Fieldwork in Jiemuxi National Nature Reserve,Hunan,China,discovered morphologically indistinguishable groups of brown frogs that bred at different,exclusive times of the year.A principal components analysis of morphometric data,molecular analyses,and the exclusive breeding season suggested the occurrence of two species.The population that breeds during the winter was found to be an undescribed species and was subsequently described as Rana jiemuxiensis sp.nov.The new cryptic species can be identified from its congeners at the same locality by having a different breeding season and a divergent DNA barcode.Six major lineages of brown frogs were resolved with high nodal support.Rana japonica,R.chaochiaoensis,R.omeimontis,and R.jiemuxiensis have independent maternal lineages.Rana hanlunica and R.maoershanensis have essentially identical maternal lineages and they appear to represent the same species.The validity of the species R.longicrus,R.zhenhaiensis,and R.culaiensis and their genealogical relationships are not resolved and deserve further study.The genealogy indicates that sympatric cryptic lineages probably widely exist in the R.longicrus group.This highlights the importance of future more fine-scaled samplings and the inclusion of nuclear genes.
