Study On The Difference Of Hipposideros Pomona From China Based On Morphology And Molecular Sequences Data

Hipposideros pomona(Mammalia: Chiroptera: Hipposideridae)is distributed in 9 provinces in South China.And the differences and subspecies differentiation of H.pomona from different regions of China is still uncertain.At present,there is no a wide range of comprehensive comparison and research about this species.Therefore,the research on infraspecific classification of H.pomona is very necessary.To explore morphological and molecular difference and classification status of H.pomona,We conducted a series of bat surveys from 2012 to 2015 in 9 Provinces in South China.We surveyed 88 bat-inhabiting roosts,of which 6 roosts were inhabited by H.pomona.Then we collected the specimen of H.pomona and recorded the relevant data.We performed multivariate morphometric analyses using morphological measurements and compared the DNA sequence divergence.And the reasons for the differences and subspecies classification between H.pomona have been discussed.1.In South China,100 H.pomona specimens were captured from the Min-Guang coastal subregion,the South Yunnan subregion,and the Hainan Island subregion.Sixteen external variables were measured for all specimens.Thirty of the 100 bats were brought back to the laboratory for cranial samples,21 skull variables were measured and the rest of the 100 captured bats were released immediately after taking samples and measurements.Statistical analyses were run in SPSS 20.0 using the 16 external and 21 cranial variables of H.pomona from 3 geographical subregions.(1)The one-way analysis of variance showed that there is no significant difference between male and female in H.pomona.But,there are distinct differences in H.pomona from different subregion.(2)Discriminant analysis of external and cranial measurements showed significant differences between the 3 geographical subregions.All individuals were separated into 3 groups in a scatter plot.The correct discriminate rates was high,and the cross validation were similar with the original accuracy,which showed the discriminate functions were stable and valid.(3)Principal component analysis results showed that H.pomona differ markedly in morphology from different subregions.The first 3 principal components extracted from external measurements explained 63.154% of the total variation,and the first 3 principal components extracted from cranial measurements explained 54.894% of the total variation.And H.pomona from different subregions was separated from one another in the scatter plots.According to the distinct differences,H.pomona from different subregions belonged to different subspecies.2.The Cytb and CO? genes have been compared and analyzed.The sequence of Cytb gene was 1119 bp,6 haplotypes(H1–H6)were observed from 3 geographical subregions.The Cytb genes sequences divergences were computed using MEGA5.0.The divergences was ranged from 6.0% to 8.5% in the different subregions and not exceeding 1% in the same region.The sequence of CO? gene was 645 bp,6 haplotypes(H7–H11)were observed.The 8 sequenced samples from the Min-Guang coastal subregion shared 1 haplotype,4 sequenced samples from the Hainan Island subregion shared 1 haplotype.The CO? gene divergence among the 3 sampled sites in the South Yunnan subregion ranged from 0.2% to 0.3%.The ML and BI trees constructed based on the Cytb and CO? genes showed that H.pomona individuals from 3 different subregions were clustered in distinct subclades.The results were consistene with morphological results.3.The specimen from the Min-Guang coastal subregion were similar to H.p.sinesis from Fuzhou in morphological measurements.We hypothesized that the H.pomona from the Min-Guang coastal subregion should be H.p.sinesis.Specimens from northern India,northern Myanmar,and Thailand were referred to H.p.gentilis,H.pomona from the South Yunnan subregion were similar to the type specimens of H.p.gentilis in some morphological measurements,but the ear length was inconsistent with H.p.gentilis.In phylogenetic tree,the population from the South Yunnan subregion clustered with the samples from Myanmar in this study.At present,there was no molecular data of H.pomona from Thailand and India,so the specimen from the South Yunnan subregion whether to H.p.gentilis need further research.The morphological measurements of H.pomona from the Hainan Island subregion were not consistent with H.p.pomona or H.p.gentilis.In phylogenetic tree,the samples from the Hainan Island subregion and Laos clustered together.So,H.pomona from Hainan Island may be evolving into a unique subspecies.