Biosystematic studies on the spittlebug genus Philaenus with the description of a new species

Populations of the genus Philaenus were collected during the last 12 years at several places in Greece ranging in altitude from sea level to 2000 m. Within some of these populations the species P. spumarius (Linnaeus, 1758), which is widely distributed in Greece, and the Mediterranean species P. signatus (Melichar, 1896) occurred together. These two species can be easily distinguished by the size of the adults, and more precisely by internal genital structures. Surprisingly, some mountainous populations of P. spumarius also contained a new species. Externally the new species closely resembled P. spumarius , but careful examination of the male genitalia provided clear identification characteristics. The new species comprised one bisexual common colour morph similar to populi of P. spumarius and P. signatus and two rare unisexual morphs which are lacking both in P. spumarius and P. signatus. The new species occurred only at altitudes above 1200 m on Eryngium spp. (Umbelliferae) between June and August.     

Localized industrial melanism in the spittlebug Philaenus spumarius (L.) (Homoptera: Aphrophoridae) in Cardiff docks, south Wales

Twenty-four large adult samples of the meadow spittlebug, Philaenus spumarius , were collected from within a 4 times 3 km area containing Cardiff docks, south Wales. All samples are highly polymorphic for colour and pattern and show marked geographic variation in the combined frequency of the eight melanic morphs (from <15% to > 75%). Maximum melanic phenotype frequencies in both sexes coincide with dock wharfs currently containing large accumulations of pulverized coal and these samples are heavily contaminated with small coal particles. However, interpretation of this association is complicated by the existence, until 1976, of a smokeless fuel factory causing local air pollution immediately adjacent to the area of highest melanic frequency. It is suggested that the association of high levels of melanism in P. spumarius with areas heavily contaminated with particulate pollutants is due to direct selective effects of small particles favouring melanies.     

Biochemical genetics and taxonomy in Plumatella fungosa and P. repens (Bryozoa: Phylactolaemata)

SUMMARY. Some authors have recently concluded that, using morphological criteria, the freshwater bryozoans Plumatella repens and P. fugosa cannot be separated and should be regarded as conspecific. To test this conclusion, electrophoretic techniques have been used to examine genetic differences between the two nominate species at several enzyme loci. Significant variation at a malate dehydrogenase locus and three aminopeptidase loci establish beyond doubt that P. repens and P. fugosa are separate but related species. No significant variation was found between two populations of P. repens . Morphological characters for the distinction of the two species are discussed and evaluated.     

Elytral polymorphism in spittlebugs: Philaenus spumarius (L.) from the Inner Hebrides

The frequencies of colour and pattern morphs of the elytra of Philaenus spumarius are similar on three of the Small Isles of the Inner Hebrides (Eigg, Canna and Muck); the population on the fourth island (Rhum: is somewhat distinct but is like that at sea level on the nearby mountainous mainland. These similarities and differences suggest that these morph frequencies are regulated by natural selection.     

Genetic comparison between Pinus sylvestris and P. mugo using isozymes and chloroplast DNA

Pinus sylvestris and P. mugo populations from Poland and Czechoslovakia were compared using genetic variability at isozyme markers, chloroplast DNA variation, and mating system measurements. Two isozyme loci were found to differ between the species. P. mugo was as variable at isozyme loci as P. sylvestris. Diagnostic cpDNA fragments were found using the restriction enzyme Bcl-I. Populations that were morphologically classified as hybrids were found to be pure species, based both on isozyme and cpDNA results.     

Study of closely related species within the Physalaemus cuvieri group (Anura): contribution of microsatellite markers

Various species of the Physalaemus cuvieri group of frogs are difficult to distinguish morphologically, making molecular analysis an attractive alternative for indentifying members of this group, which is considered to be at risk because of loss of habitat. The genetic structure of natural populations of P. ephippifer and P. albonotatus species was investigated and analyzed, together with that of five previously studied populations of P. cuvieri. Nine microsatellite loci were used in the analyses. The overall G(ST) value (0.46) revealed high genetic variation among the populations, as expected for different species. Bayesian analysis implemented by the STRUCTURE software clustered the seven populations into seven groups (K = 7). All the P. albonotatus and P. ephippifer specimens were grouped into a single cluster, both species showing clear differentiation from P. cuvieri. The different grouping based on these microsatellites of some P. cuvieri individuals from Porto Nacional and from Passo Fundo suggests that they could be a new species, indicating a necessity for taxonomic reevaluation. Despite the intrinsic difficulties in analyzing closely related species, the nine microsatellite loci were found to be adequate for distinguishing these three species of the P. cuvieri group and their populations.     

The spittlebug Philaenus spumarius feeds from mature xylem at the full hydraulic tension of the transpiration stream

Feeding dynamics in the spittlebug Philaenus spumarius were analysed to determine the food source. Evidence from previous reports indicates that this species and similar insects feed from the xylem. However, it is not clear whether they extract sap from the main transpiration stream, or from some subsidiary compartment of xylem, such as immature or cavitated vessels. In feeding from such subsidiary compartments, the insects could avoid the large hydraulic tensions thought to prevail in xylem on the main transpiration stream. Philaenus , for example, is shown here to initiate feeding, and to feed rapidly, from plants in which the full xylem tension is estimated at −10 bar. From measurements of feeding rates and feeding patterns in stressed and unstressed plants, it is shown that Philaenus cannot feed from subsidiary xylem compartments. It is concluded that this insect feeds directly from the main transpiration stream, and at the full hydraulic tension which prevails there. The anatomy of the feeding pump in Philaenus appears consistent with this conclusion. Spittlebugs therefore offer an experimental tool for quasi- non-invasive sampling of the transpiration stream.     

Interspecific hybridization and genetic variability of Phlebotomus sandflies

The first successful hybridization is reported between Phlebotomus papatasi and P. duboscqi, two important Old World sandfly vectors of leishmaniasis and other diseases. Laboratory strains of P. papatasi and P. duboscqi were separable by six diagnostic enzyme loci: Est-3, Idh-1, Mdh-2, Mpi, Tre-1 and Tre-3. Hybrids between the two species were verified by the recovery of heterozygous isozyme patterns for the diagnostic loci. No F2 or backcross progeny were obtained. P. papatasi was separated from P. bergeroti by three diagnostic enzyme loci: Est-3, Mpi and Pgd. The isozyme patterns of P. bergeroti contain elements of both P. duboscqi and P. papatasi, although seven diagnostic loci (Est-3, Idh-1, Me, Mpi, Pgd, Tre-1 and Tre-3) separated P. bergeroti from P. duboscqi. Genetic variability profiles of the three species were established for 20 enzyme loci. Three geographically distant strains of P. papatasi from Calcutta, Maharashtra and Israel had isozyme genetic distances of < 0.05. The recently established Calcutta strain showed an unexpectedly low genetic variability with only one (Idh-2) of 20 loci being polymorphic (average heterozygosity of 1.9%) in contrast to 5-8 polymorphic loci (10-12% heterozygosity) in the Maharashtra and Israel strains. Mass and single pair crosses between the three P. papatasi strains were fertile with normal progeny numbers. Thus we found no signs of speciation in P. papatasi.     

A biochemical genetic analysis of populations of the northern pike, Esox lucius L., from Europe and North America

Genetic variation as identified by starch gel electrophoresis and enzyme staining was studied in seven populations of the northern pike, Esox lucius L. Samples derived from populations in northern U.S., Canada, Swedish fresh water, Swedish Baltic, the Netherlands, England and Ireland were analysed for variation at between 10 and 26 enzyme-coding loci. Polymorphism was found to be restricted to three loci: Sod-1 (mitochondrial superoxide dismutase), Ldh-2 (heart-type lactate dehydrogenase) and Got-1 (cytoplasmic glutamateoxalate transaminase). Mean heterozygosity ( H ) over all seven population samples was estimated to be 0·019. This extremely low variability, relative to reported data for other fish species, is discussed in terms of the structure and possible origins of pike populations.     

Inter- and intrapopulation genetic variation in species ofPorphyra (Rhodophyta: Bangiales) from British Columbia and adjacent waters

Starch gel electrophoresis of 17 proteins has provided data on inter- and intrapopulation genetic variation in 20 species ofPorphyra occurring in British Columbia and adjacent areas.P. cuneiformis andP. nereocystis showed no within species variation, even over ranges of more than 1000 km. Populations ofP. abbottae, P. fallax, P. fucicola, P. gardneri andP. schizophylla were characterized by fixation for certain alleles. The number of polymorphic loci in a population ranged from zero to nine, depending on the species. Six species had populations that were polymorphic at just a single locus. Only two species (P. mumfordii andP. pseudolanceolata) had populations that were polymorphic at more than three loci. These levels of genetic variation are lower than those reported for populations of JapanesePorphyra species. Eleven taxa were polymorphic for 6-phosphogluconate dehydrogenase, the most variable enzyme. No within species polymorphisms were detected for bromoperoxidase, lactate dehydrogenase, superoxide dismutase or phycoerythrin. Possible evidence for the chimeric nature of the thallus was observed only inP. mumfordii.     

High genetic diversity in southwest Asian populations of Philaenus spumarius (Hemiptera: Auchenorrhyncha)

The genus Philaenus is one of the best investigated among Auchenorrhyncha, and several morphological, ecological, karyological, and molecular data have led to a designation of up to 10 species, distributed mainly in the Mediterranean and south-western Asia. The only widespread Palaearctic species, P. spumarius, is known to be structured phylogeographically as it consists of two highly divergent mitochondrial clades (northeast, NE and southwest, SW), with several subclades. This study contributes to the species phylogeography through the study of the genetic diversity and affinity of P. spumarius populations from southwestern Asia. Mitochondrial DNA (cytochrome B) show a high level of genetic diversity within Turkish and Iranian populations, the majority of which belong to the SW clade, and only single populations from northeastern Turkey are found to be substantially highly divergent lineages within the NE clade. One of the NE populations also showed significant differences in the distribution and amount of heterochromatin compared to other populations. According to the results of this study and previous phylogenetic and phylogeographic works on this species, we conclude that Southwestern Asia is probably the place of origin of the Philaenus spumarius.     

Disparity in population differentiation of sex-linked and autosomal variation in sibling species of the Jaera albifrons (Isopoda) complex

The genetic variation at four enzyme loci is described for 22 populations of three Jaera species--J. albifrons, J. ischiosetosa, and J. praehirsuta--in the J. albifrons complex (Crustacea, Isopoda) in Denmark. The variation at three of the loci is similar, with the allele frequency spectra close to each other in all three species. An evolutionary tree based on the variation at these three loci revealed that the populations from the different species are completely intermixed in the tree. This was supported by hierarchical F-statistics where the between-species component was zero. At a fourth locus, Gpi (glucose phosphate isomerase), the species differ substantially. This locus is sex linked in J. ischiosetosa, but in the two other species, J. albifrons and J. praehirsuta, it is either found on autosomes or is sex linked with a high recombination rate between the locus and the centromere. An evolutionary tree for this locus partitions the populations into separate groups and a hierarchical F-statistic has a between-species component of about 50%. The results are attributed to introgression with a higher rate for autosomes than for sex chromosomes.     

Isozyme variation and the reproductive biology of Hydrocharis morsus-ranae L. (Hydrocharitaceae)

Electro-phoretically detectable isozyme variation was studied for 17 enzyme systems in several N American populations of the introduced aquatic plant Hydrocharis morsus-ranae and in two European populations. Twenty-nine loci were inferred from progeny, adult and turion enzyme banding patterns with 28 of these loci homozygous in all individuals studied. Malate dehydrogenase-1 (MDH-1) was the only locus which could be interpreted as multi-allelic and heterozygous. Twenty-seven of 76 seedlings assayed showed an age specific expression for an alcohol dehydrogenase locus (ADH-2) never seen in adults or turions. Since all adults sampled were phenotypically identical at all loci assayed, it is possible that only one isozyme genotype of this species is present in N America. European turion data further indicated that the populations studied were identical to N American plants sampled at all loci except EST. Therefore, although H. morsus-ranae is dioecious, outcrossing appears to involve substantial inbreeding. Connections between extensive inbreeding and the failure of effective sexual reproduction are considered.     

Electrophoretic evidence supporting a theory of allopolyploid origin of the peatmoss Sphagnum jensenii

Based on morphological evidence it has been hypothesised that Sphagnum jensenii is an allopolyploid, with S. annulatum and S. balticum as progenitors. Analysis of nine putative enzyme loci carried out on populations of these three species from central Norway, strongly corroborate this hypothesis. Sphagnum jensenii showed fixed heterozygosity at four of the loci suggesting that it is an allopolyploid. At each of the loci screened, extant populations of S. annulatum and S. balticum shared alleles with S. jensenii . The taxonomic interpretation is that S. jensenii should be recognised as a distinct species different from S. annulatum . Implications of allopolyploidy on ecological tolerances and niche breadth are briefly discussed.     

Genetic variation in Minuartia (Caryophyllaceae) in Svalbard

Based on a survey of eight enzyme systems, genetic variation in three Minuartia species was analysed and the diversity in 15 populations in Svalbard (78°N) compared to seven populations in Norway, Iceland, and Greenland. In the two sexual diploids, M. biflora and M. rubella , genetic diversity was as high in populations from Svalbard as in populations from more southerly latitudes. In M. biflora , eight out of 15 loci were polymorphic (P=53.3 %); in M. rubella eight out of 18 (P=44.4 %). The mean number of alleles per locus (A) and the number of multilocus genotypes (MG) were also higher in M. biflora than in M. rubella , A=1.60 and MG=21 vs. A=1.44 and MG=9. The proportion of genetic diversity due to variation among populations was much lower in M. biflora than in M. rubella , FST=0.249 vs. FST=0.895, and the estimated gene flow much higher, Nm=0.745 vs. Nm=0.029, indicating that M. biflora is a mixed mater and M. rubella a selfer. In the vegetatively reproducing tetraploid, M. rossii , 13 (65 %) out of 20 putative loci showed fixed heterozygosity, the mean number of alleles per 'locus' was A=1.70, and two multilocus phenotypes were observed, proving that at least two clones occur in Svalbard. No allele was shared by all three species and Nei's genetic identity for biflora-rubella was extremely low, I=0.07. The results suggest that the three species, which are currently placed in separate sections, represent lineages that diverged a long time ago.     

Studies on genetic diversity in Changium smyrnioides Wolff (Umbelliferae)

Changium smyrnioides Wolff is an endangered species of the monotypic genus Changium (Umbelliferae). For a better understanding of its genetic diversity, four populations of this species were cytologically and allozymically investigated. The karyotypic analysis of C. smyrnioides indicated that the chromosomal number was stable (2n = 20) in all populations. Karyotypes of the populations were all “2A” type. Karyotypical polymorphism was shown as the chromosomal heterozygosity, and position shift of the satellite. Genetic diversity of 4 populations was analysed with starch gel electrophoresis. A low level of allozymic diversity was estimated based on 19 loci of 10 enzyme systems. Genetic diversity within 4 populations was relatively low with the proportion of polymorphic loci (P) ranging from 19% to 42.9%, average number of alleles per locus (A) from 1.2 to l. 7, mean expected heterozygosity ( H_e ) 0. 073 and mean observed heterozygosity ( H_o ) of O.030. In general, about 58% of genetic variation were attributed to the differentiation among populations, while the remaining 42% resided within populations. The genetic diversity of the eastern populations was higher than that of the southwestern populations. On the basis of this studies, we thought that the habitat of C. smyrnioides should be protected and most of the populations should be sampled soas to retain as much genetic diversity as possible in ex-situ conversation.     

Isozyme variation in Desmodium nudiflorum

Five populations of Desmodium nudiflorum were assayed for genetic variability in eight enzyme systems encoding thirteen genetic loci. The proportion of polymorphic loci for the species as a whole is 46.2% while the proportion of polymorphic loci within populations averages 13.5%. Average individual heterozygosity is 2.3%. D. nudiflorum shows significant differences in gene frequencies between populations, and genotypic frequencies within populations conform to Hardy-Weinberg expectations.     

栗属中国特有种居群的遗传多样性及地域差异

采用超薄平板聚丙烯酰胺等电聚焦技术对栗属（Ｃａｓｔａｎｅａ（Ｔｏｕｒｎ．）Ｌ．）３个中国特有种即中国板栗（Ｃ．ｍｏｌｉｓｉｍａＢＬ．）、茅栗（Ｃ．ｓｅｇｕｉｎｉＤｏｄｅ．）和锥栗（Ｃ．ｈｅｎｒｙｉ（Ｓｋａｎ）Ｒｅｈｄ．ｅｔＷｉｌｓ．）的３０个居群,１２个酶系统的２０个位点进行了遗传多样性与遗传结构分析。结果表明：中国板栗在种和居群水平都具有较茅栗、锥栗高的遗传多样性（Ｐ分别为９０．０％和８４．７％,Ｈｅ分别为０．３１１和０．２９５）,尤以长江流域居群表现显著,揭示了长江流域的神农架及周边地区为中国板栗的遗传多样性中心。Ｎｅｉ的遗传一致度测量结果显示中国板栗和茅栗的亲缘关系最近,并且地理距离和遗传距离有一定的相关性。中国板栗、茅栗和锥栗的遗传分化程度逐渐增大,Ｇｓｔ分别为７．５％、１０．９％和２２．１％;基因流（Ｎｍ）分别为３．２０、２．０５和０．８８。研究结果为探明栗属起源、系统进化及制定保育策略提供了科学依据。     

栗属中国特有种居群的遗传多样性及地域差异

Genetic variability in 30 populations of three endemic Castanea (Tourn.) L. species (C. moUissima BI., C. seguinii Dode. and C. henryi (Skan) Rehd. et Wils. ) in China was investigated using isoelectric focusing in thin-layer polyacrylamide slab gels at 20 loci coding for 12 enzyme systems. C. mollissima was found to possess a significant higher value of genetic variability than that of the other two species. The percentage of polymorphic loci (P) and the expected heterozygosity (He) were 90.0% and 0. 311 at the species level, respectively; while P = 84.7 %, He = 0.295 at the population level. Comparison of the population genetic variability of C. mollissima in three regions of China, the populations from the Yangtze River valley showed a markedly higher mean expected gene heterozygosity. The results provided additional evidence that the Yangtze River valley, particularly in Shennongjia area, was the center of the genetic diversity of C. moUissima. Genetic relationships among populations and species were assessed by Nei genetic identity (I) and standard genetic distance (D), suggesting that C. mollissima and C. seguinii have a closer relationship, and genetic distances were correlated with geographical distances among populations. Genetic differentiation between populations in C. mollissima, C. seguinii and C. henryi was 7.5%, 10.9% and 22.1%, respectively, and the gene flow rate (Nm) was 3.20, 2.05 and 0.88 respectively. The study provides useful imfonnation for understanding of the origin and evolutionary events in genus Castanea and planning an effective conservation strategy.     

Ecologically and evolutionarily important SNPs identified in natural populations

Evolution by natural selection acts on natural populations amidst migration, gene-by-environmental interactions, constraints, and tradeoffs, which affect the rate and frequency of adaptive change. We asked how many and how rapidly loci change in populations subject to severe, recent environmental changes. To address these questions, we used genomic approaches to identify randomly selected single nucleotide polymorphisms (SNPs) with evolutionarily significant patterns in three natural populations of Fundulus heteroclitus that inhabit and have adapted to highly polluted Superfund sites. Three statistical tests identified 1.4-2.5% of SNPs that were significantly different from the neutral model in each polluted population. These nonneutral patterns in populations adapted to highly polluted environments suggest that these loci or closely linked loci are evolving by natural selection. One SNP identified in all polluted populations using all tests is in the gene for the xenobiotic metabolizing enzyme, cytochrome P4501A (CYP1A), which has been identified previously as being refractory to induction in the three highly polluted populations. Extrapolating across the genome, these data suggest that rapid evolutionary change in natural populations can involve hundreds of loci, a few of which will be shared in independent events.