaretae in the previous and present study may be attributed to the environmental differences between the sampling locations and the genetic divergence among O. margaretae populations. Besides, 11 of the 17 HDPs identified in the present study were novel for ranids. In order to understand their roles in host defense reactions, three HDPs （odorranain-H-OM1, odorranain-M-OM and ranatuerin-2-OM）, which possess low sequence similarity with the known amphibian HDPs, were selected for further chemical synthesis and functional analysis. Odorranain-H-OM1 showed direct antimicrobial activity against bacteria and fungi. Odorranain-M-OM exhibited concentration-dependent anti-oxidant activity. Ranatuerin-2-OM showed lectin-like activity and could strongly hemagglu -tinate human intact erythrocytes with or without the presence of Ca2＋. The diverse activities of HDPs implied that they may play different roles in host defense reactions of O. margaretae.
基金supported by the National Natural Science Foundation of China to ZC (Grants U1404306 and 31601848), XHC (Grant 31572245, 31372164 and 31872220)the Project funded by China Postdoctoral Science Foundation to ZC (2016M600580)+1 种基金the Excellent Young Scholars Fund of HNNU to ZC (YQ201706)the Young Backbone Teachers Fund of HNNU to ZC
文摘Acoustic communication is essential for anuran survival and reproduction, and masking background noise can affect the effective acoustic communication. The larger odorous frog(Odorrana graminea) inhabits noise montane streams, and it has shown an ultrasound communication adaptation. However, the molecular mechanism underlying their ultrasonic hearing adaptation remains unknown. To characterize and investigate the molecular characteristics and evolution of the high-frequency hearing-sensitive gene(KCNQ4) in O. graminea, termed as OgKCNQ4, the rapid amplification of cDNA ends(RACE) was performed to amplify the cDNA of OgKCNQ4. Different bioinformatics analyses were used to investigate the molecular characteristics. Multiple nucleotide and amino acid sequence alignment were conducted, and phylogenies were reconstructed under the maximum likelihood and Bayesian approaches. The full-length cDNA of OgKCNQ4 was 2065 bp, and the open reading frame(ORF) was 2046 bp encoding for a putative protein with 681 amino acids. The relative molecular weight of OgKCNQ4 was 76.453 kD and the putative PI was 9.69. Secondary structure prediction analyses suggested 42.29% alpha helixes and 43.76% random coils in OgKCNQ4. Gene homology and Phylogenetic analyses revealed the closest relationship between OgKCNQ4 and KCNQ4 of Nanorana parkeri with 96.9% similarity and 95.0% identity. We first determined the full-length cDNA of OgKCNQ4 and the results here could provide foundations for further study on the evolution of KCNQ4 and its relationship to ultrasonic communication in amphibians.
基金a grant from the Chinese Natural Science Foundation to Fang ZHANG(NSFC grants 3187223031640073)Anhui Provincial Key Laboratory of the Conservation and Exploitation of Biological Resources(No.591601)。
文摘Nonlinear phenomena are commonly shown in the vocalization of animals and exerts different adaptive functions.Although some studies have pointed out that nonlinear phenomena can enhance the individual identification of male Odorrana tormota,whether the nonlinear phenomena play a specific role in the sexual selection of O.tormota remain unclear.Here we presented evidence that there was a significant negative correlation(Pearson:n=30,r=0.65,P<0.001)between the nonlinear phenomena content and snout-vent length in the male O.tormota,and two-choice amplexus experiments showed that female O.tormota preferred male with smaller body size containing higher nonlinear phenomena content in its calls.Phonotaxis experiments also revealed that females preferred calls with higher nonlinear phenomena content.Additionally,compared to the calls with lower nonlinear phenomena content and higher fundamental frequency,there was shorter response time in phonotactic behaviour of female induced by the calls with higher nonlinear phenomena content and lower fundamental frequency.We argue that the nonlinear phenomena content in the calls of male O.tormota can convey its body size information and may provide important clues for female frogs in darkened surroundings to identify males’body size during mate choice,meanwhile,higher nonlinear phenomena content in males’calls may increase the attractiveness of males to females.The results of this study provide confirmation that,for O.tormota,nonlinear phenomena have specific function in mate choice.
文摘The green odorous frog(Odorrana margaretae)has an interesting ring-shaped divergence pattern around the Sichuan Basin and documenting its morphological variations is essential in understanding its evolutionary history.Using curvilinear models,we detected significant geographical clinal variations in morphological traits,particularly sizes,of female O.margaretae.Males had significantly smaller sizes than females,and also had smaller variation ranges than females.One major trend of morphological variations was clinal:populations from the west tended to have a larger size with wider head and longer posterior limbs than populations from the east.Species history,with an early extended isolation and two subsequent secondary contacts,may explain most of the geographical clinal variations of O.margaretae.Bioclimatic factors may also contribute in explaining the variance of morphology.
基金supported by National Natural Science Foundation of China (30800100)Science and Technology Offi ce of Guiyang, China (2012204-28)
文摘Body surface area（BSA）was regarded as a more readily quantifiable parameter relative to body mass in the normalization of comparative biochemistry and physiology.The BSA prediction has attracted unceasing research back more than a century on animals,especially on humans and rats.Few studies in this area for anurans were reported,and the equation for body surface area（S）and body mass（W）：S=9.9 W 0.56,which was concluded from toads of four species in 1969,was generally adopted to estimate the body surface areas for anurans until recent years.However,this equation was not applicable to Odorrana grahami.The relationship between body surface area and body mass for this species was established as：S=15.4 W 0.579.Our current results suggest estimation equations should be used cautiously across different species and body surface area predictions on more species need to be conducted.
