广西融水县发现雷山琴蛙

广西境内的琴蛙有多个地理种群,主要分布于桂东北地区的龙胜、灌阳,北部地区的融水,中部地区的金秀和南部地区的上林及武鸣,现己证实在灌阳都庞岭的种群为湘琴蛙(Nidirana xiangica),在大瑶山的种群为瑶琴蛙(N.yaoica),在大明山的种群为广西琴蛙(N.guangxiensis).雷山琴蛙(N.leisha...

Nidirana leishanensis Found in Rongshui County, Guangxi

Objectives]The genus Nidirana species was endemic group of classes in Oriental realm,widely distributed in subtropical areas of East and Southeast Asia.Among the species in genus Mdirana,Mdirana adenopleura had the widest distribution area and only distributed in Taiwan,northern of Fujian,southern of Zhejiang,central Jiangxi and other regions in China.The recent contribution to the phylogeny of genus Nidirana reconsidered the previously recorded as N.adenopleura populations in southern of China were actually the genus Nidirana new species.There are several geographical population of genus Nidirana mainly distributed in the northeast,north,central and southern regions of Guangxi,had been certified the genus Nidirana population form Dupang mountain was N.xiangica,the genus Nidirana population form Dayao mountain was N.yaoica,and the genus Nidirana population form Darning mountain was N.guangxiensis.Methods]Seven male and three female specimens collected form Rongshui were examined and measured with digital calipers.The measurements were as follows:snout-vent length,head length,head width,snout length,intemasal distance,interorbital distance,eye diameter,tympanum diameter,tympanum-eye distance,hand length,radio-ulna length,foot length,tibial length,and compared characters with the N.leishanensis specimens obtained from Guizhou.All specimens were fixed in 10%buffered formalin,transferred to 70%ethanol,and deposited in NHMG.Four muscular samples attained from euthanasia specimens and then preserved in 95%ethanol and stored at-40℃were used for molecular analysis.All samples were 36 sequences from all known Nidirana species and two sequences from the out-group Babina were obtained from GenBank and incorporated into our dataset(Detail information of these materials was shown in Table 1).Genomic DNA were extracted from muscle tissue samples using DNA extraction kit.A mitochondrion genes namely partial cytochrome C oxidase I gene(COI)were amplified,and the primers were dgLCO and dgHCO.PCR amplifications were processed with the cycling conditions that initial denaturing step at 95℃for 4 min,35 cycles of denaturing at 94℃for 40 s,annealing at 52℃for 40 s and extending at 72℃for 60 s,and a final extending step at 72℃for 10 min.PCR products were sequenced with both forward using reverse primers.The Clustal W algorithm was used in MEGA 6.0 software to compare all the COI gene sequences and calculate the genetic distance between specimens and other species by the p-distance model.The comparison sequences were calculated to get the optimal nucleotide substitution model in jmodeltest v2.1.2.Sequenced data analyzed using maximum likelihood(ML)in RaxmlGUI 1.3 to structure the phylogenetic tree.Advertisement calls of the specimens were recorded in the field at the air temperature 22℃using a SONY PCM D100 digital sound recorder.Praat 6.0.27 was used to obtain the oscillogram,sonogram,and power spectrum.Raven pro 1.5 was used to quantify the acoustic properties.Results]We obtained four mitochondrial COI gene sequences length 639 bp and the optimal nucleotide substitution model for phylogenetic analysis was GTR+G+I.The Maximum likelihood phylogenetic tree of genus Nidirana showed these samples and A.leishanensis samples in Guizhou together formed a monophyletic group(Fig.1).Genetic distance estimation based on p-distance model showed the intraspecific genetic distance was 0.2%,and the genetic distance was 0.5%between the specimens and the N.leishanensis samples(Table 2).The following principal characters of genus Nidirana specimens:a largebody size(Snout-vent length,SVL,males:47.5 mm