Chloroplast DNA assorts randomly in intraspecific somatic hybrids of Nicotiana debneyi

Summary Plants were regenerated following intraspecific fusion of leaf protoplasts from two naturally occurring genotypes of Nicotiana debneyi. The two genotypes differed in the EcoRl fragmentation pattern of chloroplast DNA and in the nuclear-coded phosphoglucomutase (Pgm) isozymes. There was no conscious selection for hybrid genotypes during protoplast culture or plant regeneration. Among 225 plants screened for Pgm, six were identified as nuclear hybrids. Restriction endonuclease and filter hybridisation analysis revealed that the cytoplasms of the hybrids contained one or other but never both parental chloroplast DNAs. The sorting out of chloroplasts was random and complete; the limit of detecting a rare chloroplast-DNA type in a mixture was 0.1%.     

The effect of different exogenous kisspeptins on sex hormones and reproductive indices of the goldfish (Carassius auratus) broodstock

Despite several studies on fish hormone therapy, finding new candidates may provide more reproductive efficiency in artificial propagation. Kisspeptins, being upstream of the hypothalamic–pituitary-gonadal axis, appear to play a key role in the reproduction process. In the present study, the effect of different variants of kisspeptide, including goldfish (Carassius auratus) kiss1 kisspeptin (Kiss1), human kisspeptin (Hkiss), and their combination (Kiss1 + H), on the reproductive indices of goldfish broodstock in comparison to Ovaprim (a typical synthetic Gnrh hormone) was investigated. Peptides (Kiss1 and Hkiss) were synthesized using a solid-phase synthesis approach. Kiss1 and Hkiss were injected at a dose of 100 μg kg⁻¹ body weight, blood samples were taken 6 h after injection and sex hormones (E2, Dhp, and 11-Kt), gonadotropins (Lh and Fsh), cortisol and reproductive indices (fecundity, fertilization and hatching percentage) were measured. The results showed a significant increase of plasma sex hormones and gonadotropins in fish treated with kisspeptins. In addition, the cortisol and lipoprotein lipase in Kiss1, Hkiss and Kiss1 + H were remarkably increased compared to Ovaprim. In conclusion, kisspeptins could be a more suitable candidate than Ovaprim for accelerating and synchronizing oocyte maturation in the fisheries industry.     

Genetic variability in Plantago species in relation to their ecology

Summary Twelve Plantago major plants, good representatives of their populations, appeared to be genetically different for several characters which are important for adaptation to the respective habitat conditions. These characters are: juvenile growth, leaf morphology, production of secondary rosettes, flowering time, seed production, seed size and adult leaf production. The adaptive value of some of these characters was investigated by transplantation experiments in the field and by intraspecific competition experiments. The roadside type of ssp. major was adapted to trampling by being erect and elastic. The lawn type of ssp. major was adapted to a short, frequently cut, vegetation by being prostrate and by producing leaves with short petioles throughout the growth season. In the natural situations in which ssp. pleiosperma occurs, growth rate and first-year seed production of this subspecies were considerably higher than that of ssp. major. In a number of experiments, F₁s and F₂s were included, derived from crosses between the original plants. The F₁s were generally rather well adapted to both parental habitats, whereas the F₂s appeared to be less fit. The various alternatives in spending resources relevant for fitness optimization in different habitats are discussed.Grassland Species Research Group Publication No. 93     

Genetic Regulation of Liver Microsomal CYP2E1 Activity among Strains of the Viviparous Fishes Poeciliopsis occidentalis and Poeciliopsis lucida

The use of CYP2E1 (cytochrome P4502E1) expression as a biomarker for xenobiotic contamination may be useful, but for full understanding of the response, components of inheritance need to be understood. CYP2E1 expression was examined in P. lucida (M61-31), two strains of P. occidentalis (V87-15 and AV76-7), and their F1, F2, and backcross progeny. Phenotypic expression of CYP2E1 in Poeciliopsis has two distinct components – maximal activity and temperature optimum (T0) (Kaplan et al. 1991). In all P. occidentalis and P. lucida crosses, CYP2E1 activity segregated in up to three phenotypic groups. In F1 individuals (P. lucida×P. occidentalis V87-15), an overdominant phenotype was generated in 50% of the progeny. The P. lucida phenotype was absent suggesting a maternal effect on the F1 phenotype. Backcrosses of these F1 progeny to P. lucida females resulted in a recovery of the P. lucida phenotype while maintaining expression of an overdominant phenotype in 50% of the progeny. Among F2 progeny, an overdominant phenotype was expressed less often as compared to the F1 generation, and parental phenotypes were observed in equal numbers. Phenotypic expression patterns of CYP2E1 activity among F1 fish changed significantly with replacement of P. occidentalis V87-15 by P. occidentalis AV76-7. The P. lucida and P. occidentalis phenotype was equally distributed among progeny without evidence of an overdominant phenotype. Backcrosses of F1 progeny to P. lucida females resulted in equal amounts of the parental phenotypes. Backcross of an F1 male to a P. occidentalis AV76-7 female increased the P. occidentalis phenotype at the expense of the P. lucida phenotype in resulting progeny. Backcross of a F1 female to a P. lucida male resulted in equal distribution of the parental phenotypes. In addition, a greater number of F2 fish expressed the P. occidentalis phenotype as compared to the P. lucida phenotype. No significant difference in the sex ratios was seen in any of the crosses with the exception of one backcross.     

Mitochondrial DNA cytochrome b gene variation reveals different largemouth bass genealogies

A mitochondrial DNA character set based on cytochrome b gene variations is described that may be used in conjunction with established nuclear gene differences to assess largemouth bass Micropterus salmoides population structure and dynamics. Alignment of known cytochrome b sequences revealed two regions where the functional constraints of the molecule appear to be sufficiently reduced to allow several amino acid substitutions. Restriction analysis was used to probe nucleotide variation within one of these evolutionary hotspots, resulting in the discovery of fixed largemouth bass subspecific cytochrome b haplotypes. Application of two molecular character sets, one founded on fixed allelic variation in nuclear genes and the other on mtDNA cytochrome b diversity, revealed that in several Texas populations, where the Florida subspecies was introduced into extant populations of northern largemouth bass, hybridization was evident. The direction of the introgression vector could be determined through the combination of both character Sets.     

A simple plant polytene chromosome system,and its use for in situ hybridisation

When the cotyledons of Pisum sativum are cultured in the presence of 2–4D, large populations of cells with polytene chromosomes are stimulated to enter prophase; the use of these chromosome for studies of in situ hybridisation with nucleic acids is described.     

Changes in the Georgian Bay fish community,with special reference to lake trout (Salvelinus namaycush) rehabilitation

We describe and explain some of the changes in the fish community as reflected by the catches of the commercial fishery, and evaluate the efforts to restore the trout (Salvelinus spp.) in Georgian Bay. The changes in the fish community were caused in part by excessive fishing for lake trout (S. namaycush), lake whitefish (Coregonus clupeaformis) and deepwater ciscoes (Coregonus spp.), and the introduction of new species such as sea lamprey (Petromyzon marinus), alewife (Alosa pseudoharengus), rainbow smelt (Osmerus mordax) and salmonids (Oncorhynchus spp.). The most striking changes were the near extinction of the lake trout, reductions in lake herring (Coregonus artedii), deepwater ciscoes and lake whitefish, and the increase in abundance of smelt and alewife. In an effort to replace the loss of yield from the cold-water habitat, a fast growing and early maturing hybrid trout (splake [S. namaycush × S. fontinalis]) and its backcross to lake trout were selectively bred for stocking in Georgian Bay. Splake did poorly because of low survival immediately after planting. The survival of backcross after planting was better, but adult mortality was still high. Adult survival of backcross improved considerably after restrictions were placed on the commercial fishery in 1984. The first naturally produced backcross larvae were collected in 1985.     

Temporal analysis of tone pulses within the courtship songs of two siblingDrosophila species,their interspecific hybrid,and behavioral mutants ofD. melanogaster (Diptera: Drosophilidae)

Temporal analyses were applied to the tone pulses within the courtship songs of Drosophila melanogaster, D. simulans,their interspecific hybrid, and behavioral mutants of D. melanogaster.Linear regression was performed on various parameters of the song pulses (cycles per pulse, absolute peak amplitude, intrapulse frequency, number of peaks in fast Fourier transform, width of the primary frequency peak, and interpulse interval), as a function of their positions within pulse trains. Significant differences in the slope values of these two species and of the mutant genotypes allowed for discriminative quantification of temporal changes within trains. These results are discussed in relation to previous kinds of temporal analyses of Drosophilacourtship songs and also with regard to the mechanisms of song production.     

Further data on chromosomal assignments of pig enzyme loci LDHA, LDHB, MPI, PEPB and PGM1, using somatic cell hybrids

Summary. Clear interspecies differentiation between the chromosomes in pig-mouse somatic cell hybrids was achieved by using the THA-technique for the cytogenetic analysis. The assignments of LDHB and MPI to pig chromosomes nos 5 and 7 respectively, reported previously, were confirmed by analysis of 34 hybrid clones. The LDHA, PEPB and PGM1 genes were assigned to pig chromosomes nos 2, 5 and 6 respectively. Both LDHB and PEPB were indicated to be located on the long arm, except the most proximal part, of pig chromosome no. 5. The proposed synteny between LDHB and PEPB in pigs is in accordance with the synteny observed between these two loci in several other mammalian species.     

Asymmetric protoplast fusions between wild species and breeding lines of potato – effect of recipients and genome stability

 The objective of this study was to investigate if in asymmetric protoplast fusion experiments the ploidy of the recipient line (di-haploid and tetraploid) has an influence on the extent of the asymmetry of the regenerating fusion products. Nineteen different experiments with the wild species Solanum bulbocastanum and Solanum circaeifolium as donors (irradiated with 210 Gy) and different breeding lines (di-haploid and tetraploid) were carried out. The degree of genome elimination was determined by measuring the relative DNA content using flow cytometry. The data showed that the loss of DNA in hybrid plants was significantly higher for 4x, compared to 2x, plants as recipients. In addition, the stability of asymmetry in the fusion products was studied. For this purpose differences in asymmetry in individual shoots originating from the same callus were analysed. A large variation in the DNA content of individual shoots was detected. Of the 4x to 6x shoots 44% had the same DNA content as another shoot originating from the same callus, 19% had a DNA content between 4x and 6x but different from any other analysed shoot originating from the same callus, 2% were chimeras and 35% had a completely different DNA content (eutetraploid, euhexaploid, eupolyploid or asymmetric with a ploidy level above 6x). RFLP-analysis with single-copy probes of 12 regenerates from six calli (two regenerates per callus) confirmed the assumption that the different regenerates of one callus originate from the same single cell. The analysis of selected regenerates cultivated for a period of more than 1 year demonstrated that the genome of asymmetric regenerates might change during cultivation. Received: 30 April 1998 / Accepted: 24 July 1998