MATSUI ET AL.- IM l\ l OGENY Ol BROWN l R()(iS 81 



lo /\. zhengi. Tlic oiclci o\ divergence 

 among the rcinaiiniiy Asian lnowii frogs 

 was the same as ihat reported In laiiaka- 

 Ucno et al. (199X1-)). llouc\ci, most ol 

 these blanches were supporied in \ei\ low 

 itcratiotrs ol' less ilum and ilic 



relationships within the brown frogs were 

 viewed as polytomous including five clus- 

 ters, l iist, two European species formed a 

 cluster (dmup 1) supported in iterations of 

 85.1"'ii. I hen, /\. ^////;/r('///v diverged from 

 all ihc icnianiniL' hmwn hogs (Cirouj-) 1). 



1 he laiici consisted ol two niajoi elus^ 

 ters, of whieli ime, suppoiied h> iieiaiions 

 of 85.9%, eoniained A', jiifxui n-(/ , R . 

 pirica, and A', or/ialiveniris (Group }). 

 Within this group, sister relationships of A'. 

 />iruii and A', ornalivcturis, both eharaeter- 

 i/ed hy 2n 24 ehromosomes against 

 2n 2d in nmsi oihei bitnvn frogs including 

 A', lu/'onuti (lanaka el al., 1996), was 

 sn|ipoMed ni hiL'h iieraiions of 98. S^/'o. 



1 he oihei niaior clusiei contained two 

 clusieiN, one vonNisimg ol A', iw^oi ami A'. 

 saiilcri from Saiui (Clrinip 4: suiiported m 

 61. 5% iterations), and the t)ther of A'. 

 okinavana, R. isiis/iinwfisi.s. and A', suiiicn 

 from W iilai ((iionp .s; supported in 82.4''/o 

 iiei.Uhinsi. In Cuouj") 5, a srsier relation- 

 ship ol A' isi/s/iirm'n.sis and R. sauleri from 

 \\ nl.n was supiniiled in \ei\ hi;jh ilcialions 

 ol 99.9" n. 



In this way. A', zlwiiiii was lound to be 

 greatly diverged from the brown frogs 

 including liuropean species, and was not 

 close to A', saiticri Uom Sanyi or Wulai. 



In I he m,i\nmnn likelihood tree (Fig. 

 2B). boi)tsiiap sui^ports of nionophylics of 

 R. zheniii and the brown frogs (75..'>''o). 

 ami of (he brown frogs (65.6'''o) were 

 weaker than in the neighbor-joining tree. 

 Brown frogs exhibited polyiixnous rela- 

 tionships and the nionophyly of R. lagoi 

 and A*, saiiliri from Sanyi (Ciroup 4 of the 

 neighbor-joining tree) were supported in 

 low Iterations of AU'/o. Supports of 

 nionophylics of Groups .1 (70.0"'o) and ^ 

 (6.VI"o) recognized in the ncighbor-joinujg 



tree were also weaker, while those of the 

 other groups were nearly the same as in the 

 neighbor-joining tree. 



In ilie maxinnun-parsimony tree (I'ig. 

 2( ), nu)n(iph\ lies ol A', z/iefigi and the 

 blown lu)L's, and ol ilic brown frogs were 

 supporied as in the neighbor-joining tree, 

 although iterations were lower (Sy.P'o and 

 66.i)°/o, respectively). This analysis resulted 

 in a polytomous consensus tree encompass- 

 ing seven clades, and nionophylics were 

 supported only for Cin)np I (supported by 

 7().2"o iterations), (iroup 5 (65.3%), and 

 I he A'. pirica-R. ornuiiveiilris elade of 

 CiKuip .'^ (96.3'''o). The sister relationship 

 of R. tsushiiuensis and R. sauleri from 

 Wulai was supported by IOO"'o iterations, 

 whereas that of R. jupunicu with the 

 species with 24 chromosomes (R. pirica 

 and R. ornativeniris) was not supported, 

 unlike in the neighbor-joining tree. 



Disci'SSlON 



1 Yom the results of the present study, 

 monophylies of A', zhengi and the brown 

 frogs, and of the brown frogs were sup- 

 poried. Nevertheless, relationships among 

 the latter were not sufficiently resolved, 

 and only sister relationships of (1) two 

 lunopean species (A', tcntporaria and R. 

 (hihruilina), (2) R. tsiishi/ncnsis and R. 

 siiiiieri from Wulai, (3) R. okinavuna and 

 the (2) species, and (4) R. pirica and R. 

 ornalivi'niri.s were unequivocally eonfirmcU. 



Also, it is highly probable that (5) R. 

 japonica and the species with 24 cluomo- 

 somes (A', pinca and R. orntitivt'Hins) arc 

 |ihylogenetieally very close, and that M. 

 lagoi is not remote from K. sutiwn from 

 Sanyi. On the other hand, relationships of 

 R. atntirensis with other brown frogs remain 

 unclear, although the order of divergence 

 among the .Vsian brown frogs in the ncigh- 

 Ixir-joining tree «as identical with our picM- 

 ous results ( Tanaka Ueno el al.. l*WSb) 



In all the ihicc ireei* obtained. K. zhvngi 

 was located outside of the brown frog> 



