Mitochondrial DNA variation in geographic populations of Colorado potato beetle,Leptinotarsa decemlineata (Coleoptera; Chrysomelidae)

Summary This study demonstrates variability in restriction enzyme cleavage sites of mitochondrial DNA (mtDNA) among four popalations of Colorado potato beetle (CPB). A suite of three enzymes (EcoRI,HpaI,PstI) was sufficient to discriminate among the populations tested. Individuals heteroplasmic for restriction enzyme patterns were found in some populations. Variability in CPB mtDNA should prove useful in efforts to trace the origin and dispersal of the species in North America.     

Mitochondrial DNA evidence for the 19th century introduction of African honey bees into the United States

Since the introduction of an African subspecies into Brazil in the mid-1950's¹, descendent Africanized honey bees (Apis mellifera L.) have spread throughout the Neotropics and into temperate North America. Restriction enzyme analysis of 422 feral honey bee colonies collected from non-Africanized areas in the southern United States revealed that over 21% of them had mitochondrial DNA (mtDNA) derived from a European race established in North America by the 17th century, 77% of them had mtDNA common in honey bees maintained by beekeepers and about 1% exhibited African mtDNA. Further analysis revealed that the African mtDNA was derived from a north African subspecies imported to the US in the 19th century.     

An analysis of allozyme,mitochondrial DNA and morphological variation in mussel (Mytilus galloprovincialis) populations from Greece

Three populations ofM. galloprovincialis from northern Greece were investigated using isozyme analysis, discriminant analysis of morphological characteristics and analysis of restriction fragments of mtDNA. For all three types of analysis significant intra- and interpopulation differentiation was found. This differentiation is very noticeable at the mtDNA genotype frequencies. Furthermore, the restriction patterns of mtDNA were different from those reported for Atlantic populations of this species.     

Genetic variation in the New World: Ancient teeth,bone, and tissue as sources of DNA

Examination of ancient and contemporary Native American mtDNA variation via diagnostic restriction sites and the 9-pb Region V deletion suggests a single wave of migration into the New World. This is in contrast to data from Torroni et al.³⁴ which suggested two waves of migration into the New World (the NaDene and Amerind). All four founding lineage types are present in populations in North, Central, and South America suggesting that all four lineages came over together and spead throughout the New World. Ancient Native American DNA shows that all four lineages were present before European contact in North America, and at least two were present in South America. The presence of all four lineages in the NaDene and the Amerinds argues against separate migrations founding these two groups, although admixture between the groups is still a viable explanation for the presence of all four types in the NaDene.     

Gel-electrophoretic description of European populations ofTerellia virens (Loew) (Diptera,Tephritidae); implications for its use as an agent for the biological control ofCentaurea spp. (Asteraceae) in North America

Allozyme frequencies of 15 enzyme loci, 14 of which were polymorphic, were used to characterize sevenTerellia virens populations originating from three allopatrically distributedCentaurea species. The two populations whose origins were geographically furthest apart, from Israel (onC. iberica) and from Switzerland (onC. vallesiaca), showed relatively high values of genetic distance from the 5 populations sampled in Austria and Hungary (onC. maculosa) (Nei's D>0.07). The latter five displayed a high degree of genetic similarity. No diagnostic (fixed) allelic differences were observed between these three groups ofT. virens populations, but they could be well characterized by significant differences in allelic frequencies at 9 enzyme loci. Independently of this study, the populations from Switzerland (C. vallesiaca) and eastern Austria (C. maculosa) were selected as potential source populations for future introductions into North America for the biological control of introducedC. maculosa andC. diffusa. Based on the observed genetic differences and results from field experiments on the host specificity of these two potential source populations, it is argued that host specificity screening tests should be conducted separately for local (host plant) populations, as such populations might accept a different set of hosts. Biotype mismatch and the risk of spill-overs to native species could thus possibly be reduced.     

Concordant change in mitochondrial and nuclear genes in a hybrid zone between two frog species (genusBombina)

Summary In contrast to results in other studies, nuclear and mitochondrial genes were found to change concordantly in a transect across the hybrid zone betweenBombina bombina andBombina variegata. mtDNAs of both species are found in populations in the central part of the zone, whereas populations at its margins contain mtDNA corresponding to nuclear genomes.     

Rapid isolation of mitochondrial DNA. Mitochondrial DNA fromDrosophila serrata

A simple and rapid method for isolation of high quality mitochondrial DNA (mtDNA) is presented in this report. Using this method, isolation and restriction site maps for 10 enzymes of the mtDNA ofDrosophila serrata were established.     

Isolation of biologically active alkaloids from Korean mistletoeViscum album,coloratum

Summary Anticancer activity of certain highly cytotoxic alkaloids present in Korean mistletoe has been demonstrated in experimental animals. Unlike European mistletoe, no cytotoxic proteins were found in the Korean mistletoe.This work was presented at ACS/CSJ Chemical Congress, Honolulu, Hawaii, April 1–6, 1979.We are thankful to Jerome Foundation, Los Angeles, California for a generous grant for the support of this work.     

Clinal variation and selection of MDH allozymes in honey bee populations

Latitudinal clines of malate dehydrogenase-1 (MDH-1) allozymes occur within honey bee populations on three continents: Europe, North America and South America. The North and South American populations are introduced and demonstrate that Mdh allelic clines were established within the last 150 years. The frequency of the medium electrophoretic allele increases in frequency with increasing latitude while the fast allele decreases with latitude on all the three continents. The clines are best explained by the average daily high temperature for July on all continents. These parallel clines provide evidence for selection on Mdh alleles in honey bees.     

Naja siamensis, a cryptic species of venomous snake revealed by mtDNA sequencing

Because of possible variation in venom composition, an understanding of venomous snake systematics is of great importance for the optimization of antivenom treatment of snakebite patients. Intraspecific variation in the morphology of many venomous snakes complicates the definition and indentification of some species when allopatric populations are involved. Selectively neutral or near-neutral mtDNA sequences can reveal evolutionary relationships obscured by ecogenetically-caused morphological variation. We use comparative sequencing of the cytochrome oxidase subunit 1 gene to reveal the existence of a widespread, cryptic species of spiting cobra from southeast Asia. This species,Naja siamensis, is widely sympatric with other Asiatic cobra species. This may be of considerable medical significance, and calls for further research into venom composition in Asiatic cobras.     

Pheromone components of the female elephant hawk-moth,Deilephila elpenor,and the silver-striped hawk-moth,Hippotion celerio

By means of gas chromatographic and mass spectroscopic methods, and combined GC-electroantennogram and electrosensillogram techniques, (E)-11-hexadecenal and (10E, 12E)-10,12-hexadecadienal [(E,E)-bombykal is also the main constituent of the pheromone of the silver-striped hawk-mothHippotion celerio. The biological activity of the substances was demonstrated with electroantennogram and single cell recording, and the physiological efficacy of the different hexadecadienal isomers compared.Pheromones, 79; as Pheromones, 78 is taken: Wu, Cai-Hong, and Bestmann, H. J., Chinese Science Bulletin34 (1989) 1475; pheromones, 77: Attygalle, A. B., Steghaus0Kovac, S., Ahmed, V. U., Maschwitz, U., Vostrowsky, Ol, and Bestmann, H. J., Naturwissen-schaften78 (1991) 90.     

A mitochondrial DNA polymorphism in honeybees (Apis mellifera L.)

Summary Mitochondrial DNA (mtDNA), isolated from worker honeybee larvae, was digested by each of seven 6-base restriction enzymes. Only one enzyme (BglII) showed a mtDNA difference between the three tested races (Apis mellifera carcia, A. m. ligustica, A.m. caucasica). BothA.m. carnica andA.m. ligustica showed the same pattern, differing fromA.m. caucasica. The degree of fragment pattern similarity revealed that there is only a small level of mtDNA variation between the three races tested. This is in line with previous investigations of enzyme polymorphisms.     

Mixed life cycles in the transitional zone between voltinisms in the fall webworm,Hyphantria cunea

The fall webworm,Hyphantria cunea Drury, was introduced from North America into Japan at half a century ago. At present, the populations north of 36° N are bivoltine and those south of 36° N are trivoltine. In the present study, the life cycle in the transitional zone between voltinisms was clarified in Tsukuba (36.1° N). When fourth instar larvae of the second generation were collected in the field and maintained in the laboratory, the incidence of pupal diapause increased when the sampling date was delayed from 16 August to 5 September. The fact that some developed without diapause inidcates the occurrence of a trivoltine life cycle in this locality, where a bivoltine life cycle is predominant. The critical photoperiod for diapause induction at 20 and 25°C was 14 h 13 min and 14 h 10 min, respectively. The photoperiodic response may explain the rapid increase in diapause incidence in late summer. In laboratory-reared diapause pupae, the time and period of chilling (5°C) greatly influenced the time required for adult emergence at the final incubation temperature of 25°C. However, diapause pupae chilled for a sufficiently long period developed to adulthood rapidly, irrespective of the conditions before chilling. Therefore, the difference in timing of entering diapause between pupae of the second and third generations would not result in their temporal reproductive isolation in the following spring. It is concluded that the mixed voltinism in Tsukuba is not a result of a mixture of genetically distinct bivoltine and trivoltine populations, but a result of the phenotypic plasticity in a genetically rather homogeneous population.     

Ancient DNA from Bronze Age bones of European rabbit (Oryctolagus cuniculus)

The European rabbit (Oryctolagus cuniculus) is now widely distributed throughout the world as a result of transportation by man. The original populations, however, were confined to southern France and Spain. In order to investigate the role of human intervention in determining the genetic diversity of rabbit populations, we are studying the origin of rabbits introduced onto a small Mediterranean island (Zembra) near Tunis over 1400 years ago, by examining ancient DNA extracted from rabbit bones found both on Zembra and on the European mainland. Ancient DNA was successfully extracted from rabbit bones found at two archaeological sites dated to at least the Early Bronze Age (more than 3500 years ago) in south-central France, and compared to that found in modern mainland and island populations using a small variable region of the cytochromeb gene. The results confirm that the Zembra Island population is descended from that present over 1400 years ago. The technical aspects of DNA extraction from bones and the implications of this type of research for determining the origin of introduced rabbit populations are discussed.     

A simple test using restricted PCR-amplified mitochondrial DNA to study the genetic structure ofApis mellifera L

The COI-COII intergenic region ofApis mellifera mitochondrial DNA contains an important length polymorphism based on a variable number of copies of a 192–196 bp sequence (Q) and the completer or partial deletion of 67 pb sequence (Po). This length variability has been combined with a restriction site polymorphism to produce a rapid and simple test for the characterization of mtDNA haplotypes. This test included the amplification by the polymerase chain reaction of the COI-COII region followed by aDraI restriction of the amplified fragment. In a survey of 302 colonies belonging to 12 subspecies, 21 different haplotypes have been found which have been unambiguously allocated to one of the 3 mtDNA lineages of the species. Although all colonies of lineage C exhibit the same pattern (C1), each one of lineages A and M presents up to 10 different haplotypes, opening the way to studies on the genetic structure and the evolution of a large fraction of the species. This test also differentiates southern Spanish and South African colonies, which can be of great interest for the Africanized bee problem.     

Isolation of isoflavonoid phytoalexins from seeds ofPhaseolus vulgaris L.

Summary Bean seeds challenged withRhizoctonia solani or exposed to natural microflora yielded the isoflavonoid phytoalexins: phaseollin, phaseollidin, phaseollinisoflavan, coumestrol and kievitone. The data suggest that seeds may have the major host-defence factors of a plant.The author thanks Dr Suresh S. Patil of University of Hawaii for his guidance and provision of laboratory facilities. Gift of phytoalexin samples by Dr S. Pueppke and Dr N.T. Keen is gratefully acknowledged. East-West Center, Honolulu, Hawaii, USA, is thanked for the financial assistance.     

Polymorphisms in mitochondrial DNA of european and Africanized honeybees (Apis mellifera)

Summary This study demonstrates polymorphisms in both the length and in the restriction enzyme cleavage sites of honeybee mitochondrial DNA (mtDNA). The levels of variation are typical of those found in other metazoan species. These polymorphisms are potentially useful for the identification of Africanized bees in the western hemisphere and for study of honeybee phylogenetics.     

From model to mimic: Age-dependent unpalatability in monarch butterflies

Monarch butterflies (Danaus plexippus) are unpalatable to various vertebrate predators because their larvae sequester bitter and emetic cardiac glycosides (CGs) from milkweed plants (Asclepias spp.). Here we show that the concentration of the defensive CGs decrease as individual butterflies age, regardless of the CGs' initial amounts or specific chemical structures. Consequently, individual monarch butterflies can change from being unpalatable models to palatable mimics during their lifetime. Since monarchs breed continuously over the spring and summer in North America, freshly emerged adult butterflies may serve as noxious models for older individuals which become automimics as they age.     

The Dead Sea — alive again

In the thirteen years of quantitative studies on the microbiology of the Dead Sea from 1980 onwards three distinct periods can be discerned. Mass development of the green unicellular algaDunaliella parva (up to 8,800 cells/ml) and red archaeobacteria (2×10⁷ cells/ml) was observed in 1980, following a dilution of the upper water layers by rain floods. This bloom disappeared at the end of 1982 as a result of a complete mixing of the water column. During the period 1983–1991 the lake was holomictic, and noDunaliella cells were observed. Viable bacteria were present during this period in very low numbers. Heavy rain floods during the winter of 1991–1992 caused a new stratification as the upper five meters of the water column became diluted to 70% of their normal salinity. In this upper water layerDunaliella reappeared (up to 3×10⁴ cells/ml at the beginning of May, rapidly declining to less than 40 cells/ml at the end of July), and a bloom of red archaeobacteria (3×10⁷ cells/ml) once more imparted a red coloration to the lake.     

Abnormalities of meioses in male reciprocal hybrids between the Hiroshima and Ichinoseki populations ofRana japonica

In order to elucidate cytogenetically the cause of male sterility in intraspecific hybrids ofRana japonica, the behavior of chromosomes in the first meiosis was observed in spermatocytes from male reciprocal hybrids between two populations from Hiroshima and Ichinoseki. In the parental Hiroshima and Ichioseki populations, 2530 (96.7%) meiotic spreads had 13 bivalents and 78 (3.0%) contained 12 bivalents and two univalents, whereas in reciprocal hybrids only 337 (7.0%) contained 13 bivalents and the other 4445 (93.0%) had 2–26 univalents. A total of 31647 (93.4%) bivalents was ring-shaped and the other 2234 (6.6%) were rod-shaped in both parental populations, whereas in reciprocal hybrids 26352 (57.1%) and 19819 (42.9%) bivalents were ring- and rod-shaped, respectively. These results show that meiotic chromosomes of reciprocal hybrids are characterized by a remarkable increase in univalents and rod-shaped bivalents.