The radiation of microhylid frogs (Amphibia: Anura) on New Guinea: a mitochondrial phylogeny reveals parallel evolution of morphological and life history traits and disproves the current morphology-based classification

Microhylidae account for the majority of frog species on New Guinea and have evolved an extraordinarily wide range of ecological, behavioural, and morphological traits. Several species are known for their unique paternal care behaviour, which includes guarding of clutches in some and additional froglet transport in other species. We sampled 48 out of 215 New Guinean microhylid species and all but two (Mantophryne and Pherohapsis) of 18 New Guinean genera and analysed a concatenated data set of partial sequences of the mitochondrial genes 12S and 16S, which comprises 1220 aligned nucleotide positions, in order to infer the phylogenetic relationships within this diverse group of frogs. The trees do provide resolution at shallow, but not at deep branches. Monophyly is rejected for the genera Callulops, Liophryne, Austrochaperina, Copiula, and Cophixalus as currently recognized. Six clades are well supported: (1) Hylophorbus and Callulops cf. robustus, (2) its sister taxon comprising Xenorhina, Asterophrys turpicola, and Callulops except for C. cf. robustus, (3) Liophryne rhododactyla, L. dentata, Oxydactyla crassa, and Sphenophryne cornuta, (4) Copiula and Austrochaperina, (5) Barygenys exsul, Cophixalus spp., and Oreophryne, (6) Cophixalus sphagnicola, Albericus laurini, and Choerophryne. The phylogenies provide evidence for the parallel evolution of parental care modes, life styles, and morphological traits that have thus far been emphasized in recent classifications.     

Biochemical polymorphism in the red partridge (Alectoris rufa)]

Egg white proteins, hemoglobin, serum proteins and enzymes from Alectoris rufa have been examined by starch gel electrophoresis. All three genetic parameters, mean proportion of loci polymorphic per population (P), mean numbers of alleles per locus (A), and mean proportion of loci heterozygous per individual (H) are given for Alectoris rufa, and comparison were made between the proteins of the related species Coturnix coturnix japonica.     

Enzyme polymorphism and genetic population structure in Escherichia coli and Shigella

Electrophoretically demonstrable variation in 12 enzymes was studied in more than 1 600 isolates of Escherichia coli from human and animal sources and in 123 strains of the four species of Shigella. All 12 enzymes were polymorphic; and the number of allozymes (mobility variants), which were equated with alleles, averaged 9.3 per locus in E. coli. For Shigella species, the mean number of alleles was 2.9 per locus. Some 77% of the allozymes recorded in Shigella were shared with E. coli. A total of 302 unique genotypic combinations of alleles over the 12 loci (electrophoretic types, ETs) was distinguished, of which 279 represented E. coli and 23 were Shigella. Among electrophoretic types, mean allelic diversity per locus was 0.52 for E. coli and 0.29 for Shigella. It was estimated that there are, on the average, about 0.3 detectable codon differences per locus between pairs of strains of E. coli and Shigella, which is roughly equivalent to 1.2 amino acid differences per enzyme. Evidence that the enzyme loci studied are a random sample of the genome is provided by a significant positive correlation between estimates of genetic divergence between pairs of strains obtained by DNA reassociation tests and estimates of genetic distance between the same strains based on electrophoresis. A principal components analysis of allozyme profiles revealed that the 302 ETs fall into three overlapping clusters, reflecting strong non-random associations of alleles, largely at four loci. Each of the four ETs of E. coli that have been most frequently recovered from natural populations has an allozyme profile that is very similar to, or identical with, the hypothetical modal ET of one of the groups. ETs of Shigella fall into two of the groups. No biological significance can at present bbe attributed to the genetic structure revealed by Multilocus electrophoretic techniques. The electrophoretic data are fully compatible with other molecular and more conventional evidence of a close affinity between E. coli and Shigella, and they raise questions regarding the present assignments of certain strains to species. In support of evidence from DNA reassociation tests and serotyping, the present study suggests that S. sonnei is homogeneous in chromosomal genotype.     

Phylogenetic relationships among fish of the subfamily Sparinae (Perciformes: Sparidae) in the coastal waters of Taiwan

The phylogenetic relationship of five species belonging to the subfamily Sparinae was investigated using starch-gel electrophoretic analysis of enzymes in skeletal muscle, heart, eye, liver, and sarcoplasmic protein in skeletal muscle. A total of 38 loci with 105 alleles coding for 25 enzymes and general protein were analysed and scored to calculate the proportion of polymorphic loci at the 95% level (P₉₅), mean heterozygosity over loci (H), average number of alleles per locus (Na), and genetic distance (D). P₉₅ values obtained ranged from 13.16 to 18.42%, H values ranged from 0.042 to 0.057, Na values ranged from 1.21 to 1.42, and an average D value of 0.305 (0.216-0.386) between congeneric species and 0.801 (0.614-0.896) between two consubfamilial genera, Acanthopagrus and Sparus, was found. A phenogram constructed on the basis of genetic data was similar to that based on morphometric characters at the genus level. However, discordance does exist among species of Acanthopagrus between the two sets of phenograms constructed. The results of this study support the niche-width-variation hypothesis.     

Analysis of isozyme loci in the face fly,Musca autumnalis DeGeer

Vertical polyacrylamide gel electrophoresis was used to resolve enzymes at 50 putative loci in Iowa face flies, a recently arrived, colonizing, Palearctic species. Sixty-two percent of the 50 loci were polymorphic. The mean number of alleles per polymorphic locus was 3.2 +/- 1.38; the mean among all loci was 2.38 +/- 1.62 alleles. The effective number of alleles among 50 loci was 1.4 +/- 0.62. Mean observed and expected heterozygosities for the 50 face fly loci were 0.167 +/- 0.037 and 0.186 +/- 0.031, respectively. Comparison of the electrophoretic data for Musca autumnalis and for M. domestica L. showed similar high levels of gene diversity. A survey of gene diversity at 12 loci among six geographically independent laboratory colonies demonstrated statistically significant genetic differentiation that was probably due to drift after colonization.     

Genetic Variation and Relationships Within Taxus and Between the Genus and Pseudotaxus in China

In the present paper, genetic diversity of 12 populations of four Taxus taxa and one species of monotypic genus Pseudotaxus in China were studied by horizontal starch gel electrophoresis. Seven enzymes encoded by nine loci were assessed. Genetic diversity within 5 taxa were relatively high with the proportion of polymorphic loci ( P ) ranging from 44.4 % to 77.8 %, average number of alleles per locus (A) from 1.6 to 2.1; mean expected and observed heterozygosity ( He, Ho) per locus from 0. 065 to 0.152 and 0. 068 to 0.111, respectively. Combined with the analysis of other characters, the results from allozyme analysis suggest that there be very close relationships between the three species and one variety in Taxus, with high genetic identities (I) between them(from 0. 727 to 0. 995) except those between T. yunnanensis and other taxa in Taxus (0. 727～ 0. 804). Consequently, we propose that these taxa in Taxus be treated as different geographical races within a single species. Also the taxonomic position of Pseudotaxus as a distinct genus is supported by ourallozyme data.     

Gene Polymorphism in Natural Populations of DROSOPHILA PERSIMILIS

Genetic variation at 43 loci has been studied in six different populations of Drosophila persimilis by electrophoresis of enzymes and proteins. In D. persimilis the mean proportion of polymorphic loci is 0.362, the mean proportion of heterozygous loci per individual is 0.100 and the average number of alleles per locus is 1.651. In all populations, the loci coding for the hydrolytic and other nonspecific enzymes are much more variable than the loci coding for the enzymes of the glycolytic pathway, Kreb's cycle, other specific enzymes and larval proteins. Most loci have similar allele frequency in all populations except the two loci, Amylase and Pt-12, which show a pattern of associations of different alleles with different third chromosome inversions.     

Polymorphisme biochimique chez le faisan commun (Phasianus colchicus)

Egg white proteins, hemoglobin, serum proteins and enzymes from Phasianus colchicus have been examined by starch gel electrophoresis. All three genetic parameters, mean proportion of loci polymorphic per population (P), mean numbers of alleles per locus (A), and mean proportion of loci heterozygous per individual (H), are given. Comparison were made between the proteins of the related species Coturnix coturnix japonica, and the variation of pheasant proteins is discussed in connexion with the genetic structure of the population studied.     

Electrophoretic variability in natural populations of Drosophila melanogaster and Drosophila simulans

Genetic variation at 59 gene loci coding for enzymes (50) and larval proteins (9) has been studied in sympatric populations of Drosophila melanogaster and D. simulans from insular and continental origin. The average number of alleles per locus, the mean proportion of polymorphic loci and the mean heterozygosity are similar both within and between species. There are however some significant differences between D. simulans populations in the genotypic frequencies for four polymorphic loci.     

High genetic variability in an ecologically variable vertebrate,Bufo viridis

Allozymic variation in proteins encoded by 26 loci was analyzed electrophoretically in 517 specimens of green toads from 11 populations from Israel and one population from Vis Island in the Adriatic Sea. Genetic variation in this toad is the highest yet reported in any vertebrate. All three genetic parameters, mean number of alleles per locus (A), mean proportion of loci polymorphic per population (P), and mean number of heterozygous loci per individual (H), are very high (A = 1.65, range 1.38-2.04; P = 0.423, range 0.346-0.615; H = 0.133, range 0.108-0.159). Central and marginal mainland populations are only slightly more variable than desert isolates, but much more variable than the Vis Island population. Genetic similarity is very high between mainland populations (S = 0.951, range 0.93-0.97). Frequencies of two alleles (Icd-lc and Tfa) are correlated with an ecological gradient of increasing aridity. Regulatory enzymes appeared to contribute more to overall polymorphism than non-regulatory enzymes. The genetic variation observed suggests that selection for heterozygosity as an adaptive strategy is operating in the ecologically variable environment in which green toads live.     

Patterns of Gene Variation in Central and Marginal Populations of DROSOPHILA ROBUSTA

The central and marginal populations of D. robusta differ greatly in the level of inversion polymorphism; the marginal populations are monomorphic or nearly so and the central populations are highly polymorphic. This paper presents the frequencies of alleles at forty gene loci in various populations of D. robusta, studied by electrophoresis of proteins and enzymes. Population samples were obtained from eight widely separated populations of D. robusta which included the central, the extreme marginal and the intervening populations between the center and the margins. We find that the proportion of polymorphic loci and average heterozygosity per individual is slightly higher in the marginal populations than the central populations. In D. robusta on an average, 39% of the loci are polymorphic and the average proportion of loci heterozygous per individual is 11%. A breakdown of loci in three categories, viz, hydrolytic enzymes and some other enzymes, larval proteins and glycolytic and Kreb's cycle enzymes, shows that in all populations the level of polymorphism is highest in the hydrolytic enzymes, intermediate in larval proteins and least in the glycolytic and Kreb's cycle enzymes. On the average, the proportion of loci heterozygous per individual for three groups of loci is: hydrolytic enzymes and others (.164), larval proteins (.115) and glycolytic and Kreb's cycle enzymes (.037). We also observe that in all populations the level of polymorphism on the X chromosome is far less than the expected 38%; in salivary gland cells the euchromatic length of the X chromosome is 38% of the entire genome. Lower levels of polymorphism for the X chromosome loci are explained due to low probability of balanced polymorphisms for the X-linked loci since the conditions for establishment of balanced polymorphism for X-linked loci are more restrictive than for the autosomal loci.-The polymorphic loci can be grouped according to pattern of allele frequencies in different populations as follows: (1) The allele frequencies are similar in all populations at the XDH, Pep-1 and Hex-1 loci. (2) The alleles at the Est-1, Est-2, Amy loci and the AP-4(1.0) and the LAP-1(.90) alleles show north south clinal change in frequency. (3) There is north south and east west differentiation at the Pt-5, Pt-8 and Pt-9 loci and the allele AP-4(.81). (4) Polymorphism at loci such as Fum, B.Ox, Hex-8, Pep-2 and Pep-3 are restricted to only one or two of the populations. (5) Allele frequencies at the MDH and ODH loci fluctuate between populations. (6) Allele frequencies at many polymorphic loci such as Est-1, Est-2, LAP-1, AP-4, Pt-5, Pt-8, Pt-9, Pt-16, MDH, Fum change clinally within a gene arrangement. The pattern of gene variation in D. robusta is very complex and cannot be easily explained due to migration of neutral alleles between once-isolated populations or to semi-isolation of neutral alleles. The observations of the pattern of allele variation in different populations, high levels of polymorphism in the marginal populations which have small population size and low levels of polymorphism of the X chromosome loci all support the argument in favor of balancing selection as the main mechanism for the maintenance of these polymorphisms. Environmental factors must play a role in the maintenance of a great deal of these polymorphisms, since we observe clinal allele frequency changes even within a given inversion type.     

Development and testing of 13 polymorphic microsatellite markers in Larimichthys polyactis (Sciaenidae) using 5' anchored PCR

Larimichthys polyactis is a commercially important marine fish species in southeast Asia. The population crashed due to overfishing in the 1970s, but has since recovered. We developed 13 novel polymorphic microsatellite markers in L. polyactis using 5' anchored PCR. The characteristics of these loci were estimated by analyzing a sample of 30 individuals. A total of 74 alleles were detected, with a mean of 5.7 alleles per locus. There were 2 to 12 alleles, 0.2760 to 0.8247 polymorphism information content, and 0.3214 to 1.000 observed and 0.3097 to 0.8567 expected heterozygosity per locus. The mean observed and expected heterozygosity was 0.6816 and 0.6724, respectively. Three loci deviated significantly from Hardy-Weinberg equilibrium after Bonferroni's correction, and no significant linkage disequilibrum between pairs of loci was found. This information will be useful for the analysis of population genetic diversity, and the management of this important fish resource.     

Evaluation of genetic variability of the Kamchatka crab Paralithodes camtschaticus (Tilesius) using allozyme markers

Interspecific genetic variation in populations of red king crab Paralithodes camtschaticus Tilesius (Litholidae, Decapoda: Crustacea) was examined using allozyme markers. The activity of 57 enzymes and the general protein presumably encoded by 92 loci was detected. The level of allozyme variability was low: the expected heterozygosity and the proportion of polymorphic loci were respectively 0.027 +/- 0.008 and 6.5%. This level of heterozygosity is three times lower than the average value for 122 crustacean species (0.082 +/- 0.007). Although genetic variants were found at 22 loci, their frequencies were generally low: only in loci 6-Pgd, Alp-1, and Pep-1 did the frequencies of the most common alleles not exceed 0.9. All polymorphic loci except one had two alleles; the exception was 6-Pgd, which had three alleles. The possible reasons for the low level of allozyme variability in red king crab are discussed.     

福清黑松醇溶蛋白的群体遗传分析

采用聚丙烯酰胺凝胶电泳技术,对福清黑松群体进行醇溶蛋白的多样性分析,共获得40种图谱,9个等位基因位点。对这些位点的统计分析表明,福清黑松的多肽位点百分率P=55.56%, 等位基因平均数A=3.00,平均等位基因有效数目Ae=2.28,预期杂合度He=0.533,平均实际杂合度Ho=0.402,固定指数F仅为0.246>0。该群体表现出遗传多样性水平较高,但该群体偏离了Hardy-Weinberg平衡定律,原因可能是取样偏差、群体中个体密度分布不均和个体生长状况不好,不能产生充分的随机交配,出现遗传漂移导致的。     

Ecological-genomic diversity of microsatellites in wild barley,Hordeum spontaneum,populations in Jordan

We analyzed the ecological-genomic diversity of microsatellites of wild barley, Hordeum spontaneum (C. Koch) Thell., at 18 loci in 306 individuals of 16 populations from Jordan across a southward transect of increasing aridity. The 18 microsatellites revealed a total of 249 alleles, with an average of 13.8 alleles per locus (range 3-29), with nonrandom distribution. The proportion of polymorphic loci per population averaged 0.91 (range 0.83-1.00); gene diversity, He, averaged 0.512 (range 0.38-0.651). We compared the number of alleles of the 18 loci to those found in Israel populations by Turpeinen et al. Out of the 280 alleles, 138 (49.3%) were unique (i.e. occurred in only one of the countries). The percentage of unique alleles in Jordan and Israel populations was 43.0% and 17.9%, respectively, suggesting that Jordan is an important center of origin and diversity of wild barley. Estimates of mean gene diversity were highest in the populations collected near the Golan Heights, such as Shuni North, Shuni South and Jarash. Sixty nine percent of the microsatellite variation was partitioned within populations and 31% between populations. Associations between ecogeographical values and gene diversity were established for eight microsatellite loci. The cluster produced by simple sequence repeat (SSR) data is mostly coincidence with the result of the dendrogram of the Spalax ehrenbergi superspecies of subterranean mole rats in Jordan based on allozyme gene loci. The major soil type in the wild barley habitat of each ecological group was different. Stepwise multiple regression analysis indicated that the variance of gene diversity was explained by altitude (R(2) = 0.362**). These observations suggest that microsatellites are at least partly adaptive and subject to natural selection.     

秦川母牛群体遗传特性的微卫星标记研究

为了从DNA分子水平揭示秦川牛群体遗传多态性和群体遗传结构,寻找可用于秦川牛的微卫星标记,本研究选择了12个普通牛（Bos taurus）微卫星标记检测了90头秦川母牛各微卫星位点的遗传变异及多态性。结果表明,在秦川母牛群体中,12个微卫星位点共检测到了247个等位基因,各位点的等位基因数在13（INRA005）～33个（HEL13）之间,平均每个微卫星位点的等位基因数为21个;总有效等位基因数和平均每个位点平均有效等位基因数（Ne）分别分为142．6229和11．8852。各位点平均基因频率取样方差（V（pij））为2．6036×10^-4。12个微卫星位点平均观察杂合度（Ho）和平均期望杂合度（He）在0．7842（INRA005）～0．9775（BM315）和0．7952（BM315）～0．9446（HEL13）之间。12个位点平均多态信息含量（PIC）在0．7653（INRA005）～0．9420（HEL13）之间,平均为0．8965．12个微卫星位点均属于高度多态位点,这表明秦川母牛群体中所检测各微卫星位点具有丰富的遗传多态性,具备较大的选择潜力。12个微卫星位点的平均固定指数（F）为-0．0076,即各位点杂合子的缺陷度不高,即偏离Hardy—Weinberg平衡的程度不大。     

Microsatellite diversity and fitness in stranded juvenile harp seals (Phoca groenlandica)

A positive relationship between genetic diversity at neutral markers and juvenile survival has been demonstrated for many vertebrate populations, although the correlation is typically weak and the explanation for it remains controversial. We assessed variation at 9-12 microsatellite loci in 65 juvenile harp seals (Phoca groenlandica) that stranded in poor condition around Long Island, NY, from 2001 to 2004. Compared with seals that died, surviving individuals had slightly higher measures of mean d(2), which reflects the size difference between alleles within an individual and provides an index of outbreeding. In contrast, there were no significant differences between survivors and nonsurvivors in heterozygosity or estimates of internal relatedness. This pattern is attributed to the fact that these microsatellite markers were exceptionally variable in this species (9-22 alleles per locus), and all individuals were heterozygous at most loci. Under these circumstances, mean d(2) may provide a powerful measure for assessing diversity-fitness correlations.     

Development of SSR Markers and Genetic Diversity in White Birch (Betula platyphylla)

In order to study genetic diversity of white birch (Betula platyphylla), 544 primer pairs were designed based on the genome-wide Solexa sequences. Among them, 215 primer pairs showed polymorphism between five genotypes and 111 primer pairs that presented clear visible bands in genotyping 41 white birch plants that were collected from 6 different geographical regions. A total of 717 alleles were obtained at 111 loci with a range of 2 to 12 alleles per locus. The results of statistic analysis showed that polymorphic frequency of the alleles ranged from 17% to 100% with a mean of 55.85%; polymorphism information content (PIC) of the loci was from 0.09 to 0.58 with a mean of 0.30; and gene diversity between the tested genotypes was from 0.01 to 0.66 with a mean of 0.36. The results also indicated that major allele frequency ranged from 0.39 to 1.00 with an mean of 0.75; expected heterozygosity from 0.22 to 0.54 with a mean of 0.46; observed heterozygosity from 0.02 to 0.95 with a mean of 0.26; Nei''s index from 0.21 to 0.54 with a mean of 0.46; and Shannon''s Information from 0.26 to 0.87 with a mean of 0.66. The 41 white birch genotypes at the 111 selected SSR loci showed low to moderate similarity (0.025-0.610), indicating complicated genetic diversity among the white birch collections. The UPGMA-based clustering analysis of the allelic constitution of 41 white birch genotypes at 111 SSR loci suggested that the six different geographical regions can be further separated into four clusters at a similarity coefficient of 0.22. Genotypes from Huanren and Liangshui provenances were grouped into Cluster I, genotypes from Xiaobeihu and Qingyuan provenances into Cluster II, genotypes from Finland provenance into Cluster III, and genotypes from Maoershan into Cluster IV. The information provided in this study could help for genetic improvement and germplasm conservation, evaluation and utilization in white birch tree breeding program.     

Identification and properties of microsatellite markers in tsetse flies Glossina morsitans sensu lato (Diptera: Glossinidae)

Genomic libraries enriched for simple sequence repeats were constructed for Glossina morsitans morsitans, G. m. submorsitans, and G. m. centralis. Sixteen microsatellite markers were isolated from the libraries and evaluated on flies from natural G. m. morsitans populations and other Glossina species in the Morsitans and Palpalis species groups. The primers amplified appropriate sized DNA fragments in the Morsitans and Palpalis groups. In G. morsitans s.l., eight of 12 dinucleotide repeats and four of 12 trinucleotide repeats were polymorphic. The polymorphic loci showed a mean 7.5 +/- 4.8 alleles per locus and their mean heterozygosity was 55.8 +/- 7.7%.     

Genetic variation and hybridization in SwedishSchoenus (Cyperaceae)

Schoenus ferrugineus andS. nigricans have restricted distributions in Sweden and are almost exclusively confined to calcareous fen habitats. AtS. nigricans sites,S. ferrugineus is usually also present, and hybrids are frequently found. In this report, I used allozymes to estimate the amount of gene flow between the two species, and to compare the partitioning of genetic diversity in each of them. Thirteen loci were analysed at eight different enzyme systems. Seven loci were variable between or within the species. The two species had completely different alleles at two of the seven variable loci, whereas there was overlap at five loci. In all, 22 different alleles were found. Six of these alleles were confined toS. nigricans, and five alleles were confined toS. ferrugineus. Nei's genetic identity was 0.55.—InS. ferrugineus, three loci (23%) were polymorphic, and the average number of alleles per polymorphic locus was 2.0 (each polymorphic locus had two alleles). InS. nigricans, three loci (23%) were polymorphic, and the average number of alleles per polymorphic locus was 2.3.—The proportion of genetic diversity due to variation among sites (G _ST) was fairly similar in the two species, mean over loci = 0.12 inS. ferrugineus and 0.15 inS. nigricans. However, the proportion of genetic diversity due to variation among individuals within sites (G _IS) differed markedly between the two species, mean over loci = 0.54 inS. ferrugineus and 0.17 inS. nigricans. Accordingly, there was a much higher individual heterozygosity inS. nigricans than inS. ferrugineus. — Most hybrids were interpreted as F₁ hybrids. However, a small proportion, 0.5–1.6 %, were F_n hybrids or back-crosses.—On the Swedish mainland, all former occurrences ofS. nigricans are extinct, but viable hybrids are still present at a few sites in southernmost Sweden.