Cop. 1992(?). pp. 375-392

Evolution and Biogeography of the Clinidae (Teleostei: Blennioidei)

CAROLA. STEPIEN

The systematic and biogeographic relationships of the teleost family Clinidae (commonlyknown as kelpfishes or klipfishes),an antitropically distributed group of nearshore fishes characterized by widespread ecological variation, were an- alyzed using allozyme and morphological characters. The Clinidae comprises three tribes: the matritrophic (ovoviviparous) Clinini and Ophiclinini, and the oviparous tribe Myxodini. Emphasis of the present study was in resolving the relationships among the eastern Pacific Myxodini, which include the South American genus Mpodes and the North American genera Heterostichur and Gibbonsia. Allozyme data were analyzed from 40 presumptive gene loci for all four North American myxodin species, one species of the Australian chid tribe Clinini (Heteroclinus whiteleggei), 10 species (representing all tribes) of the hy- pothesized sister blennioid family, the Labrisomidae, and two species of the blennioid family Chaenopsidae (also believed to be closely related; the latter two families were used as outgroups). Several different phylogenetic approaches for analyzing allozyme data were used for purposes of comparison, including two frequency character methods, a genetic distance method, a discrete character parsimony method (using only fixed alleles), and a multistate character parsimony analysis (the latter two used presumptive gene loci as characters and selected alleles as character states). Discrete morphological characters were also analyzed both separately and in- cluded with allozyme data in parsimony analyses. Results from morphological and allozymedata, as well as the different methods of phylogenetic analysis, were congruent; yielding identical parsimonious re- lationships among taxa, except within the genus Gibbonsia. Two most parsi- monious trees were consistently obtained, which differ in whether G. metzi or G. elegans is the basal clade within the genus (and the sister group to the other two species). The Clinidae is distinguished by a suite of allozyme character state synapomorphies identified in the present study as well as three morphological synapomorphies. The present study indicates that the cryptotremin labrisomids are the sister group to the Clinidae. The trophodermic matritrophic (ovovivip arous) clinids (comprising the tribes Clinini and Ophiclinini) and the Myxodini are sister taxa, and the South American myxodin genus Myxodes is sister to the northern myxodin genera. The North American genera Heterostichus and Gib- bonsia are sister groups.

HE systematics of the eastern Pacific mem- guished by the morphological synapomorphy of T bers of the teleost family Clinidae, sub- a cordlike ligament extending from the cera- order Blennioidei, were last examined by C. tohyal to the dentary symphysis (Springer et al., Hubbs (1 952), and the genus Gibbonsia was re- 1977). The Clinidae contains three tribes cently revised by Stepien and Rosenblatt (1991). (Myxodini, Clinini, and Ophiclinini), 20 genera, Clinids are relatively small (less than 300 mm and 7 1 species, most of which are restricted to SL, except Heterostichus rostratus) benthic fishes, Australia and Africa (Nelson, 1984).The family the majority of which inhabit intertidal and has a mostly antitropical distribution, shown in shallow subtidal waters (Springer, 1982; Nel- Figure 1A, and all but five (tropical) species are son, 1984). George and Springer (1980) rede- limited to warm temperate waters (Springer, fined the Clinidae, excluding the Tripterygi- 1982). idae, Labrisomidae, and Chaenopsidae, and The present study primarily examined rela- adding the tribe Ophiclinini. Clinids are distin- tionships among the eastern Pacific myxodin

0 1992 by the American Society of Ichthyologists and Hcrpctologirts Fig. 1. Distribution maps of the families Clinidae (A) and Labrisomidae (B), based on C. Hubbs (1952), Penrith (1969), Springer (1950), Stephens and Springer (1973), George and Springer (1980). STEPIEN-CLINID EVOLUTION 377

TABLE1. PRESENTSTATUS OF THE CLINIDTRIBE MYXODINI. (After C. Hubbs, 1952; Springer, 1970;Stephens and Springer, 1973; George and Springer, 1980; and Stepien and Rosenblatt, 1991.)

Component taxa Common distribution A. Gibbonsia (Cooper) 1. G. elegans (Cooper) Point Conception, California, through central Baja California, Mexico 2. G. metzi C. L. Hubbs North of Point Conception along U.S.Pacific coast and in areas of cold-water upwelling south through central Baja California, Mexico 3. G. monttrejensis C. Hubbs North of Point Conception and in subtidal waters and areas of cold-water upwelling south through central Baja California, Mexico, including Guadalupe and San Benito islands B. Heterostichus rnstratus Girard Point Conception through central Baja California, Mexico C. Mjxodes Cuvier 1. M. viridis Valenciennes Southern Peru through central Chile 2. M. cristatus Valenciennes Central through southern Chile 3. M. ornatus Stephens Chile, range unknown and Springer D. Ribeirnclinus eigenmannt Western Atlantic from southern Brazil through central Argen- (Jordan) tina E. Clinitrachus argentatus (Risso) Mediterranean Sea

clinids. All species in the tribe Myxodini are is a Mediterranean endemic (Springer, 1970; found in shallow temperate waters of the West- Nelson, 1984; see Table 1 for summary of gen- ern Hemisphere, except for one monotypic ge- era, species, and distributions and Figure 1A nus limited to the Mediterranean Sea (Springer, for a map of their distributions). 1970; Stephens and Springer, 1973). The The Labrisomidae has been regarded as ei- Myxodini is characterized by ovuliparous ovi- ther the probable sister group to the Clinidae parity and the lack of an intromittent organ in (C. Hubbs, 1952) or a possible sister group males; both features are regarded as plesiomor- (George and Springer, 1980; Springer, pers. phies (C. Hubbs, 1952; Springer, 1970; George comm.), and 10 species of labrisomids (includ- and Springer, 1980). The other clinid tribes, ing all of the six tribes) were used as outgroups Clinini and Ophiclinini (which are primarily in the present study. Evolutionary relationships found in South Africa, temperate Australia, and of the Labrisomidae, based on allozyme and New Zealand), share the synapomorphies of ma- DNA sequence data, are further examined in tritrophy (Wourms et al., 1988) and a male in- Stepien et al. (Stepien et ai., 1992). Labrisomids tromittent organ (George and Springer, 1980). are distinguished from clinids by the following Five genera are contained in the Myxodini: morphological characters: scales with radii only Myxodes, Heterostichus, Gibbonsia, Ribeiroclinus, on anterior margins, and never small and em- and Clinitrachus. Myxodins are not found in bedded, but sometimesabsent (C. Hubbs, 1952; Central America or other tropical waters Stephens and Springer, 1973; George and (Springer, 1970; Stephens and Springer, 1973; Springer, 1980). The Labrisomidae contains 102 Nelson, 1984). Mjxodes ranges along the west species and is widely distributed throughout the coast of South America, from southern Peru New World tropics, shown in Figure 1B (Nel- through southern Chile. Heterostichus and Gib- son, 1984). A fossil labrisomid is known from bonsia share a distribution from central Baja Miocene deposits in the Mediterranean, where California, Mexico, to British Columbia, Can- the family is no longer represented. There are ada (C. Hubbs, 1952). Ribeiroclinus (monotypic) no known fossil clinids (Springer, 1970: George is found along the southwestern Atlantic coast and Springer, 1980). of South America, and Clinitrachus (monotypic) The blennioid family Chaenopsidae (tube 378 COPEIA, 1992, NO. 2 blennies) was also employed as an outgroup. 1). The diversity of sample sites examined in The Chaenopsidae is restricted to the New Stepien and Rosenblatt (1991) represented both World tropics and traditionally has been re- the centers of the ranges of each species and garded as most closely related to the neoclinin areas of infrequent occurrence and sympatry labrisomids (C. Hubbs, 1952; Stephens, 1963). with related species. Allozyme data from the Allozyme data from 40 presumptive gene loci South American myxodin M. viridis; the Aus- of all North American myxodin species and sub- tralian clinin H. whiteleggei; 10 species of labri- species, as defined by C. Hubbs (1952), were somids, including rluchenionchus tnicrocirrhis(Va- analyzed in the present study. Results from the lenciennes) (tribe Cryptotremini), Alloclinus subspecies and population analyses are present- holderi (Lauderbach) (tribe Cryptotremini), Pur- ed and discussed in Stepien and Rosenblatt aclinus integripinnis (Smith) (tribe Paraclinini), (1991). Allozyme data were also collected from P. sini Hubbs (tribe Paraclinini), Exerpes asper the South American myxodin iM. iiridis Valen- (Jenkins and Evermann) (tribe Paraclinini), ciennes and the Australian chin H. whiteleggti Starksia spinipenis (AI-Uthman) (tribe Starksiini), Ogilby. Labrisomus xanti Gill (tribe Labrisomini), Ma- There are several approaches for the phylo- lucoctenus gigas Springer (tribe Labrisomini), genetic analysis of allozyme data, including ge- .Veoclinus blanchardi Girard (tribe Neoclinini), netic distance, frequency character, discrete and Mnierpes macrocephalus (Giinther); and two character, and multistate character parsimony species of chaenopsids, Emblemaria hjpacanthus methods (Buth, 1984, pers. comm.; D. L. Swof- (Jenkins and Evermann) and Acanthemblemaria ford, pers. comm.). In the present study, four mucrospilus Brock were analyzed. different phylogenetic programs (encompassing all four methods) were used to generate and Enzyme electrophoresis. -Specimens were collect- compare trees. ed by netting intertidally with use of the anes- The relationships of the Myxodini and its sis- thetic quinaldine or subtidally by scuba diving ter group, the matritrophic tribes Clinini and and were immediately frozen and stored at -40 Ophiclinini, as well as the relationships of the C. Separate extracts of eye, liver, and muscle South and North American myxodin genera were prepared from each specimen. Tissues and the North American myxodin species, were were homogenized in a 1 : 1 volume: volume examined in the present study. Phylogenetic mixture of tissue and 0.1 M potassium phos- trees based on the allozyme data were compared phate buffer (pH 7, Waples and Rosenblatt, with those based on morphological characters. 1987) and centrifuged at 20,000 g for 10 min. Allozyme and morphological data were also The supernatant fraction was subjected to hor- combined in parsimony analyses. Several allo- izontal starch electrophoresis in 12.5% starch zyme character state synapomorphies were gels (Sigma starch; Sigma Chemical Co., St. identified which, in addition to morphological Louis, Missouri 63178). Enzymes and tissues characters, distinguish members of the family surveyed, loci scored, and buffers are given in Clinidae, the eastern Pacific members of the Stepien and Rosenblatt (1991). Staining meth- tribe Myxodini, and the North American myxo- ods and recipes were adapted from Selander et din clinids. al. (197 l),Waples (1986), and Buth and Murphy (1990). Enzyme nomenclature follows recom- MATERIALSAND METHODS mendations of the International Union of Bio- chemistry (1 984). Species and populations analyzed.-Data for the Relative migration distances were compared individual clinid subspecies and populations are with the most common allele (assigned a value compared and discussed in Stepien and Rosen- of 100) from the population of G. elegans at Bird blatt (1991), in which G. eqthra and G. norae Rock, San Diego, California, from which 63 in- were placed in the synonymy of G. montereyensis. dividuals were surveyed (Stepien and Rosen- Species, collection locations, and collection blatt, 1991). Other alleles were labeled accord- numbers of formalin-preserved voucher speci- ing to the mobility of their products relative to mens are given in Materials Examined. The this standard. North American Myxodini, as defined by Ste- pien and Rosenblatt (1991), include H.rostratus, Datu una1jsis.-Genetic distance, frequency G. metzi, G. eleguns, and G. monterejensis (Table character, discrete character, and multistate STEPIEN-CLINID EVOLUTION 379

character parsimony procedures were used to lineages. All of the clinid species and the three analyze the allozyme data in phylogenetic com- labrisomid species closest in genetic distance to puter programs to test for method-dependent the Clinidae (Auchenionchus microcirrhis, Allocli- variation in the generated trees. Different labri- nus holderz, and P. integripinnis) were analyzed. somid tribes and the two species of chaenopsids Global branch swapping and 10 separate trials were designated as the outgroup in separate, using the random-number jumble option were independent computer analyses. used in CONTML. Genetic distance clustering procedures are the A discrete character parsimony analysis using most widely used approach for analyzing allo- Phylogenetic Analysis Using Parsimony (PAUP zyme data but are also the most controversial 3.0; Swofford, 1990) was also used, in which the (Buth, 1984). Genetic distances measure the de- loci served as the characters and fixed (fre- gree of genetic divergence between each pair quencies = 1.OO) and predominant alleles (fre- of taxa. They lose information by reducing ar- quencies >0.85, >0.90, and >0.95) werecoded rays of allozyme frequency data to a single val- as discrete character states (in separate com- ue. BIOSYS-1 vers. 1.7 (Swofford and Selander, puter analyses; Tables 2-3). Using only fixed 198 1, 1989)was used to calculate modified Rog- alleles is more acceptable to some cladists who ers’ (Rogers, 1972; Wright, 1978) genetic dis- do not regard variable characters as providing tances between all pairwise combinations of taxa. proper phylogenetic data (Crother, 1990). Genetic distances were based on data from one However, this approach has the disadvantage population for each species chosen from the of discarding the information contained in al- geographic midpoint of their respective ranges. lelic variation. Allelic frequencies less than the These large population samples included H. ros- cut-off level w’ere not coded, and, if there was tratus and G. eleguns from La Jolla, California, not an allele with a given frequency at a partic- and G. metzi and G. monterepsis from San Sim- ular locus, the character state data for that spe- eon, California. Data from several other pop- cies were coded as missing. Exhaustive searches ulations for each species, representing other ar- were employed, and trees were rooted with the eas of their ranges, were analyzed in Stepien labrisomid A. microcirrhis. and Rosenblatt (1991).BIOSYS-1 (Swofford and A multicharacter state parsimony analysis us- Selander, 1981,1989) was also used to construct ing PAUP 3.0 (Swofford, 1990) was also em- distance Wagner trees (Farris, 1972),which uti- ployed, in which, as above, alleles were coded lized the modified Rogers’ genetic distances and as unordered character states (with multiple the Wagner clustering procedure. Data from character states analyzed as polymorphisms) and 10 species of labrisomids and two species of loci as the characters (Table 3). This approach chaenopsids were included in this analysis. has the advantage of including allelic variation Frequency character methods compare but, as in the discrete character analysis, dis- changes in the frequencies of individual alleles regards allelic frequency information. Exhaus- and their gain, loss, and/or fixation among taxa. tive searches were used and trees were rooted Thephylogenetic trees are based on minimizing to A. microcirrhis. the number of genetic changes between taxa. Phylogenetic hypotheses generated by the Swofford and Berlocher’s (1987, 1988) Fre- four programs were compared with each other quency parsimony (FREQPARS vers. 2.0) and and with those based on morphological data Felsenstein’s (198 1, 1984, 1988) Continuous from C. Hubbs (1952), Springer (1970), Ste- character-Maximum likelihood (CONTML in phens and Springer (1973), George and Spring- PHYLIP Phylogeny Inference Package vers. er (1980), and Stepien and Rosenblatt (1991) 3.1) methods were used. (see characters 41-53, Table 3). Discrete mor- FREQPARS generates a tree on which each phological data were analyzed using PAUP allele undergoes the least amount of frequency (Swofford, 1990), using unordered character change, while at the same time ensuring that states and an exhaustive search (rooted with the allelic frequencies in the hypothetical ancestors cryptotremin labrisomids Auchenionchus micro- add to 1.OO. This program was modified for use cirrhis and Alloclinus holderi). These morpholog- on this particular data set by D. Swofford. ical data were also added to the discrete and CONTML models evolutionary change at a lo- multistate data sets in separate PAUP analyses, cus as a Brownian motion process without re- using unordered character states and exhaus- quiring the assumption of rate uniformity across tive searches (as above) and exhaustive searches 380 COPEIA, 1992, NO. 2

TABLE2. GENOTYPICDISTRIBUTIONS OF Loci FOR CLINIDSAND LABRISOMIDS.

Number of specimens of each genotype (and relative allelic mobiliues’F spec’er sAat-A bb(090):32 bb(090):09 aa( 100):04 bb(090): 10 bb(090):21 ab(lO0):OI sAat-B aa( 100):39 aa( 100):10 cc(095):04 cc(095):43 cc(095):42 mAat-A aa( 100):25 aa( 1OO):OS aa( 100):04 aa(100):60 aa( 100):21 ab(200):O1 sAcoh-A ee( 1 15):40 dd(07 5): 04 cc( 105):04 aa(100):38 ff( 125):18 ad(075):04 Acp- 1 aa( 100):22 aa( 100):09 cc(140):04 bb(ll5):10 bb( 1 15):2 1 bc( 140):Ol Acp-A cc(090):36 cc(O90):09 cc(090):04 ee( 135):10 hh( 120):12 ef( 155):03 eh( 135):04 Adh-A aa( 100):32 aa( 100):07 aa( 100):04 bb(090):3 1 aa(100):12 Adh-B aa(lOO):30 aa(l00):OS bb(050):03 aa(100):30 aa( 100):12 bc(O70):O1 ac(070):05 ac(070):02 Ak-A bb(200):42 bb(200): 16 bb(200):04 bb(200):45 bb(200):20 bc(230):Ol Ck-A bb( 1 15):44 bb( I 15):09 cc( 105):04 cc( 105):42 cc(105):21 Ck-B bb( 105):41 cc(095):09 aa( 100):04 aa( 100):37 bb( 105):13 ab( 105):O1 Ck-c bb(1 17):42 bb( 1 17):09 cc(l25):04 cc( 125):07 dd( 140):1 3 cd( 140):O 1 Est-1 cc(O90):1 1 bb( 1 1O):OS dd( 120):04 aa( 1OO):OS aa( 100):07 ac( 1OO):O 1 ab( 1OO):O 1 Est-2 bb(090): 1 1 cc(l10):09 cc(l10):04 cc( 1 10):09 dd(l50):07 Est-3 bb(090):20 bb(090):09 dd( I 10):04 cc( 105):42 dd(085):12 Fumh-A bb(l20):33 bb( 120):09 aa(100):04 aa( 100):09 bb(120):20 ab( 100):02 ac( 150):03 G6pdh-1 bb( 120):42 bb( 120):14 bb( 120):04 bb( 120):10 bb(l20): 10 be(130):02 G6pdh-2 cc(090):37 bb(l1O):OS ee( 12 0): 04 ee( 120):10 ee( 120):10 bc(l10):02 bd(O8O):O 1 ef(090):02 Gpi-A dd(070):26 aa( 100):10 aa( 100):03 dd(070):40 aa(100):IS ae( 1 15):O1 df( 1 15):03 ad(070):02 Gpi-B bb( 1 15):1 1 aa( 100):04 aa( 100):04 dd(200):35 ee(065):30 df(2 15):05 Gtdh-A aa(lOO):42 bb(l10):09 bb( 1 10):04 bb(l10):24 bb(l10):20 ac(07 5):0 1 Gapdh-A aa( 10 0):4 3 aa(100):09 cc(200):03 cc(200):45 dd(080):21 bc( 120):O 1 ad( 100):02 Gapdh-C aa( 100):42 bb(120): 10 bb(120):04 bb( 120):40 bb(120):20 ab( 120):O 1 G3pdh-B ee( 120):27 bb(080): 14 ff(060):03 ff(060):42 gg( 1 15):07 af(100):Ol gf(060):Ol Iddh-A bb(060):OS cc( 125):04 cc(125):04 dd(200):20 ee( 106):06 ab( 1OO):O 1 de( 105):02 ce( 125):02 sldh-A cc(095):39 bb(l15):09 bb( 1 15):04 bb( 1 15):42 bb( 1 15):05 bc(095):02 mldh-A dd( 1 10):40 cc( 120):07 cc(l20):04 cc( 120):25 cc( 120):14 Ldh-A dd(080):43 aa( 100):05 aa( 100):04 aa( 100):45 ee( 120):10 ac(090):O 1 Ldh-B cc( 105):42 aa( 100):07 aa( 100):04 aa( 100):32 aa( 100):12 Ldh-C aa( 100):24 aa( 1OO):OS aa( 100):04 aa(100):45 aa( 100):12 STEPIEN-CLINID EVOLUTION 38 1

TABLE2. CONTINUED.

Number of rpenmenr of each genotype T species (and relative allelic rnobiliriesl~

sMdh-A aa(100):42 cc(l 10):14 bb(070):02 aa( 100):10 ee(095):06 ac( 1OO):O 1 ab(100):02 sMdh-B dd(l10):30 bb(090):07 ff(060):04 cc(080):40 gg(105):lO bd(090):03 be(070):Ol Mpi-A aa( 100):31 aa( 100):04 bb(085):04 cc( 120):05 cc(120):05 ac( 100):03 PepA dd(080):44 aa( 100):13 ee(l15):03 dd(080):25 dd(080):03 ad(080):O1 de(080):O 1 ad( 100):03 df(085):03 PepB cc(095):30 bb( 120):12 bb(120):04 cc(095):40 aa( 100):22 cd(l10):05 ad( 1 10):02 Pep-3 cc(050):28 bb( 125):12 bb(125):04 bb( 125):40 ee(035):07 cd(080):O 1 bc(050):02 ef(070):02 Pgm-A cc(070):41 bb(l10):06 bb( 1 10):04 ee(080):39 ee(080):20 bd(050):O 1 ef(060):03 Pgdh-A cc(120):37 dd(080): 10 cc(120):04 cc( 120):45 cc(l20):21 sSod-A dd(030):41 ee( 120):06 ff(060):04 ee(120):40 hh(110):19 eg(145):lO fh(050):Ol Xdh-A cc(080):40 bb(l10):09 cc(080):04 cc(080):20 bb(l10):08 ac( 1OO):O 1 bc(080):02

Relative allelic mobilities were measured against the standard of the most common allele in the population of Gibbonno elcganr from La Jolla. California (see Materials and Methods). For heterozygotes. the relative allelic mobility of the rarer allele is given in parentheses.

(Table 3). Strict (Rohlf, 1982), Adams (1972, the chaenopsids and remaining labrisomids are 1986), and 50% majority rule (Rohlf, 1982) con- given in Stepien et ai. (1992). Zymograms illus- sensus trees were run on the shortest trees from trating similarities and differences between taxa the discrete and multistate PAUP analyses, and are given in Figure 2. Genetic distances for all 1000 bootstrap replications (Felsenstein, 1985) individual population samples of the North on the data sets were performed using the American chid taxa are presented and ana- branch-and-bound algorithm (Hendy and Pen- lyzed in Stepien and Rosenblatt (1 99 l). Popula- ny, 1982). Consistency indices (C.I.; Swofford, tions sampled in Stepien and Rosenblatt (1991) 1990), g-1 skewness statistics (Sokal and Rohlf, were in Hardy-Weinberg equilibrium and were 1981; Hillis, 1991; Huselsenbeck, 1991), and thus pooled in the present study. lengths of most parsimonious trees are reported All methods of analysis of allozyme data, in- for all PAUP analyses. The g-1 statistic mea- cluding frequency character, genetic distance, sures degree of skewness of tree length distri- discrete character, and multistate character butions, providing a test of the relative strength parsimony approaches yielded one or both of of the phylogenetic signal (Hillis, 199 1 ; Husel- two most parsimonious trees. These two trees senbeck, 1991). were identical except among relationships with- in the genus Gibbonsia (Fig. 3) and were obtained RFSULTS when the trees were rooted with different spe- cies of labrisomids and chaenopsids. Allozyme data for 40 presumptive gene loci The two most parsimonious phylogenetic trees from the clinids H. rostratus, G. elegans, G. metzi, obtained from the various types of data analyses and G. monterejensis are given in Stepien and are presented in Figure 3A-B. Branches and Rosenblatt (199 1) and those for M. viridis, H. nodes are lettered, and allozyme character state whiteleggei, and the labrisomids Auchenionchus synapomorphies and autapomorphies delin- microcirrhis, Alloclinus holderi, and P. integripinnis eating taxa at each of these, for both trees, are are summarized in Table 2. Allozyme data for given in Tables 4 and 5. Possible homoplasies 382 COPEIA, 1992, NO. 2

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1 2 j 4 56 78 91Oi1121314 15 7 234567 39 10 aa abbbcccdddee e aaerhdc cd c 5. Pep-A and Pep- 3 D sMdh-A Fig. 2. Examples of zymograms of isozyme expressions from tissue extracts from chid and labrisomid taxa. o = origin of gel. Taxa are lettered as follows: a = Heterostichus rostratus, b = Gibbonsia elegans, c = Gibbonsia metzi, d = Gibbonsia monterejensis, e = Mjxodes yiridis, f = Heterociinus whiteleggei, and g = Auchenionchus mtcrocirrhzs. Other loci are visible on some of the photographs, e.g., sIdh-A on gel C and sMdh-B on gel D. Heterozygotes include gel A, fish 3-a, alleles a/b; gel B, Pep-A (top bands), fish 12-d, alleles b/d; Pep-3 (lower bands), fish 13-e, alleles d/c; and gel D, fish 6d,alleles a/b. of the two trees are compared in Table 5. Mor- (Felsenstein, 1988), a frequency character ap- phological synapomorphies at each branch are proach, yielded a single tree (Fig. 3A) in all 10 summarized in Table 3 and the Discussion. independent analyses. Gibbonsia elegans is de- Goodness-of-fit statistics for the Wagner dis- picted as the basal clade within the genus and tance tree varied with the number of taxa in- is the sister group to G. monterejensis and G. cluded (number of labrisomid tribes and inclu- elegans. This tree had Ln likelihood values (Fel- sion of the Chaenopsidae). Cophenetic senstein, 1988) of 127.26. correlation (Swofford and Selander, 198 1) val- Frequency parsimony (Swofford and Berloch- ues ranged from 0.87, when all 10 labrisomid er, 1988) yielded two most parsimonious trees species were included, to 0.93 when rooted to (Fig. 3A-B), which differ in whether G. metzi the two most similar labrisomid species (Au- (tree A) or G. elegans (tree B) is the basal clade chenionchus microcirrhis and Alloclinus holden], and in the genus. The lengths of these trees are 0.99 when only the eastern Pacific myxodin 134.44 and 134.65, respectively. The next clinid species were analyzed. Cophenetic cor- shortest tree places G. montereyensis as the basal relation of the three family data set (Clinidae, clade in the genus and has a length of 136.14. Labrisomidae, and Chaenopsidae) was 0.80 The shortest tree (Fig. 3A) obtained with Fre- (Stepien et ai., 1992). quency parsimony was thus identical to the only The tree derived from the Wagner genetic tree obtained with Continuous character-Max- distance method (Fig. 3B) is identical to the most imum likelihood. parsimonious tree from the frequency charac- Analysis of fixed character state data (allelic ter programs (Fig. 3A) in all but the last two frequencies = 100%) using PAUP 3.0 (Swof- branches (the relationships among species in the ford, 1990) yielded three most parsimonious genus Gibbonsia). The North American chid trees, which included each of the three species taxa are separated into two lineages, the genera of Gibbonsia as sister to the other two (57 steps, Gibbonsia and Heterostichus. C.1.s = 0.93, and g-1 skewness = -1.20 with Continuous character-Maximum likelihood allozyme data only and 74 steps, C.1.s = 0.91, STEPIEN-CLINID EVOLUTION 385

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e m -

-mY

and g-1 skewness = - 1.28 with both allozyme taxa (characters 41-53 of Table 3) were also and morphological data included). At the 0.95 analyzed separately using PAUP. Two equally cut-off level of allelic frequency, trees 3A and parsimonious trees were obtained (15 steps, C.1.s 3B were equally parsimonious (65 steps, C.1.s = = 0.93, g-1 skewness = - 1.12)-tree 3A and a 0.92, and g-1 skewness = -1.12 with allozyme tree with the positions of H. rostrutus and M. data only and 82 steps, C.1.s = 0.90, and g-1 uiridis reversed. A bootstrapping consensus tree skewness = - 1.16 with both allozyme and mor- using branch-and-bound (Hendy and Penny, phological data). At the 0.90 cut-off level, tree 1982), 50% majority-rule, and 1000 replica- 3B was the most parsimonious (78 steps, C.I. = tions resulted in confidence intervals of 92% for 0.89, and g-1 skewness = -0.92 with allozyme node “e” (of Fig. 3A), 45% for node “g,” 68% data only and 95 steps, C.I. = 0.91, and g-1 for node “i,” and 87% for node “k.” skewness = -0.98 with both allozyme and mor- Multistate character PAUP analysis of allo- phological data). Tree 3B was also the most par- zyme data (see Table 3) resulted in three most simonious at the 0.85 cut-off level (85 steps, C.I. parsimonious trees, as in the 100% fixed = 0.93, and g-1 skewness = -0.79 with allozyme discrete analysis (63 steps, C.1.s = 0.92, g-1 data only and 102 steps, C.I. = 0.91, and g-1 skewness = -0.95). Bootstrapping with branch- skewness = -0.87 with inclusion of morpho- and-bound (Hendy and Penny, 1982), 80% ma- logical data). jority-rule, and 1000 replications yielded con- Discrete morphological characters for these fidence intervals (Felsenstein, 1985) of 98% for 386 COPEIA, 1992, NO. 2

TABLE4. ALLOZYMECHARACTER STAT= LINKINGAND DISTINCU~SHINGTAXA AT BRANCHESAND NODESA THROUGH F OF TREESSHOWN IN FIGURE3A-B.

Branch or node

~ ~ ~~ a b C d e f sAat-A(a) sAat-B(c) sAat-A(b) *sAat-A(a) Est-B(b) m Aat-A(b) m Aat-A(a) AcP- sAat-B(a) sAcoh-A(d) *Est-2(c) sAcoh-A(e) AcP-.~(c) Adh-B(b) m Aat-A(b) *Ck-B(a) Gadph-C(a) *Ck-B(a) Adh-A(a) .i\dh-B(c) Acp-1 (a) Ck-B(c) Gtdh-A(a) Ck-B(b) Ak-A(b) sAcoh-A(c) Adh-B(a) Est- 1(b) *Gtdh-A(b) Est-I (c) Ck-B(a) Ck-A(c) Ck-A(b) *Fumh-A(a) Gpi-A(d) *G3pdh-B(a) Est-Z(c) Ck-C(c) Ck-C(b) *G3pdh-B(a) *mIdh-A(c) GSpdh-B(e) Fumh-A(a) Est-1 (d) Est- 1(a) GJpdh-B(c) Iddh-A(a) G6pdh-2(~) Gapdh-C(b) Est-3(d) Est-S(b) GGpdh-P(d) *Iddh-A(c) Gtdh-A(c) GSpdh-B(a) Gapdh-A(b) Fumh-A(b) Ldh-A(c) Ldh-A(d) *Gpi-A(a) GGpdh-l(b) Gapdh-A(c) Gapdh-A(a) sMdh-A(c) sMdh-B(d) Gpi-A(d) Gtdh-A(b) GSpdh-B(t) GGpdh-P(b) sMdh-B(e) *Pep-A(d) *Gpi-B(a) Gpi-A(a) GGpdh-P(e) sMdh-B(b) *Pep-A(a) PegW Gpi-B(b) Gpi-B(a) GGpdh-P(f) Mpi-A(a) *Pgdh-A(c) Xdh-A(a) *sIdh-A(b) sIdh-A(b) Gpi-A(e) Pep-A(a) Pgdh-A(d) sIdh-A(c) mldh-A(c) sMdh-A(b) PeP-3(4 Pgm-A(d) mIdh-A(d) Iddh-A(c) sMdh-B(f) sSod-A(e) Iddh-A(b) Ldh-A(a) Mpi-A(b) Xdh-A(b) *Ldh-A(a) Ldh-B(a) PeP-A(e) *Xdh-A(c) Ldh-A(d) Ldh-C(a) sSod-A(f) * Ldh-B(a) sMdh-A(a) Ldh-B(c) Pep-A(d) * Pep-A(a) PeP-B(b) * PepB(b) Pep3(b) * Pep3(b) Pgdh-A(c) PeP-J(d) Pgm-A(b) * Pgm-A(b) Xdh-A(c) Pgm-A(c) sSod-A(d)

* = Character state lor= node “e,” 92% for node “g,” and 87% for node “g,” ‘5,” and “k” (18 vs 16 at node “g,” 15 vs. “i” (of trees in Fig. 3). Tree 3B was the most 16 at node “i,” and 1 vs 2 at node “k”). Tree parsimonious obtained with a combination of 3A has six apparent homoplasies and 67 char- morphological and multistate allozyme data acter state losses, and tree 3B has five apparent (characters 1-53 of Table 3: 102 steps, C.I. = homoplasies and 68 character state losses, listed 0.91, g-1 skewness = -0.87). Bootstrapping with in Table 5. branch-and-bound (Hendy and Penny, 1982), 80% majority-rule, and 1000 replications yield- DISCUSSION ed confidence intervals (Felsenstein, 1985) of 99% for node “e,” 99% for node “g,” and 92% Four different approaches for the phyloge- for node “i” (of trees in Fig. 3). netic analysis of allozyme data yielded parsi- In all PAUP analyses, strict (Rohlf, 1982), monious relationships that were identical, ex- Adams’ (1972, 1986), and 50% majority-rule cept within the genus Gibbonsia (Fig. 3). This (Rohlf, 1982) consensus analyses of the shortest set of phylogenetic hypotheses is further sup- trees yielded unresolved trichotomies for the ported by the known morphological characters genus Gibbonsia. Allozyme character state syn- distinguishing these taxa and largely corrobo- apomorphies linking taxa at the branches of the rates the relationships proposed by C. Hubbs two trees are compared in Tables 4 and 5, which (1952), Penrith (1969), Springer (1970), and show that trees 3A and 3B are supported by George and Springer (1980). Analysis of allo- equal numbers of total synapomorphies at nodes zymes yielded a large data set that provided an STEPIEN-CLINID EVOLUTION 387

TABLE5. ALLOZYMECHARACTER STATES LINKINGAND DISTINGUISHINGTAXA AT BRANCHESAND NODESG THROUGH M OF TREESSHOWN IN FIGURE3A VERSUS 3B.

g h j k I m Tree A *sAat-A(b) xs Aat-B(c) *Acp-A(c) *sAat-B(b) *Acp-1 (b) xrnAat-A(a) *sAat-B(a) sAat-B(b) *Acp- 1 (a) Acp-A(d) *Acp-1(a) xG3pdh-B(c) *rnAat-A(b) *Adh-A(b) *rnAat-A(a) AcP=W Ak-A(a) sAcoh-A(f) GSpdh-B(d) sAcoh-A(c) *Ck-C(a) sAcoh-A(a) *Ck-C(b) *Ak-A(b) *Adh-A(b) *Mdh-B(d) *Ck-C(b) *Est-3(~) s Acoh-A(b) Fuhrn-A(a) Ck-A(a) *Ck-C(a) *Pep-B(b) *mIdh-A(b) *Gpi-A(b) Acp- 1(b) *G3pdh-B(b) *Cli-A(b) *G3pdh-B(b) *Pgrn-A(b) xGpi-A(d) *Gpi-A(c) AcP-A(a) *Gpi-A(d) Est-2(a) Gpi-A(b) sMdh-B(c) XMdh-A(b) AcP-A(b) Ldh-A(b) *Est-2(b) *rnIdh-A(a) *PepB(c) *Pep-A(a) Adh-A(b) Ldh-B(b) Est-3(a) *Pep-A(a) xsSod-A(e) * Pep-B(a) Ck-C(a) *sMdh-B(d) Est-3(~) xPep-3(d) Gapdh-C(a) Pep-A(c) Gpi-A(c) *Pgdh-A(b) * Ga pd h-C@) *Pep-B(c) Gpi-B(c) GGpdh-1 (a) sSod-A(b) mIdh-A(b) *GGpdh-l(b) *sIdh-A(b) GGpdh-2(a) Pep-A(b) *CGpdh-2(b) Pep-B(a) Gpi-A(b) Pep-S(a) sIdh-A(a) * Pep3(b) rnIdh-A(a) Pgrn-A(a) sMdh-B(a) sSod-A(a) * Pep-A(d) sSod-A(c) *Pep-3(c) Pgdh-A(a) Pgdh-A(b) Pgdh-A(c) *Xdh-A(c) Tree B *sAat-A(b) xsAat-B(c) *AcpA(c) *mAat-A(a) *mAat-A(a) *sAat-B(b) *sAat-B(a) sAat-B(b) *mAar-A(a) AcP-A(d) sAcoh-A(c) *Adh-A(b) sAcoh-A(f) *sAcoh-A(c) s Acoh-A(a) *Acp-](a) Ak-A(a) *Acp- 1(b) *Ck-C(a) *Acp-l(a) *Acp-l(b) sAcoh-A(b) Acp-A(b) *Ak-A(b) *Ck-C(b) Gpi-A(d) *G3pdh-B(b) *Est-3(c) Acp-l(b) * C k-C(b) Ck-A(a) sMdh-B(c) rnIdh-A(b) *GSpdh-B(c) *Gpi-A(b) AcF- 44 *Fumh-A(a) *Ck-A(b) *sMdh-B(d) *Pep-A(a) *GBpdh-B(d) *Gpi-A(c) Adh-A(b) *G3pdh-B(b) Est-2(a) * Pep-B(b) *rnldh-A(a) xsMdh-A(b) Ck-C(a) *Gpi-A(d) *Est-2(b) *Pep-B(c) *sMdh-B(d) Gapdh-C(a) Ldh-A(b) Est-3(a) * Pgm-A(b) *PepB(a) "Gapdh-C@) Ldh-B*b Est-3(~) xsSod-A(e) *PegB(b) GGpdh-l(a) *Mdh-B(d) GSpdh-B(b) xPep-3(d) *GGpdh-I(b) *PepA(a) xGSpdh-B(c) *Pgdh-A(b) GGpdh-2(a) PepA(c) GSpdh-B(d) *Pgm-A(b) *GGpdh-2(b) * PepB(c) Gpi-A(c) Gpi-A(b) sSod-A(b) Gpi-B(c) sldh-A(a) *sIdh-A(b) mIdh-A(a) PeP-A(b) sMdh-B(a) Pep-B(a) *Pep-A(d) Pep-3(a) *Pep-3(c) *Pep-3(b) Pgdh-A(a) Pgm-A(a) Pgdh-A(b) sSod-A(a) *Pgdh-A(c) sSod-A(c) *Xdh-A(c)

* = Character state losses. x = Possible homoplasy of eleclromorphs with the same net electric charge 388 COPEIA, 1992, NO. 2

independent test for the identification of syn- The subfamily Clininae (tribes Clinini and apomorphies and monophyletic relationships Ophiclinini; C. Hubbs, 1952) is distinguished among the Clinidae. from the tribe Myxodini by the synapomorphies The family Labrisomidae, which was tested of matritrophy and a male intromittent organ as the sister group in the present study, was (characters 44 and 45 in Table 3). Present-day recently distinguished from the Clinidae by distributions of the matritrophic tribes and the George and Springer (1980). Labrisomids are Myxodini do not overlap (Fig. 1 A). The present widely distributed throughout the New World study supports the monophyly of each group tropics and are also represented by two species and the hypothesis that they have long been in the eastern Atlantic, on the coast of Africa separated, proposed by C. Hubbs (1952) and (Nelson, 1984; see Fig. 1B). They are distin- Penrith (1 969). guished from the Clinidae by a few morpholog- Myxodins examined in the present study share ical characters (see Introduction), and results of nine allozyme character state synapomorphies the present study support their familial sepa- (node “e” of Fig. 3 and Table 4). Sister rela- ration. tionships of the North and South American Results of the present study suggest that the Myxodini suggest that they may have shared a cryptotremid labrisomids, including the South continuous ancestral distribution, extending American Auchenionchus microcirrhis and the through the tropics and were later separated by North American Alloclinus holderi are sister to the formation of a barrier to dispersal and gene the Clinidae (Table 3) and indicate that the Lab- flow, which led to genetic divergence (a vicar- risomidae may be paraphyletic (Stepien et al., iant hypothesis; see Humphries and Parenti, 1992). In the present study, the cryptotremin 1987). A vicariant event which may explain this labrisomid Auchrnionchus microcirrhis and clinids antitropical pattern of relationship and distri- share 27 allozyme character state synapomor- bution is the Miocene warming of the tropics phies, listed in Table 4. DNA sequence data proposed by White (1986) in his analysis of the from nuclear ribosomal and mitochondrial DNA silverside fish subfamily Atherinopsinae. Alter- from additional blennioid families and tribes natively, such antitropical relationships have (including the labrisomid tribes) are being used traditionally been explained by long-distance to further elucidate these relationships (Stepien dispersal (C. L. Hubbs, 1952). et al., 1992; Stepien, unpubl.). Briggs’ (1 974, 1987) competitive exclusion Allozyme data from the clinid species ana- hypothesis offers another explanation for these lyzed in the present study support Springer’s antitropical biogeographic patterns. He regards hypothesis (Springer, 1970; George and Spring- antitropically distributed taxa as older, relict er, 1980) that the Clinidae is monophyletic. Ri- groups which were displaced from the tropics bosomal DNA sequence data from the chid by more recently evolved and competitively su- tribes Clinini and Myxodini and the families perior species (Briggs, 1974, 1987; Newman and Labrisomidae, Chaenopsidae, Tripterygiidae, Foster, 1987). and Blenniidae also support monophyly of the Myxodes viridis and the North American Clinidae (Stepien et al., 1992). The present study myxodins share the behavioral synapomorphy identifies 16 allozyme character state synapo- of depth segregation of mature males and fe- morphies that unite the clinids examined (see males, males being found deeper (character 46 branch “c” of Fig. 3A and Table 4). In addition of Table 3; Williams, 1954; Stepien, 1987; Ste- to the morphological synapomorphies previ- pien et al., 1988). The North American Myxo- ously mentioned (see Introduction and char- dini are united by 16 allozyme character state acters 41 and 42 of Table 3), all clinids whose synapomorphies and are unequivocally dem- color patterns have been described share simi- onstrated to be a monophyletic clade, as all spe- liar variable patterns, unlike most labrisomids cies and subspecies were analyzed in the present (E. asprr, however, occurs in red and green study (node “g” of Fig. 3 and Table 5). Unlike morphs; C. Hubbs, 1952). These clinids are ca- M ziridis (Stepien, 1990), the North American pable of slow, extensive color changes with species share the synapomorphies of sexual dif- background (character 43, Table 3), which in- ferences in size, adult females attaining greater volve gain and loss of carotenoid pigments, and lengths than adult males (character 47 of Table often occur in red, green, and brown morphs 3; Stepien, 1986b, 1987; Stepien et al., 1988). that match their plant habitats (Stepien 1986a, Heterostichus rostratus is distinguished from the 1987, 1990; Stepien et al., 1988). other northern myxodins by the morphological STEPIEN-CLINID EVOLUTION 389

apomorphies of a pointed snout and a forked ature boundary at Point Conception is the most caudal fin, in addition to five allozyme character important geographic feature affecting the state apomorphies (Table 5). The genus Gib- distributions of the California ichthyofauna, bonsia is a monophyletic clade defined by a suite separating the warm-temperate Californian of 15 allozyme character state synapomorphies biogeographic province to the south from the (node “i” of Fig. 3 and Table 5). Its reduced cold-temperate Oregonian province to the north dorsal and anal fin and vertebral elements are (C. L. Hubbs, 1952; Horn and Allen, 1978; Da- morphological synapomorphies further distin- vis et al., 198 1 ; Waples and Rosenblatt, 1987), guishing it from Heterostichus. which is also shown in clinid distributions (Ste- The phylogeny in which G. elegans is the sister pien and Rosenblatt, 1991; Stepien et al., 1992). species of the other two species in the genus The three species of Gibbonsia have been (Fig. 3B) contains one less apparent allozyme shown to differ significantly in temperature tol- character state homoplasy than the tree de- erances (Davis, 1974, 1977), which appears to picting G. metri as the basal clade (Fig. 3A; see be directly related to their present-day ranges. Table 5 and Results). However, morphological Although G. monterejensis and G. metzi appear characters distinguishing the species of Gibbon- capable of dispersal and a considerable degree sia best support tree 3A (Table 3). Gibbonsia of gene flow between populations north of Point montereyensis and G. elegans share the apparent Conception, California, and off Baja California, synapomorphies of reduced fin and vertebral Mexico, has been shown (Stepien and Rosen- counts and are smaller in total length than G. blatt, 1991), these species are restricted to cool- metii (C. Hubbs, 1952). Gibbonsia metii has rel- er waters throughout their ranges. Davis (1974, atively equally spaced posterior dorsal soft fin 1977) found that G. metzi has the widest tem- rays and scalation extending onto the caudal perature tolerance of the three species but has fin. This scale pattern is shared with G. elegans, a lower critical thermal maximum than G. ele- whereas G. rnonterqrensis exhibits the apomor- guns. Gibbonsia elegans is more tolerant of the phic condition of a naked caudal fin (C. Hubbs, warmer temperatures characteristic of its pres- 1952; Stepien and Rosenblatt, 1991). The latter ent-day range and least tolerant of cooler tem- two species also share the synapomorphies of peratures. unequally spaced posterior dorsal fin rays and Speciation in the North American Mvxodini the presence of prominent dorsal ocelli on the may be related to shifts in coastal temperatures body (characters 48 and 49 of Table 3; C. Hubbs, during the Pliocene and early Pleistocene, which 1952: Stepien and Rosenblatt, 1991). may have served as vicariant events. In the hy- Gibbonsia elegans and G. monterepsis share one pothesized relationships shown in the tree in allozyme character state synapomorphy at Figure 3A, the first event may have separated branch “k” of the tree shown in Fig. 3A. Gib- the ancestral North American clinid stock into bonsia elegans is distinguished from G. rnonterej- two groups: Heterostichus rostratus subtidally in ensis by two allozyme character state apomor- the warmer waters south of Point Conception, phies (Table 3, branch “I” of Table 5.). California, and G. metzi intertidally to the north. Heterostichus and Gibbonsia have planktonic A second temperature-related event may have larval stages lasting approximately two months resulted in another north/south separation of (Stepien, 1986b). The late larval stages, as well ancestral Gibbonsia, G. elegans in southern wa- as juveniles and adults, are thigmotactic and ters and G. montereyensis in the northern waters. have been known to raft long distances from Variation in coastal current and temperature shore with pieces of drift algae. They thus ap- patterns may maintain gene flow between pop- pear to be capable of some relatively long-dis- ulations semi-isolated by temperature barriers tance dispersal. Gibbonsia metzi and G. monte- (Stepien and Rosenblatt, 1991). There is strong rejensis share primary distributions north of support for dispersal and gene flow between Point Conception (latitude 34.5”N),California, widely separated populations of clinids, which reappearing in northern Baja California, Mex- are apparently due to their relatively long larval ico, in areas of nearshore cold-water upwelling period, although some clinal variation is also (C. Hubbs, 1952: Stepien and Rosenblatt, 1991). evident. Significant differences in temperature Heterostichus rostratus and G.elegans share a com- tolerances between the species of Gibbonsia (Da- mon distribution, primarily south of Point Con- vis, 1977) and the reappearance of northern ception through central Baja California. Data California clinids in areas of cold-water up- from several studies indicated that the temper- welling off the coast of Baja California, Mexico 390 COPEIA, 1992, NO. 2

(C. Hubbs, 1952: Stepien and Rosenblatt, 1991; substantially from critical reviews by R. H. Ro- Stepien et al., 1991), indicate that distributions senblatt, D. M. Hillis, V. G. Springer, R. c. of these species are determined by temperature. Brusca, D. C. Cannatella, J. T. Williams, and B. Also, El Niiio patterns may periodically restrict R. Crother. D. G. Buth, C. Hubbs, W. A. New- the ranges of the cooler-water species (G. rnetzi man, M. H. Horn, G. C. Wilson, R. R. Wilson, and G. monttrejensis) and expand those of the Jr., and R. J. Lavenberg also contributed valu- warmer-water species (G. elegans and H. rostra- able suggestions in discussions of this work. I tus). It is thus likely that speciation events in the especially thank D. L. Swofford, D. G. Buth, Clinidae are related to coastal temperature pat- and G. C. Wilson for help with the various com- terns. puter analyses and R. H. Rosenblatt, R. R. Wil- son, Jr., and R. S. Waples for assistance with electrophoretic techniques. R. S. Waples con- MATERIALSEXAMINED tributed some allozyme data for clinids from Guadalupe Island and some of the labrisomids, SPPOPIuvd /or olla:?me onalw$and voucher numbers -HeLero.hrhu~ TO,- lrnlus 35 from Mission Bay. San Diego, California (SI0 90-80). IO from which were also done in the laboratory of R. Santa Catalina Island. California (SI0 90-81). 6 from Puma Clara. Baja H. Rosenblatt. A. Wheeler, R. J. Lavenberg, California. Mexico (SI0 90-82). 5 from the San Benito Islands. Baja W. N. Eschmeyer, and V. G. Springer lent study California. Mexico (SI0 90-83). and 6 from Guadalupe Island. Baja Califorma, Mexico (SI0 90-84): Gibbonria rlqoganr 63 from Bird Rock, specimens. The following persons helped to col- La Jolla, San Diego, California (SI0 90-85). IO from Punta Clara, Baja lect specimens: R. McConnaughey, R. Rosen- California. Mexico (SI0 90-82). IO from Santa Barbara. California (SI0 90-87). 3 from San Sirneon. California (SI0 90-88). IO from blatt, K. Dickson, R. Lea, S. Naffziger, D. Hoese, Middle Coronado Island. northern Baja California, Mexico (SI0 90- R. Thresher, T. Gill, R. Waples, V. Radeke 89). 4 from Pelican Bay, Santa Cruz Island. California (SI0 90-90). 20 Clark, M. Brooks, R. Snodgrass, S. Hendrix from the San Benito Islands, Baja California. Mexico (SI0 90-83) and 13 from Guadalupe Island, Baja California. Mexico (SI0 90-84): Gib- Kramer, A. Block, N. Jones, L. Bookbinder, J. bonsta ~CI:I 14 from Soberanes Point. Carmel, California (SI0 90-91). O’Sullivan, S. Anderson, J. Adler, G. Rosen- 55 from San Sirneon. California: and 21 from Puma Clara, Baja Cali- fornia. Mexico (SI0 90-82): Gibbonsio monlrrr2pnrir IO from Soberanes blatt, L. Fullan, A. Fink, K. Verhoffstadt, R. Point. Carmel. California (SI0 90-82). 38 from San Smeon, California Martin, P. Hedberg, R. Pawlan, M. Burkholder, (SI0 90-88). 2 from Sanra Barbara. California (SI0 90-87). 4 from L. Badzioch, R. Schilling, C. Jones, C. Zucca, middle Coronado Island, Baja California. mexico (SI0 90-89). 18 from Punta Clara. Baja California, Mexico (SI0 90-88). and I2 from Gua- D. Perry, F. Balbontin, and F. Alcazar. dalupe Island. Mexico (SI0 90-84): Mpdrr wndia 46 from Montemar, Viia del Mar. Chile(SI0 87-132): Hprprorlinur~l.hil~l~~~~,17 from Peb- ble Beach.Sydney. hustralia(S1090-158). Ten labrisamidrpecies were employed as outgroups. including Alloclznus holdrri 38 from Santa Cat- LITERATURECITED ahaIsland. California, 6 from the Coronado Islands. Baja California. Mexico. and 2 from La Jolla, San Diego. California (SI0 90-159): ADAMS,E. N., 111. 1972. Consensus techniques and Pororhnus tnkgnpznnrs 21 from Bird Rock. La Jolla. San Diego. Cali- the comparison of taxonomic trees. Syst. 2001. 21: fornia (SI0 90-157): .4uchmionrhur mtcroorrhzr 5 from Montemar. Viia del Mar. Chile (SI0 87-139): Paraclinus sm12 from San Carlos Bay. 390-397. Gulf of California. Mexico (SI0 90-160): SIorksra sptnipnu 7 from the -. 1986. N-trees as nestings: complexity, simi- Gulf of California, Mexico: EXPT~PSnrprr 7 From Kino Ba) in thc Gulf larity, and consensus. J. Classif. 3:299-3 17. of California. hlexico (SI0 90-160); Lobniomur xnnli IO from Puerto Priaxo. Mexico (SI0 90-162); Maloroclrnur hubbsi IO from Puerto BRIGGS,J. C. 1974. Marine zoogeography. McGraw- Peiaxo. Gulf of California. Mexico (SI0 90-162); .Vwrlinur blonchardi Hill, New York, New York. 2 from La Jolla. San Diego. California (SI0 90-161) and 9 from Mon- -. 1987. Antitropical distribution and evolution terey. California, and Mnirrptr macrocephalus 5 from Azuero Peninsula, in the Indo-West Pacific Ocean. Syst. Zool.36:237- Panama. Twospecies of chaenopsids were also employed as outgroupr. including Acanlhrmblrmann macrorphr 4 from central Gulf of Califor- 247. nia. Mexico and Emblrmana hjpacanlhur 4 from Puerto Peiaxo, Gulf BUTH, D. G. 1984. The application of electropho- of California. Mexico. retic data in systematic studies. Ann. Rev. Ecol. Syst. 15:502-522. -, AND R. W. MURPHY.1990. Appendix 1: En- ACKNOWLEDGMENTS zyme staining formulas p. 99-126. In; Molecular systematics. D. M. Hillis and C. Moritz (eds.). Sin- This research was supported by a National auer Associates, Sunderland, Massachusetts. Science Foundation postdoctoral fellowship in CROTHEX,B. I. 1990. Is “some better than none” or Systematic Biology, Biotic Systems and Re- do allele frequencies contain phylogenetically use- sources, grant #BSR-8600180 and was con- ful information? Cladistics 6:277-281. DAVIS, B. J. 1974. Temperature adaptations, and ducted in the laboratory of R. H. Rosenblatt. distributions, in the fish genus Gibbonsia. Unpubl. Collections of Chilean fishes were supported by Ph.D. diss., Univ. of California, San Diego. National Geographic grant #36 15-85 and the -. 1977. Distribution and temperature adap University of California Research Expeditions tation in the teleost fish genus Gibbonsia. Marine program (UREP). This manuscript benefited Biol. 42:315-320. STEPIEN-CLINID EVOLUTION 39 1

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