The Role of Hybrid Vigor in the Replacement of Pecos Pupfish by Its Hybrids with Sheepshead Minnow

Abstract:  Many species are jeopardized by hybridization and genetic introgression with closely related species. Unfortunately, the mechanisms that promote or retard gene flow between divergent populations are little studied and poorly understood. Like many imperiled fish species, the Pecos pupfish ( Cyprinodon pecosensis ) is threatened with replacement by its hybrids with a close congener. We examined swimming performance and growth rate of hybrid pupfish to determine the role of hybrid vigor in the genetic homogenization of C. pecosensis by its hybrids with sheepshead minnow ( C. variegatus ). The F1 hybrids, backcross hybrids, and purebred C. variegatus displayed greater swimming endurance than purebred C. pecosensis . In addition, F1 hybrids and C. variegatus grew more rapidly than C. pecosensis . The ecological superiority of hybrids probably promoted their rapid spread through and beyond the historic range of C. pecosensis . These results indicate that eradication of hybrids and restoration of C. pecosensis to its native range is unlikely. Extinction of unique species via genetic homogenization can result from human activities that increase gene flow between historically fragmented populations; conservation managers must weigh the potential for such a catastrophe against the presumed benefits of increased interpopulation gene flow. This example illustrates how, after hybridization has occurred, conflict may arise between formerly complementary conservation goals.     

Prenatal diagnosis of the derivative chromosome 22 associated with cat eye syndrome by fluorescence in situ hybridization

Cytogenetic studies of cultured amniocytes demonstrated a karyotype of 46, XX/47, XX,+mar. A bisatellited, dicentric, distamycin-DAPI negative, NOR-positive marker was present in 76 per cent of the metaphases examined. Similar markers have been associated with cat eye syndrome (CES). We report on the utilization of fluorescence in situ hybridization (FISH) with a 14/22 a-satellite probe and a chromosome 22-specific cosmid for locus D22S9 to determine the origin of the prenatally detected supernumerary marker chromosome. FISH studies demonstrated that the marker is a derivative of chromosome 22 and enabled us to provide the family with additional prognostic information.     

Demonstration of spontaneously dividing male fetal cells in maternal blood by negative magnetic cell sorting and fish

We investigated a case of massive feto-maternal bleeding by using negative magnetic cell sorting (MACS) and fluorescent in situ hybridization (FISH). A 37-year-old pregnant woman had an uncomplicated amniocentesis for advanced maternal age at 16 weeks' gestation. The fetal karyotype was 46, XY. At 19 weeks' gestation, she had a minor car accident and slight vaginal bleeding. A subsequent Kleihauer-Betke test showed a 140 ml feto-maternal haemorrhage. Serial sonographic examinations indicated a normal fetus and placenta. We performed FISH analysis on maternal peripheral blood at 25 weeks. Anti-CD45 and MACS were used to deplete maternal leucocytes, enriching the proportion of fetal nucleated erythrocytes present. The isolated cells were analysed by using dual-colour FISH with X and Y specific probes. Approximately 65 800 nucleated cells were obtained after MACS depletion. A total of 234 cells were analysed by FISH. The results revealed that 70 of the nucleated cells (30 per cent) were male with one X and one Y signal. Among these cells, six male metaphases were observed in spontaneously dividing cells.     

Application of fluorescent in situ hybridization for ‘de novo’ anomalies in prenatal diagnosis

Fluorescent in situ Hybridization (FISH) was carried out for three cases of abnormal karyotypes in prenatal studies. Two concerned de novo structural anomalies and the third a marker chromosome. The origin of the extra material could be defined in all three cases, which gives a better insight into the relationship between genotype and phenotype and makes more adequate genetic counselling possible.     

Prenatal diagnosis of tetrasomy 12p by in situ hybridization: Varying levels of mosaicism in different fetal tissues

Prenatal diagnosis of tetrasomy 12p is complicated by the discrimination of the 12p isochromosome from the duplication 21q as well as the level of mosaicism demonstrated in the particular tissue sampled. In this disease, a high percentage of chromosomally abnormal cells are generally found in fibroblastic cells, but lymphocyte karyotypes from the same individual may be normal. We report on the pregnancy of a 37-year-old female who presented to our centre at 16 weeks' gestation for genetic amniocentesis. Sonography of the fetus revealed dextrocardia and diaphragmatic hernia. Chromosome analysis of amniocytes demonstrated mosaicism of a 47,XY,+i(12p) line in 80 per cent of cells and a normal male line (20 per cent), consistent with the Pallister-Killian syndrome. Following termination, a 220 g male fetus of 18 weeks was examined. A flattened nose and low-set ears were noted. In situ hybridization with a chromosome 12 centromeric probe in lymphocytes and skin cells unequivocally confirmed the karyotype and showed the presence of a single centromere in the abnormal chromosome, suggesting a true isochromosome. Chromosome analysis of various fetal tissues was performed and the following percentages of abnormal cells were found: skin 100 per cent, chorion 50 per cent, placenta 30 per cent, and blood 80 per cent. The high frequency of tetrasomic cells in fetal blood at this early gestational age is noteworthy, since most reports of this syndrome show a very low percentage of abnormal cells postnatally.     

Adaptive introgression as a resource for management and genetic conservation in a changing climate

Current rates of climate change require organisms to respond through migration, phenotypic plasticity, or genetic changes via adaptation. We focused on questions regarding species’ and populations’ ability to respond to climate change through adaptation. Specifically, the role adaptive introgression, movement of genetic material from the genome of 1 species into the genome of another through repeated interbreeding, may play in increasing species’ ability to respond to a changing climate. Such interspecific gene flow may mediate extinction risk or consequences of limited adaptive potential that result from standing genetic variation and mutation alone, enabling a quicker demographic recovery in response to changing environments. Despite the near dismissal of the potential benefits of hybridization by conservation practitioners, we examined a number of case studies across different taxa that suggest gene flow between sympatric or parapatric sister species or within species that exhibit strong ecotypic differentiation may represent an underutilized management option to conserve evolutionary potential in a changing environment. This will be particularly true where advanced‐generation hybrids exhibit adaptive traits outside the parental phenotypic range, a phenomenon known as transgressive segregation. The ideas presented in this essay are meant to provoke discussion regarding how we maintain evolutionary potential, the conservation value of natural hybrid zones, and consideration of their important role in adaptation to climate.     

Screening differentially expressed genes in an amphipod (Hyalella azteca) exposed to fungicide vinclozolin by suppression subtractive hybridization

Vinclozolin, a dicarboximide fungicide, is an endocrine disrupting chemical that competes with an androgenic endocrine disruptor compound. Most research has focused on the epigenetic effect of vinclozolin in humans. In terms of ecotoxicology, understanding the effect of vinclozolin on non-target organisms is important. The expression profile of a comprehensive set of genes in the amphipod Hyalella azteca exposed to vinclozolin was examined. The expressed sequence tags in low-dose vinclozolin-treated and -untreated amphipods were isolated and identified by suppression subtractive hybridization. DNA dot blotting was used to confirm the results and establish a subtracted cDNA library for comparing all differentially expressed sequences with and without vinclozolin treatment. In total, 494 differentially expressed genes, including hemocyanin, heatshock protein, cytochrome, cytochrome oxidase and NADH dehydrogenase were detected. Hemocyanin was the most abundant gene. DNA dot blotting revealed 55 genes with significant differential expression. These genes included larval serum protein 1 alpha, E3 ubiquitin-protein ligase, mitochondrial cytochrome c oxidase, mitochondrial protein, proteasome inhibitor, hemocyanin, zinc-finger–containing protein, mitochondrial NADH-ubiquinone oxidoreductase and epididymal sperm-binding protein. Vinclozolin appears to upregulate stress-related genes and hemocyanin, related to immunity. Moreover, vinclozolin downregulated NADH dehydrogenase, related to respiration. Thus, even a non-lethal concentration of vinclozolin still has an effect at the genetic level in H. azteca and presents a potential risk, especially as it would affect non-target organism hormone metabolism.     

降解剩余污泥中纤维素/半纤维素微生物富集培养实验研究

为促进剩余污泥中木质纤维素能源转化,首先需要探知污泥中是否存在可以降解纤维素/半纤维素的微生物.以木聚糖作为单一碳源分别从剩余污泥和厌氧消化污泥中富集培养可降解纤维素/半纤维素的微生物.实验结果显示,两种污泥源中均存在可降解纤维素/半纤维素的微生物,并可成功富集培养获得该类微生物.FISH与LIVE/DEAD实验证实,富集微生物为梭状芽孢杆菌属和芽孢杆菌属的混合物,并以梭状芽孢杆菌为主;富集微生物均具有很高活性.     

Effect of plant harvest on methane emission from two constructed wetlands designed for the treatment of wastewater

The emission of methane from two constructed wetlands [a free water surface flow system (FWS) and a subsurface flow system (SF)], constructed for the treatment of waste water, was evaluated at different sites inhabited by reeds (Phragmites communis), to test the effects of plant harvest. High methane emission was recorded immediately after harvesting in both wetlands. Several days after harvesting, the emission decreased in the FWS but remained high in the SF. The variation was significantly influenced by temperature, with lower emission and higher dissolved CH(4) in water occurring at lower temperatures. Both the emission and concentration of dissolved CH(4) were also influenced significantly by water quality, wetland design, level of stalk butt left above the water level, etc. The methane flux was explained on the basis of rizhospheric methanogenic and methanotrophic microbial populations. FISH analysis indicated the presence of Type A and Type B methanotrophs in both wetlands, and the methane flux was directly influenced by the quantitative variation in methanogenic and methanotrophic bacteria in both wetlands.