Genetic distinction of roe deer {Capreolus pygargus Pallas) sampled in Korea

5 Pages·2001·2.6 MB·English

by  H S Koh

Home ränge and habitat utilization ofmale Felissilvestris 369
showed avoidance of fields/meadows in venting wildcat recovery from low popula-
spring{x2 = 28.63,df=4,p< 0.001)andused tion numbers.
habitats in proportion to their availability
during summer (j2 = 1-75, df=4, p = 0.78).
The average annual home ränge size of Acknowledgements
1662 ha in this study was larger than ranges
reported for radiocollared male wildcats in The presented project was part of a study con-
previous studies (Corbett 1979; Stahl et ducted at the Institute of Biogeography at the
al. 1988). However, MCP seasonal ränge University of the Saarland, Germany. I thank
sizes in this study were smaller than seaso- Dr.P.Müller,InstituteofBiogeography,Univer-
nal MCP ränge sizes recorded for male sity of the Saarland for financial support and
addedtelemetrydataandH.Cariusfortelemetry
wdiilcadtciantgs itnhaStwithzoemrelanrdän(gLeibesriezeks19i9n9)m,ailne- fliaenl,d wDorr.k.UrIsaBrmeittheannmkfouslert,o Darn.dBrRuocgeerMcRLaeml--
European wildcats vary under different charita for comments on earlier drafts of this
ecological conditions. Use of home ranges manuscript. Dr.Anne Olson and Dr.Dan
is believed to be exclusive in areas where Weary provided helpful comments on Statistical
lagomorphs dominate the diet as opposed analyses.
to exclusiveness only for animals of the
same sex in areas where rodents dominate
the diet (Stahl et al. 1988). As the ener- References
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Mamm. biol. 66 (2001) 371-375 ^Ittfe. Mammalian Biology
© Urban & FischerVerlag
^^P"
http://www.urbanfischer.de/journals/mammbiol Zeitschriftfür Säugetierkunde
Short commurrication
Genetic distinction of roe deer (Capreolus pygargus
Pallas) sampled in Korea
By H. S. KOH and E. Randi
Department of Biology, Chungbuk Um'versity, Korea and Istituto Nazionale per La Fauna Selvatica, Ozzano
deU'Emilia, Bologna, ItaLia
ReceiptofMs. 17. Ol. 2001
AcceptanceofMs. 30.05. 2001
Key words: Capreoluspygargus, mitochondrial DNA, moleculartaxonomy
The roe deer (genus Capreolus, Artiodacty- eastern Siberia, and C.p. tianschanicus,
la, Cervidae) includes twospecies: the smal- from Tian Shan and eastern Asia, including
ler European (C. capreolus) and the larger Korea. Other authors referred to roe deer
Siberian (C. pygargus) roe deer (Groves populations from Korea as a distinct sub-
and Grubb 1987; Sokolov and Gromov species, e.g., C.p. bedfordi (Sokolov and
1990; Hewison and Danilkin 2001), which Gromov 1990), or C.p. ochracea (Barclay
have widespread distributions in central- 1935). The genetic basis and taxonomical
western Europe, and in Asia and east Eu- significance of morphological Variation
rope, respectively. They show a number of among Siberian roe deer populations are
diagnostic morphologic, chromosomal and still unclear. Aim of this study is to assess
DNA traits (Groves and Grubb 1987; the genetic distinction and clarify the taxo-
Heptner et al. 1989; Douzery and Randi nomic Status of the C.pygargus population
1997; Randi et al. 1998a). Populations of from Cheju Island in Korea.
the two species occur in putative contact Three specimens ofroe deer from Cheju Is-
zones in the Caucasus, where C capreolus land in Korea were collected and muscle
is distributed at the southern slopes and sampleswere storedin a deep-freezer. Total
DNA
C.pygargus at the northern slopes of the cellular was extracted from muscle
mountainranges, and alongthe riversVolga samples digested for 2hours at 55°C in
and Don (Heptner et al. 1989; Danilkin 500 ul of STE buffer (0.1 M NaCl, 10mM
1996). However, most probably European Tris-HCl, ImM EDTA, pH8.0), 25 ul of
and Siberian roe deer populations do not 10 mg/ml Proteinase K stock Solution, and
hybridize in nature where they overlap in 25 ul of 20% SDS Solution. DNA was ex-
distribution (Danilkin 1996; Hewison and tracted with equal volumes of phenol-
Danilkin 2001). chloroform and Chloroform, precipitated
Populations of the Siberian roe deer show with 2 volumes of ethylalcohol, and resus-
extensive body size and coat colour varia- pended in 50 ul ofdistilled water. The Solu-
bility, and up to six subspecies were recog- tion was incubated at 37°C for 30min with
nized (Grubb 1993). However, recentlyDa- 5 ul of 10 mg/ml RNase stock Solution, and
nilkin (1996) and Hewison and Danilkin DNAwas extracted again.
(2001) recognized only two subspecies: The entire mtDNA CR was PCR-amplified
C.p.pygargus, from western and part of using primers L-Pro and H-Phe (Douzery
1616-5047/01/66/06-371 $15.00/0.
372 H.S. KoHetal.
and Randi 1997; Randi et al. 1998b) and AJ311188 and AJ311189). The average
the following thermal cycle: 94°C for TN93 genetic distance between the Cheju
5 min; 94°C for 1min, 72°C for 1min, samples and European or Siberian roe deer
54°C for 1 min (32 cycles); 72°C for 5 min. was 4.5%, and 2.5%, respectively. Roe deer
PCR-amplified products were purified from Cheju showed two distinct mtDNA
using the DNA PrepMateTM kit (Bioneer CR haplotypes, which diverged at 0.2% of
Co., Cheongwon-gun, Korea), and se- their sequences. Moreover, we have aligned
quenced using an automated DNA sequen- partial mtDNA CR sequences, 679 nucleo-
cer (Perkin Elmer 377) at the Korea Basic tide long, obtained from 23 samples of Eu-
Science Institute (Daejeon). ropean, Siberian, and Korean roe deer.
The new mtDNA CR sequences were Equally weighted MP analysis of the CR
aligned with homologous complete (Dou- alignment (with 47 parsimony-informative
zery and Randi 1997) and partial (Randi characters) produced 90 trees (length
et al. 1998b) sequences, using the Computer L= 81; consistency index CI = 0.617; reten-
program ClustalX (Thompson et al. 1997). tion index RI = 0.881). The 50% majority-
Phylogenetic analyses were performed by rule consensus tree is equivalent to the NJ
maximum parsimony (MP), using paup* phylogenetic tree, which is shown in
4.0b2a (Swofford 1998), with unordered Fig. 1.
character states (uninformative nucleotide MP and NJ trees concordantly showed that:
positions excluded), 10 replicates of ran- (1) European and Siberian roe deer se-
dom addition of terminal sequences and quences split into two divergent evolution-
TBR branch swapping, and by neighbor- ary lineages (BP = 100%); (2) Siberian roe
joining procedure (NJ; Saitou and Nei deer sequences split into three significantly
1987), with Tamura and Nei (1993) DNA different evolutionary lineages (BP= 70%-
distances (TN93), using PAUP*. Majority- 100%), including roe deerfrom east Siberia
rule (50%) consensus trees were con- (Amur Region), Korea, and west Siberia
structed when multiple equally parsimo- (Kurgan Region); and (3) roe deer se-
nious topologies resulted from MP analyses. quences from Korean individuals joined
Robustness ofthe phylogenies was assessed into a monophyletic dade (supported by
by bootstrap percentages (BP; Felsenstein BP = 91%), which was nested within the Si-
1985), with 1000 random resamplings. berian roe deer clade which is the sister
Clades can be considered significantly sup- clade ofthe roe deer sampledinthe Kurgan
ported when BP values are >70%. Region (west Siberia).
PCR products were clean Single bands of Thus, mtDNA sequences support a genetic
the expected molecular weight, and the se- distinction between west Siberian and far
quencing allowed unambiguous nucleotide eastern roe deer, in accordance with both
identifications. Regional Organization ofthe Sokolov and Gromov (1990) and Danilkin
CR from Capreolus was concordant with (1996) taxonomical listings. The new
data from other cervid CRs (Douzery and mtDNA sequences analysed in this study
Randi 1997; Randi et al. 1998b; Nagata et strongly suggest that roe deer from Cheju
al. 1998; Cook et al. 1999; Randi et al. island are more closely related to popula-
2001), thus suggesting that they represent tions sampled from the geographically dis-
true mtDNA sequences and not nuclear co- tant Kurgan Region in west Siberia (C p.
pies. The CR of Capreolus includes a cen- pygargus) thantothose from the geographi-
tral conserved region (CR-II), a left domain cally close Amur region in south-east Sibe-
(CR-I, on the tRNAs Thr and Pro side), ria (Fig. 1). The mtDNA sequences from
and a right domain (CR-III, on the tRNA Cheju Island form a significantly distinct
Phe side; not shown). clade within C. pygargus, thus indicating
A complete alignment 926 nucleotide long that roe deer from Korea belong to distinct
was obtained (the new sequences are avail- populations, and supporting Sokolov and
able from the GenBank with accession nos. Gromov's (1990) view that roe deer from
Systematics ofroe deerin Korea 373
the far east including Korea, should be kept gion, and it is bound on the north by north-
distinct from the other Siberian roe deer eastern China andfar eastern Russia. Cheju
populations. The Korean peninsula is a part Island, the largest ofthe southern Korea is-
of eastern Asia within the Palaearctic re- lands, originated by a series of volcanic ac-
91 ^ iK\oorreeaarn_1.1 Cheju Island
(Korea)
KorearM.2and1.3
^
i—
Siberian_1.1
Ii—
Siberian_1.2
West Siberia
IL
Capreolus pygargus ]_ (Kurgan Region)
1 Siberian_1.3
100 —— Siberian_1.4
|qq i— Siberian_2.1 East Siberia
(Amur Region)
Siberian 2.2
European_l.l
European_3.2
European_4.1
50
European_1.2
European^5.1
i—
European_2.1
L European_3.1
Capreolus capreolus
European_7.1
European_3.3
European_5.2
European_6.1
European_6.5
76
- European_6.4
0.001 nucleotidesubstitutions
Fig. 1. Neighbor-joining tree (rooted using homologous C. elaphus mtDNA CR sequences as outgroups; not
shown) dustering mtDNA CR haplotypes from Capreolus, computed using Tamura and Nei (1993) DNA distances.
Atinternodes are reportedthe bootstrapvalues>50%. FortheIdentificationofC. pygargus (Siberian) and C. ca-
preolus(European) haplotypes, see: Randietat. 1998b.