中华水韭(Isoetes sinensis)等位酶分析的初步研究

采用超薄平板聚丙烯酰胺等电聚焦技术对珍稀涉危植物中华水韭（Isoetes sinensis Palmer）72个样品的等位酶遗传变异进行了初步研究。从24种酶筛选,得到了13种酶系统可用于该植物的遗传变异分析,其中7种酶系统在中华水韭的迁地保护区群中检测出16个位点,共27个等位基因,12个位点表现出多态性。遗传多样性初步分析表明：中华水韭具较低的等位酶遗传多样性。研究结果为中华水韭的综合保育及居群恢复、重建提供了初步的依据。中华水韭遗传变异的全面研究将在进一步改善等位酶方法的基础上,采用其它更为灵敏的分子标记方法做深入检测。     

Allozyme variation within and between populations inLolium (Poaceae)

Isozyme analysis was used to determine genetic variation within and between populations of sevenLolium species. All populations from the inbreeding species (L. temulentum, L. remotum, L. loliaceum, andL. persicum) were completely fixed for all enzymes scored. They also contained, for four of the five enzyme systems studied, exactly the same allelic variant. The three cross-breeding species showed large within-population variation and much less between-population variation. The great similarity of the allozymic variants found in all species, made the division of the genusLolium into species on basis of allozymic data difficult. It was not possible to separate the different inbreeding species from each other. Within the cross-breeding groupL. multiflorum andL. rigidum could be distinguished fromL. perenne. L. multiflorum, andL. rigidum could, with more difficulty, also be separated from each other. Allelic variation could have more relation with the provenance of the populations than with taxonomic classification.Genetic variation inLolium spp. II. For first part see Pl. Syst. Evol. 188: 87–99.     

Frequencies of null alleles at enzyme Loci in natural populations of ponderosa and red pine

Pinus ponderosa and P. resinosa population samples have mean frequencies of enzymatically inactive alleles of 0.0031 and 0.0028 at 29 and 27 enzyme loci, respectively. Such alleles are rare and are apparently maintained by selection-mutation balance. Ponderosa pine have much higher amounts of allozymic and polygenic phenotypic variation than red pine, yet both species have similar frequencies of null alleles. Thus, null alleles apparently do not contribute to polygenic variation, as has been suggested. The concordance between allozymic and polygenic variation adds support to the view that allozyme studies may be valuable in predicting the relative amount of polygenic variation in populations.     

Early stages and the classification of the milkweed butterflies (Lepidoptera: Danainae)

The Danainae, or milkweed butterflies, are a moderate-sized group of around 150 species that are of considerable ecological, physiological and behavioural interest. There are two currently accepted classifications for the subfamily and the aim of this study was to try to resolve this conflict using characters derived from the eggs, larvae and pupae. The 130 new characters (of which 85 are illustrated) are analysed by both phenetic methods (average linkage cluster analysis and principal coordinate analysis) and by cladistic techniques (Wagner tree, character compatibility analysis and manually derived cladograms). Overall, the results corroborate the more recent classification of the Danainae. However, some points of difference are found and, after component analysis, an alternative classification is presented in accordance with the conventions for constructing annotated Linnaean hierarchies.     

ELECTROPHORESIS IS MODIFYING OUR CONCEPTS OF EVOLUTION IN HOMOSPOROUS PTERIDOPHYTES

Analyses of electrophoretically detectable enzyme variants in homosporous pteridophytes are facilitating the development of new insights into their genetics and evolution. The number of isozymes per enzyme indicates that homosporous pteridophytes are genetic diploids, in spite of the fact that they have high chromosome numbers. High levels of heterozygosity and genetic variability in sporophytic populations indicate that many diploid species are outcrossing with inbreeding representing a derived character state. Because the congeneric homosporous pteridophyte species analyzed to date have low genetic identities, allozymic characters are also proving to be useful as genomic markers for elucidating patterns of reticulate evolution. The accumulated data suggest that the genetic system of homosporous pteridophytes differs fundamentally from that of seed plants. The present genomic constitution of extant taxa may be the result of repeated cycles of allopolyploidy followed by gene silencing and extinction of progenitor taxa. Alternatively, the original homosporous pteridophytes may have had high chromosome numbers. Although current species probably evolved recently, their phylogenetic roots may be difficult to trace because even closely related pteridophytes are genetically distant and extinction has obliterated the ancestral intermediates between lineages. These hypotheses can and should be tested using a combination of molecular, phylogenetic, and population biology methods.     

Allozymic variation and divergence in three species of Antirrhinum L. (Scrophulariaceae-Antirrhineae)

An allozymic study of three wild species of Antirrhinum L. A. lopesiamum Rothm., A. mollissimum Roihni. and A. microphyllum Rothm. is described. All are members of subsection Kickiella Rothm., and are narrow-range endemics of the Iberian Peninsula. The variability of the different loci, as well as the number and mobility of the alleles, differ among the three species, a demonstration of the usefulness of allozymes for the systematics of the genus. The finding of alleles unique to each species indicates high divergence among species suggesting ancient diversification, and supports the hypothesis of a geographical model of speciation. All three species show high levels of within-species variability, mainly partitioned within populations, while between populations genetic differentiation is low. Correlations between population size, sample size and genetic variability, and the usefulness of allozymic data for conservation purposes, are discussed.     

Testing hypotheses of speciation in the Plethodon jordani species complex with allozymes and mitochondrial DNA sequences

Salamander populations of the Plethodon jordani species complex form a challenging system for applying the general lineage concept of species to diagnose population-level lineages. The present study reports and analyses mitochondrial-DNA haplotypes (∼1200 nucleotide bases from the genes encoding ND2, tRNA^Trp, and tRNA^Ala from 438 salamanders) from 100 populations representing six species of the P. jordani complex ( Plethodon amplus , Plethodon cheoah , Plethodon jordani , Plethodon meridianus , Plethodon metcalfi , and Plethodon montanus ) with comparative analyses of previously published allozymic data to reconstruct the evolutionary history of this group and to diagnose species lineages. Analyses of mitochondrial haplotypic data include nested-cladistic analysis of phylogeography, analysis of molecular variance, hierarchical analysis of nucleotide-diversity measures, and likelihood-based estimates of recent temporal changes in population size. New analyses of allozymic data include multidimensional scaling and principal component analyses, and both data sets are analysed and compared for congruent genetic structure using Mantel correlation tests. These analyses in combination identify the six named species as distinct evolutionary lineages despite sporadic genetic exchanges among them and some discordance between mitochondrial DNA and allozymic markers. Sexual isolation is not complete for any pair of these six species, but they replace each other geographically and appear to block the geographical spreading of their neighbours. The P. jordani complex is a strong study system for investigating the genetic and ecological processes responsible for vicariant speciation.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 89 , 25–51.     

Correlation of ecological factors and allozymic variations with resistance toErysiphe graminis hordei inHordeum spontaneum in Israel: Patterns and application

The association of ecological factors and allozymic markers of wild barley,Hordeum spontaneum, with genotypes varying in resistance to 3 cultures of the pathogenErysiphe graminis hordei, which incites the disease powdery mildew of barley, were explored theoretically and practically. The study involved 275 accessions comprising 16 populations largely representing the ecological range ofH. spontaneum in Israel. From earlier studies of allozymic variation and disease resistance it now becomes apparent that genetic polymorphisms for resistance toE. graminis hordei are structured geographically, and are predictable by climatic as well as allozymic variables. Three-variable combinations of temperature and water factors explain significantly 0.32 of the spatial variance in disease resistance between localities. Also, several allozyme genotypes, singly or in combination, are significantly associated with disease resistance. A high correlation was found between the standard deviation of infection types of the culture of the pathogen from Israel, and allozymic polymorphism,P (r_s = 0.86, p < 0.001). Consequently, the IsraelH. spontaneum populations, growing in the center of diversity of the species, contain large amounts of unexploited disease resistance polymorphism. These could be effectively screened and utilized for producing resistant barley varieties by using ecological factors and allozymic variants as guidelines.     

The evolution of cardenolide-resistant forms of Na⁺,K⁺ -ATPase in Danainae butterflies

Cardenolides are a class of plant secondary compounds that inhibit the proper functioning of the Na⁺, K⁺‐ATPase enzyme in susceptible animals. Nonetheless, many insect species are able to sequester cardenolides for their own defence. These include butterflies in the subfamily Danainae (Family: Nymphalidae) such as the monarch (Danaus plexippus). Previous studies demonstrated that monarchs harbour an asparagine (N) to histidine (H) substitution (N122H) in the α subunit of Na⁺, K⁺‐ATPase (ATPα) that reduces this enzyme’s sensitivity to cardenolides. More recently, it has been suggested that at ATPα position 111, monarchs may also harbour a leucine (L)/glutamine (Q) polymorphism. This later amino acid could also contribute to cardenolide insensitivity. However, here we find that incorrect annotation of the initially reported DNA sequence for ATPα has led to several erroneous conclusions. Using a population genetic and phylogenetic analysis of monarchs and their close relatives, we show that an ancient Q111L substitution occurred prior to the radiation of all Danainae, followed by a second substitution at the same site to valine (V), which arose before the diversification of the Danaus genus. In contrast, N122H appears to be a recent substitution specific to monarchs. Surprisingly, examination of a broader insect phylogeny reveals that the same progression of amino acid substitutions (Q111L → L111V + N122H) has also occurred in Chyrsochus beetles (Family: Chrysomelidae, Subfamily: Eumolpinae) that feed on cardenolide‐containing host plants. The parallel pattern of amino acid substitution in these two distantly related lineages is consistent with an adaptive role for these substitutions in reducing cardenolide sensitivity and suggests that their temporal order may be limited by epistatic interactions.     

Alcohol dehydrogenase thermostability variants in Drosophila melanogaster: Comparison of activity ratios and enzyme levels

Representatives of five allozymic classes of Drosophila alcohol dehydrogenase have been compared with respect to their activity levels on two alcohol substrates, quantities of ADH protein, and stability in crude extracts. Within each allozymic class, strains from widely diverse geographic locations differ in their enzyme activity levels but are identical for a measure known as "activity ratio," which is obtained by dividing the average activity reading on isopropanol by that obtained with ethanol. They are also similar in the rate at which ADH activity declines in crude extracts held at 25 degrees C. For several of the fast-resistant and fast-moderate strains, differences in ADH activity are associated with differences in the amount of enzyme present. The catalytic efficiencies of the fast-resistant forms are considerably lower than those of the fast-moderate allozymes. The origin and persistence of the rare but ubiquitous fast-resistant allozyme is discussed.     

Allozymic and morphometric variation inLemna minor (Lemnaceae)

Allozymic and morphometric variation was studied in 28 clones ofLemna minor. This variation was compared with the corresponding variation in four clones ofLemna gibba and four clones ofSpirodela polyrrhiza. A high level of allozymic variation was observed among the clones, despite having been grown under uniform laboratory conditions for several years and despite its quasi-exclusive clonal means of propagation. Based on degree of allozymic similarity,Spirodela polyrrhiza was distinguished from the twoLemna species but the latter species were genetically indistinguishable. Allozymic similarity among clones ofLemna minor was not related to morphometric similarity, nor was it related to the degree of geographic separation or climatic similarity of their sites of origin. The results suggest that allozymic variation among these clones ofLemna minor may be largely neutral and not a consequence of differential selection.     

Phylogeographic analysis of mitochondrial gene flow and introgression in the salamander, Plethodon shermani

Plethodon shermani comprises a series of geographically disjunct populations occupying high-elevation mountain isolates. These populations hybridize at their borders with salamanders of the Plethodon glutinosus species complex, and past range expansions inferred from Pleistocene climatic cycles may have increased the possible genetic interactions between P. shermani and species of the P. glutinosus complex. Because mitochondrial DNA haplotypes often show introgression across species borders, we survey mtDNA variation for evidence of past and ongoing genetic interactions between P. shermani, its close relative Plethodon cheoah, and species of the P. glutinosus complex. Ongoing hybridization with the P. glutinosus-complex species Plethodon teyahalee is accompanied by extensive mitochondrial introgression in some Unicoi populations of P. shermani, but it has little genetic impact on P. shermani populations outside hybrid zones at three other isolates (Tusquitee, Wayah Bald, Standing Indian). Some Unicoi populations of P. shermani exhibit mtDNA evidence of past hybridization with diverse lineages from P. aureolus and P. glutinosus. The Tusquitee isolate of P. shermani is also characterized by mtDNA haplotypes most closely related to Plethodon aureolus and P. glutinosus, presumably introduced by past genetic contact with these species or with introgressed populations of Unicoi P. shermani. The mtDNA variation in sampled populations of the Wayah Bald and Standing Indian isolates of P. shermani appears largely unaffected by ongoing hybridization. Principal components analyses of allozymic data indicate that P. shermani isolates collectively form a genetically homogeneous unit clearly demarcated from species with which they have had current or past genetic interactions. Rapid mtDNA introgression associated with transient contacts between P. shermani and other species permits a fine-level resolution of evolutionary lineages not evident from allozymic data.     

Estimation of Genetic Variability in Natural Populations of DROSOPHILA SIMULANS by Two-Dimensional and Starch Gel Electrophoresis

Genic variation in natural populations of Drosophila simulans was surveyed using allozymic and two-dimensional electrophoretic techniques. Consistent with some previous reports, allozymic heterozygosity appeared lower than in the sibling species D. melanogaster (0.07 vs. 0.16). No variation was detected by two-dimensional electrophoresis of 19 lines scored for 70 abundant proteins. This is consistent with reported reductions in estimates of genic heterozygosity by two-dimensional electrophoresis in D. melanogaster, Mus musculus, and man. Although the amount of intraspecific variation detected in abundant proteins was lower than that detected for allozymes in D. simulans and D. melanogaster, the genetic distances between the sibling species calculated from the two data sets are not significantly different (0.35 and 0.20). The allozyme and two-dimensional electrophoresis data confirmed the impression from other measures of genetic variation (mitochondrial DNA restriction maps and inversion polymorphisms) that D. simulans is substantially less variable than D. melanogaster.     

A study of genetic variation in the threatened hepatic Petalophyllum ralfsii (Wils.) Nees. and Gottsche (Fossombroniaceae)

Genetic diversity of the dioicous thallosehepatic Petalophyllum ralfsii wasinvestigated and its significance for thespecies' conservation assessed. This globallythreatened hepatic apparently reaches itsgreatest abundance in the British Isles.Samples were taken from throughout the Britishrange; 178 individual thalli from 24 coloniesin nine localities were scored for variation in11 enzyme systems and 16 putative loci. Novariation was observed. Many studiesdemonstrating allozymic monomorphism inbryophytes have been of bisexual, orpredominantly vegetatively propagating taxa.Lack of variability in an obligatelyoutbreeding taxon, with no specialisedvegetative dispersal mechanism, lends supportto the view that hepatic taxa, at least withinEurope, routinely support little allozymicvariation. Unlike flowering plants, lifehistory and breeding system do not appear tohave significant effects on the extent andpartitioning of allozymic diversity.     

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。     

ALLOZYMIC VARIATION IN LOCAL APOMICTIC POPULATIONS OF LEMNA MINOR (LEMNACEAE)

Flowering occurrence and allozymic variation were studied in eight local populations of Lemna minor in eastern Ontario, Canada. After 2 years of survey, not a single flower was observed. This absence of flowering suggests the possibility of loss of sexual reproduction. This may have had no net adverse effect on fitness given the simple life history and prolific vegetative propagation in L. minor. However, the absence of sexual reproduction may limit genotypic diversity. The allozymic analysis detected 18 loci from 13 enzyme systems. Large deviations from Hardy-Weinberg equilibrium were common because of an excess of homozygotes for several enzyme systems. The genotypic diversity within these eight populations had a mean D value of 0.973 with an average number of genotypes per population of 19.6. These results suggest that genotypic diversity within these populations is not severely limited by the rarity of sexual reproduction. The mean genotypic distance index (D₁₄ = 0.801) suggests a high degree of differentiation between populations. The mean number of populations per genotype was 1.78. Using a Mantel test, the genotypic distance matrix was not significantly related to the population-to-population distance matrix (t = -0.161, P = 0.413). Although rare events of sexual reproduction may help maintain genetic variation, somatic mutations and multiple origins of clones may be important factors maintaining genetic diversity both within and between populations of L. minor.     

A comparative analysis of allozyme variability in vertebrate animals]

Comparative analysis of levels of the allozymic variation in the vertebrates is conducted using evaluation of a) within population heterozygosity of ferment coding homologous loci, b) average (per each species) allele frequencies of homologous loci, and c) overall sample of the loci being analyzed. It is established that amphibians and reptiles are characterized by the highest, birds and mammals--by the lowest, and fishes--by the middle level of genetic diversity. The diversity of genetic systems, if the constant heterozygosity of duplicated loci is taken into consideration, decreases from fishes to mammals and from cold-blooded to warm-blooded vertebrates. This tendency could be considered as an extrapolation of the "progressive specialization" rule on molecular level. Three basic factors determining certain level of genetic diversity of a species are acknowledged position in the phylogenetic system, population structure, and level of introgressive hybridization. From methodological viewpoint, evaluation of the genetic diversity by homologous loci seems to be most valid in comparative studies.     

Allozymic variation in tubificid oligochaetes from the Laurentian Great Lakes

Aquatic oligochaetes are an important component of the benthos in many freshwater habitats. Yet, virtually nothing is known about the population genetics of this group. Electrophoretic surveys of allozymic variation of selected members of the Family Tubificidae were conducted at six locations in the Laurentian Great Lakes of North America. Branchiura sowerbyi was the dominant member of the oligochaete fauna at two sites, and was found to be monomorphic at all enzyme loci that were examined. In contrast, members of the genus Limnodrilus, which included L. cervix, L. claparedianus, L. hoffmeisteri, L. maumeensis, and L. udekemianus showed considerable allozymic variation at several enzyme loci. L. udekemianus exhibited tri- and tetra-allelic heterozygous electromorph banding patterns at the monomeric Pgm locus, along with unbalanced heterozygous patterns at both Pgm and the dimeric Pgi locus.Genetic distance analyses suggest that L. cervix, L. claparedianus, and L. maumeensis are closely-related (genetic identifies ranged from 0.92–0.85), and may represent subspecies rather than distinct species. Breeding studies need to confirm this assertion. This survey represents the first attempt to characterize allozymic variation of aquatic oligochaetes in North America. Additional work should focus on elucidating taxonomic ambiguities within this group via both morphological and biochemical genetic studies.     

Correlation between variability and body size in vertebrates

An analysis of allozymic variation carried out in the main groups of vertebrate animals revealed a tendency towards the increased level of genetic polymorphism in the species of small animals compared to the large ones. This tendency was clearly followed in caudate amphibians, fishes, and mammals. The data are discussed in terms of the integration of monogenic and polygenic systems in the populations. It is hypothesized that this relationship between heterozygosity and body size confirms more general regularity consisting in highly statistically significant correlation between polygenic heterozygosity, maturation rate and life span. It is suggested that high rate of development in small animals resulting in early sexual maturation, can serve as a mechanism determining correlation between heterozygosity and body size at the species level. As a result, compared to large animals, small animals display higher rates of generation change, resulting in accelerated growth of population size and faster accumulation of genetic variability.     

Correlation between Genetic Variability and Body Size in Vertebrates

An analysis of allozymic variation carried out in the main groups of vertebrate animals revealed a tendency towards the increased level of genetic polymorphism in the species of small animals compared to the large ones. This tendency was clearly followed in caudate amphibians, fishes, and mammals. The data are discussed in terms of the integration of monogenic and polygenic systems in the populations. It is hypothesized that this relationship between heterozygosity and body size confirms more general regularity consisting in highly statistically significant correlation between polygenic heterozygosity, maturation rate and life span. It is suggested that high rate of development in small animals resulting in early sexual maturation, can serve as a mechanism determining correlation between heterozygosity and body size at the species level. As a result, compared to large animals, small animals display higher rates of generation change, resulting in accelerated growth of population size and faster accumulation of genetic variability.