Genetic divergence in the snorkel snail,Rhiostoma housei,a species complex in Thailand (Caenogastropoda: Cyclophoridae)

Allozyme variation was examined in 292 samples of the snorkel snail, Rhiostoma housei, collected from 12 localities within four regions of Thailand. Ten allozyme loci were screened across all the samples using horizontal starch gel electrophoresis, and eight of these loci, from eight enzyme systems, were found to be polymorphic. Within these eight loci the expected heterozygosity (H_exp) was moderate to high ranging from 0.015 to 0.148 with an overall mean of 0.081 ± 0.041, whilst a high genetic heterogeneity among samples was found (F_st = 0.798). R. housei exhibited a weak pattern of isolation by distance over the entire tested species range in Thailand. The high F_st and moderate to high H_exp suggest gene flow amongst populations is partially restricted resulting in fragmentation into local gene pools. Based on allele frequencies and an allozyme dendrogram, six groups were identified, and these entities may represent separate biological species. The large values of genetic distances and fixed allelic differences obtained for each combination of allopatric samples imply the presence of cryptic species under the name R. housei.     

Comparison of allozyme,RFLP, and RAPD markers for revealing genetic variation within and between trembling aspen and bigtooth aspen

We examined genetic variation in allozyme loci, nuclear DNA restriction fragment length polymorphisms (RFLPs), and random amplified polymorphic DNAs (RAPDs) in 130 trembling aspen (Populus tremuloides) and 105 bigtooth aspen (P. grandidentata) trees. In trembling aspen 10 out of 13 allozyme loci assayed (77%) were polymorphic (P), with 2.8 alleles per locus (A) and an expected heterozygosity (H_e) of 0.25. In contrast, bigtooth aspen had a much lower allozyme genetic variability (P=29%; A=1.4; H_e=0.08). The two species could be distinguished by mutually exclusive alleles at Idh-1, and bigtooth aspen has what appears to be a duplicate 6PG locus not present in trembling aspen. We used 138 random aspen genomic probes to reveal RFLPs in HindIII digests of aspen DNA. The majority of the probes were from sequences of low copy number. RFLP results were consistent with those of the allozyme analyses, with trembling aspen displaying higher genetic variation than bigtooth aspen (P=71%, A=2.7, and H_e=0.25 for trembling aspen; P=65%, A=1.8, and H_e=0.13 for bigtooth aspen). The two species could be distinguished by RFLPs revealed by 21 probes (15% of total probes assayed). RAPD patterns in both species were studied using four arbitrary decamer primers that revealed a total of 61 different amplified DNA fragments in trembling aspen and 56 in bigtooth aspen. Assuming a Hardy-Weinberg equilibrium, estimates of P=100%, A=2, and H_e=0.30 in trembling aspen and P=88%, A=1.9, and H_e=0.31 in bigtooth aspen were obtained from the RAPD data. Five amplified DNA fragments were species diagnostic. All individuals within both species, except for 2 that likely belong to the same clone, could be distinguished by comparing their RAPD patterns. These results indicate that (1) RFLPs and allozymes reveal comparable patterns of genetic variation in the two species, (2) trembling aspen is more genetically variable than bigtooth aspen at both the allozyme and DNA levels, (3) one can generate more polymorphic and species-specific loci with DNA markers than with allozymes in aspen, and (4) RAPDs provide a very powerful tool for fingerprinting aspen individuals.     

Genetic variability of tall junipers (Juniperus excelsa Bieb.) on the northern and southern boundaries of their natural distribution

The genetic structure, subdivision, and differentiation of six populations of tall junipers (Juniperus excelsa Bieb.) in the Crimean Mountains and one population in Lebanon were investigated using 18 polymorphic allozyme loci as genetic markers. A high level of genetic variability of J. excelsa was established at the northern and southern boundaries of their natural habitats. The mean values of the main indicators of genetic polymorphism were P₉₉ = 1.000, A = 3.167, H_E = 0.370, and H_O = 0.405. The subdivision and differentiation of populations were low (F_ST = 0.032, D_N = 0.026), indicating the similarity of their gene pools.     

GEOGRAPHIC DISTRIBUTION OF MOLECULAR VARIANCE WITHIN THE BLUE MARLIN (MAKAIRA NIGRICANS): A HIERARCHICAL ANALYSIS OF ALLOZYME,SINGLE-COPY NUCLEAR DNA,AND MITOCHONDRIAL DNA MARKERS

This study presents a comparative hierarchical analysis of variance applied to three classes of molecular markers within the blue marlin (Makaira nigricans). Results are reported from analyses of four polymorphic allozyme loci, four polymorphic anonymously chosen single-copy nuclear DNA (scnDNA) loci, and previously reported restriction fragment length polymorphisms (RFLPs) of mitochondrial DNA (mtDNA). Samples were collected within and among the Atlantic and Pacific Oceans over a period of several years. Although moderate levels of genetic variation were detected at both polymorphic allozyme (H = 0.30) and scnDNA loci (H = 0.37), mtDNA markers were much more diverse (h = 0.85). Allele frequencies were significantly different between Atlantic and Pacific Ocean samples at three of four allozyme loci and three of four scnDNA loci. Estimates of allozyme genetic differentiation (θ_O) ranged from 0.00 to 0.15, with a mean of 0.08. The θ_O values for scnDNA loci were similar to those of allozymes, ranging from 0.00 to 0.12 with a mean of 0.09. MtDNA RFLP divergence between oceans (θ_O = 0.39) was significantly greater than divergence detected at nuclear loci (95% nuclear confidence interval = 0.04–0.11). The fourfold smaller effective population size of mtDNA and male-mediated gene flow may account for the difference observed between nuclear and mitochondrial divergence estimates.     

Evolutionary implications of allozyme and RAPD variation in diploid populations of dioecious buffalograss Buchloë dactyloides

Buffalograss, Buchloë dactyloides, is widely distributed throughout the Great Plains of North America, where it is an important species for rangeland forage and soil conservation. The species consists of two widespread polyploid races, with narrowly endemic diploid populations known from two regions: central Mexico and Gulf Coast Texas. We describe and compare the patterns of allozyme and RAPD variation in the two diploid races, using a set of 48 individuals from Texas and Mexico (four population samples of 12 individuals each). Twelve of 22 allozyme loci were polymorphic, exhibiting 35 alleles, while seven 10-mer RAPD primers revealed 98 polymorphic bands. Strong regional differences were detected in the extent of allozyme polymorphism: Mexican populations exhibited more internal gene diversity (H_e= 0.20, 0.19) than did the Texan populations (H_e= 0.08, 0.06), although the number of RAPD bands in Texas (n= 62) was only marginally smaller than in Mexico (n= 68). F-statistics for the allozyme data, averaged over loci, revealed strong regional differentiation (mean F_RT=+ 0.30), as well as some differentiation among populations within regions (mean F_PR=+ 0.09). In order to describe and compare the partitioning of genetic variation for multiple allozyme and RAPD loci, we performed an Analysis of Molecular Variance (AMOVA). AMOVA for both allozyme and RAPD data revealed similar qualitative patterns: large regional differences and smaller (but significant) population differences within regions. RAPDs revealed greater variation among regions (58.4% of total variance) than allozymes (45.2%), but less variation among individuals within populations (31.9% for RAPDs vs. 45.2% for allozymes); the proportion of genetic variance among populations within regions was similar (9.7% for RAPDs vs. 9.6% for allozymes). Despite this large-scale concordance of allozyme and RAPD variation patterns, multiple correlation Mantel techniques revealed that the correlations were low on an individual by individual basis. Our findings of strong regional differences among the diploid races will facilitate further study of polyploid evolution in buffalograss.     

Evaluation of Genetic Variation in the Clown Knifefish, Chitala chitala, Using Allozymes, RAPD, and Microsatellites

Twenty-seven enzyme systems, six random amplified polymorphic DNA (RAPD) primers, and two microsatellite loci were tested to determine intraspecific divergence in the natural population of the endangered Indian featherback fish, Chitala chitala, for the first time. The 262 samples of C. chitala were collected from six riverine locations in India: the Satluj, Ganga (Ghagra, Bhagirathi, and Brahmaputra), Mahanadi, and Narmada river systems. The analysis revealed population subdivisions, with an F_ST value from 0.1235 (95% confidence 0.0868–0.1621) for RAPD and a combined F_ST of 0.0344 (95% confidence 0.0340–0.0350) for microsatellite loci. An analysis of 38 allozyme loci did not reveal any polymorphism in the samples from any of the riverine localities; a possible explanation for this could be that the ancestors of Chitala could have faced a population reduction in prehistoric periods, as low allozyme variation is also reported for other species of Chitala from south Asia.     

Characterization of polymorphic microsatellite loci in cultivated and wildPanax ginseng

Ginseng (Panax ginseng) is one of the most important herbal remedies used in East Asia. The present study investigated six polymorphic microsatellite markers (PG29, PG281, PG287, PG668, PG1319, and PG1481) in samples of cultivated and wildP. ginseng collected in Korea. Total allelic number observed in this study was 27 (average allelic numbers per locus: 4.5). All examined loci exhibited deviation from the Hardy-Weinberg equilibrium and deficiency of heterozygosity in both cultivated and wild groups. Although the wild ginseng group exhibited slightly more polymorphic behavior (mean PIC=0.392, GD=0.454 and H_obs=0.129), compared with the cultivated group (mean PIC=0.383, GD=0.438 and H_obs=0.105), no significant differences of allele frequencies and genotype distributions were revealed. By combined analysis of six loci in 100 cultivated ginsengs, 71 different types were observed. The analyzed microsatellite loci in this study will be helpful for understanding genetic variation, QTL mapping and phylogenic studies inPanax species.     

Genetic diversities of four little-known species of Malesherbia (Malesherbiaceae) endemic to the arid inter-Andean valleys of Peru

Few studies of genetic variation have been conducted on plants of the Pacific coastal desert and neighboring Andes of Peru, although the region has many endemic taxa. Enzyme electrophoresis was employed to examine allozyme diversities of four species of the family Malesher-biaceae, an endemic to the arid Andes and coastal desert. One population ofMalesherbia splendens and two ofM. tubulosa, both endemics of the department of Lima, one population ofM. weberbaueri var.weberbaueri, an endemic of the Andean department Huancavélica, and the two Lima populations ofM. scarlatiflora were studied. Fifteen loci were examined for all populations and an additional seven loci were resolved forM. weberbaueri andM. splendens. Malesherbia splendens, which is known from three populations, has a low mean number of alleles per locus (A), proportion of polymorphic loci (P), and expected heterozygosity (H_s) (A=1.214, P=0.214, H_s=0.057).Malesherbia weberbaueri (A=1.231, P=0.154, H_s=0.079) andM. scarlatiflora (A=1.364, P=0.273, H_t=0.083) both have average expected heterozygosities and relatively low mean numbers of alleles per locus and proportions of polymorphic loci. InM. tubulosa, all measures of genetic diversity are high in comparison with other endemics (A=1.818, P=0.364, H_t=0.206).Malesherbia tubulosa has high interpopulation differentiation, whereasM. scarlatiflora has low among-population diversity. The relatively low allozyme diversities, restricted habitats, narrow ranges ofM. splendens andM. weberbaueri, and the morphological similarities of all four species suggest that they evolved recently by founder events. Greater allozyme diversities inM. tubulosa could be attributable to its maintenance of larger populations in a greater variety of habitats.     

High Polymorphism and Autotetraploid Origin of the Rare Endemic SpeciesOxytropis chankaensisJurtz. (Fabaceae) Inferred from Allozyme Data

Using starch electrophoresis, we examined 19 enzyme systems presumably controlled by 35 loci in the rare endemic tetraploid speciesOxytropis chankaensisJurtz. (Fabaceae). Electrophoretic patterns and their genetic interpretation are presented. The isozyme data suggest tetrasomic inheritance in O. chankaensis. Three or four alleles at a particular locus were found in a number of individual plants, which indicate the autotetraploid origin of this species. Seventeen loci were shown to be polymorphic. As reliable gene markers for population systems, we recommend highly active polymorphic systems showing good allozyme separation (Ce-2, Gpi-2, Gpt-2, Idh-2, Lap, Mdh-2, and Mdh-3). Parameters of allozyme variability proved to be very high for a rare species with a restricted range: P = 48.6%, A = 2, A _p = 3.06, H _ob = 0.173.     

Polymorphic microsatellite markers in Thrips hawaiiensis (Thysanoptera: Thripidae)

Thrips hawaiiensis (Morgan, 1913) is an important insect pest of fruits and vegetables. At present, it is primarily distributed in tropical and subtropical area. For a better understanding of the genetic makeup and migration patterns of T. hawaiiensis throughout the world, we isolated 11 novel polymorphic microsatellite loci from an enriched genomic library based on a biotin/streptavidin capture protocol. Genetic parameters were estimated on 80 individual thrips from two natural populations. The results showed that all 11 loci were highly polymorphic; the number of alleles ranged from six to 37, and nine loci demonstrated polymorphic information content (PIC) > 0.5. The observed (H _O) and expected (H _E) heterozygosities ranged from 0.350 to 0.925 and 0.307 to 0.952, respectively. Furthermore, only four locus/population combinations significantly deviated from Hardy–Weinberg equilibrium (HWE). These microsatellite markers have potential utility in population structure and gene flow studies of this species.     

Allozyme diversity among local populations of the pacific mussel <Emphasis Type="Italic">Mytilus trossulus</Emphasis> (Bivalvia: Mytilidae) from polluted areas of Peter the Great Bay (Sea of Japan)

Allozyme variability was assessed, using starch gel electrophoresis, at 15 polymorphic loci in two samples of the Pacific mussel Mytilus trossulus collected from local populations in heavily (Golden Horn Bay) and slightly polluted areas (Sukhoputnaya Bay) of Peter the Great Bay. Significant differences between samples were found in the genotypic frequencies at nine loci and in allele frequencies, at six loci. The results are suggestive of the differential survival of individuals having different genotypes and alleles at some of the surveyed loci under conditions of pollution. Our data are not in conflict with the hypothesis of the adaptive significance of allozyme polymorphism.     

Allozyme variation among Rana balcanica,R. levantina,and R. ridibunda (Amphibia: Anura)1

The allozyme variation among water frogs of the species R. balcanica, R. levantina and R. ridibunda, all formerly considered as one species (R. ridibunda Pallas, 1771), was studied using horizontal starch gel electrophoresis. Blood samples (N # 63) of frogs were collected from five populations in Greece and Israel. Samples (N = 9) of the hybrid frog R. esculenta collected from a locality in Germany were used as an outgroup for phylogenetic analyses. Fifteen enzymes controlled by twenty presumptive loci were identified. Thirteen loci were polymorphic within or among the studied populations. Genetic differentiation among the species was considerably greater than among populations of the same species. Even at Nestos River where R. ridibunda and R. balcanica occur in the same habitats, individuals could be assigned to either species due to characteristic differences of the genotypes (GPI1). This indicates reproductive isolation among these species. The reconstruction of phylogenetic relationships among the three species based on the allozyme data corroborated the model presented on the basis of bioacoustic data: R. ridibunda and R. balcanica (Nei's genetic distance D = 0.0820) are sibling species pertaining to an Eurasian lineage, whereas R. levantina (distance to the European species D = 0.1780 - 0.1955) together with R. perezi represent an independent afroasian lineage.     

Allozyme differentiation and systematics of the endemic subterranean mole rats of South Africa

We have analyzed allozyme differentiation, encoded by 20 putative gene loci, of three genera and four currently accepted species of Southern African mole rats, Bathyergidae (Cryptomys hottentotus hottentotus, C. hottentotus natalensis, C. hottentotus damarensis, Georychus capensis, Bathyergus suillus and B. janetta). Two major results are indicated: (a) genetically the family reflects distinct homozygosity (H=0.019, range 0.00–0.05) as is typical for underground fossorial mammals and (b) remarkable large genetic distances exist between the six taxa analyzed (D= 1.865, range 0.547–2.917). These results suggest that homoselection operates in the underground ecotope in accordance with the niche width variation hypothesis, and that bathyergid systematics needs urgent revision, particularly the genus Cryptomys. The three subspecies of Cryptomys appear to be at least three different species, possibly even falling into two genera. The adaptive radiation of bathyergids started at least in Miocene times, but probably began earlier in the Oligocene as indicated by the fossil record.     

Genetic variation and differentiation in Nordic populations of Elymus alaskanus (Scrib. ex Merr.) Löve (Poaceae)

 To gain information on the extent and nature of genetic variation in Elymus alaskanus, levels and distribution of genetic variation were assessed within and among 13 populations originating from Iceland, Norway, Sweden and Russia using allozymes. The results showed that four (30.7%) of the 13 loci were polymorphic within the species, while the mean percentage of polymorphic loci within the populations was 1.9%. The mean number of alleles per locus for the species was 1.8 and 1.02 across the populations. Genetic diversity at the species level was low (H _es=0.135), and mean population diversity was notably lower (H _ep=0.005). A high degree of genetic differentiation was observed among populations. The salient points emerging from this study are: (1) statistically significant differences were found in allele frequencies among populations for every polymorphic locus (P<0.001), (2) the high mean coefficient of gene differentiation (G _ST) showed that 95% of the total allozyme variation was attributable to differences among populations, and (3) relatively high genetic distances between the populations were obtained (mean D=0.16). The Norwegian populations had the highest genetic diversity as compared with the other populations. Geographical comparisons revealed three different groups of populations clearly differentiated, i.e. Scandinavia (Norway and Sweden), Iceland and Russia. Cluster and principal coordinates analyses revealed the same genetic patterns of relationships among populations. Generally, this study indicates that E. alaskanus contains low allozymic variation in its populations. The implications of these results for the conservation of the species are discussed. Received: 23 October 1998 / Accepted: 19 December 1998     

珍稀濒危植物长叶红砂种群遗传多样性的ISSR分析

采用ISSR分子标记技术,对濒危小灌木长叶红砂(Reaumuria trigyna) 集中分布的5个种群的遗传多样性水平和遗传结构进行了研究。14条引物共检测到114个位点,其中99个为多态位点,多态位点比率为86.84％,长叶红砂种群具有较高的遗传多样性。物种水平上Shannon多样性指数(I)为0.468 8,Nei基因多样性指数(H)为0.308 4;种群水平上,多态位点比率P为77.89%,I为0.410 6,H为0.260 9,基因分化系数Gst为0.106 9,揭示了长叶红砂种群遗传变异多存在于种群内,种群间的遗传分化较小,占10.69%。 基因流(Nm)为4.178 7＞1,说明种群间的基因交流,防止了由于遗传漂变导致的遗传分化。聚类分析表明长叶红砂种群遗传距离与地理距离之间无显著的相关性。研究结果说明遗传多样性水平与物种本身特性和所处不同群落有关,濒危植物并不一定表现为遗传变异水平的降低。     

Genetic structure and differentiation of populations of the tetraploid species <Emphasis Type="Italic">Oxytropis chankaensis</Emphasis> (Fabaceae)

The population genetic variation of the tetraploid species Oxytropis chankaensis Jurtz. (Fabaceae), a local endemic of the western coast of Khanka Lake (Primorye), was examined. Five populations were analyzed using 28 isozyme loci encoding 16 enzyme systems. Significant allelic heterogeneity among the populations was found for six out of twelve polymorphic loci. The heterozygosity of the samples (total sample size 294 plants) H _e = 0.301 was considerable higher than the mean values in populations of endemic species (0.076). Based on the results of this study, we identified two groups of O. chankaensis populations (southern and northern), in spite of the absence of marked hiatus between them. Of special interest is the population from Przhewalski Spit, which is a natural reserve of genetic diversity of the species and the putative center of formation of the autotetraploid O. chankaensis.     

A second case of genetic host races in Rhagoletis? A population genetic comparison of sympatric host populations in the European cherry fruit fly,Rhagoletis cerasi

Despite an increasing acceptance in the biological community for sympatric speciation as a mode of species formation, well documented examples of sympatrically evolved ‘incipient species’ remain rare. The sympatric host races of apple maggot, Rhagoletis pomonella (Walsh), represent one of the most prominent case studies for sympatric speciation via a host shift. The European cherry fruit fly, R. cerasi (L.), shows strong ecological similarities to R. pomonella: (1) infestation of two different host plants, Lonicera xylosteum L. and Prunus avium L., and (2) divergent phenological and behavioral adaptations of flies on different hosts. The population genetic study presented here addresses whether the host associated populations of R. cerasi also represent genetically differentiated true host races. Out of a total of 29 allozyme loci examined, six were polymorphic and used to analyze six sympatric pairs of R. cerasi populations on Lonicera and Prunus from Switzerland and Germany. A direct comparison of allele frequencies between sympatric sites showed no pattern indicative of host races in R. cerasi. However, the hierarchical F‐statistic for one locus, mannose 6‐phosphate isomerase (Mpi), showed significant population differentiation that was in accordance with host race differentiation. Mpi is one of several loci that are also diagnostic for host race differentiation in R. pomonella. Results from Mpi suggest the formation of sympatric host races in R. cerasi, but additional polymorphic markers are necessary.     

Linkage Disequilibrium in Natural Populations of DROSOPHILA MELANOGASTER. Seasonal Variation

Linkage disequilibrium among ten polymorphic allozyme loci and polymorphic inversions on chromosomes 2 and 3 in a natural population of Drosophila melanogaster was examined early and late in the annual season. Similar to previous studies, little linkage disequilibrium was observed among allozymes. The two significant cases that were observed in the first sample behaved in a contradictory way. One declined much more rapidly than expected due simply to recombination; the other declined slowly as expected. There was little change in allozyme or inversion frequencies during the season.     

Low level of allozyme polymorphism in relict aquatic plants of the Far East Nelumbo komarovii Grossh. and Euryale ferox Salisb.

Using allozyme analysis, genetic variation of two relict aquatic plants from Primorsky krai, Komarov lotus (Nelumbo komarovii Grossh.) and Gorgon plant (Euryale ferox Salisb.), was examined. The absence of allozyme variation in the Primorye populations of Nelumbo komarovii along with low polymorphism level in the population of Euryale ferox (P ₉₅ = 7.69; A = 1.07; H _o = 0.072; H _e = 0.038) was demonstrated. Since the data for the species examined are reported for the first time ever, the phenotypes and genetic interpretation of the enzyme systems tested are presented. The isoenzyme profiles of N. komarovii were compared with the data reported for N. nucifera from China. The absence of allozyme variation in N. komarovii, along with extremely low level of variation revealed for E. ferox, is discussed in association with the evolutionary histories of these species, their dispersal after the Pleistocene-Holocene cooling, and survival on this territory in range boundaries.     

Molecular biogeography of the arctic-alpine disjunct burnet moth species Zygaena exulans (Zygaenidae, Lepidoptera) in the Pyrenees and Alps

Aim The phylogeography of ‘southern’ species is relatively well studied in Europe. However, there are few data about ‘northern’ species, and so we studied the population genetic structure of the arctic‐alpine distributed burnet moth Zygaena exulans as an exemplar. Location and methods The allozymes of 209 individuals from seven populations (two from the Pyrenees, five from the Alps) were studied by electrophoresis. Results All 15 analysed loci were polymorphic. The mean genetic diversities were moderately high (A: 1.99; H_e: 11.5; P: 65%). Mean genetic diversities were significantly higher in the Alps than in the Pyrenees in all cases. F_ST was 5.4% and F_IS was 10%. Genetic distances were generally low with a mean of 0.022 between large populations. About 62% of the variance between populations was between the Alps and the Pyrenees. The two samples from the Pyrenees had no significant differentiation, whereas significant differentiation was detected between the populations from the Alps (F_ST = 2.8%, P = 0.02). Main conclusion Zygaena exulans had a continuous distribution between the Alps and the Pyrenees during the last ice age. Most probably, the species was not present in Iberia, and the samples from the Pyrenees are derived from the southern edge of the glacial distribution area and thus became genetically impoverished. Post‐glacial isolation in Alps and Pyrenees has resulted in a weak genetic differentiation between these two disjunct high mountain systems.