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D S Haymer

Publications and source records attributed to D S Haymer.

16 recordsLinked to original sources

Microsatellite analysis of medfly bioinfestations in California.

The Mediterranean fruit fly, Ceratitis capitata, is a destructive agricultural pest with a long history of invasion success. This pest has been affecting different regions of the United States for the past 30 years, but a number of studies of medfly bioinfestations has focused on the situation in California. Although some progress has been made in terms of establishing the origin of infestations, the overall status of this pest in this area remains controversial. Specifically, do flies captured over the years represent independent infestations or the persistence of a resident population? We present an effort to answer this question based on the use of multilocus genotyping. Ten microsatellite loci were used to analyse 109 medflies captured in several infestations within California between 1992 and 1998. Using these same markers, 242 medflies from regions of the world having 'established' populations of this pest including Hawaii, Guatemala, El Salvador, Ecuador, Brazil, Argentina and Peru, were also analysed. Although phylogenetic analysis, amova analysis, the IMMANC assignment test and geneclass exclusion test analysis suggest that some of the medflies captured in California are derived from independent invasion events, analysis of specimens from the Los Angeles basin provides support for the hypothesis that an endemic population, probably derived from Guatemala, has been established.

Agriculture↗

Repetitive A-T rich DNA sequences from the Y chromosome of the Mediterranean fruit fly, Ceratitis capitata.

Copies of a repetitive DNA sequence distributed over 90% of the length of the long arm of the Y chromosome of the Mediterranean fruit fly, Ceratitis capitata (medfly), have been characterized. Sequencing reveals that these repeats, ranging in size from approximately 1.3 to 1.7 kb, are A-T rich overall (67%). In most cases the repeat units appear to occur in tandemly linked arrays. The repeat copies also all contain a highly similar internal region, approximately 200 bp in length, with a more extreme A-T content bias. This internal region, designated as the AT element, exhibits an A-T content of at least 83%. This exceeds what has been described for any comparable element among invertebrates. Using primers designed from the DNA sequence, PCR amplification of an internal region encompassing the AT element also reveals that these sequences are present only in the male genome in different strains of the medfly.

AT Rich Sequence↗

Genetic relationships of populations and the origins of new infestations of the Mediterranean fruit fly.

A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) approach based on the variation in intron sequences of the glucose-6-phosphate dehydrogenase (Zw) gene was used to assess genetic variability in 26 populations and infestations of the Mediterranean fruit fly (medfly), Ceratitis capitata. Beginning with the exon-primed intron-crossing PCR (EPIC-PCR) method to amplify introns of this gene, five alleles were identified on the basis of DNA sequence variants. Several of these variants affect recognition sites for the restriction enzymes RsaI and TaqI. Using these enzymes in successive digestions of the EPIC-PCR products, each of these alleles can be identified directly from individuals. From this, surveys were conducted to document genotypes and allele frequencies in these samples. The relationships of existing populations and the invasion process represented by new infestations of the medfly were analysed using a principal coordinate analysis and the amova method to quantify the distribution of genetic diversity at different levels in a hierarchical manner. From these results, a framework of genetic relationships among the populations and infestations is presented. In addition, for at least some of the infestations, populations that are probably acting as sources of origin have been identified.

Animals↗

A member of the hsp70 gene family from the Mediterranean fruit fly, Ceratitis capitata.

A gene from the Mediterranean fruit fly (Medfly) Ceratitis capitata, belonging to the hsp70 family of genes, has been cloned and characterized. This gene appears to be an example of a heat shock cognate (hsc) gene based on a variety of structural and functional criteria. The heat shock cognate genes serve critical roles in chaperoning of cellular proteins, and they are expressed in a more constitutive manner compared to the inducible heat shock genes. We determined the sequence of the entire coding region of this gene (designated Cerhsc1) together with 5' and 3' flanking regions. The coding region exhibits specific structural features that are characteristic of the subclass of heat shock genes producing proteins localized to the cytosol. The 5' regulatory region of Cerhsc1 contains onlythe minimal array of canonical heatshock element sequences, a phenomenon commonly seen in heat shock cognates. Abundant transcripts are detected from this gene in adult tissue compared to the larval stage. In larval tissue, heat shock appears to produce only a slight increase in the level of transcripts detected. Based on overall structure and function, this Medfly gene appears to be an ortholog of heat shock cognate gene 4 (Drohsc4) of Drosophila melanogaster.

Amino Acid Sequence↗

Codon bias in actin multigene families and effects on the reconstruction of phylogenetic relationships.

Codon usage patterns and phylogenetic relationships in the actin multigene family have been analyzed for three dipteran species--Drosophila melanogaster, Bactrocera dorsalis, and Ceratitis capitata. In certain phylogenetic tree reconstructions, using synonymous distances, some gene relationships are altered due to a homogenization phenomenon. We present evidence to show that this homogenization phenomenon is due to codon usage bias. A survey of the pattern of synonymous codon preferences for 11 actin genes from these three species reveals that five out of the six Drosophila actin genes show high degrees of codon bias as indicated by scaled chi 2 values. In contrast to this, four out of the five actin genes from the other species have low codon bias values. A Monte Carlo contingency test indicates that for those Drosophila actin genes which exhibit codon bias, the patterns of codon usage are different compared to actin genes from the other species. In addition, the genes exhibiting codon bias also appear to have reduced rates of synonymous substitution. The homogenization phenomenon seen in terms of synonymous substitutions is not observed for nonsynonymous changes. Because of this homogenization phenomenon, "trees" constructed based on synonymous substitutions will be affected. These effects can be overt in the case of multigene families, but similar distortions may underlie reconstructions based on single-copy genes which exhibit codon usage bias.

Actins↗

The actin gene family in the oriental fruit fly Bactrocera dorsalis. Muscle specific actins.

The actin protein is a critical protein in eukaryotic cells. Four actin genes, constituting what appear to be a set of muscle specific actin genes, have been isolated from the genome of the oriental fruit fly Bactrocera dorsalis. DNA sequences have been determined for the coding as well as 3' and 5' flanking regions for each of these genes. These genes have also been characterized in terms of RNA expression patterns, and comparisons have been made to actin genes from other species. Consistent with other actins, there is a high degree of amino acid sequence conservation in the coding regions of these genes. However, even within the coding regions codon usage patterns in the oriental fruit fly are quite different from some other well characterized species. In addition, the DNA sequences in the intermediate 3' and 5' flanking regions exhibit virtually no detectable sequence homology both within and between species. In terms of introns, three of the four actin genes from the oriental fruit fly described here have a single intervening sequence. Two of these genes share the same intron position with the two muscle specific actin genes act79B and act88F from Drosophila melanogaster and with one muscle specific actin gene CcA1 from the Mediterranean fruit fly, Ceratitis capitata. Another gene from the oriental fruit fly shares the same intron position as the muscle specific actin gene act57B from D. melanogaster. Such conservation of intron positioning between species is highly unusual among previously characterized actin genes. Using unique sequences found in the 3' untranslated regions, gene specific probes have also been constructed. These have been used to detect the expression patterns of individual genes in a temporal and spatial manner. Each of the four genes examined here show differential patterns of expression. The patterns indicate that all four genes are most likely to encode muscle specific actins.

Actins↗

Resolution of populations of the Mediterranean fruit fly at the DNA level using random primers for the polymerase chain reaction.

We have used the polymerase chain reaction (PCR) and the random amplified polymorphic DNA (RAPD) method to identify DNA polymorphisms that can be used as genetic markers to characterize populations of the Mediterranean fruit fly, Ceratitis capitata. In this study, RAPD markers have been used to resolve genetic variability between populations of this major agricultural pest species. The populations analyzed represent either laboratory stocks or wild collections originating from different geographic localities. Using the same set of individual flies from each of several populations, we show that the use of different primers in the RAPD method permits detection of different levels of population differentiation. We show results from RAPD primers (e.g., primer 14) that identify regions of the genome (through PCR amplification) that are essentially monomorphic in all flies originating from a particular geographic locality. We also show RAPD primers (e.g., primer 67) that identify what appear to be highly variable regions of the genome. We have used primers of this type to produce genetic markers that can distinguish even between laboratory versus wild populations as well as subpopulations of flies from more broadly defined geographic localities, such as within the Hawaiian islands. These results show that the RAPD method is a broadly applicable, high resolution method for documenting genetic variability within and between populations of insect pest species.

Animals↗

The role of the transposable element hobo in the origin of endemic inversions in wild populations of Drosophila melanogaster.

Evidence from in situ hybridizations of DNA from the transposable element hobo to polytene salivary gland chromosome squashes reveals that hobo occupies both cytological breakpoints of three of four endemic inversions sampled from natural populations of Drosophila melanogaster in the Hawaiian islands. The fourth endemic inversion has a single hobo insert at one breakpoint. Cosmopolitan inversions on the same chromosomes do not show this association. Frequencies of both endemic and cosmopolitan inversions in Hawaiian populations fall in ranges typical for natural populations of D. melanogaster sampled worldwide, suggesting that these results may be typical of other regions besides Hawaii. This appears to be the first direct demonstration that transposable elements are responsible for causing specific rearrangements found in nature; consequently, it is also the first direct demonstration that chromosome rearrangements can arise in nature in a manner predicted by results of hybrid dysgenic crosses in the laboratory. Possible population genetic and evolutionary consequences are discussed.

Animals↗

Y enriched and Y specific DNA sequences from the genome of the Mediterranean fruit fly, Ceratitis capitata.

DNA sequences that are enriched or specific to the genome of the male medfly, Ceratitis capitata, have been isolated using a differential hybridization approach. Twelve phage clones from a genomic library have been identified that consistently display more intense hybridization with a genomic DNA probe from males as opposed to one from females. Southern DNA blot analysis reveals that these recombinant clones contain at least one EcoRI fragment that is either specific to the male genome, or more highly represented in it, as compared with the female genome. These EcoRI fragments, when used as probes, all generate a similar pattern of intense multiple bands in genomic DNA of males. This suggests the presence of repetitive sequences that are at least partially homologous in these regions of the genome that are specific to or enriched in males. In situ hybridization to mitotic chromosomes confirms a Y chromosomal origin for the male specific repetitive sequences. Data on the genomic organization, representation and evolutionary conservation of these sequences that are specific to or enriched in males are presented. Studies of the genomic organization and representation of flanking sequences that are not male specific are presented as well.

Animals↗

A muscle-specific actin gene from the Mediterranean fruit fly, Ceratitis capitata.

A characterization of an actin gene isolated from the genome of the Mediterranean fruit fly, Ceratitis capitata, including the complete sequencing of the coding, 3' and 5' flanking regions of this gene and a partial cDNA was carried out. The partial cDNA was derived from the 3' untranslated region of the actin gene described here, and has been used to identify this gene uniquely. The DNA sequence data presented here, together with the pattern of expression exhibited by this gene during development, strongly support the interpretation that this is a muscle-specific actin gene. Peaks of expression are seen in tissues and during temporal phases of development where muscle differentiation is occurring. The derived protein sequence of the Medfly acting gene shows the highest degrees of similarity, 98.4 and 96.6% respectively, with the two muscle-specific actin genes 79B and 88F from D. melanogaster. The Medfly actin gene also has a single intervening sequence, and an intron is found at the same position in the 79B and 88F actin genes. In the coding region at the DNA level, 17.2 and 16.4% nucleotide differences, respectively, are observed between the Medfly actin gene and these same two D. melanogaster actin genes. The disparity between the amino acid and nucleotide comparisons can be explained, in part, by a high level of synonymous changes in the DNA sequence. In addition, despite the many similarities, codon usage appears to be very different between the actin genes of these species.

Actins↗

Genetic variation between strains of the Mediterranean fruit fly, Ceratitis capitata, detected by DNA fingerprinting.

DNA fingerprinting has been used to detect genetic variation in the Mediterranean fruit fly, Ceratitis capitata. Three different probes have been identified that can be used to detect DNA restriction fragment length polymorphisms between strains of this species. The strains used in this study differ only in terms of their geographic origin or genetic background. One of the probes used is the bacteriophage vector M13, and the other two are repetitive sequences derived from the medfly genome based on a weak homology to M13. Within a strain, each probe produces a consistent restriction fragment profile that is not affected by the method or timing of DNA extraction. Between strains, when M13 is used as a probe, an average of 10% of the observable bands are polymorphic. Use of the medfly genomic sequences as a probe increases the proportion of polymorphic bands between strains up to 30%. The fact that genetic differences between even such closely related strains can be reliably detected by this method holds great promise for studies of insect pests including the ability to monitor the movements of pest species, determining the extent of genetic variation in pest populations, and in making identifications from otherwise unidentifiable material.

Animals↗

Actin genes in the Mediterranean fruit fly, Ceratitis capitata.

We have undertaken the study of actin gene organization and expression in the genome of the Mediterranean fruit fly (medfly), Ceratitis capitata. Actin genes have been extensively characterized previously in a wide range of eukaryotic organisms, and they have valuable properties for comparative studies. These genes are typically highly conserved in coding regions, represented in multiple copies per genome and regulated in expression during development. We have isolated a gene in the medfly using the cloned Drosophila melanogaster 5C actin gene as a probe. This medfly gene detects abundant messages present during late larval and late pupal development as well as in thoracic and leg tissue preparations from newly emerged adults. This pattern of expression is consistent with what has been seen for actin genes in other organisms. Using either the D. melanogaster 5C actin gene or the medfly gene as a probe identifies five common cross reacting EcoRI fragments in genomic DNA, but only under less than fully stringent hybridization conditions.

Actins↗

Germ line and somatic instability of a white mutation in Drosophila mauritiana due to a transposable genetic element.

A spontaneous white mutation recovered in Drosophila mauritiana is unstable and reverts to normal eye color at a frequency greater than 4 per 1,000 X-chromosomes. Germ line reversion occurs at a high rate in D. mauritiana males and in interspecific hybrid females, while the rate is depressed in D. mauritiana females. These events are not restricted to the germ line, as cases of variegated patterns of eye pigmentation, indicating somatic reversion, are recovered at a frequency comparable to that of the male germ line reversion rate. Germ line reversion events are genetically stable, while the somatic variegation patterns are not heritable. The patterns of eye pigment variegation produced suggests that reversion events are occurring throughout development. Whole genome DNA digests blotted and probed with the cloned D. melanogaster white gene indicate that this unstable white mutation in D. mauritiana is associated with an insertion of DNA that is lost upon reversion to wild type, indicating that this DNA insert is in fact a transposable element.

Animals↗

Molecular structure of yoyo, a gypsy-like retrotransposon from the mediterranean fruit fly, Ceratitis capitata.

We have isolated and characterized a new LTR-retrotransposon in the genome of the Mediterranean fruit fly (Medfly), Ceratitis capitata. This retrotransposon, which we named yoyo, appears to be a member of the gypsy/Ty3 class of elements. The yoyo element was originally discovered on the Y chromosome of the Medfly. Although the Y chromosome copy appears to be truncated, at least two other apparently complete copies of yoyo from other genomic locations have been isolated and characterized. The complete element is approximately 7.7 kb in size. In addition to fairly typical GAG and POL coding regions, the yoyo element contains a potential ENV gene. The presence of an ENV gene is a key feature distinguishing potential retroviral-like elements, such as gypsy (and possibly yoyo), from many other invertebrate retrotransposons previously described. In addition to the structural features of yoyo, evidence is provided to show that yoyo is capable of movement in the genome, including RFLPs showing variability in genomic localization of copies of yoyo between strains, and differences among individuals in the presence of yoyo at a specific site in the genome.

Amino Acid Sequence↗

Eye color pigment granules in Drosophila mauritiana: mosaics produced by excision of a transposable element.

Compound eyes of the white-peach (wpch) mutant strain of Drosophila mauritiana have some pigment and receptor cells with wild-type eye color pigmentation. These eyes are mosaic, because excision of a transposable element reverts wpch to wild type during the development of somatic cells. Wild-type patches have three types of pigment granule residing in three respective cell types: primary pigment cells, secondary pigment cells, and retinula (visual receptor) cells. Most aspects of these granules, as well as all other aspects of compound eye ultrastructure, are exactly as in the better studied sibling species D. melanogaster. In the wpch parts of the eye, small and giant unpigmented "pigment granules" reside in secondary pigment cells. These white granules are just like the corresponding granules of w mutant D. melanogaster. Small vs. large patches of pigmented cells likely represent excision events occurring late vs. early respectively during development. Mosaics of eye color markers have been important in developmental analyses; the ease of constructing mosaics of D. mauritiana gives this preparation advantages for mosaic analyses.

Animals↗