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MOLECULAR GENETIC RELATIONSHIPS OF
THE ZOKORS (RODENTIA,
MYOSPALACINAE): ANALYSIS OF D-LOOP
REGION POLIMORPHISM
Tsvirka Marina1, Pavlenko Marina1,
Korablev Vladimir1, Pang Junfeng2
1Institute
of Biology and Soil Science, Far Eastern Branch
Russian Academy of Sciences, Vladivostok, 690022, Russia
2Kunming
Institute of Zoology, Chinese Academy of
Sciences, Kunming, 650223, Yunnan, China
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.
PROBLEMS
Genus Myospalax
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)
Genus Myospalax
M. myospalax
M. aspalax
M. psilurus
Genus Eospalax
M. rothschildi
M. smithii
M. fontanieri
M. cansus
M. rufescens
M. baileyi
(Zheng, 1994 )
Map of Zokors Distribution in the Russia
M. myospalax
M. psilurus
2n=64
2n=44
M. aspalax
2n=62
M. armandii
2n=62
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.
Electrophoretic patterns of transferrin (TF)
of zokors (Pavlenko & Korablev, 2003; 2005)
1
2
3
4
5
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
Map of Zokors Distribution in the China
M. fontanieri
M. cansus
M. baileyi
2n=62
M. rufescens
M. smithii
2n=62
M. rothschildi

C. Zhou and K. Zhou. The validity of different zokor
species and the genus Eospalax inferred from
mitochondrial gene sequences / Integrative Zoology
2008; 3: 290–298.
Myospalax and Eospalax genera
Phylogenetic relationships of zokors based on the RAPDPCR analysis (Tsvirka et al., 2009)
M. psilurus
100
54
M. smithii
74
M. aspalax
M. armandii
100
M. myospalax
4 species
groups within
genus
Myospalax
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 groups with taxonomic ranks of the genera
or subgenera:
•
•
•
M. myospalax;
M. aspalax and M. armandii;
M. smithii and M. psilurus
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.
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
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.
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.
NJ and UPGMA trees of Zokors based 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
Interspecific relationships within the genus Myospalax
1
2
3
4
5
6
7
1. M. cansus
-
2. M. smithii
0.120
-
3. M. rufescens
0.123
0.062
-
4. M. aspalax
0.190
0.167
0.164
-
5. M. armandii
0.185
0.185
0.181
0.070
-
6. M. myospalax
0.225
0.226
0. 224
0.177
0.190
-
7. M. psilurus (П)
0.158
0.154
0.154
0.076
0.084
0.159
-
8. M. psilurus (З)
0.164
0.164
0.160
0.108
0.104
0.181
0.063
Estimate value of Kimura two-parametric distance matrix for 8 taxa of
Myospalax
8
-
Genetic distances for nuclear (RAPD) and mitochondrial
(D-loop) DNA
RAPD
D-loop
0.05
0.003-0.028
0.04-0.05
0.003
0.08
0.0620.07
between two forms of M.
psilurus
0.08
0.063
between distant species
(Myospalax – Eospalax)
0.25-0.35
0.154-0.226
0.31
0.238-0.304
Distances
within species
within forms of M. psilurus
between species
(M. aspalax - M. armandii
M. smithii - M. rufescens)
between genera
(Myospalax – Spalax)
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 .
The Authors
Marina Tsvirka
Vladimir Korablev
Marina Pavlenko
Junfeng Pang
ACKNOWLEDGMENTS
The study was supported by the RFBR (grant nos. 06-0439015) and FEBRAS (grant nos. s 09-III-В-138).