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Russian Journal of Genetics, Vol. 38, No. 4, 2002, pp. 434438. Translated from Genetika, Vol. 38, No.

4, 2002, pp. 534538.


Original Russian Text Copyright 2002 by Malyarchuk, Derenko, Denisova, Nassiri, Rogaev.

HUMAN
GENETICS

Mitochondrial DNA Polymorphism in Populations


of the Caspian Region and Southeastern Europe
B. A. Malyarchuk1, M. V. Derenko1, G. A. Denisova1, M. R. Nassiri2, and E. I. Rogaev3
1

Institute of Biological Problems of the North, Russian Academy of Sciences, Magadan, 685000 Russia;
fax: (41322)34-463; e-mail: ibpn@online.magadan.su
2 Meshhed University, Meshhed, Iran
3 National Research Center of Mental Health, Russian Academy of Medical Sciences, Moscow, 113152 Russia
Received June 27, 2001

AbstractMitochondrial DNA (mtDNA) restriction polymorphism was examined in Turkmens, Eastern Iranians, and Ukrainians. The gene pools of all populations studied were characterized by the presence of European mtDNA lineages. Mongoloid component observed in Turkmen and Iranian populations with the frequencies of about 20% was represented by groups C, D, and E/G in Turkmens, and by M*, D, A, and B in Iranians.
The relative positions of the populations studied, of populations from the Caucasus, Western Iran, and Russian
populations from the Krasnodar krai and Belgorod oblast in the space of principal components revealed a geographically specific pattern of the population clustering. The data on mtDNA polymorphism indicated pronounced differentiation of Eastern and Western Iranians. The latter were characterized by a mtDNA group composition similar to that in Eastern Slavs. The historical role of the Caspian populations in the formation of the
population of Southeastern Europe is discussed.

INTRODUCTION
Since ancient times, the population inhabiting western regions of the Central Asia and Iranian upland
played the key role in the peopling of East European
territories. It is suggested that one of the routes of the
Homo sapiens sapiens penetration to Europe passed
though the Caspian regions. Later, during the
Mesolithic, the Caspian regions and refuges at the south
of Eastern Europe were the starting points for recolonization of the European territories [1, 2]. During the
Bronze Age, cattle-breeder tribes from the Caspian
regions settled in the European territories [3, 4].
According to paleoanthropological data, during Scythian time (the early Iron Age) Eastern European steppes
were the territories of activity of tribes belonging to the
Scythian confederation [4]. Apparently, exactly Iranian
tribes played an important role at the first stages of
Scythian ethnogeny. Later Scythians spread over wide
territories of Europe, Central Asia, and Southern Siberia, accumulating many ethnic components. The Iranian substrate also played a significant part in the formation of Eastern Slavs, specifically, Ukrainians and
southern ethnic territorial groups of Russians. According to anthropological and archaeological data, in early
Middle Ages Iranian-Slavic symbiosis (Chernyakhovskaya culture) was a typical feature of the population of East European steppes [5, 6]. In view of this, it
is likely that Iranian, or Iranianized tribes could have
been affiliated with Eastern Slavs.
The data on biochemical and molecular genetics
generally agree with the ideas of anthropologists,
archaeologists, and linguists considered above. Gene

mapping analysis of classical markers showed that the


distribution of alleles of protein loci reflected several
trends of variability that can be interpreted on the historical basis. For instance, according to the data of Cavalli-Sforza et al. [7], the distribution of the third principal component in the populations of Europe suggests
that during the Bronze Age the populations migrated
from the Caspian regions to the east of Europe. Based
on the polymorphism of autosomal DNA loci, analogous variability trend was also described for the second
principal component of European population gene pool
[8]. Analyses of biallelic and microsatellite Y-chromosomal loci variability revealed more evident similarity
between the gene pools of East European and West
Asian populations. Examination of Y-chromosomal
polymorphism pointed to the existence of distinct differentiation between the populations of Western and
Eastern Europe. At the same time, the populations of
Eastern Europe (specifically, Slavic populations,
including Western Slavs) demonstrated substantial similarity with the populations of Western Asia, and Iranian populations in particular, in respect of the distribution of Y-chromosomal markers [9]. Detailed examination of the populations of Southwestern Asia showed
that distribution of Y-chromosomal type group, HG3,
marked with the SRY-1532-A variant, favored the
hypothesis that during the Bronze Age cattle-breeder
tribes of Western Asia have spread in various directions,
namely, to India, Central Asia, Siberia, and Eastern
Europe [10]. Maximum HG3 frequencies are typical of
the populations of Eastern Iran and Western Asia [10],

1022-7954/02/3804-0434$27.00 2002 MAIK Nauka /Interperiodica

MITOCHONDRIAL DNA POLYMORPHISM IN POPULATIONS

435

Slavic populations of Europe [11], and the populations


of Altai-Sayan region of Siberia [12].

Table 1. Polymorphic restriction variants determining


the groups of mtDNA types in the populations of Eurasia

The studies of variability of another polymorphic


system, maternally inherited mitochondrial DNA
(mtDNA), can also provide useful information for the
investigations of the genetic history of the populations
of the Caspian regions and adjacent territories. These
populations, however, still remain unexplored in
respect of mtDNA polymorphism. Here we present the
data on mtDNA polymorphism in the populations of
Iran, Turkmenistan, Southern Russia and Ukraine. The
data on mtDNA variability obtained were compared to
those for adjacent territories by use of phylogeographic
approach. Application of the given method shed the
light on the peculiarities of the distribution of mtDNA
markers in the populations studied.

mtDNA
group

MATERIALS AND METHODS


Total DNA from biological tissues (blood and hair
bulbs) was extracted by use of standard methods. The
sample of Iranians (Persians; n = 25) from the western
regions of Iran (province Khorasan) was studied. On
the northeast the province is bordered by Turkmenistan.
The sample of Turkmens (n = 26) was mostly comprised of Turkmens-Tekke (the cities of Ashgabat and
Chardzhou). The sample of Ukrainians (n = 36) was
formed from the inhabitants of Magadan and Nikolaev
oblast, originating from different parts of the Ukraine.
According to the questionnaire data, 11 of these were of
eastern Ukrainian origin, 8 persons originated from
western, and 10, from southern Ukraine.
Screening of polymorphic sites, determining the
main groups of mtDNA types distributed in the populations of Eurasia, was performed with the help of restriction analysis of mtDNA fragments amplified in polymerase chain reaction (Table 1). Restriction fragments
were fractionated in 8% polyacrylamide gels. The DNA
fragments were detected by staining with ethidium bromide with subsequent visualization in UV light.
Polymorphism was scored by the presence (+) or
absence () of the restriction endonuclease recognition
sites. The mtDNA types were identified in accordance
with the classification of mtDNA type groups in the
populations of Eurasia [13, 15]. Interpopulation differences relative to the frequencies of mtDNA type groups
were estimated using factor analysis (STATISTICA,
v. 5.0). Euclidean distances between the populations
(based on the data on the frequencies of mtDNA type
groups) were calculated using cluster analysis (tree
clustering; STATISTICA,v. 5.0). For comparative analysis the data on mtDNA polymorphism in Iranians
(Western Iran; n = 101), Azerbaijanians (n = 48), Armenians (n = 191) [14], Adygeians (n = 50) [13], and Russians from Krasnodar krai (n = 49) and Belgorod oblast
(n = 69) [17] were used.
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H
V
HV*
U
K
J
T
I
W
X
M:
C
D
E
G
A
B
F

Group-specific restriction variants


14766 MseI, 7025 AluI
14766 MseI, +15904 MseI, 16297 MseI,
4577 NlaIII
14766 MseI
+12308 HinfI
+12308 HinfI, 9052 HaeII, +10394 DdeI
13704 BstNI, +10394 DdeI
+13366 BamHI, +15606 AluI
4529 HaeII, +8249 AvaII, +16389 BamHI,
+10032 AluI
+8249 AvaII, 8994 HaeIII
14465 AccI
+10394 DdeI, +10397 AluI
+10394 DdeI, +10397 AluI, 13259
HincII/+13262 AluI
+10394 DdeI, +10397 AluI, 5176 AluI
+10394 DdeI, +10397 AluI, 7598 HhaI
+10394 DdeI, +10397 AluI, +4830 HaeII/+4831
HhaI
+663 HaeIII
9-bp deletion
12406 HpaI/HincII

Note: Groups of mtDNA types were denoted according to the classification suggested in [1315]. Positions of polymorphic
restriction sites are indicated relative to the Cambridge Reference Sequence of human mtDNA [16].

RESULTS AND DISCUSSION


Table 2 demonstrates the results of the screening for
polymorphic variants determining the main groups of
Eurasian mtDNA types in the populations of Iranians,
Turkmens, and Ukrainians examined. The search for
European (H, V, HV*, J, T, U, K, I, W, and X) and Mongoloid (M*, C, D, E, G, A, B, and F) markers was performed. For comparison, the data on the frequencies of
mtDNA groups in the populations of Western Iran (Iranians), Transcaucasia (Azerbaijanians and Armenians),
Northern Caucasus (Adygeians), and also in the populations of Russians from Krasnodar krai and Belgorod
oblast are presented. Nearly all populations were characterized by the presence of a substantial European
component, represented by the corresponding mtDNA
types. Low frequencies (from 0.52 to 6%) of the Mongoloid component were observed in Russians from Belgorod oblast, Adygeians, Armenians, and Azerbaijanians. These mtDNA types mostly belonged to group C.
Among the populations studied, the most pronounced
Mongoloid component with similar frequencies of 20
and 19.2% was revealed in Eastern Iranians and Turk2002

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MALYARCHUK et al.

Table 2. Distribution (in %) of mtDNA type groups among the populations of Southeastern Europe, Caucasus, and the Caspian region
Population
mtDNA
group

Russians
(Krasnodar)1

Russians
Western
Ukrainians
(Belgorod)1
Iranians2

Eastern
Iranians

Turkmens

Azerbaijanians2

Armenians2

Adygeians3

30.36

30.0

38.78

37.68

41.7

43.56

19.23

29.17

8.16

7.25

5.6

7.92

3.85

4.17

HV*

4.08

2.9

1.98

8.0

7.69

4.17

7.33

18.37

11.59

8.3

14.85

8.0

7.69

4.17

8.38

4.0

12.24

8.69

8.3

8.91

16.0

15.38

16.67

11.52

14.0

10.2

18.84

16.7

8.91

24.0

3.85

16.67

20.94

32.0

1.45

8.3

6.93

4.0

2.08

7.33

2.0

1.45

2.8

2.97

4.0

7.69

4.17

1.05

2.0

2.04

4.35

2.97

4.0

2.08

1.57

1.45

2.8

0.99

4.17

2.62

M*

4.0

1.45

7.69

4.17

6.0

4.0

7.69

E/G

3.85

4..0

8.0

0.52

2.9

5.6

12.0

8.33

8.38

10.0

6.13

Note: The data are cited from:

1 [17]; 2 [14]; 3 [13]. The

15.38

sign ? denotes mtDNA types unidentified by restriction endonuclease analysis.

mens. The structure of the Mongoloid component in these


populations was somewhat different: mtDNA found in
Turkmens, was from group M (C, D, E/G), and in Iranians,
from M (M*, D), A and B. Turkmens had an unusual
mtDNA type marked by polymorphic variants determining two mtDNA groups, E (7598HhaI) and
G (+4830HaeII/+4831HhaI). According to the literature data, these mtDNA types were never described in
human populations before. However, we detected low
frequencies of E/G mtDNA types among indigenous
populations of Southern Siberia (Derenko et al., unpublished results).
As follows from Table 2, different Iranian populations display substantial genetic differences, mostly
resulting from differences in the frequencies of groups
H, V, and the Mongoloid component [14]. The population of Western Iranians was characterized by a typical
European gene pool composition accompanied by the
complete absence of the Mongoloid component. Note
that the data on Y-chromosomal polymorphism also
point to substantial differentiation of the populations of
Western and Eastern Iran [10]. The observed westward
decline of the frequency of the Mongoloid component
in the gene pools of the Central Asian populations from
60% in Kyrgyzes, Kazakhs, and Uigurs [18] to 20% in

Turkmens and Eastern Iranians is most likely associated with the processes of settling of the Turkic tribes.
These tribes were initially formed in the Central Asia
among the mixed EuropeanMongoloid forms [4]. The
data on genetic differentiation of Iranian populations do
not conflict with historical data, because it is known
that only central and eastern parts of Iran were included
into the ethnic territories of Persians, but the western
and southeastern parts of Iran since ancient times were
inhabited by the populations of another origin (Azerbaijanians, Kurds, Lurs, Balochi, etc.) [19]. For these
reasons, the revealed differences in the structure of
mtDNA types in Iranians can be explained not only by
the presence of the Mongoloid component in Eastern
Iranians, but also by interethnic differences in the structure of the European component in Western and Eastern
Iranians. This proposal, however, requires detailed
examination of mtDNA variability in different ethnic
territorial groups of Iran.
The figure illustrates the distribution of the examined populations in the space of the principal components. Factor analysis revealed two groups of related
populations. The first group is comprised of Armenians
and Azerbaijanians (Adygeians are located somewhat
apart from them), while the second group is represented

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Table 3. Euclidean distances between populations of southern Eastern Europe, the Caucasus, and Caspian region based on
mtDNA polymorphism data
Population

1. Russians (Krasnodar)1

0.13

0.17

0.12

0.46

0.29

0.21

0.21

0.30

2. Russians (Belgorod)1

0.13

0.11

0.14

0.42

0.31

0.16

0.15

0.22

3. Ukrainians

0.17

0.11

0.13

0.46

0.33

0.18

0.16

0.24

0.12

0.14

0.13

0.51

0.35

0.24

0.23

0.33

5. Eastern Iranians

0.46

0.42

0.46

0.51

0.32

0.33

0.33

0.35

6. Turkmens

0.29

0.31

0.33

0.35

0.32

0.22

0.27

0.34

4. Western

7.

Iranians2

Azerbaijanians2

0.21

0.16

0.18

0.24

0.33

0.22

0.12

0.18

8. Armenians2

0.21

0.15

0.16

0.23

0.33

0.27

0.12

0.17

Adygeians3

0.30

0.22

0.24

0.33

0.34

0.18

0.17

9.

0.35
1

Note: Data on mtDNA polymorphism from the following studies were used: [17], [14], [13].

by Eastern Slavic populations. Interestingly, Western


Iranians are located in close proximity of Eastern
Slavic populations. The populations of Eastern Iranians
and Turkmens are most distant from each other as well
as from other population groups. Table 3 presents the
data on Euclidean distances calculated from the data on
the frequencies of mtDNA groups in the populations
examined. Quantitative estimates of the differences
between the population gene pools obtained also point
to genetic differentiation of these populations. Minimal
differences were observed between the populations of
Western Iranians, Russians, and Ukrainians, on the one
hand, and between Caucasian populations, on the other.
The genetic similarity between Eastern Slavs and
Western Iranians described in the present study
deserves special interest. A more detailed examination
of the Russian population from the southern regions of
Principal component 2
1.2
1.0
1
0.8
0.6
0.4
0.2
0.0
0.2
0.1
0.1
0.3

2
3
5

4
8

0.5

7
9

0.7
0.9
1.1
Principal component 1

Distribution of the populations in the space of the principal


components. Principal component 1 reflects 76.3% of the
total variability of mtDNA, while principal component 2
reflects 13.8% of general variability of mtDNA. Population
designations: 1, Eastern Iranians; 2, Adygeians; 3, Armenians; 4, Azerbaijanians; 5, Turkmens; 6, Russians from
Belgorod oblast; 7, Ukrainians; 8, Russians from Krasnodar
krai; 9, Western Iranians.
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European Russia along with the examination of other


populations of the Caspian region would probably provide identification of the desired Iranian component
involved into ethnogeny of Eastern Slavs.
ACKNOWLEDGMENTS
We thank the Corresponding Member of the Russian
Academy of Sciences I. A. Zakharov (Vavilov Institute
of General Genetics, Russian Academy of Sciences) for
his help in this study.
This work was supported in part by the Russian Foundation for Basic Research (grants no. 00-06-80448 and
99-06-80430) and the Russian State Program Frontiers in Genetics (grant no. 99-04-30).
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