Taxonomy And Molecular Phylogeny Of Earthworms(Oligochaeta:Megascolicidae) From Wuliang Mountains,Yunnan

Wuliang mountains are the key areas of biodiversity research in Yunnan Province,China.But there is no record of earthworm before this research.In order to reveal taxonomy and phylogeny of the earthworm from this region both morphologic and phylogenetic methods were used to identify certain species.We analyzed both composition and p-distance base on mitochondrial genes CO?,CO?,12S rRNA,16S rRNA,ND1,12S rRNA-16S rRNA sequences and CO?-CO?-12S rRNA-16S rRNA-ND1 combied sequences.Phylogenetic trees and ancient distribution were also based on the combined genes.As results,a total of 3 families,8 genera and 28 species of earthworm species were classified,including 21 species of Megascolecidae,4 species of Lumbricidae,and 3 species of Moniligastridae.3 new species were identified,they are Amynthas wuliangmontis sp.nov.,Amynthas jingdongensis sp.nov.and Amynthas lacustris sp.nov.The mitochondrial genes COI,CO?,12S rRNA,16S rRNA,and ND1 were sequenced,and the average inter-species p-distance of combined sequences was determined to be 18.6%±1.1%,while the average intra-species p-distance was 0?2.7%.Both Maximum Likehood tree and Bayesian Inference tree shows the same phylogenetic relationship of these specimens.We classified the combined tree into 8 clusters.Cluster 1 are constructed by all 4 Lumbricidae species.However,cluster 2?8 are constructed by all Megascolecidae specimens.According to cluster 8,it is verified that Amynthas and Metaphire are in a paraphyletic group.Moreover,we surmised that the dispersal direction of Megascolecidae is from south to north according to the reconstruction of ancient distribution area,hence species may spread from Mt.Wuliang&Mt.Ailao to Mt.Cang This is the first attempt for taxonomy and phylogeny of earthworm in Wuliang Mountains.