Isolation and characterization of eight polymorphic microsatellite loci for the coconut pest, Brontispa longissima (Coleoptera: Hispidae)

Brontispa longissima is one of the most serious insect pests of coconut in Southeast Asia; it was first discovered on Hainan Island in June 2002. Despite the economic risk associated with this pest, genetic aspects of the invasion process have remained relatively unexplored. Using microsatellite markers, we investigated the population structure, genetic variability and pattern of invasion in various geographic populations. The methodology was based on a modified biotin-capture method. Eight polymorphic microsatellite loci were isolated and characterized for the pest. The allele number per locus varied from 2 to 3 (N = 30). The expected and observed heterozygosities of the eight loci ranged from 0.042 to 0.509 and from 0.042 to 0.963, respectively. Although the frequency of polymorphisms was not very high in this population, the microsatellite loci that were isolated will be useful for investigating the genetic diversity and migration routes of B. longissima populations.     

海南岛中华蜜蜂遗传多样性的微卫星DNA分析

为了解海南岛中华蜜蜂Apis cerana cerana的遗传多样性和遗传结构及其与大陆种群的关系, 本研究应用10个微卫星DNA标记对海南岛11个地点627个蜂群的627头工蜂样本和大陆2个地点102个蜂群的102头工蜂样本进行了分析。结果表明： 海南岛中华蜜蜂遗传多样性较高, 单个位点检测到等位基因5～17个; 各种群平均等位基因数为4.5～7.0个, 平均杂合度为0.59～0.65。海南岛中华蜜蜂在10个位点上表现出相似遗传结构, 文昌和屯昌种群在AT101位点的等位基因频率较特殊。岛内 岛外中华蜜蜂的遗传分化系数F_ST范围为0.06～0.13; 文昌、 屯昌种群分别同海南岛内其他9个种群的F_ST（0.06～0.12）大于这9个种群间的FST（0～0.05）。海南岛中华蜜蜂同邻近大陆种群发生了明显的遗传分化; 除文昌、 屯昌种群发生中等程度的分化外, 海南岛内其他种群之间遗传分化较小。本研究结果对海南岛中华蜜蜂资源的保护和合理利用具有重要的指导意义。     

Eight microsatellite loci for the Irukandji syndrome-causing carybdeid jellyfish, Carukia barnesi (Cubozoa, Cnidaria)

Microsatellites are high-resolution genetic markers that may be applied to examine parentage, population structure and the direction and extent of dispersal. Here we present eight polymorphic microsatellite loci developed for the carybdeid jellyfish, Carukia barnesi. The loci were developed from a microsatellite-enriched, partial genomic DNA library and tested for polymorphism on animals from each of two geographically distinct populations, Lizard Island and Double Island, from the Great Barrier Reef. The number of alleles observed for each locus ranged from 7 to 19.     

Characterization of 10 microsatellite loci in the spiny‐tailed iguana Ctenosaura hemilopha

Ten microsatellite loci were developed for the Baja California spiny‐tailed iguana Ctenosaura hemilopha, using an enriched genomic DNA library. In the Cerralvo Island population, seven loci were polymorphic and presented moderate levels of variation. Number of alleles ranged from two to eight (average 4.43), and observed heterozygosity from 0.150 to 0.857 (average 0.492). Polymorphism was detected at six loci on C. hemilopha individuals from the southern Baja California Peninsula. These markers will be useful to study familiar relationships and behaviour on the Cerralvo Island population.     

Isolation and characterization of microsatellite loci in the whale shark (Rhincodon typus)

In preparation for a study on population structure of the whale shark (Rhincodon typus), nine species-specific polymorphic microsatellite DNA markers were developed. An initial screening of 50 individuals from Holbox Island, Mexico found all nine loci to be polymorphic, with two to 17 alleles observed per locus. Observed and expected heterozygosity per locus ranged from 0.200 to 0.826 and from 0.213 to 0.857, respectively. Neither statistically significant deviations from Hardy-Weinberg expectations nor statistically significant linkage disequilibrium between loci were observed. These microsatellite loci appear suitable for examining population structure, kinship assessment and other applications.     

Isolation and characterization of microsatellite loci in endangered Cycas changjiangensis (Cycadaceae)

Eight microsatellite loci were isolated from repetitive DNA enriched libraries for Cycas changjiangensis, an endangered endemic species in Hainan Island, China. The number of allele ranged from three to seven. The observed (H _O) and expected (H _E) heterozygosities ranged from 0.0000 to 0.8750 and from 0.2359 to 0.7582, respectively. These microsatellite loci will enrich our scientific understanding for C. changjiangensis conservation.     

Phylogeography of the Japanese pipistrelle bat,Pipistrellus abramus,in China: the impact of ancient and recent events on population genetic structure

The influence of Pleistocene climatic oscillations on shaping the genetic structure of Asian biota is poorly known. The Japanese pipistrelle bat occurs over a wide range in eastern Asia, from Siberia to Japan. To test the relative impact of ancient and more recent events on genetic structure in this species, we combined mitochondrial (cytochrome b) and microsatellite markers to reconstruct its phylogeographic and demographic history on continental China and its offshore islands, Hainan Island and the Zhoushan Archipelago. Our mitochondrial DNA tree recovered two divergent geographical clades, indicating multiple glacial refugia in the region. The first clade was mainly confined to Hainan Island, indicating that gene flow between this population and the continent has been restricted, despite being repeatedly connected to the mainland during repeated glacial episodes. By contrast, haplotypes sampled on the Zhoushan Archipelago were mixed with those from the mainland, suggesting a recent shared history of expansion. Although microsatellite allele frequencies showed clear discontinuities across the sampling range, supporting the current isolation of both Hainan Island and the Zhoushan Archipelago, we also found clear evidence of more recent back colonization, probably via post‐glacial expansion or, in the latter case, occasional long distance dispersal. The results obtained highlight the importance of using multiple sets of markers for teasing apart the roles of ancient and more recent events on population genetic structure. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 99 , 582–594.     

Development and characterization of ten new microsatellite markers in a mangrove tree species Bruguiera gymnorrhiza (L.)

Bruguiera gymnorrhiza is an ecologically and somewhat economically important mangrove tree species. We isolated 10 polymorphic microsatellite loci from B. gymnorrhiza using a dual‐suppression polymerase chain reaction (PCR) technique. These loci provided microsatellite markers with polymorphism of two to five alleles per locus within 216 individuals from nine natural populations of B. gymnorrhiza on Iriomote Island, the Sakishima Islands, Japan. The expected and observed heterozygosities ranged from 0.220 to 0.720 and from 0.104 to 0.447, respectively.     

A panel of microsatellite loci for population studies of Sakhalin taimen Parahucho perryi (Brevoort)

Using DNA samples of Sakhalin taimen and a set of microsatellite loci, earlier reported for other salmonid fishes (Salmonidae), successful cross-species amplification was performed. A total of 56 Sakhalin taimen (Parahucho perryi) samples from the Daga, Nabil, Poronai, and Agnevo rivers (Sakhalin Island) were examined at 36 microsatellite loci, most of which were described for other species and first tested in taimen. Among the 21 loci first tested in taimen, two loci produced no amplification products. The remaining 19 loci were successfully amplified (for some loci, new primers were generated). Thirteen of these loci were monomorphic, while six loci were polymorphic and used in further population genetic analysis. In addition, with the purpose of modification of the allele sizes and optimization of the research technique, new primers for the already known 12 loci of Sakhalin taimen were designed. Three more loci were included in analysis without changes. As a result, a joint panel consisting of 21 polymorphic microsatellite markers was suggested for analysis of Sakhalin taimen. This panel was tested with four population samples from the rivers of Sakhalin Island.The results showed that this panel of markers could be used in detailed population studies for evaluation of the level of genetic differentiation, inbreeding, and migrations in Sakhalin taimen, an endangered species with a fragmented range. Using this approach in further studies will make it possible to isolate basic populations, which is necessary for conservation of this rare species.     

Sex-linked and autosomal microsatellites provide new insights into island populations of the tammar wallaby

The emerging availability of microsatellite markers from mammalian sex chromosomes provides opportunities to investigate both male- and female-mediated gene flow in wild populations, identifying patterns not apparent from the analysis of autosomal markers alone. Tammar wallabies (Macropus eugenii), once spread over the southern mainland, have been isolated on several islands off the Western Australian and South Australian coastlines for between 10 000 and 13 000 years. Here, we combine analyses of autosomal, Y-linked and X-linked microsatellite loci to investigate genetic variation in populations of this species on two islands (Kangaroo Island, South Australia and Garden Island, Western Australia). All measures of diversity were higher for the larger Kangaroo Island population, in which genetic variation was lowest at Y-linked markers and highest at autosomal markers (θ=3.291, 1.208 and 0.627 for autosomal, X-linked and Y-linked data, respectively). Greater relatedness among females than males provides evidence for male-biased dispersal in this population, while sex-linked markers identified genetic lineages not apparent from autosomal data alone. Overall genetic diversity in the Garden Island population was low, especially on the Y chromosome where most males shared a common haplotype, and we observed high levels of inbreeding and relatedness among individuals. Our findings highlight the utility of this approach for management actions, such as the selection of animals for translocation or captive breeding, and the ecological insights that may be gained by combining analyses of microsatellite markers on sex chromosomes with those derived from autosomes.     

Development and characterization of microsatellite markers for Hibiscus glaber Matsum. ex Nakai, an endemic tree species of the oceanic Bonin Islands, Japan

Polymorphic microsatellite markers were developed for Hibiscus glaber, an endemic tree of the Bonin Islands. Eighty-seven of the 208 sequences from an enriched library were unique and containing microsatellites. Ten loci were proved to be highly polymorphic among 78 individuals from the Nishi-jima Island. Total exclusionary powers for the first and the second parents were 99.989% and 99.999%, respectively. Nine loci also amplified single fragment from genomic DNA of H. tiliaceus, a related and widespread congener. Our markers can be reliably used for the estimation of current gene flow within/among populations of the two woody Hibiscus species.     

Polymorphic microsatellite loci for population studies of the Japanese scallop,Patinopecten yessoensis

Fifteen microsatellite loci were developed to examine the population structure of Japanese scallop, Patinopecten yessoensis. These markers were tested in samples from two geographically distant populations: the Sakhalin Island (Russia) and Dalian City (China). The mean numbers of alleles, observed and expected heterozygosities between the two populations were summarized by locus, but none of them show significant divergence between the two populations. Most loci conformed to Hardy–Weinberg expectation with the exception of loci HLJX‐06, HLJX‐12, HLJX‐13 and HLJX‐28, which had heterozygote deficits.     

Isolation and characterization of eight microsatellite loci for the vulnerable Hainan Eld’s deer (<Emphasis Type="Italic">Cervus eldi hainanus</Emphasis>) in China

We report on the isolation and characterization of eight microsatellite markers in the Hainan Eld’s deer (Cervus eldi hainanus) from genomic DNA-enriched libraries. Thirty-three microsatellites were screened from the libraries, and 8 of the screened microsatellites were polymorphic. The number of observed alleles for each locus in 47 individuals ranged from 2 to 9, and the expected and observed heterozygosity was 0.141–0.792 and 0.128–0.957, respectively. Three loci (CEH-2, CEH-6 and CEH-8) of eight deviated from Hardy-Weinberg expectation and no significant linkage association was found among all these loci. These microsatellite markers provide useful tool for population genetic studies of the Eld’s deer.     

家养水牛30个微卫星标记的多重PCR体系建立及其多态性检测

利用变性聚丙烯酰胺凝胶电泳结合硝酸银染色方法, 从30个微卫星标记中筛选出9组多重PCR体系, 其中4组三座位组合, 5组二座位组合; 在此基础上进一步检测了这些标记在中国海南兴隆水牛中的遗传变异性。结果显示其中26个标记具有多态性, 而其余4个标记(CSSM045, ILSTS008, RM099和HMH1R)为单态, 群体平均等位基因数4.7, 平均期望杂合度为0.534。所筛选出的多重PCR组合为家养水牛的群体遗传多样性 检测和亲子鉴定等研究提供了技术基础。     

Isolation and characterization of 11 microsatellite loci in Scotophilus kuhlii (Lesser Asiatic Yellow House Bat)

We used enriched genomic library method to isolate and characterize 11 di-nucleotide microsatellite loci in the Lesser Asiatic Yellow House Bat, Scotophilus kuhlii. The polymorphism of these loci was tested on a natural population of 36 individuals from Hainan Province, South China. All loci revealed the polymorphism ranging from 5 to 18 alleles. The observed heterozygosity values were from 0.694 to 1.000 and expected heterozygosity values were from 0.558 to 0.929. No significant linkage disequilibrium was detected between any pairs of loci. These polymorphic markers will be used to assess population structure in S. kuhlii.     

黑头酸臭蚁微卫星标记的分离与开发(英文)

【目的】本研究旨在分离黑头酸臭蚁Tapinoma melanocephalum基因组微卫星标记,确定这些微卫星位点的多态性。【方法】使用454 GS-FLX焦磷酸测序技术开发来自中国华南陆地和岛屿的11个黑头酸臭蚁地理种群基因组微卫星位点。从随机设计的100对微卫星引物中筛选出10对引物,用于确定黑头酸臭蚁4个地理种群［东澳岛(DAD)、荷包岛(HBD)、梅州(MZ)和山咀(SJ)］10个微卫星位点的多态性,分析种群遗传多样性和种群分化。【结果】从11个黑头酸臭蚁地理种群基因组中成功开发和分离10对微卫星引物。在DAD, HBD, MZ和SJ 4个地理种群中,10个微卫星位点中7个有高多态性,这10个位点均显著偏离Hardy-Weinberg平衡;每个位点的等位基因数量(A)是3.50~9.00个,每个地理种群每个位点等位基因丰富度(A_R)在1.992~12.938之间。岛屿地理种群(DAD和HBD)的AR和预期杂合度(H_E)与大陆地理种群(MZ和SJ)的相比差异不显著。4个地理种群均显示高水平遗传分化(F_ST=0.15969);HBD和MZ种群与其他配对地理种群相比,遗传分化较高(F_ST=0.185),基因流较低,说明这两个种群基因流被限制。此外,遗传变异来自种群内个体之间。【结论】筛选新的微卫星位点能够为研究黑头酸臭蚁种群结构和繁殖结构提供有效工具,以深入了解其传播机制。     

Eight new polymorphic microsatellite DNA markers for Sakhalin taimen Parahucho perryi

We developed eight polymorphic microsatellite markers for the endangered salmonid Parahucho perryi from genomic libraries enriched for (GACA)n and (GATA)n repeat sequences. Emphasis was placed on developing highly polymorphic, perfect repeats that could be scored with confidence. Six tetra- and two di-nucleotide loci were screened in 49 individuals from two populations on Sakhalin Island, Russia. Allelic variation was high with eight to 23 alleles per locus and expected heterozygosity ranging from 0.48 to 0.93. These highly variable markers should prove useful in evaluating inbreeding, gene flow and population structure in Sakhalin taimen throughout its range.     

Variability of allozyme and cpSSR markers in the populations of Siberian spruce

The variability of 21 allozyme and three microsatellite loci of chloroplast DNA (cpDNA) was studied in the populations of Siberian spruce (Picea obovata Ledeb.) from Irkutsk oblast, Magadan oblast, Buryatia, and Mongolia. It was demonstrated that the highest level of genetic diversity among the examined populations at both allozyme and microsatellite loci was observed in the Tulyushka population from Irkutsk oblast. The lowest level of genetic diversity was observed in marginal isolated populations of Bogd Uul and Magadan. In the relict spruce population from Olkhon Island, differing from the other populations in the lowest allelic diversity of both types of markers, no expected decline of expected heterozygosity and haplotype diversity was observed. In this population, the variability parameters mentioned were close to the population mean. The obtained intrapopulation and intraspecific variability parameters of allozyme and microsatellite loci of chloroplast DNA and the data on the population differentiation at these loci indicate that the given markers can be used for the analysis of the population structure of Siberian spruce.     

Nuclear microsatellite DNA markers for New Zealand kiwi (Apteryx spp.)

Kiwi (Apterygidae) is an endemic New Zealand avian family comprising five species whose conservation is actively managed. We present five polymorphic microsatellite DNA loci isolated from North Island brown kiwi (Apteryx mantelli). In addition, we demonstrate cross‐amplification, and in some cases, polymorphism, of these microsatellite DNA loci in four other kiwi species. Therefore, these markers may be broadly applicable to conservation genetic studies within this family.     

Microsatellite loci in the stable fly,Stomoxys niger niger (Diptera: Muscidae) on La Réunion Island

Six polymorphic microsatellite markers have been isolated from a microsatellite‐enriched DNA library from the stable fly, Stomoxys niger niger. These loci exhibited five to 10 alleles per locus and an expected heterozygosity ranging from 0.57 to 0.81 in the studied populations from La Réunion Island (Indian Ocean). They should therefore be useful for the study of genetic diversity and population structure of S. niger. In addition, cross‐species amplification of four of these six loci in the closely allied species Stomoxys calcitrans produced interpretable results, two of which would be useful for population biology studies.