Lucanus

datunesis Hashimoto 1984 (Coleoptera: Lucanidae)

1.

58

DNA -80C

2.

-80C

 Lucanus swinhoei Parry, 1874

Lucanus miwai Y. Kurosawa, 1996 , ,

DNA ,
universal insect mtDNA primers
primers PCR kits
DNA EditSeq MegAlign program
, PAUP* MrBayes program
Neolucanus doro
Modeltest Likelihood Ratio Test

3.

haplotypes DnaSP program

( ) s (number of segregation sites) h (number of
haplotypes) Hd (haplotype diversity)

 Ne coalescent method BEAST program

4. DNA

DNA
DNA , Rsa I Alu I enzyme
, 500 bps DNA , SNX linker,
PCR , GATA
GAAA oligo-nucleotides , TA cloning kit
vector ,

32 P

, 400-850 bps

PCR DNA

1.


DNA , COI (Cytochrome Oxidase I)
(AT-rich) Wg ( Wingless ) universal
insect primers 1300 bps ( COI ) 600 bps (AT-rich)
500 bps ( Wg ) DNA EditSeq
MegAlign program , PAUP*
(maximum parsimony), (maximum likelihood),
(Bayesian phylogenetic analysis) ,
(nonparametric bootstrapping)
(posterior probability) Likelihood Ratio Test
GTR+G+I COI ,

COI : 1) COI
, Lucanus ,
2) Lucanus

 3)
4)
4)
Lucanus fortunei 5)
, Lucanus swinhoei
. 6)

Lucanus miwai Lucanus szetschuanicus

szetschuanicus ,
Lucanus formosanus ,
Lucanus hermani Lucanus planeti
(Bayesian relative rate) Lucanus

2.
10 COI , DNA

10

COI

(haplotype),
( Hd , haplotype diversity = 0),
Lucanus Lucanus formosanus
( Hd = 0.995) , AT-rich
DNA ,
DNA

3. DNA
DNA enriched library
DNA 40
8

 ( ),

Loci

Microsatellite Core Repeat

Length

33 (TATC)10

168

34 (GATA)8

149

8 (AAAG) 10

125

19 (TATC) 9

332

22 (TATC) 12

297

23 (TATC) 10

175

27 (TATC) 8

197

31 (TATC) 10 AATCATGTCCCTC(TATC) 8

156

( Lucanus swinhoei
Lucanus miwai )
Lucanus fortunei

 , Lucanus formosanus

, ,
,

2006 54: &

160
Avise,

J.C.,

2004.

Molecular Markers,

Natural History,

and

Evolution. Sinauer Associates, Sunderland, MA, pp. 684.

Felsenstein, J., 1985. Confidence limits on phylogenies: an
approach using the bootstrap. Evolution 39, 783 791.
Frankham, R., J. D. Ballou, and D. A. Briscoe. 2002. Introduction
to Conservation Genetics. Cambridge University Press,
Cambridge, UK. 617 pp.

Huelsenbeck, J.P., Ronquist, F.R., 2001. MrBayes: bayesian

inference of phylogeny. Biometrics 17, 754 755.
Posada, D., Crandall, K.A., 1998. MODELTEST: testing the model
of DNA substitution. Bioinformatics 14, 817 818.
Simon, C., Frati, F., Beckenbach, A., Crespi, B., Liu, H., Flook, P.,
1994.

Evolution,

mitochondrial

weighting,

gene

and

sequences

phylogenetic
and

utility

of

compilation

of

conserved polymerase chain reaction primers. Ann. Entomol.

Soc. Am. 87, 651 701.
Swofford, D.L., 1998. PAUP*, version 4.0b10. Sinauer Associates,
Sunderland, MA.