Developmental genetics in larvae,pupae and adults ofSepedon fuscipennis fuscipennis [Dipt.: Sciomyzidae]

An analysis of 28 enzyme loci throughout developmental stages ofSepedon fuscipennis fuscipennis Loew indicated that 16 were polymorphic and 4 were monomorphic in all stages. Nine loci were differentially expressed among the stages: EST-1, EST-2, MDH-2, MDH-3 and PGI-1 occurred only in larvae, AK-3 mainly in pupae, and AK-1, AO and HK-1 only in adults. The average heterozygosity ofS. f. fuscipennis was 0.146 (±0.028) across all stages.      

不同地区库蚊复组群体的同工酶遗传多样性研究

采用水平切片淀粉凝胶电泳的方法,对分布于我国5省的8个库蚊复组（Culex pipiens complex）野生群体的遗传多样性进行研究,分析了4个酶系统7个基因座(ME、MDH-1、MDH-2、MDH-3、GPD、EST-2、EST-3)的酶谱资料。结果显示：（1）群体内存在不同程度的遗传变异（He为0.098～0.41）;（2）较低的基因流水平（Nm=0.64）使遗传漂变起主要作用,造成群体之间的遗传分化（Gst=0.303）,而总群体的遗传多样性相对富集于群体之内(Hs/Dst=2)。（3）库蚊群体的遗传结构属于距离隔离模式。（4）群体间的遗传一致性(或遗传距离)反映出群体间的遗传分化程度,也表明与地理位置存在对应关系。Abstract: Eight field populations of Culex pipiens complex collected from five provinces (Guangdong, Henan, Shandong, Beijing and Yunnan) in 2001 were used to study genetic diversity by starch gel electrophoresis. Data from seven loci (ME、MDH-1、MDH-2、MDH-3、GPD、EST-2、EST-3) of four isozymes were analyzed by software Biosys2.0 and FSTAT(Version 2.9.3). The results were as follows: (1) The values of He (from 0.098 to 0.41) indicated genetic variabilities of different degree in populations.(2)The low level of gene flow (Nm=0.64) could not prevent genetic drift to cause the gene differentiation between populations. The genetic diversity between populations attributed to the genetic diversity of total populations is small (Gst =0.303), and the great part is accumulated within populations (Hs/Dst=2). (3) The genetic structure of Culex pipiens complex population was the isolation-by-distance model. (4) The genetic identity (or genetic distance) revealed the scale of genetic differentiation between populations which related to the collection sites.     

Genetic variation,population structure and identification of yellow catfish, <Emphasis Type="Italic">Mystus nemurus</Emphasis> (C&;V) in Thailand using RAPD,ISSR and SCAR marker

Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were used to investigate the genetic structure of four subpopulations of Mystus nemurus in Thailand. The 7 RAPD and 7 ISSR primers were selected. Of 83 total RAPD fragments, 80 (96.39%) were polymorphic loci, and of 81 total ISSR fragments, 75 (92.59%) were polymorphic loci. Genetic variation and genetic differentiation obtained from RAPD fragments or ISSR fragments showed similar results. Percentage of polymorphic loci (%P), observed number of alleles, effective number of alleles, Nei’s gene diversity (H) and Shannon’s information index revealed moderate to high level of genetic variations within each M. nemurus subpopulation and overall population. High levels of genetic differentiations were received from pairwise unbiased genetic distance (D) and coefficient of differentiation. Mantel test between D or gene flow and geographical distance showed a low to moderate correlation. Analysis of molecular variance indicated that variations among subpopulations were higher than those within subpopulations. The UPGMA dendrograms, based on RAPD and ISSR, showing the genetic relationship among subpopulations are grouped into three clusters; Songkhla (SK) subpopulation was separated from the other subpopulations. The candidate species-specific and subpopulation-specific RAPD fragments were sequenced and used to design sequence-characterized amplified region primers which distinguished M. nemurus from other species and divided SK subpopulation from the other subpopulations. The markers used in this study should be useful for breeding programs and future aquacultural development of this species in Thailand.     

Genetic divergence between two forms of a tongue sole <Emphasis Type="Italic">Cynoglossus interruptus</Emphasis>

Genetic differences between trilinear and dilinear forms of a tongue sole, Cynoglossus interruptus, were examined by use of isozymes. Unequivocal differences were detected between the two forms, including complete replacement of alleles at the FBALD-2*, MDH-3*, PROT-1*, and SOD-1* loci, almost complete replacement at the AAT-3*, AH-1*, AH-2*, EST-3*, G3PDH-3*, GPI-2*, IDHP-1*, and MDH-1* loci, and extreme differences in allelic frequencies at the GPI-1* and PGDH* loci. The genetic distances (D values) between the two forms were 0.4207–0.4353, figures predicatively significant at the specific level. The considerable genetic differences strongly suggested that the two forms represent distinct species. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Genetic variation in relation to demography of peripheral pool frog populations (Rana lessonae)

Summary Genetic variation in an isolated northern metapopulation of the pool frog (Rana lessonae) in Sweden was compared to that of Central European populations using enzyme electrophoresis and literature data. Of the 31 loci scored, two (EST-2 andIDH-2) were polymorphic while no variation occurred in seven of the eight loci which are polymorphic in Central European populations.The heterozygosity level of the Swedish pool frogs is very low compared to that of other anuran populations, but their mean proportion of fertilized eggs within egg masses (97.5%) was not lower than in more heterozygous species, and their body size-specific fecundity did not differ from that of Polish conspecifics. The low genetic variability of the Swedish pool frogs is discussed in relation to features of the local populations such as size (N), calculated effective size (N _e ) reproductive success and probable history. It is concluded that long-term strong fluctuations in population size caused by reporductive failure in cold years have contributed more to the low genetic variability than could a single founder event due to a recent introduction by man.     

Polymorphic microsatellite loci for population studies of the razor clam, <Emphasis Type="Italic">Sinonovacula constricta</Emphasis>

The razor clam (Sinonovacula constricta) is an important aquacultured bivalve in China. The natural populations of this species are decreasing quickly. To facilitate studies on genetic diversity and population structure of wild populations, microsatellites were isolated from a CA enriched genomic library. Eight microsatellite loci were polymorphic in 30 individuals from Chongming in Shanghai, China. The number of alleles per polymorphic locus varied from 6 to 13 and the values of observed heterozygosity and expected heterozygosity ranged from 0.350 to 1.000 and from 0.602 to 0.902, respectively. These microsatellites are being used in studying population differentiation and genetic diversity for effective conservation and management genetic resources of S. constricta.     

Clone identification and clinal allozyme variation in populations ofAbies alba from the Eastern Alps (Austria)

Allelic frequencies at the polymorphic enzyme loci IDH-1, IDH-2, LAP-1, and 6-PGDH-1 were determined in population samples ofAbies alba from 5 different areas in Austria. The results reveal clear genetic differentiation between the eastern and the western provenances. Gradual frequency clines are observed with all four loci. The amount of variation, measured as average heterozygosity and genotypic diversity, is significantly higher in the west. The multilocus genotypes proved to be useful markers for the identification of clones. Functional differences with respect to substrate specifity were found for the two allozymes coded by the coexisting alleles of the IDH-2 locus.     

福建泰宁野生铁皮石斛种群的ISSR亲缘关系分析

为了解铁皮石斛(Dendrobium officinale)种质间的亲缘关系,利用ISSR技术对34份铁皮石斛种质资源进行亲缘关系和遗传多样性分析。结果表明,9条ISSR引物在34份种质中共扩增出78条带,多态位点百分率达100%。UPGMA聚类分析表明,种质的相似系数为0.61~0.92,在相似系数0.626处,福建省泰宁的野生铁皮石斛与栽培铁皮石斛分为两大类。泰宁野生铁皮石斛种群的Nei’s基因多样性(H)和遗传分化系数(Gst)分别为0.3111和0.4609,均高于栽培种群(0.3056和0.4204),表明泰宁野生铁皮石斛具有较丰富的多样性和较高的种群分化系数。AMOVA分析表明,铁皮石斛种群内变异指数为74%,种群间变异指数为26%,表明不同种群间可能存在基因交流。这些为不同地域的野生铁皮石斛资源的有效保护及利用提供理论依据及技术参考。     

Protein variation in wild Atlantic salmon, with particular reference to southern Ireland

The extent of genetic variation in wild Atlantic salmon parr, Sulmo salur L., from river systems in Ireland, Iceland and eastern Canada, was investigated using starch gel electrophoresis. Within Ireland, seven polymorphic enzyme loci ( sAAT-4 *, GPI-1 *, IDDH-1 *, IDDH-2 *, IDHP-3 *, MDH-3 * and mMEP-2 *) were screened in nine different rivers and nine tributaries from the River Blackwater. Significant heterogeneity in gene frequencies occurred between riverine samples and between samples from tributaries of the River Blackwater. Variation between tributaries was as great as between rivers elsewhere in the country. Levels of population differentiation were comparable to those found in other regions throughout the range of the species, and temporal stability in gene frequencies was apparent when the results were compared with previously published data. Screening of riverine samples from Iceland and eastern Canada (Newfoundland and New Brunswick) allowed the Irish results to be considered in a broader context. Irish salmon cluster in the western European group, to which may be added Icelandic populations. Salmon from eastern Canada show a high level of genetic distinctiveness from the European group.     

Polymorphic microsatellite loci for species of Australian freshwater cod, <Emphasis Type="Italic">Maccullochella</Emphasis>

The Australian freshwater cod genus, Maccullochella is represented by three species: Murray cod, M. peelii peelii, eastern freshwater cod, M. ikei, and trout cod, M.macquariensis. Seven novel microsatellite loci from M. ikei and six previously published loci from M. peelii peelii were tested on wild populations of Murray, eastern and trout cod. Levels of polymorphism varied between species with 13 loci polymorphic in Murray cod, 9 in trout cod and 7 in eastern cod. Observed heterozygosities ranged from 0.053 to 0.842. This suite of microsatellite loci will facilitate future studies of the genetic status of wild and hatchery bred populations of Maccullochella.     

草履蚧不同寄主和地理种群遗传分化的RAPD分析

应用随机扩增多态性DNA(RAPD)技术对草履蚧保定、石家庄、邯郸16个不同寄主地理种群遗传多样性和种群分化进行研究,结果显示4个RAPD引物共扩增出41个多态性位点,多态位点比率为100%。遗传距离指数在0.701—0.4360,平均为0.2395。其中以邯郸枫杨和邯郸垂丝海棠为寄主的草履蚧种群遗传距离最小(0.0701);以石家庄紫叶李和邯郸木槿为寄主的种群遗传距离最大(0.4360)。遗传一致度系数在0.6466—0.9290。说明草履蚧不同种群遗传多样性丰富并存在遗传差异。聚类分析结果表明草履蚧种群遗传多样性同时受到寄主和地理因素的双重影响,且不同寄主草履蚧种群已产生明显的遗传分化。     

Genetic variation of wild litchi (<Emphasis Type="Italic">Litchi chinensis</Emphasis> Sonn. subsp. <Emphasis Type="Italic">chinensis</Emphasis>) revealed by microsatellites

Understanding the amount and distribution of genetic diversity in natural populations can inform the conservation strategy for the species in question. In this study, genetic variation at eight nuclear microsatellite loci was used to investigate genetic diversity and population structure of wild litchi (Litchi chinensis Sonn. subsp. chinensis). Totally 215 individuals were sampled, representing nine populations of wild litchi. All eight loci were polymorphic, with a total of 51 alleles. The expected heterozygosity in the nine populations ranged from 0.367 to 0.638 with an average value of 0.526. Inbreeding within wild litchi populations was indicated by a strong heterozygote defect. Significant bottleneck events were detected in the populations from Yunnan and Vietnam, which could be responsible for lower levels of genetic diversity in these populations. Measures of genetic differentiation (F _ST = 0.269) indicated strong differentiation among wild litchi populations. Significant correlation was found between genetic differentiation and geographical distance (r = 0.655, P = 0.002), indicating a strong isolation by distance in these populations. Bayesian clustering suggested genetic separation among three regional groups, namely, the western group, the central group and the eastern group. Some conservation strategies for wild litchi populations were also proposed based on our results.     

Seventeen novel microsatellite markers from an enriched library of the pest species Bactrocera dorsalis sensu stricto

Seventeen novel microsatellite markers were isolated and characterized from an enriched DNA library of the pest species Bactrocera dorsalis sensu stricto. The polymorphism of these loci was tested in individual fruit flies from a Thai mass‐rearing strain and three wild Asian populations: two from Thailand and one from Myanmar. Allele numbers per locus ranged from two to 15. In the natural populations, a high level of polymorphism was detected in all loci suggesting the usefulness of these markers to quantify the genetic variation in this highly invasive insect.     

Genetic diversity and environmental associations of wild wheat,Triticum dicoccoides,in Israel

Summary Allozyme variation in the tetraploid wild progenitor of wheat, Triticum dicoccoides, was studied for the proteins encoded by about 50 gene loci in 457 individuals representing 12 populations from Israel. Six spikelet morphological traits were measured in the same populations. The results indicate that: (a) 16 loci (= 32%) were monomorphic in all 12 populations, 15 loci (= 30%) were locally polymorphic, and 19 loci (= 38%) were regionally polymorphic. All polymorphic loci (but one) displayed high levels of polymorphism ( 10%). In Israel, the proportion of polymorphic loci per population, P, in wild wheat averaged 0.25 (range, 0.16–0.38), and the genetic diversity index, H_e averaged 0.07, (range, 0.03 – 0.12). (b) Altogether there were 110 alleles at the 50 putative loci tested (c) Genetic differentiation of populations included regional and local patterns: (i) The coefficients of genetic distance between populations were high (mean D = 0.10 range, 0.02 – 0.25), and indicated sharp genetic differentiation over short distances, (ii) Common ( 10%) but sporadic and localized alleles were frequent (76%), and (iii) Rare alleles were few (only 5 alleles). (d) The patterns of allozyme and spikelet variation in the wild gene pool were significantly correlated with, and partly predictable by, water factors, including those of precipitation, evaporation, and relative humidity as well as of soil type, (e) All six spikelet characters showed statistically significant variation among localities and (f) Allozymic variation was correlated with spikelet variation.These results suggest in T. dicoccoides: (i) the operation of natural selection in population genetic structure, (ii) local adaptive genetic differentiation caused by diversifying selection through climate and soil, and (iii) the guidelines for sampling these resources for use in wheat breeding programs.     

白芨SSR引物筛选及群体遗传多样性研究

白芨(Bletilla striata Rchb.)为中国珍稀濒危植物,重要的药用植物。该研究基于Illumina测序技术构建白芨基因组文库和微卫星文库,设计白芨微卫星引物,用白芨4个野生种群80个个体对引物进行多态性检测,应用4个白芨近缘种中进行引物的通用性检测,并在此基础上分析了白芨的遗传多样性和遗传分化,以探讨白芨的遗传结构和进化潜能。结果表明:(1)白芨基因组中微卫星片断丰富,共检测出17 841条微卫星片断。基于白芨微卫星库对100个位点设计了引物对,经PCR扩增和检测筛选出能够稳定扩增的多态性位点20个,每个位点的复等位基因数(Na)在2~6之间,平均为3.85;20对引物的大部分能够在4个白芨近缘种中成功扩增。(2)白芨在物种水平均有较高的遗传多样性(Na=3.85,I=1.07,H=0.614 7),白芨种群遗传分化强烈(Gst=0.43),居群间的基因流较弱(Nm=0.867 6),居群聚类分析结果均表明地理距离较近的居群具有较近的遗传关系。     

Electrophoretic study and genetic affinity in theCampanula elatines andC. fragilis (Campanulaceae) rock-plants group from Italy and W. Jugoslavia

Variability and genetic divergence of 11 field populations of seven species of theCampanula elatines andC. fragilis rock-plants group from the Mediterranean and pre-Alpine areas have been studied by starch-gel electrophoretic techniques.Campanula isophylla, C. elatines, C. elatinoides, C. fragilis subsp.fragilis and subs.cavolinii, C. garganica were collected in Italy, whilstC. fenestrellata subsp.istriaca andC. portenschlagiana came from W. Jugoslavia. Twelve enzymatic loci for each population were genetically analysed: PGI-1 and 2, PGM-2, IDH-1 and -2, SKDH, ME, ADH, GOT-2, MPI-1 and -2, SOD-1. The genetic distances among the above mentioned entities have been calculated by Nei's index and depicted in a dendrogram.     

Genetic characterization in four sciaenid species from the Arabian Sea

Four sciaenid species Johnieops dussumieri, Kathala axillaris, Pennahia macropthalmus and Otolithes ruber were analysed electrophoretically for genetic variation at 18 loci (16 in P. macropthalmus and O. ruber ). Twelve loci were polymorphic in J. dussumieri , 10 in K. axillaris , three in P. macropthalmus and 12 in O. ruber ( P <0·99). Average heterozygosity ranged from 0·033 ± 0·100 to 0·070 ± 0·122. The allele frequencies of 14 loci were used to estimate Nei's genetic distance (). The values ranged from 0·334 to 0·612. Three isozyme loci ( LDH-B*, MDH-2* and G3PDH-1* ) were found to be the most reliable species-specific markers.     

Analysis of genetic diversity in the endangered Hucho taimen from China

Hucho taimen are listed as endangered in China. The population size has declined recently, prompting an increase in the level of listing from grade three in 2002 to grade five in 2006. We analyzed the genetic diversity of wild populations using 17 microsatellite markers to establish a scientific basis for conservation of this species. We collected tissue samples from four populations in the Heilongjiang River basin: Huma River (HM), Hutou (HT), Haiqing (HQ), and Zhuaji (ZJ). A total of 21 loci were amplified, 18 of which were polymorphic. The number of alleles per locus ranged from 2 to 9 (mean: 4.1905). There were 13 highly polymorphic loci and 5 moderately polymorphic loci. Analysis of five genetic diversity parameters (Na, Ne, Ho, He, and PIC) suggested moderate levels of diversity within the populations. The populations were ranked HT > HQ > ZJ > HM, but the differences in diversity were not statistically significant (P > 0.05). A comparison of variation among all four populations suggested Hardy–Weinberg disequilibrium at 20% of the loci. Genetic differentiation (Fst) was 0.0644 and the gene flow among populations was estimated at 3.36 individuals per generation. The majority of diversity (93.88%) occurred among individuals within a population. In contrast, relatively little (6.12%) of the genetic diversity was distributed between the populations. An analysis of genetic differentiation and genetic distance between pairs of populations revealed that both parameters were higher in comparisons of the HM population to the HT, HQ, and ZJ populations than among the three latter populations. This suggests that the HM population has a distinct genetic structure. We hypothesize that habitat degradation and excessive fishing, not low genetic diversity, has caused the decline in H. taimen populations. However, this species should be protected from further declines in genetic diversity.     

Population Genetic Structure of Astyanax scabripinnis (Teleostei, Characidae) from an Urban Stream

Despite the great anthropogenic interference on urban streams, information is still scarce about the genetic variability and structure of native fish populations inhabiting such streams. In the present study, random amplified polymorphic DNA (RAPD) markers were used to analyze genetic variability and structure of populations assigned to the Neotropical fish species Astyanax scabripinnis from an urban stream located in Londrina, Paraná State, southern Brazil. Thirty individuals of this species were collected from three sites throughout the upper Cambé stream. A total of 10 primers amplified 159 loci, of which 128 (80.5%) were polymorphic. Each of the three populations showed very similar proportions of polymorphic loci, which ranged from 63.5 to 64.8%. Unbiased genetic distances varied from 0.0612 to 0.0646. Theta_p-test values indicated moderate to high genetic differentiation among individuals from different localities. The number of migrants varied from 1.34 to 1.46, suggesting a low gene flow between populations. The genetic similarity among all individuals studied ranged from 0.424 to 0.848. The results suggest that populations of A. scabripinnis in Cambé stream are undergoing genetic differentiation.     

The genetic structure of endangered indigenous populations of the amago salmon, <Emphasis Type="Italic">Oncorhynchus</Emphasis><Emphasis Type="Italic">masou</Emphasis><Emphasis Type="Italic">ishikawae</Emphasis>, in Japan

The amago salmon, Oncorhynchus masou ishikawae, is an endemic subspecies of O. masou in Japan. Owing to the extensive stocking of hatchery fish throughout Japan, indigenous populations of O. m. ishikawae are now on the verge of extinction. We examined the genetic effects of stocking hatchery fish on wild populations in the River Koza, Japan, using microsatellite and mitochondrial DNA (mtDNA) markers. For mtDNA, haplotype mt1, which is common in wild populations, was present exclusively in isolated wild populations assumed to be unaffected by previous stocking, while it was never observed in hatchery fish. Genetic diversity was much higher in wild populations in the stocked area, which shared many mtDNA haplotypes with hatchery fish, than in isolated wild populations with haplotype mt1. Pairwise F _ST estimates based on microsatellites showed significant differentiation among the isolated populations with many microsatellite loci monomorphic. Significant deviation from Hardy–Weinberg equilibrium was observed in wild populations in the area subject to stocking, where a Bayesian-based assignment test showed a high level of introgression with hatchery fish. These results suggest that wild populations with haplotype mt1, which became isolated through anthropogenic environmental change in the 1950–1960s, represent indigenous populations of O. m. ishikawae in the River Koza. They have low genetic diversity, most likely caused by genetic bottlenecks following damming and environmental deterioration, while stocking of hatchery fish over the past 30 years apparently had a large impact on the genetic structure of wild populations in the main channel of the River Koza.