Comparative analysis of diploid species of Avena L. using cytogenetic and biochemical markers: Avena pilosa M. B. and A. clauda Dur

The diploid oat species containing the Cp genome - Avena pilosa and A. clauda - were studied using C-banding, fluorescence in situ hybridization with probes pTa71 and pTa794, and electrophoresis of grain storage proteins (avenins). Species with the C genome differed considerably from the species of the A genome group in the karyotype structure, heterochromatin type and distribution, relative positions of the 45S and 5S rRNA gene loci, and avenin patterns. These facts confirmed that the C genome had diverged from the ancestral genome before the radiation of the various A genome. Presumably, further evolution of the A -and C genome species occurred separately.     

Molecular study and C-banding of chromosomes in common wheat alloplasmic lines obtained from the backcross progeny of barley–wheat hybrids Hordeum vulgare L. (2n = 14) × Triticum aestivum L. (2n = 42) and differing in fertility

We studied common wheat alloplasmic lines differing in fertility traits, which had been obtained from the backcross progeny of barley—wheat hybrids Hordeum vulgare L. (2n = 14) × Triticum aestivum L. (2n = 42), using molecular analysis and chromosome C-banding. It was found that the nuclei of all alloplasmic lines studied, regardless of their fertility traits, contained only the common wheat chromosomes (2n = 42). The formation of line L-79(10)(3)F₆, stable for self-fertility, from line L-79(10)F₃ was accompanied by changes of the proportions of simple sequence repeats of the parental common wheat varieties in the nuclear genome. The presence of barley genome fragments in line accessions with incomplete self-fertility was shown by RAPD. Heteroplasmy for mitochondrial genome loci was detected in these lines with the use of primers specific to the tMet-18S-5S repeat of mitochondrial ribosomal genes.Translated from Genetika, Vol. 40, No. 12, 2004, pp. 1668–1677.Original Russian Text Copyright © 2004 by Bildanova, Badaeva, Pershina, Salina.     

Hyperunstable (TCT/TCC) n Microsatellite Loci in Parthenogenetic Lizards Darevskia unisexualis (Lacertidae)

Using multilocus DNA fingerprinting, we have examined variability of (TCT) _n microsatellite and M13 minisatellite DNA repeats in populations, families, and tissues of parthenogenetic Caucasian rock lizards Darevskia unisexualis (Lacertidae). It has been shown for the first time that population and family DNA samples of D. unisexualis (75 samples in total) have individually specific DNA fingerprinting patterns of (TCT) _n fragments. Analysis of inheritance of (TCT) _n microsatellites in 46 first-generation progeny in 17 parthenogenetic D. unisexualis families revealed their extremely high instability. Mutant TCT fingerprint phenotypes were found in virtually each animal of the progeny. Moreover, varying fragments in the progeny and their original variants in the mothers were shown to simultaneously contain (TCT) _n and (TCC) _n polypyrimidine clusters. At the same time, no variability of (TCT) _n fragments has been detected in the tissues and organs of mature parthenogenetic lizards and in the analogous tissues of the two-week-old progeny of this year. This suggests the absence of somatic mosaicism and methylation of the corresponding loci in the samples. Along with the hyperinstability of (TCT/TCC) _n polypyrimidine clusters, we have shown that the population and family DNA fingerprinting patterns of M13 minisatellites were invariable and monomorphic in the same DNA samples of D. unisexualis.Our results indicate that mutations at loci containing polypyrimidine microsatellites significantly contribute to the total genomic variability of parthenogenetic lizards D. unisexualis.     

History of Modern Chromosomal Analysis. Differential Staining of Plant Chromosomes

Differential staining methods found extensive use in karyotype studies of many plant and animal species and provide for reliable identification of all chromosomes of the organism. Below we describe the most widespread methods and history of their advent. In addition, we discuss specific structure of the chromosomes and possible mechanisms responsible for differential segmentation.     

Localization of Human Ribosomal Gene DNA Probes on Barley Chromosomes

To estimate the possibility of plant genome mapping using human genome probes, the probes fluorescent in situ hybridization (FISH) of human 18S–28S rDNA (clon 22F9 from the LA-13NCO1 library) was carried out on chromosomes of the spring barleyHordeum vulgareL. As a control, wheat rDNA probe (clon pTa71) was taken. Hybridization of the wheat DNA probe revealed two major labelling sites on mitotic barley chromosomes 5I (7H) and 6I (6H), as well as several minor sites. With the human DNA probe, signals were detected in the major sites of the ribosomal genes on chromosomes 5I (7H) and 6I (6H) only when the chromosome preparations were obtained using an optimized technique with obligatory pepsin treatment followed by hybridization. Thus, this study demonstrates that physical mapping of plant chromosomes with human DNA probes that are 60 to 70% homologous to the plant genes is possible. It suggests principal opportunity for the FISH mapping of plant genomes using probes from human genome libraries, obtained in the course of the total sequencing of the human genomes and corresponding to the coding regions of genes with known functions.     

Manifestations of the neurotoxicity of the organochlorine pesticide dilor during the postnatal period in the rat

When pregnant rats are administered chlororganic pesticide dilor, certain ultrastructural changes are observed in neurocytes and in myelin fibers, in the spinal cord, that demonstrates some disorders in cellular and tissue metabolism. In the newborn spinal cords a retarded differentiation is observed against the background of an intense cellular metabolism. By means of the electron paramagnetic resonance technique, an increasing concentration of free radicals in the brains and spinal cords of the pregnant animals and in their one-month-old offsprings is demonstrated. The investigation on fatty acid composition of lipids in pregnant test animals demonstrates a decrease in cholysterine, steroids and some fatty acids in myelin fractions and in synaptosomic membranes.     

Manifestation of the neurotoxic effect of the organochlorine pesticide hexachlorobutadiene in the postnatal period of ontogeny in the rat

Daily peroral administration of chlororganic pesticide hexachlorobutadiene in doses 8.1 mg/kg (1/20 LD50) to pregnant rats results in certain ultrastructural changes of neurocytes and myelin fibers of the spinal cord both in the animals and their offspring (newborns and 1-2-month-old rats). By means of electron paramagnetic resonance (EPR) method, changes in intensity of the EPR-signals of free radicals in the spinal cord, ceruloplasmin of blood serum have been revealed in the experimental pregnant animals, as well as in 1-month-old rats (in the latter--in the brain, too). Gas-liquid chromatography reveals the preparation contents in the adrenals, heart, brain and spinal cord, in the uterus of the pregnant animals, as well as in corresponding organs of their offspring. Certain retardation in growth and decrease in body mass are noted in the offspring.     

Chromosome substitutions of Triticum timopheevii in common wheat and some observations on the evolution of polyploid wheat species

Whether the two tetraploid wheat species, the well known Triticum turgidum L. (macaroni wheat, AABB genomes) and the obscure T. timopheevii Zhuk. (A^tA^tGG), have monophyletic or diphyletic origin from the same or different diploid species presents an interesting evolutionary problem. Moreover, T. timopheevii and its wild form T. araraticum are an important genetic resource for macaroni and bread-wheat improvement. To study these objectives, the substitution and genetic compensation abilities of individual T. timopheevii chromosomes for missing chromosomes of T. aestivum Chinese Spring (AABBDD) were analyzed. Chinese Spring aneuploids (nullisomic-tetrasomics) were crossed with a T. timopheevii x Aegilops tauschii amphiploid to isolate T. timopheevii chromosomes in a monosomic condition. The F₁ hybrids were backcrossed one to four times to Chinese Spring aneuploids without selection for the T. timopheevii chromosome of interest. While spontaneous substitutions involving all A^t- and G-genome chromosomes were identified, the targeted T. timopheevii chromosome was not always recovered. Lines with spontaneous substitutions from T. timopheevii were chosen for further backcrossing. Six T. timopheevii chromosome substitutions were isolated: 6A^t (6A), 2G (2B), 3G (3B), 4G (4B), 5G (5B) and 6G (6B). The substitution lines had normal morphology and fertility. The 6A^t of T. timopheevii was involved in a translocation with chromosome 1G, resulting in the transfer of the group-1 gliadin locus to 6A^t. Chromosome 2G substituted for 2B at a frequency higher than expected and may carry putative homoeoalleles of gametocidal genes present on group-2 chromosomes of several alien species. Our data indicate a common origin for tetraploid wheat species, but from separate hybridization events because of the presence of a different spectrum of intergenomic translocations.     

Sucrose Breakdown in Relation to Fruit Growth of Acid Lime (Citrus aurantifolia)

The physiological capacity for sucrose breakdown in developingjuice sac cells of acid limes was estimated by assaying theactivity of the three enzymes of sucrose catabolism in additionto vacuolar acid hydrolysis. The maximum potential rates ofsucrose breakdown were compared with the observed rates of carbonutilization. Highest potential rates of sucrose breakdown (28.621mmol cm^–3 per hydrated active space d^–1) occurredat the initial stages of fruit development where carbon utilizationwas highest. As the fruit developed, the potential rates ofsucrose breakdown and carbon utilization declined to very lowlevels. At 80% of development, vacuolar acid hydrolysis becamethe only physiological mechanism for sucrose breakdown. Therelatively low amounts of sucrose hydrolysed by acid hydrolysisat this time were just sufficient to account for the measuredcarbon demands. The results suggest that carbon supplied bythis distinct sucrose catabolizing system is able to provideadequate levels of carbon skeletons for the observed levelsof respiration and dry weight deposition early in development,but becomes a limiting factor for growth in the later stages. Key words: Vacuolar acid hydrolysis, Citrus aurantifolia