MOTOKAWA ET AL.— GENETIC RELATIONSHIPS OF EUMECES 



71 



these siu'cimciis, and c\aiiiiiici.l lliciii 

 cicclrophorclically logclhcr willi lissuc 

 samples of E. iiiarjiincilus Irom almost 

 whole of its range. Our puipose was to 

 determine the taxonomie status oi the 

 lotorishima pojiulation on the basis of its 

 relal ionshiiis uiih /■'. iiuirv.iiiulus pt)pula- 

 tions rel'leelcil hv allo/yme variations. 



Matfriai.s and Mfthoos 



We used lour speeiiiieiis Ironi loloi isliima 

 Island aiul 224 l iinicccs nuir'^iiii/liis s|-)eei- 

 mens Irom eight islands of the Kvukyu 

 Arehipelago: the I'okara Cncnip ( T'akara- 

 jima and Kodakarajiina), the Amami 

 Ciioup ( Amamioshima, rokuiu)shima, Oki- 

 noerahujima, and \'oroniima), and the 

 ()kinaua Cliiuip (( )kina\\ ajima and Kume- 

 lima) dig. I). All s]->cennens iyom the 

 same island, except lor those from 

 ()kma\\anma, were combined for the anal- 

 yses iindei an a jirioi i assumption that thc\ 

 rei">rescnl a smgle landomK i^i ceding juipu- 

 lalion. Specimens liom liiiec localities 

 uithin Okmawajima were irealed as ihiee 

 separate sami">les, [lecause the island's laige 

 si/e (120 km 111 leiii'lh, ID km in breadth 

 and 1200 km in area) ma\ have enhanced 

 sulnliv isions of its li/ard population. 



lliMiiogeni/ed tissue extraets were ana- 

 Is /ed In hori/ontal siaich gel electrophore- 

 sis. Sample iMcpaialioii and elect ropluMel ie 

 IMoceduies lollovved Shaw aiul Prasad 

 lUrO) and Mains and llopkiiison ( pr^) as 

 modilied In Kalo cl al. ll'^'Ml. I n/\incs 

 examined and buffer systems used liere 

 were almost same as ilu>se in Kato el al. 

 (1W4). Nevertheless, Ah 2 (Kalo et al., 

 |W^)4) was excluded liom the an.dyses, 

 because proilucls ol this locus were not 

 consistent Iv clear enough lo permit reliable 

 scoiiui' liisieail, peptidase was exanuned 

 for I lie fiist time for the .lapanese 

 luimvcc'i, and lesuliant bands from liver 

 extracts were mieiprctcd a.s being controlled 

 In ,1 sinuie locus, |\ p I), A total of 22 loci 

 ot Id en/vmes (i c . \.it l, Aat 2. Ada 2, 



Ah 1, ('k-2, List-4. Ill, (,dh, (,pi, Idli 1. 

 ldh-2, l.dh-1, I,dh-2, I dli \Klh I , \ldli- 

 2, Mpi, Pep-I). 6Pgdli-l, I'l'in I, Pgm-2, 

 and Sod - 1 ) w ci e I li us examined . 



Alk)/yme data were treated as products 

 of codominant alleles at a given locus, lor 

 some specimens of /:'. inari;inii!iis, allo/yme 

 data, taken from our previous study (Kato 

 el al., 1994), were subjected to the analyses. 

 All loci were evaluated for genetic 

 polymorphism (mean number of alleles per 

 locus, proportion of polymorphic loci, and 

 observed mean heterozygosity). In order 

 to estimate overall genetic differentiation 

 among populations, Nei's (ly^S) unbiased 

 genetic distance (D) was calculated on the 

 basis of pcipulation allele Iretiuencies. 1 he 

 resultant pairwise D matri.x was further 

 subjected lo analyses by the UPGMA algo- 

 rithm (Sneaih and Sokal, 1973) for the 

 purpose of estimating genetic relationships 

 among iiojiulat ions. All cominiiat ions were 

 carried oui using the BlOS^S-l computer 

 program (Svvofford and Sclander, 19S1). 



Ill ai.ldiiion, we i.|iiani 1 1 led a few external 

 chaiacieis (snoui vein length, tail length, 

 cxieni ol ilic median white line on tail, and 

 the number o\ midbody scale rows), and 

 also investigated the postnasal coiulilion 

 loi the loiorisiiima specimens. 



Rl M lis 



Ol ilie 22 presumptive loci analyzed, nine 

 I :, ( k 2. I h. Cidh, I dh-l. I,dh-2. 

 \Ipi, I'gm 1 and Sod-1) were fixed in all 

 samples. .Allele frequencies at the other 13 

 lH)lymori'>hic K>ci are given in Tabic I. .-Xt 

 SIX of these (Idh l. ldh-2. I dh-3. Mdh-l. 

 Mdh-2. and F*gm-2). the predominant 

 (frei|ucncy of occurrence ^O'o) electro- 

 morph was identical in all samples. The 

 lotorishima sample had alleles common lo 

 the Tokunoshinta \ainplc al all lixi. and nil 

 but the Lst-4 locus were fixed or nearly 

 fi.xcd for these shared alleles. Wnh respect 

 to the latter locus, all of the lotorishima 

 spccuncns h.nd a homo/vt;oic of an allele. 



