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PROBLEMS

MOLECULAR GENETIC RELATIONSHIPS OF THE ZOKORS (RODENTIA, MYOSPALACINAE): ANALYSIS OF D-LOOP REGION POLIMORPHISM Tsvirka Marina 1 , Pavlenko Marina 1 , Korablev Vladimir 1 , Pang Junfeng 2.

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PROBLEMS

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  1. MOLECULAR GENETIC RELATIONSHIPS OF THE ZOKORS (RODENTIA, MYOSPALACINAE): ANALYSIS OF D-LOOP REGION POLIMORPHISMTsvirka Marina1, Pavlenko Marina1, Korablev Vladimir1, Pang Junfeng2 1Institute of Biology and Soil Science, Far Eastern Branch Russian Academy of Sciences, Vladivostok, 690022, Russia2Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, 650223, Yunnan, China

  2. PROBLEMS • Among others subterranean rodents, zokors (Myospalacinae) are still remains poorly studied in viewpoint of taxonomy and evolution. Recent molecular phylogenetic studies have demonstrated that zokors are closely related to Spalacinae and Rhyzomyinae in the family Spalacidae (Norris et. al, 2004; Jansa, Weksler, 2004) • However relationships among the species within Myospalacinae are not clearly understood.

  3. PROBLEMS GenusMyospalax Subgenus Myospalax M. myospalax M. aspalax M. psilurus Subgenus Eospalax M. rothschildi M. smithii M. fontanieri ( M. f.cansus, M. f.rufescens, M. f. baileyi) (Corbet, 1991) GenusMyospalax M. myospalax M. aspalax M. psilurus Genus Eospalax M. rothschildi M. smithii M. fontanieri M. cansus M. rufescens M. baileyi (Zheng, 1994 )

  4. Map of Zokors Distribution in the Russia M. myospalax M. psilurus 2n=64 2n=44 M. aspalax 2n=62 2n=62 M. armandii

  5. Background • Professor N. N., Vorontsov prominent Russian biologist was initiator of study of fossorial rodents in Russia and adjacent countries • L. Martynova, N. Vorontsov, 1975. Population cytogenetic of Zokors Chromosomes • L. Martynova, 1976.Chromosomal differentiation of three species of zokors. • L. Martynova, I. Fomicheva, N.Vorontsov, 1977.Electrophoretic study of blood protein of zokors.

  6. Electrophoretic patterns of transferrin (TF) of zokors(Pavlenko & Korablev, 2003; 2005) 1. Tf-A – M. aspalax, Transbaikalia 2. Tf-B – M. psilurus, Primorye 3. Tf-C – M. psilurus, Transbaikalia 4. Tf-C1 – M. armandii 5. Tf-D – M. myospalax, Altai region 1 2 3 4 5

  7. Map of Zokors Distribution in the China M. fontanieri M. cansus M. baileyi 2n=62 M. rufescens 2n=62 M. smithii M. rothschildi

  8. C. Zhou and K. Zhou. The validity of different zokor species and the genus Eospalax inferredfrom mitochondrial gene sequences / Integrative Zoology 2008; 3: 290–298. Myospalaxand Eospalax genera

  9. Phylogeneticrelationships of zokorsbased on the RAPD-PCRanalysis(Tsvirka et al., 2009) M. psilurus 100 4species groupswithingenusMyospalax 54 M. smithii 74 M. aspalax M. armandii 100 M. myospalax

  10. Puzachenko A., Pavlenko M., Korablev V.Craniological variability of the zokors (Myospalacinae) with simplified first upper molar. Abstracts of 11 International Conference "Rodens et Spatium", 24-28 July 2008, Myshkin (Russia). P.144 Puzachenko A., Pavlenko M., Korablev V.Variability of skulls in Zokors (Rodentia,Miospalacinae)// Zool. Z. 2009. V. 88. №1. P. 92-112. • 3 species groupswith taxonomic ranks of the genera or subgenera: • M. myospalax; • M. aspalax andM. armandii; • M. smithii andM.psilurus

  11. The aims of the present study were • to determine the systematic position of Myospalax and Eospalax; • to clarify the phylogenetic relationships among zokor species; • to confirm or contradict the taxonomic position of M. armandii as a distinct species; • to confirm or disprove previous data concerning genetic differentiation of Manchurian zokors M. psilurus from Khanka Plain and Transbaikalia.

  12. MATERIAL • 54 specimens of 7 species from 30 different localities in Russia and China. • 1 specimen of Spalax judaei from GenBank was used as outgroup. M. cansus

  13. METHODS Sequencing 5’ end of the D-loop • Phylogenetic reconstructions using MEGA 4 (Tamura et al., 2007): Neighbor-Joining (NJ), Maximum Parsimony (MP), Minimum Evolution (ME), unweighted pair-group method with arithmetic averaging(UPGMA) • Calculation of genetic distances based on Kimura two-parametric (K2P) estimator.

  14. RESULTS • The alignment of the mt DNA sequences of 7 taxa comprises 518 nucleotides. • 206 (40%) nucleotides were variable. • 187 (36%) nucleotides were parsimony informative. • The average ratio of transition/transvertion was 3.58.

  15. NJandUPGMAtreesof Zokorsbased on the D-loop sequences(MEGA-4.1) M. psilurus Primorye M. psilurus M. psilurus Zabaykalye M. aspalax M. armandii M. smithii M. cansus M. rufescens M. myospalax

  16. Interspecific relationships within the genus Myospalax Estimate value of Kimura two-parametric distance matrix for 8 taxa of Myospalax

  17. Genetic distances for nuclear (RAPD) and mitochondrial (D-loop)DNA

  18. Conclusions • Our results didn't confirm the hypothesis of validity of the genus Eospalax suggested by Zhou and Zhou (2009). The all studied zokor species should be divided into 3 species groups within one genus Myospalax: "myospalax", "psilurus-armandii-aspalax" and "smithii-cansus-rufescens". • The species within the "psilurus-armandii-aspalax" (M. psilurus, M. armandii, M. aspalax) as well as within "smithii-cansus-rufescens" (M. smithii, M. cansus, M. rufescens) groups were closely related among themselves.The M. myospalax, differs from the other species in chromosomal and molecular features, is more closely related with the first species group than the second one. • The hypothesis of species status of M. aspalax suggested on craniometrical analysis of zokors were confirmed (Puzachenko et al., 2009). • The hypothesis about the independent species status of two geographical populations of M. psilurus from Khanka Plain and Transbaikalia earlier suggested on t biochemical and karyological data (Pavlenko, Korablev, 2003) was also confirmed on the results of mt DNA analysis .

  19. The Authors Marina Tsvirka Marina Pavlenko Vladimir Korablev Junfeng Pang

  20. ACKNOWLEDGMENTS • The study was supported by the RFBR (grant nos. 06-04-39015) and FEBRAS (grant nos. s 09-III-В-138). THANKS FOR YOUR ATTENTION

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