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%.     

Population genetics of Glossina morsitans submorsitans (Diptera: Glossinidae)

Breeding structure of Glossina morsitans submorsitans Newstead was evaluated by using genetic markers in mitochondrial DNA where diversity was scored at two loci in five natural populations from The Gambia and two populations in Ethiopia (form ugandensis Vanderplank), countries separated by c. 5450 km. Twenty six haplotype combinations were found, of which 17 were shared among two or more populations. Nine haplotypes were found in The Gambia and 23 haplotypes in Ethiopia. There were 12 unique haplotypes. Only six haplotypes were shared between the two countries. Populations in The Gambia (he = 0.26 +/- 0.04) showed less than a third of the diversity of populations in Ethiopia (he = 0.84 +/- 0.03). This suggests recovery from an earlier reduction in population. In a nested analysis of molecular variance of haplotype frequencies, 65% of the variance was due to differences within populations, 34% to differences between populations grouped by country, and only 1% was due to differences among populations within countries. In terms of gene flow, the fixation index FST = 0.35, which leads to an estimate by Wright's island model of less than one reproducing migrant per generation exchanged between the eastern and western submorsitans populations. Nei's genetic similarity measure showed a deep division between Gambian and Ethiopian populations.     

Characterization of microsatellite markers in the tsetse fly, Glossina pallidipes (Diptera: Glossinidae)

Glossina pallidipes is a vector of African trypanosomiasis. Here we characterize eight new polymorphic microsatellite loci in 288 G. pallidipes sampled from 12 Kenya populations. The number of alleles per locus ranged from four to 36 with a mean of 20.5 +/- 10.1. Expected single locus heterozygosities varied from 0.044 to 0.829. Heterozygosity averaged 0.616 +/- 0.246. No linkage disequilibrium was found. We also report results in eight other tsetse species estimated by using the primers developed in G. pallidipes. The primers worked best in G. swynnertoni and G. austeni and worst in G. m. morsitans and G. m. submorsitans.     

Macrogeographic population structure of the tsetse fly, Glossina pallidipes (Diptera: Glossinidae)

Tsetse flies are confined to sub-Saharan Africa where they occupy discontinuous habitats. In anticipation of area-wide control programmes, estimates of gene flow among tsetse populations are necessary. Genetic diversities were partitioned at eight microsatellite loci and five mitochondrial loci in 21 Glossina pallidipes Austin populations. At microsatellite loci, Nei's unbiased gene diversity averaged over loci was 0.659 and the total number of alleles was 214, only four of which were shared among all populations. The mean number of alleles per locus was 26.8. Random mating was observed within but not among populations (fixation index FST=0.18) and 81% of the genetic variance was within populations. Thirty-nine mitochondrial variants were detected. Mitochondrial diversities in populations varied from 0 to 0.85 and averaged 0.42, and FST=0.51. High levels of genetic differentiation were characteristic, extending even to subpopulations separated by tens and hundreds of kilometres, and indicating low rates of gene flow.     

Genetic Variation at Structural Loci in the Glossina morsitans Species Group

Gene diversity was investigated in four taxa of tsetse flies (Diptera: Glossinidae) including Glossina morsitans morsitans, G. m. centralis, G. swynnertoni, and G. pallidipes. Histochemical tests were performed for 35–46 isozymes. Polymorphic loci were 20% in G. morsitans morsitans, 32% in G. m. centralis, 17.6% in G. swynnertoni, and 26% in G. pallidipes. Mean heterozygosities among all loci were 6.6% in G. morsitans morsitans, 6.0% in G. m. centralis, 7.1% in G. swynnertoni, and 6.8% in G. pallidipes. Allozyme gene diversities were considerably less than those reported for many Diptera. The low gene diversities are probably related to small effective population sizes.     

Genetic differentiation of some Glossina morsitans morsitans populations

To study the population structure of Glossina morsitans morsitans Westwood (Diptera: Glossinidae), polymerase chain reaction (PCR) and singlestrand conformational polymorphism (SSCP) methods were used to estimate mitochondrial DNA diversity at four loci in six natural populations from Zambia, Zimbabwe and Mozambique, and in two laboratory cultures. The Zambian and Zimbabwean samples were from a single fly belt. Four alleles were recorded at 12S and 16S1, and five alleles at 16S2 and COI. Nucleotide sequencing confirmed their singularities. Chi-square contingency tests showed that allele frequencies differed significantly among populations. Mean allele diversities in populations averaged over loci varied from 0.14 to 0.61. Little loss in haplotype diversity was detected in the laboratory cultures thereby indicating little inbreeding. Wright's fixation index F(ST) in the natural populations was 0.088+/-0.016, the correlation of haplotypes within populations relative to correlations in the total. A function of its inverse allows an estimate of the mean equivalent number of females exchanged per population per generation, 5.2. No correlation was detected between pairwise genetic distance measures and geographical distances. Drift explains the high degree of differentiation.     

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.     

Microsatellite genetic variation in small and isolated populations of Magnolia sieboldii ssp. japonica

Magnolia sieboldii ssp. japonica, distributed mainly in western Japan, is restricted to high elevation areas (1000-2000 m above sea level) and usually forms small isolated populations. Four microsatellite loci were assayed for 19 populations from six regions spanning the range of distribution, and the levels and distribution of genetic variation were estimated. All four loci were variable, with a total of 39 alleles, but the overall level of microsatellite genetic variation was low, especially compared with a related species, M. obovata. Genetic structure in M. sieboldii was characterised by low intrapopulational genetic variation (A = 3.74 and H(o) = 0.366 on average) and high genetic differentiation even among regional populations. Highly significant isolation-by-distance (IBD) models at the short distance were detected. Genetic drift and limited gene flow was considered to be important in determining the genetic structure within regions. Total genetic differentiation was remarkably high (F(ST) = 0.488 and R(ST) = 0.538), suggesting genetic barriers among regions. Neighbour-joining dendrograms relating the 19 populations, and further analysis on the IBD models, revealed that a stepwise mutation model was more suited than an infinite allele model to explain the genetic differentiation among regions. It is suggested that mutation at microsatellite loci might be influential in generating the genetic differentiation among regions. These results showed the potential of hypervariable microsatellite loci to evaluate the effects of genetic drift and population isolation within regions, and to detect genetic distinctiveness, in spite of the loss of overall genetic variation in M. sieboldii.     

Low genetic diversities in isolated populations of the Asian black bear (<Emphasis Type="Italic">Ursus thibetanus</Emphasis>) in Japan,in comparison with large stable populations

Populations of the Asian black bear (Ursus thibetanus) are relatively large and continuous in central Honshu, the main island of Japan, but they are isolated in western Honshu. To clarify the degree of genetic isolation of the populations in western Honshu, we compared the genetic diversities of four populations in western Honshu with that of one of the continuous populations of central Honshu. Three of the four western Honshu populations were isolated and the other was continuous with the central Honshu populations on a geographical distribution basis. The genotypes at 10 microsatellite loci of the sampled individuals were determined and the genetic structures of the populations examined. Genetic diversities were significantly lower in the isolated populations than in the continuous populations. The continuous population in central Honshu had high levels of genetic diversity, comparable to those in populations of the American black bear (Ursus americanus) and the brown bear (Ursus arctos). The genetic distances between the two continuous populations were smallest, even though their geographic distance was largest (>200 km) among all the pairs of neighboring populations examined. Low genetic diversity within the isolated populations suggested genetic drift due to the small population size; the genetic differentiation among the populations indicated low rates of gene flow among them.     

Isolation and characterization of microsatellite loci in the western pearlshell mussel, Margaritifera falcata (Gould)

Ten microsatellite loci were isolated from the western pearlshell, Margaritifera falcata (Gould, 1850) and characterized in populations from Washington and Montana, USA. We also assessed eight microsatellite loci developed in M. margaritifera, two of which showed utility. Both of our test populations showed significant heterozygote deficiencies at most loci, consistent with a hermaphroditic life history. Populations differed markedly with respect to allelic richness, allele frequencies and numbers of identical multilocus genotypes. This panel of loci should prove useful in describing gene flow and genetic diversity patterns among M. falcata populations, information that should aid future conservation efforts.     

内蒙古亚洲小车蝗种群遗传多样性的微卫星分析

亚洲小车蝗Oedaleus asiaticus Bei-Bienko是我国北方草原和农牧交错区主要的害虫之一。为了从分子水平评价内蒙古地区亚洲小车蝗种群的遗传多样性和种群间遗传分化, 本文采用8对微卫星引物对内蒙古15个地点的亚洲小车蝗种群进行遗传多样性分析。结果表明： 各位点有效等位基因数为3.4517~13.2881, 多态信息含量值为0.5601~0.8563, Shannon氏多样性指数在0.7018~4.1789之间。15个种群的平均期望杂合度为0.6836, Nei氏期望杂合度为0.5303~0.6513, 群体遗传距离为0.1092~0.4235, 群体分化率Fst平均值为0.1612, 基因流Nm平均值为1.6164。8个微卫星位点均具有较高的多态性, 各种群间的遗传分化水平较大, 基因交流程度中等, 个体间的遗传变异大于种群间的遗传变异。15个不同地点的亚洲小车蝗种群根据遗传距离共聚为6支。种群间遗传分化与地理距离呈正相关关系。高山和沙漠对不同地区亚洲小车蝗种群的迁移具有阻碍作用, 可能是形成遗传分化的主要原因。研究结果从分子水平探索不同地区亚洲小车蝗种群间的内在联系, 为制定亚洲小车蝗的综合治理策略提供了分子生物学的基础资料。     

Patterns of genetic diversity and differentiation in the tsetse fly <Emphasis Type="Italic">Glossina morsitans morsitans</Emphasis> Westwood populations in East and southern Africa

Genetic diversity and differentiation within and among nine G. morsitans morsitans populations from East and southern Africa was assessed by examining variation at seven microsatellite loci and a mitochondrial locus, cytochrome oxidase (COI). Mean COI diversity within populations was 0.63 ± 0.33 and 0.81 taken over all populations. Diversities averaged over microsatellite loci were high (mean number of alleles/locus ≥7.4; mean H _E ≥ 65%) in all populations. Diversities averaged across populations were greater in East Africa (mean number of alleles = 22 ± 2.6; mean h _e = 0.773 ± 0.033) than in southern Africa (mean number of alleles = 18.7 ± 4.0; mean h _e = 0.713 ± 0.072). Differentiation among all populations was highly significant (R _ST = 0.25, F _ST = 0.132). Nei’s G _ij statistics were 0.09 and 0.19 within regions for microsatellites and mitochondria, respectively; between regions, G _ij was 0.14 for microsatellites and 0.23 for mitochondria. G _ST among populations was 0.23 for microsatellite loci and 0.40 for mitochondria. The F, G and R statistics indicate highly restricted gene flow among G. m. morsitans populations separated over geographic scales of 12–917 km.     

微卫星标记分析中国梨木虱种群的遗传多样性

中国梨木虱Cacopsylla chinensis (Yang et Li)是梨树主要害虫之一。为了从分子水平评估中国梨木虱种群内的遗传变异和种群间的遗传分化, 本文应用7对微卫星DNA引物对中国16个地区中国梨木虱种群进行遗传多样性分析。结果表明： 各位点有效等位基因为2.2927～10.0610, 多态信息含量(PIC)值在0.5073～0.8735之间。16个种群的平均期望杂合度为0.7876, 遗传距离在0.0951～1.0139之间, Nei氏期望杂合度为0.4771～0.7892, Shannon信息指数在0.8396～1.9989之间, 群体分化率F_ST为11.61%, 基因流平均值为2.2236。结果表明, 7个微卫星位点均具有较高的多态性, 各种群间的遗传分化水平较低, 基因交流程度较高, 遗传变异主要存在于种群内的个体间。研究结果为制定针对中国梨木虱的有效防治策略提供部分分子生物学的基础资料。     

Microsatellites reveal regional population differentiation and isolation in Lobaria pulmonaria, an epiphytic lichen

Many lichen species produce both sexual and asexual propagules, but, aside from being minute, these diaspores lack special adaptations for long-distance dispersal. So far, molecular studies have not directly addressed isolation and genetic differentiation of lichen populations, both being affected by gene flow, at a regional scale. We used six mycobiont-specific microsatellite loci to investigate the population genetic structure of the epiphytic lichen Lobaria pulmonaria in two regions that strongly differed with respect to anthropogenic impact. In British Columbia, L. pulmonaria grows in continuous old-growth forests, while its populations in the old cultural landscape of Switzerland are comparably small and fragmented. Populations from both British Columbia and Switzerland were genetically diverse at the loci. Geographically restricted alleles, low historical gene flow, and analyses of genetic distance (upgma tree) and of differentiation (amova) indicated that populations from Vancouver Island and from the Canadian mainland were separated from each other, except for one, geographically intermediate population. This differentiation was attributed to different glacial and postglacial histories of coastal and inland populations in British Columbia. In contrast to expectations, the three investigated Swiss populations were genetically neither isolated nor differentiated from each other despite the long-lasting negative human impact on the lichen's range size in Central Europe. We propose that detailed studies integrating local landscape and regional scales are now needed to understand the processes of dispersal and gene flow in lichens.     

Gene flow across species boundaries in sympatric, sexually deceptive Ophrys (Orchidaceae) species

Abstract Orchids of the genus Ophrys (Orchidaceae) are pollinated by male bees and wasps through sexual deception. The Ophrys sphegodes group encompasses several closely related species that differ slightly in floral morphology and are pollinated by different solitary bee species. Populations representing different species of the O. sphegodes group often flower simultaneously in sympatry. To test whether gene flow across the species boundaries occurs in these sympatric populations, or whether they are reproductively isolated, we examined the distribution of genetic variation within and among populations and species of this group. We collected at each of five different localities in southern France and Italy two sympatric, co-flowering Ophrys populations, representing six Ophrys species in total. The six microsatellite loci surveyed were highly variable. Genetic differentiation among geographically distant populations of the same species was lower than differentiation among sympatric populations of different species. However, the strength of genetic differentiation among species was among the lowest reported for orchids. Genotype assignment tests and marker-based estimates of gene flow revealed that gene flow across species boundaries occurred and may account for the low observed differentiation among species. These results suggest that sexual deceit pollination in Ophrys may be less specific than thought, or that rare mistakes occur.     

Genetics of cattails in radioactively contaminated areas around Chornobyl

Research on populations from radioactively contaminated areas around Chornobyl has produced ambiguous results for the presence of radiation effects. More studies are needed to provide information on whether radiation exposure at Chornobyl significantly affected genetic diversity in natural populations of various taxa. Eleven and nine variable microsatellite loci were used to test for differences in genetic diversity between reference and Chornobyl populations of two cattail species (Typha angustifolia and Typha latifolia, respectively) from Ukraine. Our purpose was to determine whether radiation had a significant impact on genetic diversities of the Chornobyl Typha populations, or if their genetic composition might be better explained by species demography and/or changes in population dynamics, mainly in sexual and asexual reproduction. Populations closest to the reactor had increased genetic diversities and high number of genets, which likely were due to factors other than radiation including increased gene flow among Chornobyl populations, enhanced sexual reproduction within populations, and/or origin of the genets from seed bank. Both Typha species also demonstrated small but significant effects associated with latitude, geographical regions, and watersheds. Typha's demography in Ukraine possibly varies with these three factors, and the small difference between Chornobyl and reference populations of T. latifolia detected after partitioning the total genetic variance between them is probably due primarily to these factors. However, the positive correlations of several genetic characteristics with radionuclide concentrations suggest that radiation may have also affected genetics of Chornobyl Typha populations but much less than was expected considering massive contamination of the Chornobyl area.     

Development of microsatellite markers for the tropical moss,Acanthorrhynchium papillatum

We isolated and characterized microsatellite loci for the palaeotropic moss, Acanthorrhynchium papillatum. Eight loci tested on 98 gametophytic samples generated four to 26 alleles per locus with genetic diversities ranging from 0.578 to 0.936. These microsatellite loci are now being used as genetic markers for studies on the effects of deforestation on moss populations in South‐East Asia.     

菲律宾蛤仔EST-SSRs标记开发及不同地理群体遗传多样性

利用13对微卫星引物对大连、莆田、青岛3个地理群体蛤仔遗传多样性进行了检测。结果表明:13个基因座共检测到154个等位基因,每个座位检测到的等位基因数在2-7个之间,平均有效等位基因数为2.7657;3个群体平均观测杂合度分别为0.4387、0.4194、0.2383,平均期望杂合度分别为0.6488、0.6484、0.5526;群体间的遗传多样性差异不显著(P＞0.05)。NJ聚类结果显示大连和莆田群体的蛤仔亲缘关系较近,二者与青岛群体关系较远。3个群体均有不同程度的偏离Hardy-Weinberg平衡现象,表明各群体基因频率和基因型频率的稳定性相对较低。本研究所获得的微卫星标记的多态信息含量(PIC)>0.5,说明这些微卫星位点的多样性较高,可为下一步遗传图谱构建研究提供参考。     

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.     

Genetic variation at microsatellite loci in Spanish sheep

Genetic variation at 18 microsatellite loci was analysed in six indigenous Spanish sheep: Churra; Latxa; Manchega; Rasa-Aragonesa; Castellana and Merino. Merinos had frequently the highest number of alleles per locus, whereas Latxas showed the lowest one at many loci. Markers ordered decreasingly according to the number of variants differentiated in the whole population were: MAF70; TGLA13; CSSM66; BM143, BM6444; MAF36; MAF64; CSSM6; TGLA53; OarFCB11; MAF33; BM4621; MAF48; MAF65; BM1258; ILSTS002; ADCYC and OarCP34. Parameters of variability such as effective number of alleles and gene diversities corroborated the high level of variation frequently displayed by microsatellite markers. Comparison of allele distributions among populations and loci did not reveal consistent shapes. Distributions were centralised in some cases, whereas in others some kind of skewness was evident. Breed-specific alleles were detected at most loci, being frequent in Merinos and rare in Churras.