Allozyme variation in endangered Castanea pumila var. pumila

Allozyme genetic variation in 12 populations of the endangered Castanea pumila var. pumila (Allegheny chinkapin), sampled across the natural range of the species in the United States, was evaluated using 11 loci from seven enzyme systems. At the species level, the percentage of polymorphic loci (Ps) was 72.7 %, the mean number of alleles per locus (As) was 1.9, the mean number of alleles per polymorphic locus (APs) was 2.3, the effective number of alleles per locus (Aes) was 1.5 and the genetic diversity (Hes) was 0.296. At the population level, Pp = 49.2 %, Ap = 1.5, Aep = 1.4, APp = 2.1 and Hep = 0.21. Most of the allozyme variation (70 %) in C. pumila var. pumila occurred within populations. Wright's gene flow rate [Nm(W)] was as low as 0.57. Population differentiation along the species range was not detected. Populations of C. pumila var. pumila in Florida had the most variable levels of genetic diversity, but populations in Virginia and Mississippi also showed high levels. Based on the results of this study, conservation management strategies are discussed.     

Microsatellites in Aureliana fasciculata var. fasciculata (Solanaceae), a shrub that inhabits the Atlantic Rainforest

? Premise of the study: Microsatellite primers were developed in Aureliana fasciculata var. fasciculata, a shrub that inhabits the Atlantic Rainforest, to investigate the intraspecific genetic structure of the species. ? Methods and Results: Nine pairs of microsatellite primers were designed that were based on a simple sequence repeat (SSR)-enriched library. Seven loci were amplified successfully. Two populations of A. fasciculata var. fasciculata were genotyped and all loci were polymorphic, with two to five alleles per locus per population detected. Cross-species amplification was achieved in other members of the genus and members of the sister genus Athenaea. ? Conclusions: The development of these microsatellite markers will contribute to future population genetic studies in A. fasciculata var. fasciculata. Cross-amplification among other species in the genus and in the sister genus Athenaea might be a useful tool to establish patterns of genetic variability within the Atlantic Rainforest.     

Isolation and characterization of nine polymorphic microsatellite loci in the endangered shrub Disanthus cercidifolius var. longipes (Hamamelidaceae)

Nine polymorphic microsatellite loci were isolated and characterized from an AC-enriched genomic library of Disanthus cercidifolius var. longipes. Microsatellite polymorphism was investigated using 24 individuals from one natural population. The observed number of alleles per locus ranged from two to four. Observed and expected heterozygosities ranged from 0.17 to 0.92 and from 0.16 to 0.72, respectively. These polymorphic microsatellite loci provide useful tools for the ongoing population genetic studies of D. cercidifolius var. longipes.     

光叶珙桐的等位酶分析及其生物地理学意义

采用水平切片淀粉凝胶电泳实验方法,对光叶珙桐(Davidia involucrata var．vilmoriniana)4个居群、78个个体、11种酶系统,21个位点进行了等位酶分析。各居群的多态位点百分比P＝28.6％-47．6％,实际杂合度Ho＝0．177—0.308,平均每个位点的等位基因数A=1．3-1．6。在变种水平上,多态位点百分比P＝52．4％,平均每个位点的等位基因数A＝1．8,实际杂合度Ho＝0．272,期望杂合度Ho＝0．216。基因分化系数FST＝0．1928,说明光叶珙桐居群问分化小。各项指标表明光叶珙桐的遗传多样性水平较高,比原变种珙桐高,外部环境可能是影响珙桐分布格局的重要因素。昭通与宝兴的遗传多样性明显高于其他两个居群,遗传多样性保存较为完整,而处于分布区东、西两侧边缘(云南西北部和湖北五峰)遗传多样性则相对较低,推测四川盆地边缘山地是该种的遗传多样化中心,可能是在地质灾难中(如第四纪冰期),光叶珙桐真正的避难所。分布区东西两侧的居群可能是从四川盆地边缘山地扩散而来。     

Allozyme frequencies in New Jersey and North Carolina populations of Atlantic white-cedar, Chamaecyparis thyoides (Cupressaceae)

The present study was conducted to assess the distribution of genetic variation within and among populations of Chamaecyparis thyoides in both marginal and centrally located populations. Allozyme frequency analyses of ten loci from foliage of four New Jersey populations and two North Carolina populations of C. thyoides showed polymorphic loci = 50%, mean number of alleles per locus = 2.8, effective number of alleles per locus = 1.17, and expected heterozygosity = 0.14. Diversity was highest in two populations from southern New Jersey. The isolated population at High Point, New Jersey had only two polymorphic loci and expected heterozygosity of 0.03. There was no correlation between genetic and geographic distances among populations, implying that cedar must have possessed some means of long-distance dispersal at the end of the last glacial period, rather than advancing northward step by step.     

Fifteen polymorphic microsatellite markers for the giant spiny frog, Paa spinosa

We characterized 15 polymorphic microsatellite loci for the giant spiny frog, Paa spinosa. These loci were highly polymorphic when screened in 38 individuals from two different populations, with nine to 23 alleles per locus. The range of observed and expected heterozygosities was 0.231-0.916 and 0.296-0.944, respectively. These polymorphic loci will be useful for assessing genetic diversity, population structure, gene flow, population assignment and determining paternity in the giant spiny frog.     

Isolation and characterization of microsatellite loci from Weigela coraeensis (Caprifoliaceae)

Weigela coraeensis var. coraeensis is a deciduous shrub species distributed in Japan on the mainland, Honshu, whereas its variety W. coraeensis var. fragrans is endemic to the Izu Islands located south of Honshu. We isolated eight polymorphic microsatellite loci from the species and characterized these loci for 20 individuals from a population in Honshu. The number of alleles per locus ranged from 7 to 15 and the observed and expected heterozygosities ranged from 0.60 to 0.90 and from 0.65 to 0.90, respectively. These eight polymorphic microsatellites will be useful for examining intraspecific genetic differentiation in W. coraeensis.     

Development and characterization of microsatellite markers for the endangered Chinese yew Taxus chinensis var. mairei (Taxaceae)

We isolated and characterized 11 microsatellite loci for the Chinese yew, Taxus chinensis var. mairei, an endangered tree species in China, by constructing a (CA)(12)-enriched library. The number of alleles per locus ranged from 5 to 10. The observed heterozygosities ranged from 0.2500 to 0.8333 and the expected heterozygosities ranged from 0.5196 to 0.8680. No significant linkage disequilibrium was detected at these loci. However, four loci significantly deviated from Hardy-Weinberg equilibrium. The null alleles were found to be present at locus Tach9 and locus Tach11 by the Micro-checker test (P < 0.001). These polymorphic loci could be employed in research of gene flow and spatial genetic patterns of T. chinensis var. mairei.     

四个不同地理区域的库蚊复合组居群的遗传多样性研究

库蚊种群在农药强选择压下的生存及抗性进化是与其种群的遗传多样性密切相关的。本文主要从居群的遗传多样性角度出发 ,研究其抗性遗传行为。分别对采自 4个不同地理区域的库蚊复合组样群 ,采用水平切片淀粉凝胶电泳的方法 ,分析了 3个酶系统 (苹果酸酶Me、苹果酸脱氢酶Mdh、甘油醛-3-磷酸脱氢酶Gpd) ,获得了 5个位点共 2 7个等位基因的资料。并计算了 4个库蚊居群的遗传多样性指标为 (A ,P ,He,Ho) ,同时分析了 5种居群间的遗传距离。分析结果表明 :遗传多样性主要存在于各居群之内 (FIS=0 .5 4 37) ,但居群之间的遗传分化程度也相对较大 (Gst =0 .4 498)。     

Genetic diversity following demographic recovery in the insular endemic plant <Emphasis Type="Italic">Galium catalinense</Emphasis> subspecies <Emphasis Type="Italic">acrispum</Emphasis>

Galium catalinense (Rubiaceae) is a perennial shrub consisting of two subspecies endemic to California’s Channel Islands: Galium catalinense subsp. catalinense on Santa Catalina Island, and G. catalinense subsp. acrispum, a state-endangered taxon on San Clemente Island. A long history of overgrazing by introduced herbivores has contributed to population declines in G. catalinense subsp. acrispum. We surveyed 12 populations throughout the taxon’s range for genetic variation using eight polymorphic microsatellite loci to determine the genetic impact of this demographic bottleneck. At the taxon level, 65 alleles were identified with an average of 8.1 alleles per locus, although many alleles were rare; the effective number of alleles per locus averaged 2.6. Expected heterozygosity was 0.550. Individual populations had between six and eight polymorphic loci, with expected heterozygosities ranging from 0.36 to 0.60, and effective numbers of alleles ranging from 1.8 to 3.5 per locus. Populations fell into three or four genetic clusters, depending on type of analysis, which may represent refugia where the populations persisted during intense herbivory. There is little evidence of genetic bottlenecks or substantial inbreeding within populations. These findings, coupled with indications of recent migration between populations, suggest that G. catalinense subsp. acrispum is currently unlikely to be endangered by genetic factors, but small population sizes make the taxon vulnerable to future loss of genetic diversity. Management strategies based on these genetic data, population sizes, and the spatial distribution of populations are discussed.     

Reproductive biology and genetic structure in Lloydia serotina

Concern regarding the conservation status of small, isolated populations of the arctic-alpine plant species Lloydia serotina prompted research to establish the status and performance of this species in Wales, in comparison with large populations in its more typical alpine habitat. Relationships between reproductive strategies and genetic variation were investigated in a number of populations, representing a wide habitat, geographic and population size range. In all populations, vegetative reproduction predominates over sexual reproduction, but seed produced is viable and germinates readily under controlled conditions. Smaller, peripheral populations produced fewer flowers and seeds than the larger ones, but all populations studied supported significant percentages (>30%) of male plants, due to either the occurrence of androdioecy in this species or to a resource limited breeding system. Analysis of allozyme variation in sixteen populations from North America, the European Alps and Wales showed lower levels of genetic variation in smaller populations which averaged 1.1–1.2 alleles per locus and 10–20% of loci polymorphic, whereas larger populations averaged 1.4 alleles per locus and 30–40% polymorphic loci. This applied especially to the most northerly and southerly populations in North America, suggesting the occurrence of genetic drift in these small, peripheral populations. F-statistics suggest relatively high levels of differentiation among smaller populations, even among those closely related geographically, but genetic variation has been retained in all but one population, possibly due to infrequent sexual reproduction by long lived clones. RAPD analysis of four small populations in Wales provided further evidence of clonal growth and possible inbreeding dominating a mixed mating reproductive system with consequent genetic structuring in these populations.     

Disjunct habitats as islands: genetic variability in the Caucasian rock lizard Lacerta portschinskii

Genetic diversity at 37 allozyme loci was surveyed in Lacerta portschinskii from contiguous populations and from a disjunct population. Indices of genetic diversity (heterozygosity, number of alleles per locus, and percentage of loci polymorphic) were greater in contiguous populations than in the smaller disjunct population. In this regard, disjunct populations appear to be similar to island populations. Indices of genetic diversity in Caucasian Lacerta are less than those reported from vagile lizard taxa and more similar to those of sit-and-wait predators. This revised version was published online in July 2006 with corrections to the Cover Date.     

Patterns of allozyme diversity in the threatened plant Erigeron parishii (Asteraceae)

Thirty-one occurrences of Erigeron parishii, a narrowly endemic plant threatened by mining, were sampled for allozyme diversity. This taxon held considerable genetic variation at the 14 allozyme loci surveyed. Species (e.g., alleles per locus [A] = 4.3 and proportion of polymorphic loci [P] = 0.64) and population (e.g., A = 2.15 [SD = 0.3] and P = 0.53 [SD = 0.12]) genetic diversity measures were higher than expected for narrowly endemic plant taxa. The proportion of polymorphic loci and numbers of alleles per locus indicated that E. parishii has not experienced severe or long-lasting population bottlenecks. Within-population f indicated low to moderate levels of inbreeding. Populations were only moderately differentiated (theta-p = 0.12), suggesting either that there is substantial gene flow among populations or that populations have not been isolated long enough to detect effects of genetic drift. There was no significant differentiation among populations in different vegetation types nor was there a relationship between genetic distance and geographic distance among sites. Continued fragmentation by mining activities would isolate populations, disrupting gene flow, exacerbating loss of diversity, and increasing inbreeding in the remaining fragments. Protection of large, interconnected populations throughout the range of the taxon is warranted to maintain processes that have sustained the observed diversity.     

Development of microsatellite markers for the critically endangered Limonium dufourii (Girard) Kuntze (Plumbaginaceae)

Limonium dufourii is an endemic plant from the eastern Mediterranean coast of Spain with a triploid chromosome number and apomictic reproduction. We have isolated and characterized 13 polymorphic microsatellite loci from an enriched library in order to investigate its population genetic structure. Simple sequence repeat (SSR) loci were screened in 120 individuals from the six extant populations of this species. They show an average of 5.76 alleles per locus, ranging from 2 to 18, with seven loci exhibiting heterozygosities larger than 0.60. Three loci present one single allele in each individual, whereas one locus presents three alleles in every individual analysed.     

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.     

Isolation and characterization of polymorphic microsatellite loci in the gudgeon,Gobio gobio (Cyprinidae)

A total of seven polymorphic microsatellite loci from Gobio gobio were isolated and characterized. A preliminary population survey of 82 specimens from four populations, located in a downstream pollution gradient of cadmium and zinc, demonstrated the usefulness of these primers both in population genetic studies in general, as well as in evaluating the effects of anthropogenic pollution on genetic structure. Overall locus polymorphism ranged from two to 13 alleles. Observed heterozygosity per locus ranged from 0.39 to 0.73.     

Population genetic structure of wild Phaseolus lunatus (Fabaceae), with special reference to population sizes

To set up an in situ conservation strategy for Phaseolus lunatus, we analyzed the genetic structure of 29 populations in the Central Valley of Costa Rica. Using 22 enzyme loci, we quantified the proportion of polymorphic loci (P(p)), the mean number of alleles per locus (A), and the mean effective number of alleles per locus (A(e)), which equaled to 10.32%, 1.10, and 1.05, respectively. The total heterozygosity (H(T)), the intrapopulation genetic diversity (H(S)), and the interpopulation genetic diversity (D(ST)) were 0.193, 0.082, and 0.111, respectively. The genotypic composition of the analyzed populations showed a deviation from the Hardy-Weinberg proportions (F(IT) = 0.932). This disequilibrium was due to either genetic differentiation between populations (F(ST) = 0.497) or nonrandom mating within populations (F(IS) = 0.866). From the level of genetic differentiation between populations and the private alleles frequencies estimates, gene flow was calculated: Nm(W) = 0.398 and Nm(S) = 0.023, respectively. The results suggested that wild Lima bean maintains most of its isozyme variation among populations. Significant positive correlation was observed between population size and P(p), A, and H(o) (observed heterozygosity), whereas no correlation was observed with the average fixation index of population (F). The loss of genetic variability in populations was attributed to inbreeding and the bottleneck effects that characterized the target populations. In situ conservation and management procedures for wild Lima bean are discussed.     

Development of ten microsatellite markers from the keystone mistletoe <Emphasis Type="Italic">Tristerix corymbosus</Emphasis> (Loranthaceae) using 454 next generation sequencing and their applicability to population genetic structure studies

Tristerix corymbosus (Loranthaceae) is a keystone mistletoe from the South American temperate rainforests. As most mistletoes, T. corymbosus relies on biotic pollination and seed dispersal, which may cause population structure. For a better understanding of its ecology, we isolated and characterized ten polymorphic microsatellite loci for this species. We used 454 Next Generation Sequencing to build a microsatellite library from a high quality DNA sample. We tested 90 sequences from which we obtained ten polymorphic markers. In order to assess their variability, the novel markers were tested in 48 individuals from three locations of the Valdivian Coastal Reserve in Chile. We also estimated genetic differences between pairs of populations using the F_ST statistic. The mean number of alleles per locus in the 48 individuals studied was 7.1 (2–17 alleles per locus). The observed and expected heterozygosity ranged from 0.298 to 0.634 and from 0.310 to 0.881, respectively. There were genetic differences among the three populations assessed, according to the F_ST values (ranging from 0.048 to 0.100, all significant) and, the number of alleles per locus ranged from 3.9 to 5.1. These are the first microsatellite markers developed for T. corymbosus, and they arise as a powerful tool for studying population structure, genetic diversity and gene flow at the landscape scale, along its distribution.     

Microsatellite analysis of genetic diversity in the Chinese alligator (<Emphasis Type="Italic">Alligator sinensis</Emphasis>) Changxing captive population

Chinese alligator (Alligator sinensis) is a critically endangered species endemic to China. In this study, the extent of genetic variation in the captive alligators of the Changxing Reserve Center was investigated using microsatellite markers derived from American alligators. Out of 22 loci employed, 21 were successfully amplified in the Chinese alligator. Sequence analysis showed loci in American alligators had a bigger average size than that of the Chinese alligators and the longest allele of an individual locus almost always existed in the species with longer stretch of repeat units. Eight of the 22 loci were found to be polymorphic with a total of 26 alleles present among 32 animals scored, yielding an average of 3.25 alleles per polymorphic locus. The expected heterozygosity (H _E) ranged at a moderate level from 0.4385 to 0.7163 in this population. Compared to that in the American alligators, a lower level of microsatellite diversity existed in the Changxing population as revealed by about 46% fewer alleles per locus and smaller H _E at the homologous loci. The average exclusion power and the ability to detect shared genotypes and multiple paternity were evaluated for those markers. Results suggested that when the polymorphic loci were combined, they could be sensitive markers in genetic diversity study and relatedness inference within the Chinese alligator populations. The level of genetic diversity present in the current Changxing population indicated an important resource to complement reintroductions based on the individuals from the other population. In addition, the microsatellite markers and their associated diversity characterized in this population could be utilized to further investigate the genetic status of this species.     

Development of microsatellite markers for Aquilegia buergeriana var. oxysepala (Ranunculaceae), a vulnerable Japanese herb

Aquilegia buergeriana var. oxysepala is a vulnerable herb, for which 20 microsatellite markers were developed. The applicability of these markers was confirmed by genotyping of 66 individuals collected from four natural populations in Japan. Of the 20 loci, 15 showed polymorphism. For the 15 polymorphic markers, the number of alleles per locus ranged from 2 to 12, and the expected and observed heterozygosities ranged from 0.13 to 0.77 and 0.02 to 0.65, respectively. The markers described here are sufficiently polymorphic and informative to investigate the genetic diversity and population genetic structure of A. buergeriana var. oxysepala.