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Trichinella spiralis: genetic evidence for synanthropic subspecies in sylvatic hosts.

Isolates of the nematode genus Trichinella from sylvatic hosts differ in their potential to reproduce in domestic swine. The structure of the genomic DNA from 13 sylvatic isolates from North America and 5 pig isolates, 4 from North America and 1 from Asia, was examined and correlated with the infectivity of the isolate for domestic pigs. DNA restriction fragment length differences, identified by ethidium bromide staining and by hybridization with 32P-labeled ribosomal RNA, served as molecular markers to classify each isolate. All 5 pig isolates and 8 of 13 sylvatic isolates had a high infectivity and reproductive capacity in pigs. All isolates that were highly infectious for pigs regardless of host origin had similar DNA characteristics and were classified operationally as T. spiralis spiralis (pig) and those of the second group as T. spiralis ssp. A DNA clone of repetitive DNA from T. s. spiralis, pBP2, was selected from a library of genomic DNA in plasmid pUC8. When used as a probe, pBP2 hybridized only to the DNA of T. s. spiralis isolates, thus making it a useful diagnostic reagent to predict whether new isolates are highly infectious for pigs (i.e., T. s. spiralis). These results show that T. s. spiralis occurs in wild mammals and this should be considered a serious obstacle to efforts to eradicate trichinellosis from domestic swine.

Animals↗

Extensive chromosomal repatterning and the evolution of sterility barriers in hybrid sunflower species.

New species may arise via hybridization and without a change in ploidy. This process, termed homoploid hybrid speciation, is theoretically difficult because it requires the development of reproductive barriers in sympatry or parapatry. Theory suggests that isolation may arise through rapid karyotypic evolution and/or ecological divergence of hybrid neospecies. Here, we investigate the role of karyotypic change in homoploid hybrid speciation by generating detailed genetic linkage maps for three hybrid sunflower species, Helianthus anomalus, H. deserticola, and H. paradoxus, and comparing these maps to those previously generated for the parental species, H. annuus and H. petiolaris. We also conduct a quantitative trait locus (QTL) analysis of pollen fertility in a BC2 population between the parental species and assess levels of pollen and seed fertility in all cross-combinations of the hybrid and parental species. The three hybrid species are massively divergent from their parental species in karyotype; gene order differences were observed for between 9 and 11 linkage groups (of 17 total), depending on the comparison. About one-third of the karyoypic differences arose through the sorting of chromosomal rearrangements that differentiate the parental species, but the remainder appear to have arisen de novo (six breakages/six fusions in H. anomalus, four breakages/three fusions in H. deserticola, and five breakages/five fusions in H. paradoxus). QTL analyses indicate that the karyotypic differences contribute to reproductive isolation. Nine of 11 pollen viability QTL occur on rearranged chromosomes and all but one map close to a rearrangement breakpoint. Finally, pollen and seed fertility estimates for F1's between the hybrid and parental species fall below 11%, which is sufficient for evolutionary independence of the hybrid neospecies.

Chromosome Mapping↗

On the success of a swindle: pollination by deception in orchids.

A standing enigma in pollination ecology is the evolution of pollinator attraction without offering reward in about one third of all orchid species. Here I review concepts of pollination by deception, and in particular recent findings in the pollination syndromes of food deception and sexual deception in orchids. Deceptive orchids mimic floral signals of rewarding plants (food deception) or mating signals of receptive females (sexual deception) to attract pollen vectors. In some food deceptive orchids, similarities in the spectral reflectance visible to the pollinator in a model plant and its mimic, and increased reproductive success of the mimic in the presence of the model have been demonstrated. Other species do not mimic specific model plants but attract pollinators with general attractive floral signals. In sexually deceptive orchids, floral odor is the key trait for pollinator attraction, and behaviorally active compounds in the orchids are identical to the sex pheromone of the pollinator species. Deceptive orchids often show high variability in floral signals, which may be maintained by negative frequency-dependent selection, since pollinators can learn and subsequently avoid common deceptive morphs more quickly than rare ones. The evolution of obligate deception in orchids seems paradoxical in the light of the typically lower fruit set than in rewarding species. Pollination by deception, however, can reduce self-pollination and encourage pollen flow over longer distances, thus promoting outbreeding. Although some food deceptive orchids are isolated through postzygotic reproductive barriers, sexually deceptive orchids lack post-mating barriers and species isolation is achieved via specific pollinator attraction. Recent population genetic and phylogenetic investigations suggest gene-flow within subgeneric clades, but pollinator-mediated selection may maintain species-specific floral traits.

Flowers↗

Effects of jasmonate-induced defenses on root-knot nematode infection of resistant and susceptible tomato cultivars.

Jasmonates, such as jasmonic acid (JA), are plant-signaling compounds that trigger induced resistance against certain pathogens and a broad range of arthropod herbivores. One goal of this study was to determine the effects of JA-dependent defenses in tomato on root-knot nematodes. Another was to determine if the artificial induction of these defenses could enhance nematode control on plants that carry Mi-1.2, a nematode resistance gene that is present in many tomato cultivars. At moderate soil temperatures, Mi-1.2 can effectively suppress reproduction of most isolates of the common root-knot nematode species Meloidogyne javanica, M. incognita, and M. arenaria. Mi-mediated resistance has its limitations, however. Mi-1.2 is reported to lose its effectiveness at soil temperatures above 28 degrees C, and certain virulent nematode isolates can overcome resistance even at moderate soil temperatures. This study used a foliar application of JA to activate induced resistance in two near-isogenic lines of tomato (Lycopersicon esculentum) with and without Mi-1.2, and evaluated the effects of induced resistance at moderate soil temperatures on one avirulent nematode isolate (M. javanica isolate VW4) and two virulent isolates (M. javanica isolate VW5 and M. incognita isolate 557R). In addition, the effects of induced resistance on avirulent nematode performance were examined at a high temperature (32 degrees C). The results indicate that JA application induces a systemic defense response that reduces avirulent nematode reproduction on susceptible tomato plants. Furthermore, JA-dependent defenses proved to be heat-stable, whereas the effects of Mi-mediated resistance were reduced but not eliminated at 32 degrees C. JA treatment enhanced Mi-mediated resistance at high temperature, but did not suppress either of the virulent nematode isolates tested.

Animals↗

Concentrations and composition of glycosaminoglycans in the female bovine reproductive tract.

Our objective was to isolate glycosaminoglycans from various regions of the female bovine reproductive tract to ascertain their concentrations and composition. Oviducts, uterine horns, and cervices from 104 female bovine reproductive tracts obtained at slaughter were flushed with 5 ml phosphate buffered saline. Following proteolysis, glycosaminoglycans were isolated by alcoholic precipitation, and concentrations were quantitated by high performance liquid chromatography. Concentrations of glycosaminoglycans decreased significantly anterior to the cervix. There was an increase of chondroitin sulfates and heparin-like material in estrual cervical mucus compared with cervical mucus collected during the luteal phase of the estrous cycle.

Animals↗

Nonallopatric and parallel origin of local reproductive barriers between two snail ecotypes.

Theory suggests that speciation is possible without physical isolation of populations (hereafter, nonallopatric speciation), but recent nonallopatric models need the support of irrefutable empirical examples. We collected snails (Littorina saxatilis) from three areas on the NW coast of Spain to investigate the population genetic structure of two ecotypes. Earlier studies suggest that these ecotypes may represent incipient species: a large, thick-shelled 'RB' ecotype living among the barnacles in the upper intertidal zone and a small, thin-shelled 'SU' ecotype living among the mussels in the lower intertidal zone only 10-30 m away. The two ecotypes overlap and hybridize in a midshore zone only 1-3 m wide. Three different types of molecular markers [allozymes, mitochondrial DNA (mtDNA) and microsatellites] consistently indicated partial reproductive isolation between the RB and the SU ecotypes at a particular site. However, each ecotype was related more closely to the other ecotype from the same site than to the same ecotype from another site further along the Galician coast (25-77 km away). These findings supported earlier results based solely on allozyme variation and we could now reject the possibility that selection produced these patterns. The patterns of genetic variation supported a nonallopatric model in which the ecotypes are formed independently at each site by parallel evolution and where the reproductive barriers are a byproduct of divergent selection for body size. We argue that neither our laboratory hybridization experiments nor our molecular data are compatible with a model based on allopatric ecotype formation, secondary overlap and introgression.

Animals↗

Physiological and genetic characterisation of some new Aphanomyces strains isolated from freshwater crayfish.

Five Aphanomyces strains were isolated during suspected outbreaks of crayfish disease in Spain and Italy. Genetic and physiological evidence show that the strains isolated from the freshwater crayfish Procambarus clarkii and Pacifastacus leniusculus, do not fit into any previously identified group of Aphanomyces astaci and are not capable of killing crayfish following standardised experimental infection. RAPD-PCR and ITS sequencing analysis show a high degree of similarity between the new isolates, while they are clearly different from the A. astaci reference strains. They do, however, possess some properties, which are commonly associated with parasitic species such as repeated zoospore emergence and the lack of sexual reproduction. The five isolates share some physiological properties i.e. a high growth rate, and germination in response to nutrients and, in contrast to A. astaci, they do not express chitinase constitutively during growth or sporulation. Until their taxonomic status is fully elucidated we suggest that the new isolates be given the tentative species name Aphanomyces repetans.

Animals↗

Sperm transport, size of the seminal plug and the timing of ovulation after natural mating in the female tammar wallaby Macropus eugenii.

The distribution of spermatozoa and seminal plug in the reproductive tract and the timing of ovulation were examined at various times in a naturally mated monovular macropodid marsupial, namely the tammar wallaby (Macropus eugenii). After the first post partum (p.p.) mating, 28 females were isolated and their reproductive tracts dissected at 0.5, 6, 18, 36 and 40 h post coitum (p.c.). Each tract was ligated into 13 major anatomical sections and spermatozoa and eggs were recovered by flushing. Mating was possibly delayed by handling and occurred 21.7 +/- 2.5 h p.p. in these animals. Copulation lasted 7.8 +/- 0.7 min. Within 0.5 h after a single mating, the tract contained 25.8 +/- 10.2 x 10(6) spermatozoa and 21.6 +/- 8.8 g of seminal plug, 96% and 70% of which was lost within 6 h p.c. respectively. Spermatozoa reached the uterus, isthmus and ampulla of the oviduct on the side of the developing follicle within 0.5, 6 and 18 h p.c., respectively, and a uterine population of 26.1 +/- 12.10(3) spermatozoa was maintained for over 40 h. Sperm numbers were reduced at the cervix (up to 57-fold) and uterotubule junction (eight-fold) and only one in approximately 7500 ejaculated spermatozoa (3.4 +/- 0.9 x 10(3)) reached the oviduct on the follicle side. Differential transport of spermatozoa was not observed. Although the numbers of spermatozoa were reduced in the parturient uterus, they were highly variable and were not significantly different to those in the non-parturient uterus. Ovulation and recovery of sperm-covered eggs from the isthmus occurred 36-41 h p.c. (49-72 h p.p.). In contrast with the polyovular dasyurid and didelphid marsupials, the tammar wallaby ejaculates large numbers of spermatozoa, but transport is relatively inefficient and sperm storage in the tract before ovulation is limited.

Animals↗

Effect of isolated fractions of Barleria prionitis root methanolic extract on reproductive function of male rats: preliminary study.

Male rats treated with 100 mg/kg for 60 days of isolated fractions of the Barleria prionitis root methanolic extract (Fr. I and Fr. II) showed a significant reduction on spermatogenesis without affecting general body metabolism. Sperm motility as well density in cauda epididymides was reduced significantly. The fertility was decreased by 33.4% in Fr. I and 100% in Fr. II treated rats. The blood parameters were within the normal range. Total protein, glycogen and sialic acid contents of testes were reduced after the plant fractions treatment. Seminal vesicular fructose was decreased significantly after the treatment. The population of various spermatogenic cells such as primary spermatocytes, secondary spermatocytes and round spermatids were declined significantly in Fr. II treatment groups whereas in Fr. I treated animals preleptotene spermatocyte and spermatid number was decreased. There was no significant change in the number of Sertoli cells and spermatogonia in any of the treatment group.

Acanthaceae↗

Dynamics of some genetic marker changes of respiratory-syncytial virus strains circulating among nursery-school children.

Respiratory-syncytial (RS) virus strains circulating during several years appeared polymorphic in respect of two genetic markers: the regression coefficient of infectious activity (RCIA39) characterizing the isolates by their reproduction in tissue cultures at supraoptimal temperature (39 degrees C) and the regression coefficient of neutralization indices (RCNI) characterizing the degree of sensitivity of the strains to antibodies. High-yield RS viruses were more often isolated from children frequently afflicted by the disease, moderate-yield viruses from moderately sick children, while low-yield or none-yield (at 39 degrees C) strains were isolated from rarely afflicted children. On the other hand, RS strains of low reactivity with prototype antibodies were mainly found in often or moderately sick children and the high-reactive ones in rarely sick children. The variability of the RS virus population was continuous, which is consistent with the uninterrupted course of the epidemic process in the nursery-school community. A change of the RCIA39 marker was observed in nearly 50% of strains already after 1 1/2 to 2 months, but most frequently within 5-6 months from the end of disease. The changes of growth intensity at 39 degrees C followed the pattern: high----moderate----low----none, however, in the next epidemic season these properties showed reversion in an opposite direction.

Cell Line↗

Immunohistochemical detection of porcine reproductive and respiratory syndrome virus using colloidal gold.

Two cytopathic agents were isolated on porcine alveolar macrophages following inoculation with homogenates of lung tissues from pigs showing respiratory problems. These isolates were identified as porcine reproductive and respiratory syndrome (PRRS) virus isolates by indirect immunofluorescence using a PRRS virus (PRRSV) specific monoclonal antibody (MAb) and were designated as LHVA-92-1 and LHVA-92-2. Immunogold electron microscopy using a porcine PRRS positive serum pool and protein A-gold resulted in an intense labelling of aggregates of viral particles. Dark specific cytoplasmic staining of porcine alveolar macrophages infected with both virus isolates could be observed by immunogold silver staining (IGSS) using the specific MAb. This method proved effective in detecting PRRSV antigens in several ethanol-fixed tissues of piglets intranasally inoculated with the supernatants of macrophages infected with each isolate. Immunogold silver staining was also successfully used for the detection of PRRSV antigens on sections of formalin-fixed paraffin-embedded lung tissues and on frozen sections of lungs. The present results indicate that colloidal gold may be useful for the identification and immunohistochemical detection of PRRSV in tissues.

Animals↗

MUC1/episialin: a critical barrier in the female reproductive tract.

The female reproductive tract must resist microbial infections as well as support embryonic development, implantation and placentation. Reproductive tract mucins, in general, and Muc1/episialin, in particular, play key roles in implantation related events and in protection from microbial infection. High levels of mucin expression in the lower reproductive tract presumably affords protection against infection while down-regulation of uterine mucins has been suggested to provide access to the uterine surface. The present studies demonstrate that mucins, particularly Muc1, are effective barriers to embryo attachment. Furthermore, a strain of female Muc1 null mice in normal housing displays chronic infection and inflammation of the lower reproductive tract and markedly reduced fertility rates. This phenotype is not observed when Muc1 nulls are housed in a pathogen-free environment indicating that this phenotype results from chronic microbial exposure. Only normal endogenous flora were isolated from the reproductive tracts of affected Muc1 null mice, suggesting that these bacterial species become opportunistic with loss of the mucin barrier. Staphylococcal adherence to lower reproductive tract epithelia was found to be mediated by cell surface mucin carbohydrates. Collectively, these studies demonstrate a critical barrier role for Muc1 in various aspects of female reproductive tract physiology.

Animals↗

Identification, cloning and functional characterization of novel beta-defensins in the rat (Rattus norvegicus).

BACKGROUND: Beta-defensins are small cationic peptides that exhibit broad spectrum antimicrobial properties. The majority of beta-defensins identified in humans are predominantly expressed in the male reproductive tract and have roles in non-immunological processes such as sperm maturation and capacitation. Characterization of novel defensins in the male reproductive tract can lead to increased understanding of their dual roles in immunity and sperm maturation. METHODS: In silico rat genomic analyses were used to identify novel beta-defensins related to human defensins 118-123. RNAs isolated from male reproductive tract tissues of rat were reverse transcribed and PCR amplified using gene specific primers for defensins. PCR products were sequenced to confirm their identity. RT-PCR analysis was performed to analyze the tissue distribution, developmental expression and androgen regulation of these defensins. Recombinant defensins were tested against E. coli in a colony forming unit assay to analyze their antimicrobial activities. RESULTS: Novel beta-defensins, Defb21, Defb24, Defb27, Defb30 and Defb36 were identified in the rat male reproductive tract. Defb30 and Defb36 were the most restricted in expression, whereas the others were expressed in a variety of tissues including the female reproductive tract. Early onset of defensin expression was observed in the epididymides of 10-60 day old rats. Defb21-Defb36 expression in castrated rats was down regulated and maintained at normal levels in testosterone supplemented animals. DEFB24 and DEFB30 proteins showed potent dose and time dependent antibacterial activity. CONCLUSION: Rat Defb21, Defb24, Defb27, Defb30 and Defb36 are abundantly expressed in the male reproductive tract where they most likely protect against microbial invasion. They are developmentally regulated and androgen is required for full expression in the adult epididymis.

Amino Acid Sequence↗

Clusterin in the male reproductive system: localization and possible function.

Clusterin is a glycoprotein that was initially isolated from the male reproductive system. Subsequently, clusterin has been found to be widely distributed in a variety of tissues in mammals. One characteristic of the expression of clusterin is that it is induced as a result of cellular injury, death, or pathology. Despite the efforts of many laboratories working in diverse biological systems, the function of clusterin remains unknown. Recent studies have revealed a 'heat-shock element' in the promoter of the gene that may account for the inducible nature of the clusterin gene. Overall, the evidence suggests that function of clusterin is to protect surviving cells after damage. This protection may result from a detergent-like action of the protein.

Animals↗

Isolation and identification of N-terminally extended forms of 5-oxoprolylglutamylprolinamide (Glp-Glu-Pro-NH2), a thyrotropin-releasing-hormone (TRH)-like peptide present in human semen.

N-terminally extended forms of 5-oxoprolylglutamylprolinamide (Glp-Glu-Pro-NH2), a thyrotropin-releasing-hormone(TRH)-like peptide associated with the male reproductive system, were isolated from human semen by gel exclusion on Sephadex G50, ion-exchange chromatography on SP-Sephade C25 and QAE-Sephadex A25, and by HPLC. The peptides were located by trypsin-catalysed release of their C-terminal fragments which were detected by RIA with a TRH-specific antibody. A series of overlapping peptides containing 16, 18, 22 and 25 residues was obtained in homogeneous form and their sequences were determined by automatic Edman degradation. The peptides all terminated in -Lys-Gln-Glu-Pro-NH2 and were found to correspond to sequences occurring between residues 350-374 of semenogelin, a protein present in human semen. In semenogelin, however, the Gln-Glu-Pro sequence is followed by tryptophan and not glycine which is normally essential for formation of the C-terminal amide group. Model experiments with the synthetic peptide Glp-Glu-Pro-Trp showed that under a range of experimental conditions the tetrapeptide did not undergo conversion to Glp-Glu-Pro-NH2. This would indicate that the tripeptide and its extended forms are generated from a precursor that is related to semenogelin but in which Trp375 is replaced by glycine.

Amino Acid Sequence↗

Multiple porcine circovirus 2-associated abortions and reproductive failure in a multisite swine production unit.

Porcine circovirus type 2 was detected in several stillborn and nonviable neonatal piglets presenting with chronic passive congestion, cardiac hypertrophy, and severe diffuse myocarditis. The presence of the virus in the heart and other tissues of affected piglets was confirmed by polymerase chain reaction, immunohistochemistry, and virus isolation techniques. Other reproductive losses and associated infectious agents in the herd are discussed.

Abortion, Veterinary↗

[Long-term group isolation is a factor of dysregulation of reproductive function in rats].

In the condition of prolonged group sex starvation of rats variations were seen in behavioral reactions, fertility, extent of weight gain, litter, blood alpha-tocopherol level, eosinophil count, lipid peroxidation, evoked brain potentials. All these processes are the sign of stress development in rats. Thus, stress in mature rats influences litter and fertility, newborn rats had underdeveloped nervous system. Changes in the activity of endogenic antioxidant system in this experiment are discussed.

Animals↗