Allozyme variation and evolutionary relationships of grain amaranths (Amaranthus spp.)

Summary Allozyme studies in amaranth provided useful assays of genetic variation in order to verify the patterns inferred from morphological traits, for elucidating the genetic structure of landraces, and for the studies of evolutionary relationships among wild, weedy and crop species. Thirty-four populations of cultivated New World amaranths were surveyed along with 21 weedy New World populations for allozyme variation at nine electrophoretic enzyme loci. Eleven populations of cultivated amaranths from the Indian State of Uttar Pradesh and six from Nepal were also surveyed for a comparison. In the New World populations, heterozygosity was low, and different populations ranged from 0 to 44% polymorphic loci. Adjacent populations were often fixed for different alleles or had very different allele frequencies at certain loci, with no apparent geographical patterns. Diversity index H was partitioned into the intra- and interpopulation as well as the interspecific components of variability. The crop versus weed genetic distances were the largest, whereas the intra- and interpopulation components of H were about equal. Genetic structure of all three species of the New World amaranths together can be described as a collection of distinct populations, each more or less a heterogeneous collection of highly homozygous individuals. The North Indian populations showed relatively less allozyme variability with the most common alleles same as those of Mexican landraces. Alleles at several loci proved to be diagnostic of the crop and weed groups, and of the three individual crop species. Genetic distances based on pooled gene frequencies showed the three crop species to be generally more closely related inter se than they were to their putative weedy progenitor species, respectively (with the exception of the weed-crop pair A. quitensis and A. caudatus). This implies a single domestication event involving A. hybridus as the common ancestor rather than three separate domestication events. Close similarity between A. caudatus and A. quitensis might have resulted from transdomestication based on a weedy or semi-domesticated species having migrated from Meso-America to South America. This preliminary report must now be expanded by further ecogeographical, cytogenetic and population studies on new extensive collections from the areas of early domestication. Some evidence of recent introgression and/or segregation of crop-weed hybrids between A. caudatus and A. retroflexus is available in the form of rare individuals in crop populations with crop allozyme genotypes except for a single homozygous weedy allele.     

Genetic and morphological variation in the venus clam Cyclina?sinensis along the coast of China

Six geographic samples of Cyclina sinensis were collected from the coast along China and analyzed to reveal morphological and genetic variation by using nine allozyme loci and 11 morphological variables. The discriminant function analysis (DFA) of morphology suggested a clear separation between the southern and northern populations. Polymorphism was detected at nine loci across all six populations. The mean allele number ranged from 2.44 to 2.78, and the mean observed heterozygosity ranged from 0.218 to 0.296. High level of genetic differentiation was found between three northern populations and three southern populations. The marked genetic differentiation can be explained by the upwelling of the Zhejiang province and the freshwater outflow of Yangtze River. The results obtained in this study indicate that the northern and southern populations of C. sinensis should be treated as separate units for conservation management.     

Genetic and morphological identification of Sebastiscus tertius in the East China Sea (Scorpaeniformes: Scorpaenidae)

The allozymes and morphology of 110 specimens of three Sebastiscus species (S. marmoratus, S. tertius, and S. albofasciatus) in the East China Sea and near Japan were compared. Results of 20 allozyme loci studied showed that all three species were closely related (Nei's unbiased genetic distances, 0.057–0.133) but could be identified on the basis of informative loci with a few exceptions. Initial identification based on color patterns agreed with allozyme identification in more than 98% of Sebastiscus specimens and agreed completely in S. albofasciatus. One specimen that was initially identified as S. marmoratus because of the dark body color was actually S. tertius according to two informative allozyme loci. Number of pectoral fin rays differed between S. marmoratus (18 or fewer, 98%) and S. tertius (19 or more, 85%) in this study. The previously mentioned dark specimen had 19 pectoral fin rays, which are characteristic in S. tertius. Using seven morphological measurements, canonical discriminant analysis between S. marmoratus and S. tertius classified less than 90% of specimens into the original groups (species). Some specimens of S. tertius resembled S. marmoratus in body shape and vice versa. A combination of genetic characterization and morphological examination is necessary to identify S. marmoratus and S. tertius accurately. Distinction based on allozymes and color patterns with numbers of pectoral fin rays should provide satisfactory identification. Received: May 16, 2000 / Revised: December 19, 2000 / Accepted: January 24, 2001     

Analysis of F ST outliers at allozyme loci in Pacific salmon: implications for natural selection

Natural selection has been invoked to explain the observed geographic distribution of allozyme allele frequencies for a number of teleost species. The effects of selection on allozyme loci in three species of Pacific salmon were tested. A simulation-based approach to estimate the null distribution of population differentiation (F _ST) and test for F _ST outliers was used. This approach showed that a majority of allozyme loci conform to neutral expectations predicted by the simulation model, with relatively few F _ST outliers found. No consistent F _ST outlier loci were found across species. Analysis of population sub-groups based on geography and genetic identity reduced the number of outlier loci for some species, indicating that large geographic groups may include genetically divergent populations and/or that there is geographic heterogeneity in selection pressure upon allozyme loci. Two outlier allozyme loci found in this analysis, lactate dehydrogenase-B and malic enzyme, have been shown to be influenced by selection in other teleost species. This approach is also useful in identifying allozyme loci (or other genetic markers) that meet assumptions for population genetic study.     

Genetic differentiation and detection of cryptic species in the genus Lepismachilis (Insecta: Microcoryphia) from the Western Mediterranean region

Different species of the bristletail genus Lepismachilis were collected in 14 localities in Italy and Spain and an allozyme electrophoretic survey was carried out to estimate the degree of genetic variability and differentiation at intra- and interspecific levels. Four morphological species were initially identified (L osellai, L. y-signata, L. affinis, L. targionii), but the electrophoretic analysis demonstrated the presence of two additional species among the individuals of L. targionii (Lepismachilis spl and sp2). The validity of these species and their differentiation from L targionii were demonstrated by the fixation of alternative allelic patterns at several loci (7 in Lepismachilis spl and 8 in Lepismachilis sp2), coupled with fixed, previously undetected, morphological differences. In addition, Lepismachilis sp2 was sympatric with L. targionii in three collecting sites, where the fixation of alternative allelic patterns unequivocally demonstrated reproductive isolation. Genetic variability did not seem to be correlated with local ecological factors, and differences between species should rather be explained by different historical factors. Low levels of gene flow, estimated with two different indirect methods, were observed in L. targionii and L. y-signata, and were due to high levels of structuring among populations. Genetic differentiation among conspecific populations was not correlated to their geographical arrangement and the presence of loci fixed for different alleles among them suggested that stochastic factors (such as genetic drift) may have played a role in determining genetic differentiation of geographically isolated populations. Genetic divergence values indicated that the six species are well differentiated and allozyme profiles were diagnostic for all of them. On the other hand, allozyme data did not provide adequate information to resolve evolutionary relationships among the species, nor did they confirm the validity of the two subgenera (Lepismachilis and Berlesilis) in which the genus Lepismachilis is traditionally divided.     

Sibling species and cryptic speciation in the Brachionus plicatilis species complex (Rotifera)

An attempt is made to identify sibling species within the Brachionus plicatilis complex (Rotifera). Allozyme and morphological data for nine strains from all over the world are provided. Although the analysis of morphological data classified individuals from nine strains into two groups, cluster analysis using genetic distance data obtained from allozyme data revealed at least three groups. A male choice design is described for the first time in rotifers and was used to test for male mating preferences among sympatric strains belonging to three distinct species. The results suggest that the B. plicatilis complex is actually composed of more than three sibling species.     

Morphological and genetic diversity within Pilosella hoppeana aggr. (Asteraceae) in Italy and taxonomic implications

Morphological variation, ploidy level and genetic diversity have been studied on 10 populations of the Pilosella hoppeana aggr. from the Alps, Abruzzo, Calabria and Sicily. Chromosome counts showed that the plants from Abruzzo and those from Sicily are tetraploid (2n = 36); they are assigned to P. hoppeana subsp. macrantha. The plants from the Alps (P. hoppeana subsp. hoppeana) and those from Calabria are diploid. The Calabrian populations, previously included in P. hoppeana subsp. macrantha, are shown to belong to a separate species, P. leucopsilon. The principal component analysis, based on 25 morphological characters, allowed distinguishing clearly four groups. An allozyme study using 10 enzyme systems revealed 7 polymorphic loci with a total of 20 alleles, some of them exclusive at regional level, others shared between populations showing similar morphological features. The genetic differentiation between populations was relatively high. The obtained dendrogram supports recognition of the morphologically defined taxa.     

Biosystematics of two sympatric species ofEucharis (Amaryllidaceae)

Eucharis candida andE. formosa are two often sympatric species of bulbous geophytes restricted to neotropical rain forest understory. The species are most common in eastern Ecuador, and are the only two east Ecuadorean species of the genus found north of the Pastaza valley. Data from phenetic, karyotypic, and preliminary isozyme electrophoretic analyses of both species are represented. The species are distinguishable phenetically and karyologically, but isozyme-based relationships are more complex. Phenetic resolution of the isozyme phenotypes supports recognition of two species in Ecuador. A Peruvian isolate ofE. formosa, though not morphologically distinct, shows both allozyme and chromosomal divergence from Ecuadorean populations. Cladistic relationships based on overall allozyme data do not support species distinction, but a novel electrophoretic phenotype for glutathione reductase is shared only by individuals ofE. candida. An apparent geographic component within the monophyletic groups resolved in the cladogram suggests that some degree of gene flow between these two species has been maintained without the complete loss of morpholgoical species identity. This may have been mediated either by artificial population structures due to a probable long history of cultivation, or via Pleistocene refugia effects. Both species may have originated in eastern Ecuador from a common ancestral population which has since radiated outward, perhaps several times.     

Comparison of genetic variation and differentiation using microsatellite markers among three rare threatened and one widespread toad lily species of Tricyrtis section Flavae (Convallariaceae) in Japan

Genetic diversities were examined using six microsatellite markers amplifiable in three rare and one widespread species of Tricyrtis section Flavae, which are endemic to Japan. Contrary to a general expectation, the three rare species, Tricyrtis flava, Tricyrtis ohsumiensis and Tricyrtis perfoliata, have comparable genetic variation at the species level to that of the widespread Tricyrtis nana. This is probably because T. nana has not sufficiently recovered genetic diversity from the bottleneck at speciation or because recent range contractions have occurred in the three rare species. Genetic diversity at the population level was smaller in the putative selfing species T. nana than in the other three outcrossing species. Compared with a preceding study using allozyme markers, the genetic diversity in microsatellite loci was considerably larger, probably resulting from higher mutation rates at the microsatellite loci. Owing to the high genetic diversity of the microsatellite markers, genetic differentiation among populations could be estimated even in T. nana with little allozyme polymorphism.     

PATTERNS OF GENETIC VARIATION IN SALIX SECTION LONGIFOLIAE (SALICACEAE)

Enzyme electrophoresis was used to examine genetic relationships within a taxonomically complex group of diploid willows (Salix section Longifoliae). Forty-eight populations representing seven morphogeographic taxa were analyzed for 15 putative enzyme loci. Levels of genetic variation (A, P, H_e, and H_T) differed greatly among populations, but were generally lower than those reported for other species with similar life history traits. In populations and species from southern latitudes, where populations tend to be smaller and more isolated, genetic variation decreased and population differentiation increased. Genetic identity values suggest that the section is divisible into four major elements: 1) Salix interior, 2) S. taxifolia, 3) the S. exigua group, and 4) S. melanopsis. The first three occupy eastern, southern, and western portions of the continent, respectively. Salix melanopsis is sympatric with the S. exigua group in western North America, but is ecologically distinct. The S. exigua group is differentiated into four major geographic entities: S. hindsiana from west of the Sierras in California and southwest Oregon, S. sessilifolia from west of the Cascades in central Oregon to British Columbia, and a northern and southern race of S. exigua in the Intermountain West. The relationships suggested by allozyme data do not agree with traditional taxonomic concepts. Evidence of hybridization was obtained at five points where taxa come into contact, and Salix fluviatilis may be of hybrid origin. Taxonomic confusion appears to be largely the result of frequent hybridization events and overemphasis of a few highly variable morphological traits. The section appears to be a syngameon undergoing geographic speciation, with one taxon gaining reproductive isolation by a shift in habitat preference.     

Relationships and Hybridization among Smilax china and Its Affinities: Evidence from Allozyme Data

Smilax china L. is a widespread species in China with different ploidy levels. It is morphologically similar to S. davidiana, S. trinervula, and S. glauco-china. In this study, the chromosome number and the variation in allozyme patterns of eight enzyme systems with 25 alleles in 11 populations of S. china and three affinitive species were investigated. The allozyme data, together with morphological and cytological data, suggest that S. glauco-china is not closely related to the other taxa investigated. The diploid species S. davidiana and S. trinervula are involved as ancestor species and share great introgressions with S. china. In S. china, populations from Guilin and Guiyang are allotetraploid; their diploid progenitors probably are diploid populations of S. china and S. trinervula. The results suggest this species arose from multiple origins.     

Allozyme polymorphisms of maize populations from southwestern China

Maize (Zea mays L.) is one of the most-important food crops in southwestern China. The diversity of maize populations from southwestern China has been evaluated on the basis of agronomic and morphological data, but not on marker data. Our objectives were to evaluate the allozyme polymorphism of these populations, and group the populations on the basis of allozyme data. We analyzed 27 maize populations from southwestern China and two populations [BS13(S)C2 and Lancaster] from the USA for genetic variation at 18 allozyme loci. We found a total of 69 alleles at 18 allozyme loci with an average of 3.8 alleles per locus. Compared with inbreds, hybrids, and populations from the U.S. Corn Belt, the 27 Chinese populations had a significantly higher (p<0.01) number of allozyme alleles per locus. Maize populations from southwestern China have accumulated abundant genetic diversity, and might be valuable germplasm for broadening the genetic base of U.S. Corn Belt breeding germplasm. The analyses of allele-frequency distributions and the expected heterozygosity also reflected the differences between the Chinese and the U.S. germplasm. The Chinese populations might be valuable germplasm for complementing U.S. Corn Belt breeding germplasm. The analysis of gene diversity showed that 77% of the allozyme variation resided within populations and 23% between populations. This result suggested that breeders should identify one or a few Chinese populations with the best agronomic performance, and exploit the genetic variation within these selected populations. Cluster analysis classified the 29 populations into four main groups. Groupings based on allozyme data could be useful for classifying the populations into different heterotic groups and, consequently, exploiting them in hybrid breeding. Received: 12 October 2000 / Accepted: 13 March 2001     

Allozyme diversity and systematics inAnnonaceae—a pilot project

Five species ofAnnona and one species fromArtabotrys, Cananga, Polyalthia, andRollinia were investigated in regard to 11 different allozyme loci. Preliminary studies on small population samples ofAnnona suggest genetic uniformity in three species and variability within and between populations in two other species. The allotetraploid origin ofA. glabra is clearly shown by its hybrid enzyme bands. The genetic distance between fiveAnnona species partly corresponds with their morphological relationships; onlyA. muricata appears more separated than is suggested by morphology. A comparison of the five genera demonstrates close relationship betweenAnnona andRollinia. Two enzyme loci are identical within all taxa investigated and possibly may serve as a genetic marker for the family.     

POSTGLACIAL RANGE FLUCTUATION,GENETIC SUBDIVISION AND SPECIATION IN THE WESTERN NORTH AMERICAN SPOTTED FROG COMPLEX,RANA PRETIOSA

The western North American complex of spotted frogs (Rana pretiosa) exhibits isolation-by-distance, genetic subdivision, and speciation in association with its extensive northward range shift in postglacial times. The southern relict populations of R. pretiosa species B existing at high altitudes or in desert springs have been subjected to restricted gene flow, high inbreeding, and bottlenecks to produce significant between-population genetic diversity. The more recently established northern populations, however, show genetic uniformity and isolation-by-distance, as estimated using Slatkin's (1993) statistic M?. Middle latitude populations have higher heterozygosities than populations at either extreme. Fixed differences in allozyme variation separate 21 populations of species B from five populations of R. pretiosa species A found in southwest Washington State and the Cascades Mountains of Oregon. Morphological variation of 20 metric characters among 38 samples, examined using multiple discriminant function analysis, could partially resolve partitioning among populations but specimens from the vicinity of the type series of R. p. pretiosa could not be assigned to either species A or species B. Speciation in these frogs may not be correlated with morphological evolution since comparatively neutral allozyme changes may be established more rapidly than changes in morphology.     

Hidden diversity in the freshwater planktonic diatom Asterionella formosa

Many freshwater and marine algal species are described as having cosmopolitan distributions. Whether these widely distributed morphologically similar algae also share a similar gene pool remains often unclear. In the context of island biogeography theory, stronger spatial isolation deemed typical of freshwater lakes should restrict gene flow and lead to higher genetic differentiation among lakes. Using nine microsatellite loci, we investigate the genetic diversity of a widely distributed freshwater planktonic diatom, Asterionella formosa, across different lakes in Switzerland and the Netherlands. We applied a hierarchical spatial sampling design to determine the geographical scale at which populations are structured. A subset of the isolates was additionally analysed using amplified fragment length polymorphism (AFLP) markers. Our results revealed complex and unexpected population structure in A. formosa with evidence for both restricted and moderate to high gene flow at the same time. Different genetic markers (microsatellites and AFLPs) analysed with a variety of multivariate methods consistently revealed that genetic differentiation within lakes was much stronger than among lakes, indicating the presence of cryptic species within A. formosa. We conclude that the hidden diversity found in this study is expected to have implications for the further use of A. formosa in biogeographical, conservation and ecological studies. Further research using species‐level phylogenetic markers is necessary to place the observed differentiation in an evolutionary context of speciation.     

Character displacement in sailfin mollies, <Emphasis Type="Italic">Poecilia latipinna</Emphasis>: allozymes and behavior

Synopsis We analyzed variation in allozymes and mating preferences in 12 populations across much of the range of the sailfin molly, Poecilia latipinna. Sailfin mollies can be sympatric with its sexual parasite Amazon mollies, P. formosa. Amazon mollies must co-exist and mate with bisexual males of closely related species (including sailfin mollies) to induce embryogenesis but inheritance is strictly maternal. Where sailfin and Amazon mollies are sympatric there is evidence of reproductive character displacement as males show a significantly stronger mating preference for sailfin molly females over Amazon mollies compared to preferences of males from allopatric populations. From the allozyme data we found a moderate amount of genetic variation across all populations but this variation did not reveal significant partitioning between sympatric and allopatric populations. Additionally, we found no evidence for isolation by distance as genetic distance was not significantly correlated with geographic distance. While allozyme variation also did not significantly correlate with male mating preferences, there was a significant correlation between male mating preferences and geographic distance. This correlation between mating preferences and geographic distance may have arisen from coevolution with Amazon mollies resulting in reproductive character displacement. Taken together, the distribution of genetic and behavioral variation among sympatric and allopatric populations suggests that behavioral evolution has outpaced evolution at the allozyme loci we examined in P. latipinna.     

New variation and biosystematic patterns detected by allozyme and morphological comparisons inLimnanthes sect.Reflexae (Limnanthaceae)

Sixty-one populations ofLimnanthes sect.Reflexae were surveyed for variation at 19 allozyme loci and reassessed for their morphological diversity and biosystematic relationships. Cluster analysis of allozyme data at individual population level provided evidence for a fairly good fit with the earlier systematic work on this genus but both morphological and allozyme data together defined some new clusters as well as exceptional entities. For example, only var.rosea of the fourL. douglasii varieties described byMason (1952) on the basis of geographical distribution, flower color and leaf morphology, remained as a distinct entity; the others were realigned along new geographic boundaries.L. striata appeared to have two well-developed subgroups, one of which was closely related to theL. douglasii complex. Further ecogeographic and hybridization studies are needed to fully describe these new phenetic findings in the context of phylogenetic inference. SectionInflexae was readily distinguished from sect.Reflexae by several diagnostic loci, thus supporting the validity of this division. Interspecific variation levels and population differentiation inLimnanthes were high when compared to other plant taxa, possibly indicating highly dynamic evolutionary processes for which variable habitats and extreme subdivision within species have been suggested as explanations.     

Delimitation of morphologically similar sponge crab species of the genus Pseudodromia (Crustacea, Decapoda, Dromiidae) from South Africa

Stewart, B. A., Gouws, G., Daniels, S. R. & Matthee, C. A. (2004). Delimitation of morphologically similar sponge crab species of the genus Pseudodromia (Crustacea, Decapoda, Dromiidae) from South Africa. — Zoologica Scripta , 33 , 45–55.
Presently, three Pseudodromia sponge crab species are recognized, all of which are endemic to the continental shelf off the coast of South Africa. Two of these differ only in the morphology of their rostral teeth, making them difficult to distinguish, and can thus be considered as cryptic species. In addition they have very similar distribution ranges, thus raising doubts as to their specific status. Discriminant function analysis of morphometric data, differentiation at 10 allozyme loci, and sequence data derived from the 12S rRNA mitochondrial gene were used to test whether specimens identified as P. latens and P. rotunda are morphological forms of a single, widespread species, or represent two, distinct, reproductively isolated species, and to establish whether these two taxa are sister species, and thus form a monophyletic entity. The presence of fixed allele differences at three, and strong genetic heterogeneity at five other allozyme loci, indicating no gene flow occurring between sympatric populations, as well as the relatively high degree of 12S rRNA and allozyme genetic differentiation observed, supported the recognition of P. latens and P. rotunda as separate species. The 12S rRNA topology suggested that the genus Pseudodromia , as presently constituted, is paraphyletic, thus inferring that the morphological characters used to define this taxon might not be useful for phylogenetic inferences. It was concluded that in view of the uncertainties raised regarding the designation and composition of certain genera within the family Dromiidae, further rigorous analyses of morphological and genetic data are needed to further our understanding of the taxonomy of the sponge crabs.     

Evidence for African origins of founders of the asiatic lion species survival plan

The Asiatic lion (Panthera leo persica) exists in the wild as a single relict population of approximately 250 individuals in the protected Gir Forest Sanctuary in western India. In 1981, a species survival plan (SSP) for the Asiatic lion was established by the American Association of Zoological Parks and Aquariums to manage the 200 + descendants of Asiatic lions in captivity in western zoological facilities. This captive population was derived from seven founders. In order to compare the genetic structure of the Gir Forest population with that of the captive SSP population, a genetic survey of 46 electrophoretic allozyme systems resolved from extracts of lion blood was undertaken by using 29 SSP Asiatic lions and 28 wild-caught or captive-bred lions maintained at the Sakkarbaug Zoo in India but originally derived from the Gir Forest. The Gir lion population was found to be genetically monomorphic at each of 46 allozyme loci. This was in contrast to several African lion (Panthera leo leo) populations, which show moderate levels of allozyme variation at the same loci. The SSP lion population was polymorphic at three allozyme loci (IDHI, TF, and PTI) for alleles that were previously found only in African lion populations. Pedigree analysis of the genetic transmission of these three biochemical loci demonstrated that two of the five primary founder animals of the SSP Asiatic lion population (a breeding pair originally imported from the Trivandrum Zoo in southern India) were descendants of the African subspecies. Three other founder animals were pure Asian. A retrospective SSP pedigree analysis of two morphologic characters (prominent abdominal fold and pairing of infraorbital foramen) that are partially diagnostic for persica vs leo was consistent with this conclusion as well. The implications for the management of small captive populations of threatened species and of the Asiatic lion SSP population are discussed.