Infraspecific variability in the volatile leaf oils of Chamelaucium uncinatum (Myrtaceae)

Variation in leaf essential oils and morphometric characters among 38 locations of Chamelaucium uncinatum were used to describe infraspecific diversity and geographic patterns of variation. Four chemical types were designated: citronellal, -pinene or limonene dominant, or one in which all three monoterpenes were co-dominant. The citronellal and limonene types were geographically restricted but the -pinene and co-dominant types were widespread. The citronellal type also demonstrated higher diversity (by Shannon–Weiner index) in the oil profile than the other types. Biochemical differentiation was not strongly paralleled by morphological differences between chemotypes; only the citronellal-type differed morphologically from the other three types with respect to floral bud and leaf characters. Based on biochemical and morphological differences, and geographic localization, it was concluded that the citronellal type may represent a recently evolved ecotype of C. uncinatum.     

Africa goes Europe: The complete phylogeography of the marbled white butterfly species complex Melanargia galathea/M. lachesis (Lepidoptera: Satyridae)

Climatic oscillations influence the distribution of species in time. Thermophilic species survived the ice ages in refugia around the Mediterranean. Northern Africa is one of the possibly important refugia. In this study we test the genetic differentiation between northern African and European populations, using the marbled white butterfly species complex, Melanargia galathea/M. lachesis, as a model. We studied 18 allozyme loci in 876 individuals from 23 populations representing a major part of Europe (northern Spain to Romania) and the western part of northern Africa (Atlas Mountains). The African populations resemble the European ones in allelic richness; their genetic diversity is higher than in Europe. Cluster analysis discriminated five European genetic groups: M. lachesis, a western European lineage, and three eastern European lineages. However, the African samples did not form a separate cluster within this phenogram, but clustered randomly within the Balkan/southeastern European groups. The genetic differentiation among the African populations (F_ST 8.8%) was higher than that within any of the European lineages (F_ST 2.6–5.5%). The high genetic diversity and the relatively strong differentiation of the four African populations sampled in a comparatively limited area of the Atlas Mountains indicate that the most probable origin of M. galathea is northern Africa, with its sibling species, M. lachesis, evolving in parallel in Iberia. Most probably, M. galathea colonised Europe first during the Eem interglacial, some 130 ky ago. Since M. lachesis must have existed on the Iberian peninsula during that period already, M. galathea should have reached Europe via Italy. The genetic differentiation to distinct groups in Europe most probably evolved during the following Würm glacial period.     

Population genetic structure among populations of three predaceous nabid species: Nabis alternatus Parshley, Nabis roseipennis Reuter and Nabis americoferous Carayon (Hemiptera: Nabidae)

An electrophoretic survey of 28 presumptive loci distributed among 15 enzyme systems was conducted to determine the genetic structure among local populations of three predaceous nabid species: Nabis alternatus, N. roseipennis and N.americoferous in human-disturbed habitat. For all three species, F_IS values indicated a deficiency of heterozygotes at most loci in most populations. The amount of genetic differentiation based on the standardized variance, F_ST when combined across all poulations was 0.100 for N. alternatus, 0.189 for N. roseipennis, and 0.335 for N. americoferous. When local populations were grouped according to a set of defined geographical limits, based on the naturally occurring spatial arrangement of the sampled localities, some unexpected patterns of genetic variation were revealed. The degree of genetic differentiation detected among local populations of nabids is discussed in light of the frequent disruptions and ecological changes in their ephemeral habitats.     

Genetic diversity and population structure in Tunisian Lavandula stoechas L. and Lavandula multifida L. (Lamiaceae)

The genetic diversity and population structure of 20 Tunisian Lavandula stoechas L. and Lavandula multifida L. populations, from different bioclimates, were analysed by starch gel electrophoresis using seven isozymes. The genetic diversity within populations varied according to species. Variation in L. multifida was higher than that observed for L. stoechas, and exclusive alleles were detected for taxa.

A high differentiation among populations, for each species, estimated by Wright's F-statistics was revealed. The genetic structure of populations from the same bioclimate was substantial. Nei's, R. [1978. Estimation of average heterozygosity and genetic distance from a small number of individuals. Genetics 89, 583–590] genetic distance among pairs of populations was low. The UPGMA cluster analysis of genetic distance values revealed that populations for each species were not strictly clustered together according to bioclimate or geographic proximity.

For each species, the low genetic divergence among populations and their substantial structure indicate their recent fragmentation due to anthropic pressures. The dendrogram generated from pairwise genetic distance among all populations showed two distinct clusters each corresponding to one species. The high genetic divergence between the two species, based on isozymes, corroborates their taxonomic status, as previously reported using morphological traits. The strategy for the management and conservation of populations should be made for each taxa according to its level of diversity and bioclimate.     

Population differentiation and phylogeography of Hygrophila pogonocalyx based on RAPDs fingerprints

Genetic variation of the endangered species, Hygrophila pogonocalyx Hayata (Acanthaceae), was estimated based on RAPD fingerprints. According to the criteria of the International Union for Conservation of Nature and Nature Resources, H. pogonocalyx is on the Red List Category due to its endangered status. Entomophilous plants of H. pogonocalyx are mostly pollinated by honeybees. Gene flow between populations is constrained by the migratory capacity of the pollinators. A survey based on RAPD fingerprinting using 50 random primers revealed the distribution of genetic variation following an “isolation by distance” model. A hierarchical AMOVA analyses indicated significant differentiation between geographical regions (Φ_ct=0.934; P=0.048), among populations (Φ_st=0.945; P<0.001), and among populations within region (Φ_sc=0.169; P<0.001). The differentiation between geographic populations may be ascribed to a long isolation since the formation of the Central Mountain Range 1 million years ago. In contrast to low levels of genetic variation in many endangered species, some genetic processes avoiding selfing may have evolved in H. pogonocalyx. Somatic mutation also possibly contributed to the variability maintenance within populations with limited size.     

Geographical variation in the monoterpenes of Pinus contorta oleoresin

The monoterpene composition of the shoot cortical oleoresin allowed the natural range of Lodgepole pine (Pinus contorta) to be divided into chemically distinct regions. The trees examined were all grown in Britain from seed originally collected from 150 localities in north-west America. Those from the central area of the natural range contained a wide variety of monoterpene pattern-types, while those from more peripheral areas tended to be more exclusively characterized by their own specific pattern-types. Chemical evidence for introgression with Jack pine (P. banksiana) was detected in trees originating from central British Columbia, and there was also a highly localized and pronounced region of introgression chemotypes in coastal populations near the border of Oregon and California. Variations between trees grown in replicate plots and at different sites in Britain were small, and the results indicated that resin analysis of a sample of 25 trees was usually sufficient to determine the region of origin of an unknown population.     

Genetic structure of populations of Calomys laucha (muridae, sigmodontinae) from central Argentina

The genetic structure of populations of the Sigmodontinae rodent Calomys laucha was studied by means of allozyme electrophoresis. This highly opportunistic species is found preferably in periodically perturbed habitats of crop fields in central Argentina, where it can attain very high densities. A total of 17 enzymatic proteins assayed gave information on 25 loci; only four were monomorphic in the seven populations studied. Levels of genetic variability (H_o from 0.144 to 0.171; P_95% from 44% to 56%) were higher than mean values reported for mammals and rodents. These high levels of heterozygosity could be maintained by large populations that do not experience great fluctuations in size, or by a social structure consistent in many small breeding groups that are formed and dissappear every breeding season. Genetic differentiation at a macrogeographical scale (θ=0.018) was low but statistically significant, and showed no correlation with geographic distance between pairs of populations. The pattern of population differentiation found is compatible with a relatively recent range expansion.     

Genetic diversity and population structure of endangered Isoetes coreana in South Korea based on RAPD analysis

Isoetes coreana Chung and Choi is on the list of critically endangered species in South Korea. Using randomly amplified polymorphic DNA (RAPD) markers, we investigated the genetic diversity within and between seven local populations of I. coreana. Ten RAPD primers produced a total of 94 bands, of which 59 (62.8%) were polymorphic. A low level of genetic diversity was recognized within the populations of I. coreana: polymorphic loci (P), with values ranging from 3.4% to 33.9%, and a mean value of 15.5% being observed. Analysis of molecular variance (AMOVA) showed that genetic diversity was greater among populations (81.6%) than within populations (19.7%) or among the three regions included in this study (Han River, Youngsan River, and Nakdong River). In addition, a high degree of genetic differentiation (θ^B = 0.742) was detected among the populations. These results indicate that genetic differentiation has occurred very rapidly. However, the rate of gene flow between populations was found to be as low as 0.059, irrespective of the genetic and geographical distances between the populations, which indicates that genetic drift must have played an important role in forming the present populations of I. coreana. Because a reduction of genetic diversity as a result of genetic drift is undesirable, increasing the gene flow between populations of Korean quillwort I. coreana should be considered as a conservation strategy.     

Genetic diversity in natural Cupressus sempervirens L. populations in Turkey

Nine polymorphic out of 13 isoenzymes (Gdh-A, Lap-A, Mdh-B, Mdh-C, Ndh-A, 6Pgd-B, Pgi-B, Pgm-A and Pgm-B) were used to assess genetic diversity in seven natural Cupressus sempervirens populations in South-western Anatolia. Except for Pgi-B, all loci were in Hardy–Weinberg equilibrium. A high level of genetic diversity was observed within populations. The mean number of alleles was 2.3±0.08, the effective number of alleles was 1.53±0.10, the percentage of polymorphic loci was 82.6±8.7%, expected heterozygosity was 0.350±0.039, observed heterozygosity was 0.326±0.039, and the fixation index was 0.049. Interpopulation differentiation was moderate, with differentiation indices of G_ST=0.073 and δ=0.104. A UPGMA tree and differentiation (D_j) showed that Kumluca, Kemer and Kas were the most differentiated populations. Pgm-B was one of the loci contributing most to interpopulation differentiation (G_ST=0.119; D_i=0.201), clearly distinguishing Eastern populations, as indicated by a highly significant correlation coefficient between D_j and longitude (R²=0.88^**). No other significant correlation was found between genetic measures and geographical traits for the Turkish cypress populations. A metanalysis of these and other cypress populations from Crete and the Aegean islands, that had been characterised previously using the same isoenzymes, showed no significant relationship between altitude and genetic measures (within-population differentiation or δ_T, observed heterozygosity, fixation index and D_j). Observed heterozygosity and latitude were negatively correlated. Longitude was found to be correlated with heterozygosity, fixation index and differentiation. Genetic and geographic distances were positively correlated over loci. Deviations from selective neutrality by Ewens–Watterson test were observed for Pgm-B, Ndh-A and 6Pgd-B.     

Predation by Callinectes sapidus (Rathbun) within Spartina alterniflora (Loisel) marshes

This study examined predation by the blue crab, Callinectes sapidus Rathbun, within intertidal Spartina alterniflora (Loisel) marshes of Dauphin Island, Alabama. Species and size preferences displayed by the predator when foraging within the marsh were investigated using nektonic, epifaunal, and infaunal prey populations including Fundulus similis Baird and Girard, Littorina irrorata Say, and Geukensia demissa Dillwyn.

Short-term field experiments involving the use of predator inclusion cages, in which the relative abundances of all prey species and the density of macrophyte vegetation were manipulated, indicated that mean mortality differed significantly among species. Blue crabs exhibited a distinct species preference for Littorina, and to a lesser extent, for Fundulus. However, the predator rarely choses infaunal individuals. Within predator inclusion cages, size selection by the crabs among three size classes of each prey was evident for Littorina and Fundulus but not for Geukensia. Blue crabs tended to select intermediate-sized snails and large fish while not exhibiting a size preference for infaunal bivalves.

In the marsh, mean percentage of the Littorina population within the 14–18 mm size class exhibited an increased mortality as compared to two other size classes, which was negatively correlated with increasing tidal height. Such a relationship may have been due to a decreasing gradient of crab predation associated with increasing tidal height. Geukensia size class distributions showed little evidence of differences along the tidal height gradient. No data are available for Fundulus, a mobile species which would not experience such differential predation along a marsh gradient.

In comparing crab predation patterns among prey species, it is apparent that Callinectes utilizes prey species differentially. Such differential utilization may be based on optimization of energy yield and minimization of energy expenditure. Thus, the preference of blue crabs for nektonic and epifaunal prey is hypothesized to be the result of a smaller energy expediture as a result of the crab's visual evaluation of these prey. Infaunal prey species (e.g., Geukensia) require a greater energy investment because of the necessity of excavating the prey item. Such prey also allow little selection by size because of being cryptic.     

基于COI和ND5序列的中国南部唇鱼骨和间鱼骨群体遗传结构

唇鱼骨和间鱼骨均为分布较广的初级淡水鱼类,是理想的亲缘地理研究材料;且两者形态特征较为相似,不易鉴别,故两者的分布记述和物种有效性存在争议.为了解我国南部唇鱼骨和间鱼骨的群体遗传结构并探讨两者的物种有效性,本研究对8条水系的唇鱼骨和9条水系的间鱼骨共130尾个体的COI和ND5基因序列片段进行了测定,并对这两个基因的组合序列(2151 bp)进行了分析.结果表明: 在130尾个体的COI和ND5基因组合序列中,共有196个核苷酸变异位点,共检测出50个单倍型,单倍型多样性为0.964,核苷酸多样性为0.019,遗传多样性较高.基于COI和ND5基因组合序列构建的 NJ 树显示,所有种群可分为两支,支系Ⅰ包含了韩江和九龙江的全部单倍型以及瓯江的部分单倍型,余下的单倍型组成了支系Ⅱ.两支系间的遗传距离为0.036,而唇鱼骨与间鱼骨之间的遗传距离为0.027.单倍型网络图表明,韩江、九龙江种群和其他水系种群分化较大;漠阳江种群由海南岛种群扩散而来;海南岛各种群及漠阳江种群的单倍型分支与珠江水系单倍型的分支之间的亲缘关系较近,与长江水系单倍型分支之间的亲缘关系则较远;湘江、桂江和柳江之间的亲缘关系较近.AMOVA分析结果显示,地理区之间的变异约占71.2%,地理区内种群间变异约占16.6%,种群内的变异占12.2%,表明其遗传分化主要来自地理区之间.错配分析及中性检验结果显示,全部种群、唇鱼骨种群、间鱼骨种群、支系Ⅰ和支系Ⅱ在历史上均没有发生过明显的扩张.     

Differentiation induction of human leukemia cells (HL60) by a combination of 1,25-dihydroxyvitamin D3 and retinoic acid (all trans or 9-cis)

1,25(OH)₂D₃ and two stereoisomers of retinoic acid, all trans and 9-cis retinoic acid, are regulators of cell proliferation and differentiation. The aim of this study was to evaluate the effects of a combination of 1,25(OH)₂D₃ and retinoic acid (all trans or 9-cis) on proliferation and cell differentiation of the human promyelocytic leukemia cell line HL60, and to test the reversibility of the induced differentiation. Cell proliferation was inhibited as expected by 1,25(OH)₂D₃ and all trans retinoic acid alone (IC₅₀ of cell survival was 4 × 10⁻⁷ M, 9 × 10⁻⁶ M and 9 × 10⁻⁷ M for 1,25(OH)₂D₃, all trans and 9-cis retinoic acid, respectively). Combination of 1,25(OH)₂D₃ and either form of retinoic acid resulted in a partially additive decrease in cell proliferation. 1,25(OH)₂D₃ induced a monocytic differentiation (100% CD14+ cells with 10⁻⁷ M 1,25(OH)₂D₃), while retinoic acid led to a predominantly granulocytic differentiation (36 and 42% CD67+ cells with 10⁻⁶ M all trans and 9-cis retinoic acid, respectively). Additive effects on differentiation were observed upon combination of subtherapeutical doses of the drugs, achieving a mainly monocytic population, demonstrating the dominant role of 1,25(OH)₂D₃ in determining the direction of differentiation. The effects on proliferation and differentiation of the solitary drugs were reversible, while the proliferation arrest and differentiation induced by the combination persisted and even progressed after withdrawal of the drugs. We conclude that 1,25(OH)₂D₃ and retinoic acid (all trans or 9-cis) exert additive effects on inhibition of proliferation and induction of cell differentiation of HL60 cells, leading to a persistent differentiation, even after drug withdrawal.     

Flavonoid variability within and between natural populations of Pinus uncinata

A total of 120 individual trees representative of Pinus uncinata were analysed for their foliar flavonoid content. The ratio of two extracted anthocyanidins, prodelphinidin and procyanidin, is a constant. Among the flavonols quercetin, kaempfrerol and isorhamnetin, the percentage of quercetin discriminates the five studied populations. Its distribution conforms to the Hardy-Weinberg law, but the ratio of the two implicit alleles proves the originality of each population, and leads to the recognition of distinct chemotypes.     

Genetic variation in Haworthia pumila and H. herbacea

Horizontal starch gel electrophoresis was used to examine genetic diversity within and differences between one population each of two morphologically different species of Haworthia, namely H. pumila and H. herbacea. Twenty-five leaf samples of each species were surveyed for 24 proteins of which 13 were useful for routine analysis, and gene products of 16 protein coding loci revealed genetic variation at 7 (43.8%) thereof in both species. Values of 1.63 (± 0.20) and 1.56 (± 0.18) were obtained for the mean number of alleles per locus and the average heterozygosity per locus was calculated at 0.168 (± 0.058) and 0.144 (± 0.048) for H. herbacea and H. pumila respectively. The malate dehydrogenase-2 locus is a potential genetic marker to identify the species studied electrophoretically. The mean genotypic distance index between the populations studied was 0.184, an indication of large degree of differentiation between the species. These values are much higher than values obtained for other members of the Aloaceae, showing that normal levels of genetic variation can be expected in at least some succulent monocotyledons.     

Genetic diversity of an amphicarpic species, Amphicarpaea edgeworthii Benth. (Leguminosae) based on RAPD markers

Amphicarpaea edgeworthii Benth. is an amphicarpic legume widespread in China. Amphicarpy describes the phenomenon that a plant produces aerial as well as subterranean fruits. A. edgeworthii can reproduce via three kinds of flowers: aerial chasmogamous flowers, aerial cleistogamous flowers, and subterranean cleistogamous flowers. Although there are some studies on the population genetic structure of species with both chasmogamous and cleistogamous flowers, none has been done for that of an amphicarpic species so far. The present study uses random amplified polymorphic DNA (RAPD) to assess level and pattern of genetic variation in 15 natural populations of A. edgeworthii. A total of 131 stable and clearly scored RAPD bands were achieved from 13 primers. The average genetic diversity within populations estimated by Shannon's information index was 0.218 at the population level, but ranged from 0.119 to 0.302, which was significantly different (P < 0.01). Different statistical analyses revealed a high level of genetic differentiation among populations (G_ST = 0.473–0.527). Thus, the pattern of genetic structure of A. edgeworthii is consistent with that of an inbreeding species.     

Role of the kdr and super-kdr sodium channel mutations in pyrethroid resistance: correlation of allelic frequency to resistance level in wild and laboratory populations of horn flies (Haematobia irritans)

The kdr and super-kdr point mutations found in the insect sodium channel gene are postulated to confer knockdown resistance (kdr) to pyrethroids. Using an allele-specific PCR assay to detect these mutations in individual horn flies, Haematobia irritans (L.), we determined the allelic frequency of the kdr and super-kdr mutations in several wild and laboratory populations. Wild populations with very similar allelic frequencies had resistance levels that ranged widely from 3- to 18-fold relative to a susceptible population. Conversely, the kdr allele frequency in a lab population with 17-fold resistance was nearly double that found in a heavily pressured wild population with 18-fold resistance. We conclude that, although the kdr mutation confers significant levels of pyrethroid resistance, a substantial component of resistance in insecticidally pressured populations is conferred by mechanisms that are PBO-suppressible. High super-kdr allele frequencies were detected in two resistant lab populations, but in wild populations with equivalent resistance the super-kdr allele frequency was very low. Interestingly, in over 1200 individuals assayed, the super-kdr mutation was never detected in the absence of the kdr mutation.     

Geographical variation in the monoterpenes of Valeriana officinalis Leaf

The variation in terpene composition of the leaf essential oil was studied in three populations of Valeriana officinalis L. subsp, collina. These populations grow in Italy in environmentally very different areas. Our examination of geographically and ecologically distant populations has provided evidence for substantial Iocal intrasubspecific differentiation. Statistical treatment of chemical data revealed two distinct chemical varieties of subspecies collina.     

Chloroplast DNA and the determination of species status in the Disa tripetaloides complex (Orchidaceae), and its relationships to three species Racemosae, section Disa

Disa cardinalis and three populations within the D. tripetaloides species complex contain variation in their chloroplast DNA (cpDNA) variability. All four taxa possessed unique cpDNAs and sequence divergence values ranged from 0.34 to 1.03%. A phylogeny of these genomes was reconstructed, along with the genomes of three other species, D. racemosa, D. uniflora and D. venosa, all of which are also section Disa and series RAcemosae, to determine the relationship of these closely related species to the D. tripetaloides complex. A phylogeny of the taxa using morphological data was also reconstructed. Outgroup comparison was made with D. sagittalis, a member of section Coryphaea. Although the molecular and morphological data were not completely congruent, both data types revealed D. cardinalis, rather than D. tripetaloides ssp. aurata, to be more closely allied with D. tripetaloides ssp. tripetaloides, suggesting that D. tripetaloides ssp. aurata should be elevated to species rank. Additionally, the high sequence divergence observed between the Natal and Cape populations, coupled with their geographical isolation and alternate flowering seasons, suggests that these two D. tripetaloides ssp. tripetaloides populations may, in fact, be more appropriately ranked as subspecies.     

Environmental factors affecting the lipid metabolism in Rosmarinus officinalis L

The effect of some environmental factors on the lipid metabolism was studied in two chemotypes of Rosmarinus officinalis L. Epicuticular hydrocarbons (EH), epicuticular fatty acids (EFA), whole leaf fatty acids (WLFA) and essential oils (EO) were extracted and analysed by GC-MS during winter 1991–1992 and related to temperature and moisture variations. Leaf fresh and dry wts were determined along with some morphophysiological parameters such as specific leaf weight (SLW) and specific leaf area (SLA). Leaf areas were calculated by image analysis and statistically processed as for chemical data. The results indicated that in R. officinalis the response to some environmental factors, with particular reference to temperature and moisture, was an increase in epicuticular hydrocarbons and a decrease in epicuticular fatty acids, leaf fatty acids and essential oils. Qualitative changes in the chemical composition of the above lipid classes were found to be correlated with temperature changes. From a chemosystematic viewpoint, a clear separation between the two chemotypes was achieved only when epicuticular hydrocarbons and essential oils were considered as chemosystematic characters.