中国东亚飞蝗两个种群遗传分化的研究

采用水平淀粉凝胶电泳技术,分析了我国河北平山、辽宁葫芦岛两个蝗区东亚飞蝗自然种群的遗传结构。在酶谱分析的19个位点中,多数位点的等位基因数目较少,而位点Aat—l、Pgi和Mdh—2的等位基因数目相对较多。由于杂合子数目较少而使每个基因位点的平均杂合度降低(Ho=0．024和0．028)。对每个位点的各基因型进行x^2检验,绝大多数位点的基因型频率偏离Hardy—Weinberg平衡。较低的Fst值(Fst=0．021)表明两个种群的遗传分化程度不高。两个种群间较高的遗传一致度(I=0．991)和较小的遗传距离(D=0．092)也证实了上述结果。由此推测,该蝗虫较强的迁飞能力有可能增强种群间的基因交流,降低种群间的遗传分化。然而,在一些等位酶指标如位点Ao-2、Fbp—l、Mdh—2的等位基因频率分布、每个位点的平均等位基因数(A=2．5和2．7)和多态位点百分率(P=52．6％和57．9％)等,两个种群之间呈现出明显的差异。这也许与两个种群之间不同的生态环境条件和较远的地理距离有关。     

宽翅曲背蝗两个地理种群等位酶的比较

采用水平淀粉凝胶电泳技术及应用等位酶分析方法,研究我国河北黄骅、辽宁葫芦岛宽翅曲背蝗两个自然种群的遗传多样性和遗传分化。在检测的11种酶15个酶基因座位中,Adk-I、Fbp-I、Mdh-2和G3pd-I基因座位的等位基因少,而Fbp-2、Mdh-I和Me-I基因座位的等位基因多。对每个基因座位的各基因型进行χ^2检验,除Mdh-I在辽宁葫芦岛种群、Adk-I在河北黄骅种群分别符合Hardy-Weinberg平衡外,其余绝大多数基因座位的基因型频率显著偏离Hardy-Weinberg平衡。两个种群之间存在明显的遗传多样性和分化：多态位点百分率分别为100％和93．3％,等位基因平均数分别为3．1和2．5,平均杂合度观测值分别为0．086和0．061。与其他非迁飞性蝗虫如中华稻蝗(Oxya chinensis)比较,这种蝗虫种群的平均杂合度较低但遗传多态性较高。结果表明：该蝗虫较强的跳跃能力可使个体暴露于各种不同环境,有利于维持种内遗传多态性的动态平衡,而种群保持较高的遗传多态性能增强该物种在不同栖息地的生存和繁殖能力。F-统计量表明两个种群之间的遗传分化相对较小,但这种分化显著高于迁飞性蝗虫如东亚飞蝗(Locusta migratoria manilensis)。Nei的遗传一致度(I)和Roger的遗传距离(D)的结果分析揭示了两个种群之间较高的遗传一致度(I=0．904)和较小的遗传距离(D=0．256)。然而,在一些酶基因座位如Aep-I(Fst=0．462)和Pgi-I(Fst=0．182),F-统计值相对较大,遗传分化比较明显。     

中国东亚飞蝗四个地理种群遗传结构的比较研究

利用水平切片淀粉凝胶电泳技术,分析了不同蝗区东亚飞蝗四个地理种群的遗传结构。在检测的20个酶基因座位中,四个种群均表现出一定的遗传多态性, 多态位点的百分率普遍偏高 (P=70%～80%),但由于杂合子数目较少而使每个位点的平均杂合度观察值偏低(H_o=0.023～0.032)。对每个基因座位的各基因型进行χ²检验, 除 Adk-1、Gdh-1、G3pd-1和Pgm-1在部分种群符合Hardy-Weinberg平衡外,其余绝大多数基因座位的基因型频率显著偏离Hardy-Weinberg平衡。从F统计量看,四个种群之间的遗传分化较低(F_st=0.0606 )。它表明： 东亚飞蝗较强的长距离迁飞行为增加了种群之间的基因交流, 降低了种群之间的遗传分化。根据Nei的遗传一致度（I）和Roger的遗传距离（D）进行分析, 在山西临猗与山西永济(I=0.964, D=0.175)、河南中牟与江苏沛县种群(I=0.957, D=0.160)之间,呈现出较高的遗传一致度和较小的遗传距离。结果表明： 迁飞性蝗虫东亚飞蝗种群之间的遗传分化与地理距离呈正相关。     

柚品种的等位酶变异研究

研究了柚的48个品种的等位酶变异,利用等位酶分析技术对柚的酯酶（EST）,6－磷酸葡萄糖异的酶（PGI）,6－磷酸葡萄糖变位酶（PGM）,莽草酸脱氢酶（SKD）,超氧化物歧化酶（SOD）共5个酶系统的10个等位酶基因座进行了分析,除PGI－1,PGI－2两个基因座外,其它8个均为多态性基因座;10个等位酶基因座共观察到的等位基因25个,平均每个基因座的有效等位基因数目为1．55,基因多样度0．2805,柚的品种间具有较为丰富的等位酶标记遗传多样性,但柚类种质资源群体总的遗传多样性水平偏低。柚的较低的有效等位基因数目与基因多样度可能由于人工选择及资源流失造成。     

不同地理飞蝗种群的遗传多样性

用GenBank中飞蝗Locusta migratoriaL.的序列来设计微卫星引物,并对这些引物的有效性进行验证。结果表明,在所设计的3对引物中,只有1对为有效引物,可扩增出微卫星位点。序列分析表明本位点是一个不连续的重复微卫星位点。该多态微卫星位点含有14个等位基因,不同飞蝗地理种群的等位基因数目、大小和频率都存在较大的差异。对该位点各等位基因型进行χ2检验,基因型频率的观察和期望杂合度分别为0.4578和0.8836,该位点不属于Hardy-Weinberg平衡位点(χ2=733.12,P=0.0000)。该微卫星位点表现出高度的多态性说明是分析飞蝗种群遗传多样性的优良分子遗传标记。     

中国4种蝗虫不同种群的遗传分化

采用水平淀粉凝胶电泳技术,应用等位酶分析方法对采自我国山西、江苏、河北等地不同蝗区优势蝗虫种类3科4种8个种群的12种酶18个酶位点进行了检测,比较了4种蝗虫种群水平的等位基因频率变化和它们之间的遗传距离。等位基因频率分析表明：中华稻蝗和笨蝗各位点等位基因丰富,而长翅素木蝗和短额负蝗各位点等位基因较少。在所研究的4种蝗虫8个种群中,多态位点百分率普遍较高(P＝53．3％—100．0％),由于杂合子数目较少而使每个位点的平均杂合度观察值偏低(H。＝0．034—0．139)。受迁飞能力、繁殖方式和活动范围的限制,4种蝗虫的平均杂合度观察值表现出一定的差异：笨蝗较高(H。＝0．089—0．139),其次是中华稻蝗(H。＝0．073—0．090),而长翅素木蝗(H。＝0．0488—0．068)和短额负蝗(H。＝0．034—0．050)相对较低。除Adk-1、Ao-1、Idh-1、Ldh-1、Ldh-2和Me-1在部分蝗虫种群符合Hardy-Weinberg平衡外,其余绝大多数位点的基因型频率显著偏离H—W平衡,但短额负蝗的多个位点符合或接近H—W平衡,表明该种蝗虫在自然种群结构方面明显有别于其他种类。从4种蝗虫的F—统计量(Fst)看,笨蝗种群间基因分化程度最高(Fst＝0．32),其次是短额负蝗(Fst＝0．31),而中华稻蝗相对较低(Fsl＝0．20),长翅素木蝗种群间基因分化程度最低(Fsl＝0．18),利用非加权算术平均法(UPGMA)对I值和D值进行聚类,所得聚类树也证实了这种遗传分化差异,上述结果反映了迁飞能力、适应性和环境因子对不同蝗虫遗传分化的影响。Nei的遗传距离(D)和遗传一致度(I)进行的聚类分析基本符合传统形态学、细胞学等研究结果：即同属于斑腿蝗科的中华稻蝗和长翅素木蝗遗传距离最小(D＝0．559),遗传一致度最高(I＝0．576)。在3个科中,癞蝗科和锥头蝗科之间呈现较小的遗传距离(D＝0．776)和较高的遗传一致度(I＝0．776),而这两科与斑腿蝗科之间的遗传距离相对较大(D＝0．908),遗传一致度相对较低(I＝0．406)。等位酶分析能较好地反映蝗虫不同物种的亲缘关系和种内不同种群之间的遗传分化程度。     

天然红松林等位酶研究

利用红松种子的单倍体胚乳,采用水平切片淀粉凝胶电泳技术,对采自三个地区的天然红松林的材料进行了１０种酶系统的等位酶实验,共获得了１８个等位基因位点,对这１８个位点等位基因的统计分析表明：红松种群的多态位点百分数为Ｐ＝７７．７８％,等位基因平均数为Ａ＝２．０,预期杂合度为Ｈｅ＝０．１６４８。红松种群间遗传分化程度在松属于较低水平。在红松１８个等位基因位点的变异中,绝大部分变异９４．２％来自种群内部,     

微生境下羊草两种生态型种群的遗传多样性及遗传分化——等位酶分析

采用水平淀粉凝胶电泳技术,应用等位酶分析方法测定了松嫩平原南部微生境条件下羊草灰绿色和黄绿色两种生态型９个种群的遗传多样性和遗传分化程度。羊草种、种群和生态型水平都维持较高的遗传多样性。两种生态型之间有明显的遗传多样性差异及遗传分化。灰绿型和黄绿型的多态位点百分率分别为６７．３％和５７．１％,等位基因平均数分别是２．２和１．９,期望杂合度分别为０．３４８和０．２９８。两种生态型种群之间平均遗传距离     

苍术（Atractylodeslancea（Thunb.）DC.）种内遗传多样性分析

分析了苍术５种等位酶１２个倍点２５个等位基因的分化特征。结果表明：群体内多态位点比例平均为０．６１，每们位点的平均杂合率为０．２６，每个位点发现的平均等位基因数为１．７８。     

甘蔗绵蚜不同地理种群异柠檬酸脱氢酶（IDH）的多样性

用等位酶分析方法对三个用药背景不同的甘蔗绵蚜地理种群在9种酶（EST，G3PD，HEX，IDH，LDH，MDH，ME，PGI和PCM）上的遗传组成进行检测。结果显示：甘蔗绵蚜在9种酶共检测到9个等位酶位点，仅IDH位点具有多态性。在多态性的IDH位点共检测到3个等位基因，其中连续两年未曾用药的两院种群和用药较少的木棠种群均具有三个等位基因（a,b和c），而用药次数最多的临高种群仅存在两个等位基因（a和b）。等位基因a的频率从两院种群到临高种群逐渐升高，而等位基因b的频率却逐渐降低。说明IDH在甘蔗绵蚜的种群遗传进化过程中起着重要作用，杀虫剂的选择压力可能对甘蔗绵蚜地理种群的遗传结构具有分化作用，同时也说明IDH在甘蔗绵蚜对杀虫剂的抗性产生中具有重要作用。IDH-a频率的升高，可能导致甘蔗绵蚜对杀虫剂产生抗性，可通过检测IDH位点等位基因频率的变化来监测甘蔗绵蚜对杀虫剂的抗性。     

Genetic variation and population structure of oriental migratory locust, Locusta migratoria manilensis, in China by allozyme, SSRP-PCR, and AFLP markers

Allozyme analysis, microsatellite primer PCR (SSRP-PCR), and amplified fragment length polymorphism (AFLP) techniques were used to assess genetic diversity and population structure of the Chinese oriental migratory locust, Locusta migratoria manilensis. A total of 299 PCR markers (67 SSRPs and 232 AFLPs) were detected in eight populations, of which 98.7% were polymorphic markers. The proportion of polymorphic loci (95.5-98.8%) by SSRP+AFLP markers indicated no significant differences between populations, and all populations exhibited a similar level of variability; results of the allozyme analysis demonstrated that 19 loci gave rise to a lower level of polymorphism (55.6-66.7%). The genetic distances between the populations were relatively low. Shannon's index and Nei's gene diversity showed low differentiation among the populations. Allozyme analysis, however, reflected greater similarity and smaller differentiation between the populations than those shown by SSRP and AFLP markers. Neighbor-joining dendrograms derived from both the allozyme and SSRP+AFLP markers showed that the genetic distances among Chinese oriental migratory locust populations were not greatly influenced by geographic distance and breeding habitats.     

Genetic Variation and Population Structure of Oriental Migratory Locust, Locusta migratoria manilensis, in China by Allozyme, SSRP-PCR, and AFLP Markers

Allozyme analysis, microsatellite primer PCR (SSRP-PCR), and amplified fragment length polymorphism (AFLP) techniques were used to assess genetic diversity and population structure of the Chinese oriental migratory locust, Locusta migratoria manilensis. A total of 299 PCR markers (67 SSRPs and 232 AFLPs) were detected in eight populations, of which 98.7% were polymorphic markers. The proportion of polymorphic loci (95.5–98.8%) by SSRP+AFLP markers indicated no significant differences between populations, and all populations exhibited a similar level of variability; results of the allozyme analysis demonstrated that 19 loci gave rise to a lower level of polymorphism (55.6–66.7%). The genetic distances between the populations were relatively low. Shannon’s index and Nei’s gene diversity showed low differentiation among the populations. Allozyme analysis, however, reflected greater similarity and smaller differentiation between the populations than those shown by SSRP and AFLP markers. Neighbor-joining dendrograms derived from both the allozyme and SSRP+AFLP markers showed that the genetic distances among Chinese oriental migratory locust populations were not greatly influenced by geographic distance and breeding habitats.     

EFFECTS OF POPULATION BOTTLENECKS ON GENETIC DIVERSITY AS MEASURED BY ALLOZYME ELECTROPHORESIS

Low levels of allozyme heterozygosity in populations are often attributed to previous population bottlenecks; however, few experiments have examined the relationship between heterozygosity and bottlenecks under natural conditions. The composition and number of founders of 55 experimental populations of the eastern mosquitofish (Gambusia holbrooki), maintained under simulated field conditions, were manipulated to examine the effects of bottlenecks on three components of allozyme diversity. Correlations between observed and expected values of allozyme heterozygosity, proportions of polymorphic loci, and numbers of alleles per locus were 0.423, 0.602, and 0.772, respectively. The numbers of polymorphic loci and of alleles per locus were more sensitive indicators of differences in genetic diversity between the pre-bottleneck and post-bottleneck populations than was multiple-locus heterozygosity. In many populations, single- and multiple-locus heterozygosity actually increased as a result of the founder event. The weak relationship between a population's heterozygosity and the number and composition of its founders resulted from an increase in the variance of heterozygosity due to drift of allele frequencies. There was little evidence that selection influenced the loss of allozyme variation. When it is not possible to estimate heterozygosity at a large number of polymorphic loci, allozyme surveys attempting to detect founder events and other types of bottlenecks should focus on levels of locus polymorphism and allelic diversity rather than on heterozygosity.     

Evaluation of genetic variability of the Kamchatka crab Paralithodes camtschaticus (Tilesius) using allozyme markers

Interspecific genetic variation in populations of red king crab Paralithodes camtschaticus Tilesius (Litholidae, Decapoda: Crustacea) was examined using allozyme markers. The activity of 57 enzymes and the general protein presumably encoded by 92 loci was detected. The level of allozyme variability was low: the expected heterozygosity and the proportion of polymorphic loci were respectively 0.027 +/- 0.008 and 6.5%. This level of heterozygosity is three times lower than the average value for 122 crustacean species (0.082 +/- 0.007). Although genetic variants were found at 22 loci, their frequencies were generally low: only in loci 6-Pgd, Alp-1, and Pep-1 did the frequencies of the most common alleles not exceed 0.9. All polymorphic loci except one had two alleles; the exception was 6-Pgd, which had three alleles. The possible reasons for the low level of allozyme variability in red king crab are discussed.     

Postglacial migration and the loss of allozyme variation in northern populations of Asclepias exaltata (Asclepiadaceae)

The recent Wisconsin glaciation has provided opportunities for examining the effects of postglacial recolonization on the population genetics of plant and animal communities. In this study allozyme Variation was examined in 19 populations of the herbaceous perennial Asclepias exaltata occurring in previously glaciated regions of North America. These northern populations of A. exaltata possess significantly fewer polymorphic loci (46.31 ± 2.7; mean ± 1 SD), alleles per polymorphic locus (1.84 ± 0.24), and expected heterozygosity (0.133 ± 0.031) than populations found in the Pleistocene refugium in the southern Appalachians. Population-level allozyme diversity decreased linearly from south to north and from east to west. Nineteen uncommon alleles previously observed in southern Appalachian populations were undetected in the northern region. Seven common alleles exhibited a clinal change in allele frequency. Of these, only Pgd-1a and Mnr-1c were at low-frequency in the southern Appalachians and increased significantly with increasing latitude and longitude, respectively. Despite this loss of allozyme diversity following postglacial migration, northern populations of A. exaltata have higher allozyme diversity and lower population differentiation (G" = 0.1 17) than mean values for other long-lived herbaceous perennials. Increased habitat fragmentation in northern regions and potential habitat loss in the southern Appalachians are likely to reduce the historically rich gene pool that has provided the genetic stock for postglacial recoveries.     

Allozyme variation in cork oak (Quercus suber L.): the role of phylogeography and genetic introgression by other Mediterranean oak species and human activities

 Genetic variation in the cork oak (Quercus suber L.) was investigated using 11 loci from seven enzyme systems in 40 populations sampled over the entire distribution of this species in the western Mediterranean Basin. Mean heterozygosity values over the polymorphic loci (Ho=0.283), the percentage of polymorphic populations (M=0.76), and the total genetic diversity (Ht=0.31) from which 11% was accounted for among-population variation, are among the highest recorded in oak species. In contrast to previous results in holm oak (Q. ilex L.), another evergreen species in the same area, cork oak possessed a smaller allele pool and a lower average number of alleles per locus and per population (A=2.0). More particularly, very few low-frequency alleles were observed in cork oak except for eight populations in which allozyme polymorphism at locus Pgi 1, diagnostic between both species, indicates that these low-frequency alleles are introgressed from holm oak. On the basis of the genetic distance estimated from allozyme frequencies, 32 of the 40 cork oak populations studied were classified into two very distinct sets which also corresponded to distinct geographic areas. One set gathered together the 18 populations from the Iberian peninsula and two adjacent areas in France, i.e. the centre of origin of cork oak, according to paleobotanical data. This set was characterized by a larger allele pool, a higher within-population genetic diversity and a lower differentiation between populations than was observed in the other set, which comprised the populations from North Africa, Sicily, Sardinia, Corsica, continental Italy and the region of Provence (southeastern France). In these more southern and eastern disjunct areas, cork oak migration from Iberia may have occurred at different periods since the end of the Tertiary. The possible effect of human activity on cork oak genetic structure, i.e. the selection of good-quality cork, acorn over-use for animal food, and even human nutrition, is discussed. Received: 3 March 1998 / Accepted: 19 March 1998     

Allozyme diversity among local populations of the pacific mussel <Emphasis Type="Italic">Mytilus trossulus</Emphasis> (Bivalvia: Mytilidae) from polluted areas of Peter the Great Bay (Sea of Japan)

Allozyme variability was assessed, using starch gel electrophoresis, at 15 polymorphic loci in two samples of the Pacific mussel Mytilus trossulus collected from local populations in heavily (Golden Horn Bay) and slightly polluted areas (Sukhoputnaya Bay) of Peter the Great Bay. Significant differences between samples were found in the genotypic frequencies at nine loci and in allele frequencies, at six loci. The results are suggestive of the differential survival of individuals having different genotypes and alleles at some of the surveyed loci under conditions of pollution. Our data are not in conflict with the hypothesis of the adaptive significance of allozyme polymorphism.     

Patterns of variation within and between Greek populations of Ceratitis capitata suggest extensive gene flow and latitudinal clines

Four natural Greek populations of Mediterranean fruit fly, Ceratitis capitata (Wiedemann), was studied for genetic variability at 25 enzyme loci. The comparison of polymorphism within and between populations shows two loci with high between-population heterozygosity (HT) and very high fixation index (F(ST)) values, suggesting the presence of balancing selection. The gradual decline of common allele frequency of the polymorphic loci tested indicated that latitudinal clines are present in Greece. Indirect estimates of gene flow based both on Wright's method (Nm*) and on the Slatkin's method (Nm*), which depends on the frequencies of rare alleles found in only one population, revealed a substantial amount of gene flow (Nm = 3.493 and Nm* = 3.197). These estimates of gene flow may well explain why the "introduced" Greek populations of C. capitata, in spite of their low genetic variability, display the same polymorphic loci. Gene flow in combination with natural selection and genetic drift may have played an important role to genetic differentiation in this species in Greece.     

Genetic differentiation of pink salmon oncorhynchus gorbuscha Walbaum in the Asian part of the range

Genetic variation at 19 allozyme (including 11 polymorphic) and 10 microsatellite loci was examined in the population samples of odd- and even-broodline pink salmon from the southern part of Sakhalin Island, Southern Kuril Islands, and the northern coast of the Sea of Okhotsk. The estimates of relative interpopulation component of genetic variation over the allozyme loci, per broodline, were on average 0.43% (GST), while over the microsatellite loci it was 0.26% (the theta(ST) coefficient, F-statistics based on the allele frequency variance), and 0.90% (the rho(ST) coefficient, R-statistics based on the allele size variance). The values of interlinear component constituted 2.34, 0.31, and 1.05% of the total variation, respectively. Using the allozyme loci, statistically significant intralinear heterogeneity was demonstrated among the regions, as well as among the populations of Southern Sakhalin Island. Multivariate scaling based on the allozyme data demonstrated regional clustering of the sample groups, representing certain populations during the spawning run or in different years. Most of the microsatellite loci examined were found to be highly polymorphic (mean heterozygosity > 0.880). The estimates of interlinear, interregional, and interpopulation variation over these loci in terms of theta(ST) values were substantially lower than in terms of rho(ST) values. Regional genetic differentiation, mostly expressed at the allozyme loci among the populations from the northern and southern parts of the Sea of Okhotsk (i.e., between the Sakhalin and Kuril populations), was less expressed at the microsatellite loci. The differentiation between these regions observed can be considered as the evidence in favor of a large-scale isolation by distance characterizing Asian pink salmon. It is suggested that in pink salmon, low genetic differentiation at neutral microsatellite loci can be explained by extremely high heterozygosity,of the loci themselves, as well as by the migration gene exchange among the populations (the estimate of the genetic migration coefficient inferred from the "private" allele data constituted 2.6 to 3.4%), specifically, by the ancient migration exchange, which occurred during postglacial colonization and colonization of the range.