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.     

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.     

Low allozyme and mtDNA variability in the island endemic speciesDrosophila sechelia (D. melanogaster complex)

Summary Genetic variability ofD. sechellia is investigated at both mitochondrial and nuclear levels. The results reveal the existence of a single main type of mtDNA with very few variants and a very low enzyme polymorphism. This situation is consistent with the small population size of this specialized species.     

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.     

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.     

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.     

Preliminary observations of a bacteriophage infectingXenorhabdus luminescens (Enterobacteriaceae)

A bacteriophage infective toXenorhabdus luminescens, a bacterial symbiont of heterorhabditid nematodes, was recovered from insects that supported poor nematode development. Plaque tests showed the phage particles to be infective only to primary and not secondary colonies ofX. luminescens. The phage was not infective toX. nenatophilus primaries or secondaries. The bacteriophage particles ranged 80–90 nm in length, with the head ranging from 40 to 50 nm in diameter. Restriction analysis was performed on isolated bacteriophage DNA. This first report of a bacteriophage fromXenorhabdus species has pratical implications since it could be detrimental to cultures ofHeterorhabditis nematodes that are being produced throughout the world for the biological control of insects.     

Heterochromatin characterization and distribution in the chromosomes of two populations ofIdotea baltica basteri Audouin, 1826 (Isopoda,Valvifera)

Two mediterranean populations ofIdotea baltica basteri from Messina and Naples showed a set of chromosomes composed by 58 all-biarmed chromosomes. The heterochromatin was located in the pericentromeric region of the chromosomes, and its composition appeared heterogeneous. In fact, not all the homologs showed heterochromatin resistant to digestion with three restriction enzymes (Alu I, Hae III and Sau 3A). Moreover, the two populations showed polymorphism in a band of G+C-rich telomeric heterochromatin, which was present only in the population from Messina. It is hypothesized that chromosomal polymorphism might reflect the geographical isolation of the two populations. It is also suggested that a process of diversification is taking place.     

Selection for class IIMhc heterozygosity by parasites in subterranean mole rats

Mhc organization and polymorphism have previously been studied²⁶ in the four chromosomal species of theSpalax ehrenbergi superspecies in Israel, serologically, and at the DNA, RFLP and sequence levels of class I and class II genes. Here we demonstrate that the observed heterozygosity ofMhc class II genesP₁ with 11 alleles, andQ, with at least 14 alleles, is positively and significantly correlated with infectivities of ectoparasites (gamasid mites)¹⁷ and endoparasites (helminths)¹⁸.Mhc heterozygosity is highest in the most infected area, which is in the most humid-warm region of the superspecies range, or where two zoogeographic regions overlap. We conclude that the evolutionary forces responsible for theMhc class II two-gene polymorphisms include selection for increased heterozygosity as a defense strategy against ecto- and endoparasite infections.     

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.     

The genetics of phytotoxin production by plant pathogenic fungi

Little is known about the genetic control of phytotoxin production by plant pathogenic fungi. The production of host-selective toxins known to play a role in disease development has been genetically analyzed in three species ofCochliobolus. InC. heterostrophus, a single genetic locus with two alleles has been identified controlling the production of HMT-toxin. This locus appears to be at or near the breakpoint of a chromosome rearrangement. Single genetic loci have also been identified controlling the production of HC-toxin byC. carbonum and HV-toxin byC. victoriae. The locus inC. carbonum may be a cluster of tightly linked genes.     

Notable diversity in hemoglobin expression patterns among species of the deep-sea clam, <Emphasis Type="Italic">Calyptogena</Emphasis>

The deep-sea clams Calyptogena nautilei and C. tsubasa, which live in the cold-seep area at a depth of 3570 m in the Nankai Trough, Japan, have abundant hemoglobins (Hbs) in erythrocytes, similar to other Calyptogena species. We determined the cDNA-derived amino acid sequences of Hbs from two Calyptogena species. C. tsubasa was found to contain two dimeric Hbs, Hb I consisting of 145 amino acid residues and Hb II with 137 residues, similar to known Hbs from C. soyoae and C. kaikoi. Sequence identity was over 90% among the orthologous chains of Calyptogena Hbs. On the other hand, surprisingly, C. nautilei contained two monomeric Hbs, Hb III containing 141 residues and Hb IV with 134 residues. In addition, Hbs III and IV showed only 33–42% sequence identity with Hbs I and II from other Calyptogena species. The distal (E7) histidine, one of the functionally important residues of the heme protein, is replaced by glutamine in all Hb chains of Calyptogena species. A phylogenetic analysis indicated that C. nautilei Hb III is closer to Hb I from other Calyptogena species. We suppose that a Hb gene was duplicated at least three times in an immediate ancestor of Calyptogena and, presumably depending on physiological conditions different Hb sets are being expressed: dimeric Hbs I and II in C. soyoae, C. kaikoi and C. tsubasa, and monomeric Hbs III and IV in C. nautilei. Received 13 May 2003; received after revision 5 June 2003; accepted 12 June 2003     

Geographic populations of the medfly may be differentiated by mitochondrial DNA variation

Restriction enzyme cleavage sites of mitochondrial DNA (mtDNA) from the Mediterranean fruit fly were found to vary among introduced populations in the Neotropics. The survey included samples from 15 established natural populations and 5 laboratory cultures from Hawaii, Central America, South America and West Africa and samples from recent California infestations (1989, 1991). Based on restriction fragment length polymorphisms from 2 enzymes, Hawaii is an unlikely source for the 1989 and 1991 California infestations. Interpopulational variation in mtDNA demonstrates the potential for the technique to trace the process of colonization (geographic spread) by this insect.     

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.     

Insect hemolymph clotting

The clot’s appearance in different large-bodied insects has been described, but until recently, little was known about any insect clot’s molecular makeup, and few experiments could directly test its function. Techniques have been developed in Drosophila (fruit fly) larvae to identify clotting factors that can then be tested for effects on hemostasis, healing, and immunity. This has revealed unanticipated complexity in the hemostatic mechanisms in these larvae. While the clot’s molecular structure is not yet fully understood, progress is being made, and the loss of clotting factors has been shown to cause subtle immune defects. The few similarities between coagulation in different insect species and life stages, and the current state of knowledge about coagulation in insects are discussed.     

Pupation-temperature range in 12Drosophila species from different ecological backgrounds

Summary A comparison of pupation-temperature range was made in the laboratory on a temperature gradient (3–38°C) using 12 species ofDrosophila representing four species groups and four different ecological backgrounds (temperate-montane forest:virilis group; desert;repleta group; cosmopolitan:melanogaster group; tropical forest:willistoni group). Within groups, differences are found which usually reflect species' distributions. Comparisons of species' mating-, oviposition- and pupation-temperature ranges reveal that pupation most-often occurs at temperatures beyond those for mating and oviposition. Each species reflects a different combination of temperature effects. Individual species have different temperature-limits for mating, oviposition and pupation. Temperatures permissive for one response are not predictive of limits on other responses. Among species, temperature can affect a particular response differently. Within groups, species differences can be at high and/or low temperatures for any response, and temperature effects among closely related species can manifest themselves in one, or any combination of responses. One cannot predict which responses will be most and least limited, or at which end of the temperature scale a response will be most limited. Among groups,common, but notabsolute temperature ranges generally correspond to the geographic distributions and ecological backgrounds of the species triads. The evaluation of temperature effects on species, based on a single activity, may not be adequate for predicting adaptive strategies.     

Ontogenetic changes in isozyme patterns during seed germination of self-pollinatedSecale species

Summary Six isozymatic systems have been studied comparatively during the first week of germination of seeds of self-pollinatedSecale species (S. silvestre Host. andS. vavilovii Gross.). Isozymatic systems do not change at all, or reach their definitive adult plant pattern early during germination.Acknowledgements. This work was carried out at the Dpto. de Genética, Univ. Complutense, Madrid and supported by grants from the C.A.I.C.Y.T. (1789-82) and the P.F.P.I.     

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.     

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.     

Mapping of restriction enzyme cuts by a new two-dimensional procedure

Summary A new procedure has been worked out to establish restriction maps. The method is fast, does not in general require labeled DNA and has been applied to map the linear palindromic rDNA ofPhysarum with the restriction enzymeBstEII.Acknowledgment. This work was supported by grants from the NIH (GM-30338), the NSF (PCM 8116391) and the Swiss National Science Foundation (3.075.81).