Development and characterization of microsatellite markers in the Colorado potato beetle,Leptinotarsa decemlineata

The characterization of 11 Leptinotarsa decemlineata microsatellite loci isolated using the fast isolation by AFLP of sequences containing repeats (FIASCO) protocol is reported. Three to 15 alleles per locus were detected in 54 beetles collected from four populations of L. decemlineata. The mean number of alleles was 7.4 ± 3.2 and the level of expected heterozygosity ranged from 0.451 to 0.798. The total exclusionary probabilities using these loci for the first and the second parent were 0.982 and 0.999, respectively. These are the first microsatellite loci characterized from the Colorado potato beetles that can be used for estimating genetic diversity, population structure and parentage analysis.     

德宏水牛微卫星标记分析的群体遗传变异

德宏水牛是云南省地方水牛的优良品种之一,为了进一步阐明其群体遗传变异和遗传结构,筛选了分别位于水牛14条染色体上的15对微卫星引物,对德宏水牛81个个体进行了检测分析.共检测到62个等位基因,每个座位等位基因数目从2到6个不等,平均等位基因数为4.13,该水牛群体期望杂合度和多态信息含量分别为0.6520±0.1526和0.5863±0.1789,各座位的遗传分化系数在0～0.0919之间,平均值为0.0202.每个座位的基因流较大,平均12.1502.研究结果表明德宏水牛群体遗传多样性较丰富,亚群间的遗传分化程度低,基因流较大,且很少发生近交.     

Genetic differentiation and relationship of the dipodidsAllactaga andJaculus (Mammalia, Rodentia) in Egypt based on protein variation

Genetic differentiation among 14 populations representing all Egyptian dipodid (jerboa) species and subspecies was examined at 25 structural loci and interspecific relationships are discussed. Of the 25 loci, only 3 were monomorphic, with the same allele fixed in all taxa, 9 loci were monomorphic, but demonstrated intertaxon variation, and only 13 loci were polymorphic. The overall mean number of alleles per locus (A) was 1.23 ± 0.02 and the average percentage of polymorphic loci per taxon (P) was 23%. The overall mean of observed heterozygosity (H _o) was found significantly higher than that of expected heterozygosity (He); the overall means per taxon were 0.25 ± 0.017 and 0.085 ± 0.007. Mean levels of genetic identity (I) were 0.970 ± 0.003 among geographic populations, 0.718 ± 0.022 between subspecies, 0.590 ± 0.030 between congeneric species, and 0.368 ± 0.020 between genera. Phenetic analysis of genetic distance matrix produced a phenogram indicating a close association ofJaculus orientalis Erxleben, 1777 toJaculus jaculus (Linnaeus, 1758), particularly to its subspeciesJaculus jaculus butleri (Thomas, 1922), and indicating a distinct affinity between these latter two species andAllactaga tetradactyla (Lichtenstein, 1823). Estimates of genetic divergence demonstrated that J. orientalis appears to have shared a more recent common ancestor withJ. jaculus thanA. tetradactyla. Divergence of these species would have occurred by Miocene (ca 9.6 to 18.7 million years ago). The pattern of relationships of the dipodid species indicated in this study was closely consistent with the hypotheses derived from morphological and chromosomal data.     

HIGH LEVELS OF GENETIC VARIABILITY IN THE HAPLOID MOSS PLAGIOMNIUM CILIARE

Horizontal starch-gel electrophoresis was used to measure variability at 14 enzyme loci from 13 natural populations of the dioecious moss Plagiomnium ciliare. Overall levels of genetic polymorphism were unexpectedly high for a haploid organism. Using a 1% frequency criterion, 71% of the loci surveyed were polymorphic for the species as a whole. The number of alleles per polymorphic locus for the species as a whole was 2.82 ± 0.34 (mean ± standard error), and mean gene diversity per locus was 0.078 ± 0.035. While total gene diversity (H_T = 0.178) was similar to that observed for highly outcrossed diploid plants such as pines, the variance within (H_S = 0.098 ± 0.027) and among (D_ST = 0.080 ± 0.033) populations was more evenly distributed than that reported for populations of conifers. Genetic distances between populations ranged from 0.0002 to 0.2064, with mosses from the Piedmont region of the southeastern United States showing less differentiation among populations than did mosses from the Appalachian Mountains. Gene diversity was much reduced in populations from disturbed, secondary forests in the Piedmont (0.058 ± 0.018) relative to those from minimally disturbed, primary forests in the mountains (0.146 ± 0.048). Intensive sampling within populations revealed heterogeneity even within small (5 × 5 cm) clumps. The discovery of high levels of genetic variability in a plant with a dominant haploid life cycle challenges the traditional view of bryophytes as a genetically depauperate group. Multipleniche selection is proposed as a possible explanation for this anomaly, but the data are also consistent with the view that allozyme polymorphisms are selectively neutral.     

Isolation and characterization of microsatellite loci in Tripterygion delaisi

We isolated 49 microsatellite loci from a genomic library of Tripterygion delaisi×anthosoma enriched for CA and GA repeats. Ten loci were screened in 30 individuals with high numbers of alleles per locus (averaging 15.5 ± 2.86) and observed heterozygosity (averaging 0.765 ± 0.052). No deviations from Hardy–Weinberg expectations were detected. These highly polymorphic markers will be useful in determining the spatial patterns of genetic diversity between and within subspecies of Tripterygion delaisi.     

Genetic diversity and bottleneck studies in endangered Bhutia and Manipuri pony breeds

For studying the genetic diversity and bottleneck problem in Bhutia and Manipuri pony breeds of India, we analysed DNA samples of 34 Bhutia and 50 Manipuri, true to breed, ponies using 47 polymorphic microsatellite markers. All the microsatellites were observed to be highly polymorphic in nature in both Bhutia and Manipuri breeds with mean no. of alleles as 8.702 ± 0.0493 and 8.416 ± 0.0548 respectively. Genetic diversity values in terms of heterozygosity values within individual breeds were also high with very low inbreeding (Fis 0.102 and 0.055 in Bhutia and Manipuri ponies, respectively). Number of alleles in both the populations together ranged from 3 to 18 with an average of 10.851 ± 1.583 per locus. The mean effective number of alleles was observed to 5.34 ± 0.253. All loci except ASB017 and HTG004 showed high values of allele richness (>5.0). The mean observed and expected heterozygosities were 0.7159 ± 0.022, 0.7986 ± 0.011 (Levene’s) and 0.7936 ± 0.011 (Nei’s), respectively. The high mean values of heterozygosity indicated the presence of high genetic diversity in both the pony populations. The overall mean value of within-population inbreeding estimates (Fis) was low (0.101 ± 0.023) indicating low to moderate level of inbreeding. Bottleneck studies revealed that no recent bottleneck problem has taken place in both the populations. Both pony populations were found to be in mutation drift equilibrium. The study reveals that both the pony breeds have high diversity and timely action needs to be taken to conserve them.     

Genic Heterozygosity and Protein Polymorphism among Local Populations of OENOTHERA BIENNIS

Twenty enzyme loci were studied in 44 Illinois populations of Oenothera biennis; four were polymorphic. Most of the variation was distributed between populations. Fifty-nine percent of the populations had one genotype, 27% two genotypes and the remaining 16% from three to five genotypes; the average was 1.50. The proportion of genetic diversity present in single populations is.38 of that present in the state. Members of single populations were uniformly heterozygous for 1 to 4 loci. The mean heterozygosity per population ranged from 0 to 20%. For Illinois populations collectively, heterozygosity averaged 4.5%. There was much gene frequency heterogeneity between populations. The true standardized genetic variance among populations for alleles at polymorphic loci varied from.40 to.78. Populations from Cook County were much more similar inter se than those downstate, had fewer genotypes and polymorphic loci, and had less heterozygosity than downstate populations. The mean normalized genetic identity among Cook County populations was.987 versus.947 for downstate populations. The mean number of genotypes per population in Cook County was 1.06 versus 2.40 in downstate populations. There was only one polymorphic locus in Cook County, VLP. The genetic structure of Oe. biennis suggests that single populations are colonized by one, or at best a few individuals. Cook County populations are judged to be less variable than downstate populations because the mean age of the populations probably is less than that of those downstate.     

古丈县南方红豆杉天然群体的遗传多样性研究

通过对分布于湖南省古丈县的南方红豆杉(Taxus chinensis)天然群体中30个个体的功能叶片的过氧化物酶(POD)和酯酶(EST)同工酶谱带的分析,研究了该天然群体的遗传多样性水平.结果表明:30个个体中都存在有迁移率相同的谱带(4条),占酶谱带总数的26.7%;两个酶系统共检出15条酶谱带,等位基因位点数4个,其中多态位点数3个,单态位点数1个,多态位点百分数P=75%,平均每个位点的等住基因数A=3,平均有效等位基因数Ae=2.75,平均期望杂合度He=60.7%,平均实际杂合度Ho=22.2%,表明该群体有较丰富的遗传多样性.     

Genetic variability in a temperate intertidal Phoronid,Phoronopsis viridis

The Phoronida are a coelomate phylum consisting of only two genera and about 12–15 described species. Phoronids probably represent the common ancestral stock of all lophophorates, and may be the most primitive living deuterostomes. Using the techniques of starch gel electrophoresis, we have studied genetic variation at 39 loci in 120 individuals of Phoronopsis viridis collected in Bodega Harbor, Bodega Bay, California. Allelic variation was found at 27 (69.2%) loci. If a locus is considered polymorphic when the frequency of the most common allele is no greater than 0.99, the proportion of polymorphic loci in the total sample is 48.7%. The average number of alleles per locus is 2.23. The expected frequency of heterozygous loci per individual on the assumption of Hardy-Weinberg equilibrium is 9.4%. There is evidence of inbreeding; the mean value of F, Wright's fixation index, is 0.21±0.02. Genetic variability in P. viridis is intermediate among marine invertebrates. The tropical clam, Tridacna maxima, has on the average 20.2% heterozygous loci per individual. At the other extreme, a brachiopod from Antarctica, Liothyrella notorcadensis, has an average of 3.9% heterozygous loci per individual. Among marine invertebrates, there seems to be a gradient of decreasing genetic variability from low to high latitudes, which may reflect their different adaptive strategies.     

Variation of mitochondrial DNA in Far Eastern mullet pilengas, Liza haematocheilus Temminck and Schlegel, acclimatized in the Azov-Black Sea basin

This study continues the investigation of genetic variation in the populations of native and acclimatized in the Azov-Black Sea basin pilengas from the Sea of Japan. The previous comparison based on allozyme analysis was supplemented by analysis of restriction polymorphism of a mitochondrial DNA fragment containing the cytochrome b gene and the D-loop. Five out of fifteen tested endonucleases detected polymorphic sites. In the samples of native and acclimatized pilengas, five common haplotypes were found; ten and three “unique” haplotypes were detected in the Far Eastern and the Azov populations, respectively. The differences in haplotype distributions between these populations were highly significant (P < 0.001). The mtDNA variation was lower in the Azov than in the Far Eastern population (haplotype diversity μ respectively 6.35 ± 0.27 and 9.14 ± 0.55), which is in good agreement with the decrease in the number of polymorphic loci and the mean number of alleles per locus, found earlier for allozyme markers in this population. The reasons for these differences in the acclimatized population are discussed.     

Isolation and characterization of 18 polymorphic microsatellite markers for the critically endangered stellate sturgeon, <Emphasis Type="Italic">Acipenser stellatus</Emphasis>

The stellate sturgeon, Acipenser stellatus, is a critically endangered fish species. Knowledge on its genetic diversity and population structure is urgently needed to enable the identification of management units in order to prevent extinction. Therefore, 18 species-specific, polymorphic microsatellite loci have been isolated using GS-FLX Titanium pyrosequencing, arranged into 6 multiplex PCR sets, and characterized in 52 individuals (20 farmed and 32 wild). The total number of alleles per locus varied between 3 and 36 with an average of 8.44. The wild individuals were more diverse with an average number of 8.17 alleles per locus than the farmed ones with 3.28 alleles per locus. Observed heterozygosities ranged from 0.050 to 0.950 in the farmed and from 0.094 to 0.969 in the wild individuals. Significant deviations from Hardy-Weinberg equilibrium were found at 3 loci of the farmed and 5 loci of the wild individuals. The two sturgeon groups were significantly differentiated (F _ST = 0.118). The high sensitivity and discriminatory power of the 18 loci were proven by a very low overall probability of identity for siblings (PIsib = 8.73 × 10^?6) and a high accuracy of self-classification (98%). Thus, these newly developed markers represent a valuable genetic toolbox to identify management units for species conservation and sustainable fisheries.     

Microsatellite Polymorphisms in Cassava Landraces from the Cerrado Biome, Mato Grosso do Sul, Brazil

Using nine microsatellite loci, we investigated genetic structure and diversity in 83 Brazilian cassava accessions, including several landraces, in the Cerrado biome in Mato Grosso do Sul, Brazil. All nine loci were polymorphic, averaging 6.00 alleles per locus. Treating each of seven municipalities as a cassava group or population, they averaged 3.5 alleles per locus, with 97% polymorphic loci, high values for observed heterozygosity (0.32) and gene diversity (0.56). Total genetic variability was high (0.668), and most of this genetic variability was concentrated within municipalities (0.577). Cluster and structure analyses divided accessions into two major clusters or populations (K = 2). Also, a significant genetic versus geographic correlation was found (r = 0.4567; P < 0.0260). Migratory routes in the Cerrado are considered main contributors to the region’s high cassava diversity and spatial genetic structure, amplifying interactions between traditional farmers and the evolutionary dynamics of this crop.     

Cross-species amplification of 44 microsatellite loci developed for Chaetodon trifascialis, C. lunulatus and C. vagabundus in 22 related butterflyfish species

Forty‐four microsatellite primers developed for three species of butterflyfish were cross‐tested against 22 related confamilial species. Amplification success and cross‐species transferability of these markers were moderately high. Between 24 and 37 loci were amplified successfully in each species, with a mean success rate per species of 71.7% (± 1.8 SE). Rates of amplification success were comparable among primers designed for the three source species, ranging from a mean success rate per species of 16.9 loci (± 0.8 SE) for Chaetodon trifascialis source loci to 13.7 loci (± 1.5 SE) for C. vagabundus source loci. Polymorphism rates were high (76.1%± 3.1 SE of all successfully amplified loci), and 10 loci were polymorphic in all successfully amplified species (Tri14, B11, C5, D3, D113, D6, D117, D120, D111, D118). The number of alleles per polymorphic locus ranged from 2 to 8, and the average number of alleles across all polymorphic loci and all species was 3.6 (± 0.07 SE). Polymorphism rates were higher overall in primers designed for C. vagabundus (89.9%± 3.9 SE). Overall cross‐testing success was lowest for Heniochus chrysostomus, the most phylogenetically divergent species. The significant cross‐testing reported here provides a valuable resource that will enable population genetics studies to be undertaken on a range of butterflyfishes without the need for expensive and time‐consuming de novo microsatellite development.     

GENETIC VARIATION IN BROMUS TECTORUM (POACEAE): POPULATION DIFFERENTIATION IN ITS NORTH AMERICAN RANGE

Allelic variation in seedlings from 60 North American populations of the alien annual grass Bromus tectorum was determined at 25 loci using starch gel electrophoresis. Populations were collected from four regions; east of the Rocky Mountains, Nevada and California, the Intermountain West, and British Columbia. Compared to other diploid seed plants, genetic variation within these populations of B. tectorum is low: 4.60% of loci are polymorphic per population, with an average of 1.05 alleles per locus and a mean expected heterozygosity of 0.012. Although 2,141 individuals were analyzed, no heterozygous individuals were detected, and consequently, mean observed heterozygosity is 0.000. Extensive deviations from Hardy-Weinberg expectations were observed at every polymorphic locus due to heterozygote deficiencies. The mean genetic identity (Nei's I) between population pairs was 0.980 and indicates a high level of overall genetic similarity among populations. The among-population component of the total gene diversity is high (G_ST ⁼ 0.478), indicating substantial genetic differentiation among populations. These results are consistent with previous reports for highly self-pollinating plants of low genetic variation and substantial genetic differentiation among populations. Despite the lack of genetic variation as measured by enzyme electrophoresis, this weedy grass has become exceedingly abundant in a diverse array of arid environments throughout much of western North America, perhaps due to phenotypic plasticity.     

Genetic diversity of the cultured giant freshwater prawn (Macrobrachium rosenbergii) in China based on microsatellite markers

The Giant freshwater prawn (GFP), Macrobrachium rosenbergii, is one of the most important crustacean species cultured in China, but information on the genetic diversity of GFP in China is still limited. In this study we analyzed the genetic diversity of 6 cultured GFP populations collected in China, and 2 wild populations from Viet Nam. The wild populations were included as a reference so that the genetic diversity of the introduced cultured GFP populations could be evaluated. We used six polymorphic primer pairs to estimate genetic diversity. Results showed that the number of alleles per locus ranged from 6 to 19. The mean observed heterozygosity (0.364 ± 0.090) was less than the expected heterozygosity (0.893 ± 0.016); highly significant deficiencies in heterozygotes were detected (100%). The polymorphism information content (PIC) of 8 populations at 6 loci was greater than 0.5 (varying from 0.616 to 0.912). Pair-wise genetic differentiation (F_ST) varied from 0.006 to 0.129 among the eight populations. The dendrogram based on genetic distance showed six populations distributed on the same major branch, suggesting that they have closely genetic distance and two populations (ZJC and GDB) distributed on the other branch, so they also have closely genetic distance. The results of study provide a baseline assessment of genetic diversity in some populations in China and Viet Nam that will be useful for the development of breeding programmes in the future.     

Genetic divergence between populations of two closely related troglobitic beetle species (Speonomus: Bathysciinae,Coleoptera)

Genetic variability and divergence at 13 enzyme loci were studied in two species of troglobitic beetles (Speonomus zophosinus andhydrophilus 12 populations) collected from the Pyrenees mountains. These allopatric species exhibit a high degree of specialization to underground life and a very small geographical range. Four diagnostic loci arePgi, Est-2, Pac-1, Phi. Within each species polymorphic loci exhibited marked spatial variation of allele frequencies, sometimes over short distances (5–10 km). The levels of genetic variability are comparable to those observed in non-cave invertebrates (frequency of polymorphic loci: 0.31; average expected heterozygosity: 0.112±0.015). Moreover, a single species showed variability in the average level of heterozygosity (0.06 to 0.172). Patterns of genetic differentiation among species were also studied; mean genetic distance (D: 0.263±0.010) between the twoSpeonomus species was of the same order of magnitude as most data reported in the literature between species. A significant heterozygote deficiency occurs in local populations and for all loci. This deficit does not vary from locus to locus. Several hypotheses were examined in an attempt to account for these observations in connection with the species biology. This deficit probably is due to the breeding structure of the population (inbreeding and assortative mating) coupled with limited dispersal ability.     

Characterization of 32 microsatellite loci for the Pacific red snapper, <Emphasis Type="Italic">Lutjanus peru</Emphasis>, through next generation sequencing

We developed a set of hypervariable microsatellite markers for the Pacific red snapper (Lutjanus peru), an economically important marine fish for small-scale fisheries in the west coast of Mexico. We performed shotgun genome sequencing with the 454 XL titanium chemistry and used bioinformatic tools to search for perfect microsatellite loci. We selected 66 primer pairs that were synthesized and genotyped in an ABI PRISM 3730XL DNA sequencer in 32 individuals from the Gulf of California. We estimated levels of genetic diversity, deviations from linkage and Hardy–Weinberg equilibrium, estimated the frequency of null alleles and the probability of individual identity for the new markers. We reanalyzed 16 loci in 16 individuals to estimate genotyping error rates. Eighteen loci failed to amplify, 16 loci were discarded due to unspecific amplifications and 32 loci (14 tetranucleotide and 18 dinucleotide) were successfully scored. The average number of alleles per locus was 21 (±6.87, SD) and ranged from 8 to 34. The average observed and expected heterozygosities were 0.787 (±0.144 SD, range 0.250–0.935) and 0.909 (±0.122 SD, range 0.381–0.965), respectively. No significant linkage was detected. Eight loci showed deviations from Hardy–Weinberg equilibrium, and from these, four loci showed moderate null allele frequencies (0.104–0.220). The probability of individual identity for the new loci was 1.46^?62. Genotyping error rates averaged 9.58%. The new markers will be useful to investigate patterns of larval dispersal, metapopulation dynamics, fine-scale genetic structure and diversity aimed to inform the implementation of spatially explicit fisheries management strategies in the Gulf of California.     

Genetic diversity and population structure of Yucca filamentosa (Agavaceae)

Using 19 allozyme loci we studied genetic diversity in 18 populations of Yucca filamentosa (Agavaceae) from the southeastern United States. Of the 19 loci surveyed, 17 (89.5%) were polymorphic in at least one of the populations sampled. There was considerable variation among populations in the percentage of polymorphic loci (range = 31.6-84.2%, mean = 67.6%). Similar heterogeneity among populations was observed for mean number of alleles per polymorphic locus (range = 2.0-3.0; mean = 2.48) and mean expected heterozygosity (range = 0.113-0.288; mean = 0.213). On average, 83% of the total genetic diversity was found within populations. Duplications of three allozyme loci were detected in several populations. The life-history characteristics of Y. filamentosa (a long-lived, semiwoody, predominantly outcrossing monocot with a large geographical range) may contribute to the maintenance of such high levels of genetic diversity. These results contradict expectations of the genetic structure of Y. filamentosa based on observations of the dispersal and pollination behavior of its sole pollinator, Tegeticula yuccasella, the yucca moth.     

Isolation and characterization of 12 polymorphic microsatellite markers for the frog Pelophylax hubeiensis

Twelve polymorphic microsatellite loci were isolated from an (AC)n- and (AG)n-enriched DNA library for the endemic Chinese frog Pelophylax hubeiensis (Ranidae). The number of alleles per locus ranged from two to eight, with a mean of 5.17. The observed and expected heterozygosities ranged from 0.226 to 0.839 and from 0.204 to 0.826, with means of 0.568 and 0.656, respectively. No significant linkage disequilibrium was detected among these loci. However, two significant deviations from HWE were discovered at loci Pehu-11 and Pehu-12 (P<0.05). MICRO-CHECKER tests showed that null alleles could be present at locus Pehu-12. These polymorphic microsatellite loci can be employed for exploring mating mechanisms, population genetic structure and other relevant genetic investigations of P. hubeiensis.