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Microsatellite analysis of wheat chromosome 2D allows the reconstruction of chromosomal inheritance in pedigrees of breeding programmes.

The dwarfing gene Rht8 and the photoperiodic insensitivity gene Ppd-D1 are linked on the short arm of chromosome 2D of bread wheat and play an important role in determining the geographic adaptation of modern wheat varieties. The genes are believed to originate from the old Japanese variety 'Akakomugi' and have been distributed throughout the world by diverse breeding programmes. Twelve microsatellite loci previously mapped on wheat chromosome 2D were used for a retrospective analysis of 59 wheat varieties with known pedigree, to trace the transmittance of the chromosomal region around these genes during extended breeding programmes. Within the range of the screened varieties 100 alleles were detected at the 12 microsatellite loci. For each microsatellite locus, a screen over varieties was performed to find the alleles corresponding to the parental variety 'Akakomugi'. A comparison of wheat varieties carrying the 192-bp allele, at locus Xgwm261-2D which is diagnostic for the presence of the gene Rht8, with the varieties without Rht8, showed linkage disequilibrium of 'Akakomugi' alleles for a segment of chromosome 2D which comprised at least 28 cM. Selection was accompanied with a linkage drag of 'Akakomugi' alleles in the neighbouring loci to Rht8. A diminution of the segment of chromosome 2D originating from 'Akakomugi' during several pedigree generations was observed. Varieties of the early generations were found to carry the whole short arm of chromosome 2D of 'Akakomugi', while the varieties of further selections possessed smaller segments including the diagnostic allele at locus Xgwm261-2D. Our results demonstrate that microsatellites can be successfully used for studying the inheritance of chromosomes within pedigrees of breeding programmes.

Chromosome Mapping↗

Inheritance of self-compatibility in almond: breeding strategies to assure self-compatibility in the progeny.

To assure self-compatibility in the progenies, three different crosses were conducted for the first time in an almond breeding programme: self-pollination (266 descendants from 30 families), crosses between parents sharing an S-allele (108 descendants from five families) and crosses with homozygous self-compatible parents (62 descendants from five families). Depending on the cross, self-compatibility in the progenies was determined by observing pollen tube growth (by means of fluorescence microscopy), stylar S-RNases analysis or allele-specific PCR. The results obtained fit with the accepted hypothesis of inheritance of self-compatibility and the three crossing strategies used ensured 100% of self-compatible descendants. These strategies increase the efficiency of the breeding programme and avoid the laborious task of evaluating this characteristic. From the breeding point of view, self-fertilisation and crosses between relatives tend to produce inbreeding. Furthermore, these methods reduce the possibilities of choosing the parental combination. The use of homozygous self-compatible parents does not have any of these disadvantages. As far as we know, this is the first time that allele-specific PCR has been used for early selection of self-compatible seedlings. The advantages and disadvantages of the three methodologies used to determine self-compatibility are discussed.

Crosses, Genetic↗

Marker-evaluated selection in rice: shifts in allele frequency among bulks selected in contrasting agricultural environments identify genomic regions of importance to rice adaptation and breeding.

Conventional methods for quantitative trait locus (QTL) mapping require the selection of particular traits to be measured based on assumptions as to their importance. We have tested an alternative approach for the location of QTLs-marker-evaluated selection-that makes no prior assumptions as to which traits are important. The results of phenotype selection were evaluated in the products of modified bulk-population breeding that was replicated across a range of rice ecosystems. Selection was carried out in close collaboration with farmers in bulk populations that were all derived from a cross between an Indian upland variety (Kalinga III) and a high-yielding semi-dwarf variety (IR64).Twenty-seven diverse bulks were produced that were screened with molecular markers in order to determine whether shifts could be detected in marker allele frequency as a result of selection and if such changes varied by genomic region across ecosystems. Marker loci linked to important traits for adaptation to specific environments were identified without making any prior assumptions about which traits might be important. Genomic regions from Kalinga III were strongly selected in the upland environments and regions from IR64 in the lowland ones.However, exceptions occurred where the upland parent contributed positively to lowland adaptation and vice versa. The results can be used as a basis for the development of second-cycle varieties, using marker-assisted selection to produce genotypic ideotypes for specific target environments. The very strong selection for genomic regions from the adapted parents of the wide(upland x lowland) cross indicates that, in non-marker-assisted breeding, where genetically distant parents have been used, modified backcross breeding should be efficient. A single backcross to the adapted parent for aspecific ecosystem will result in a higher frequency of segregants with the desired high genetic contribution from the adapted parent.

Acclimatization↗

Bone response to in vivo mechanical loading in two breeds of mice.

We investigated the bone response to external loading in C57BL/6J and C3H/HeJ mice, both breeds with low and high bone density, respectively. An in vivo tibial four-point bending device previously used for application of measured external loads in rats was adapted for mice. It delivered a uniform medio-lateral bending moment to the region of the tibia located 1-5.5 mm proximal to the tibio-fibula junction. The right legs of six C57BL/6J [low bone density (LBD)] and C3H/HeJ [high bone density (HBD)] mice were externally loaded in the device for 36 cycles/day at 2 Hz, 6 days/week for 2 weeks at 9.3 +/- 0.9 N force, inducing estimated lateral periosteal surface compressive strains of 5121 +/- 1128 mu epsilon in C3H/HeJ (HBD) mice (n = 6), significantly higher than the estimated 3988 +/- 820 mu epsilon in C57BL/6J mice (n = 6) (mean +/- SD). In addition, C3H/HeJ HBD mice (n = 11) were externally sham (pad pressure or no bending) loaded in the device for 36 cycles/day at 2 Hz, 3 days/week for 3 weeks at 9.3 +/- 0.9 N force. Calcein injections for bone labeling were given at the 10th and 3rd days before sacrifice. At the end of the experiment, all mice were killed and both tibiae were removed, fixed, embedded, and cross-sectioned through the loaded region. Both tibiae were measured for marrow area (Ma.Ar), cortical area (Ct.Ar), total area (Tt.Ar), cross-sectional moment of inertia (CSMI), and periosteal and endocortical woven bone surface (Wo.B/BS), single-labeled surface (sLS), double-labeled surface (dLS), and total formation surface (FS/BS). Differences in all variables due to breed and loading (both bending and sham-bending) were tested by two-way analysis of variance (ANOVA) (P < 0.05). Ma.Ar, Tt.Ar, and CSMI were greater in C57BL/6J (LBD) than in C3H/HeJ (HBD) mice. Periosteal and endocortical woven bone and formation surface were increased significantly more by loading (bending) in C57BL/6J than in C3H/HeJ mice. Periosteal woven bone response due to sham-bending or sham-loading was significantly lower than due to bending loads in the C3H/HeJ mice. We conclude that the bone response to external loading is greater in LBD mice than in HBD mice. The high bone density of C3H/HeJ (HBD) mice is related to breed-specific factors other than the response to loading.

Analysis of Variance↗

Characterization of fecal bacterial populations in canines: effects of age, breed and dietary fiber.

The effects of age, breed, and diet on fecal chemistry, enzyme activity, and bacterial populations of dogs were studied. Eighteen dogs from two age groups (young: 2.5 +/- 0.5 years, old: 10.9 +/-0.7 years) and three different breeds (German shepherds, miniature schnauzers, and English setters) were rotated through a Latin Square design such that every dog was fed each of the diets. The test diets included a low-fiber (control) diet and a 10% fiber diet which contained 5% soybean hulls and 5% beet pulp. Inclusion of 10% fiber in the diet decreased the fecal concentration of ammonia, sulfide, and indole. Fiber inclusion significantly increased acetic, propionic, and butyric acid concentrations, while fecal pH decreased by 0.4 units. Fresh fecal samples were plated on selected aerobic and anaerobic culture media and DNA extracted for denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S ribosomal DNA fragments. Plate counts showed significant effects of breed (p < or = 0.05) and age (p < or = 0.01) on selected aerobic and anaerobic bacterial counts, while no significant effect of diet was found. Analysis of PCR-DGGE banding patterns showed there was a tendency for individual dogs to cluster together according to age (young or old dogs) and also for size (large or small dogs). However, the outstanding conclusion obtained from the DGGE analysis of fecal bacterial profiles was that individual dogs had their own characteristic banding pattern which was unique and stable. The relative stability and individuality of the patterns indicates that each individual harbored a characteristic fecal bacterial community which was independent of diet.

Age Factors↗

Impact of busy roads on breeding success in pied flycatchers Ficedula hypoleuca.

The impact of dense traffic on the breeding success in pied flycatchers (Ficedula hypoleuca) was studied using roadside nest-boxes. Nest site selection and breeding success of flycatchers were observed in relation to the distance from the road. The number of occupied territories was no higher closer to the road than it was deeper inside the forest. The distance to the road had no effect on the laying date, clutch size, or brood size. However, nests closer to the road were more likely to fail at the chick stage. The number of broods that were closer to roads and were lost completely was significantly higher than those further away. As a consequence, the number of fledglings per breeding attempt decreased closer to roads. The nestlings typically died as older chicks when the parent birds were providing them with maximum quantities of food. A possible reason for the declining reproductive success was the traffic-related mortality of parent birds, as weather conditions were not especially adverse for raising healthy offspring.

Animals↗

Prospects for whole genome linkage disequilibrium mapping in domestic dog breeds.

Linkage disequilibrium (LD) mapping is commonly used as a fine mapping tool in human genome mapping and has been used with some success for initial disease gene isolation in certain isolated in-bred human populations. An understanding of the population history of domestic dog breeds suggests that LD mapping could be routinely utilized in this species for initial genome-wide scans. Such an approach offers significant advantages over traditional linkage analysis. Here, we demonstrate, using canine copper toxicosis in the Bedlington terrier as the model, that LD mapping could be reasonably expected to be a useful strategy in low-resolution, genome-wide scans in pure-bred dogs. Significant LD was demonstrated over distances up to 33.3 cM. It is very unlikely, for a number of reasons discussed, that this result could be extrapolated to the rest of the genome. It is, however, consistent with the expectation given the population structure of canine breeds and, in this breed at least, with the hypothesis that it may be possible to utilize LD in a genome-wide scan. In this study, LD mapping confirmed the location of the copper toxicosis in Bedlington terrier gene (CT-BT) and was able to do so in a population that was refractory to traditional linkage analysis.

Alleles↗

DNA polymorphisms of the prion doppel gene region in four different German cattle breeds and cows tested positive for bovine spongiform encephalopathy.

Polymorphisms of the prion protein gene PRNP have been shown to influence the susceptibility/resistance to prion infections in human and sheep. In addition, the T174M polymorphism within the flanking prion doppel gene (PRND) was thought to be involved in susceptibility to sporadic Creutzfeldt-Jacob disease. To study a possible influence of DNA polymorphisms of the bovine PRND gene in bovine spongiform encephalopathy (BSE), previously identified and newly isolated DNA polymorphisms were genotyped in all available German cattle that tested positive for BSE. Genotypes and calculated haplotypes were compared with breeding bulls serving as controls. Analysis of the four major breeds Schwarzbunt (Holstein Friesian), Rotbunt (Holstein Red), Fleckvieh (Simmental), and Braunvieh (Swiss Brown) resulted in the isolation of the previously known polymorphisms R50H and R132Q and two novel synonymous single nucleotide polymorphisms (SNPs) C4820T and A5063T. Comparative genotype and haplotype analysis of BSE and control animals revealed a significantly different distribution of polymorphisms C4815T and R132Q in Fleckvieh animals but not in the other breeds tested. No association to BSE susceptibility was detectable for polymorphisms R50H and A5063T.

Amyloid↗

Acute and chronic effects of an aromatase inhibitor on territorial aggression in breeding and nonbreeding male song sparrows.

Many studies have demonstrated that male aggression is regulated by testosterone. The conversion of testosterone to estradiol by brain aromatase is also known to regulate male aggression in the breeding season. Male song sparrows (Melospiza melodia morphna) are territorial not only in the breeding season, but also in the nonbreeding season, when plasma testosterone and estradiol levels are basal. Castration has no effect on nonbreeding aggression. In contrast, chronic (10 day) aromatase inhibitor (fadrozole) treatment decreases nonbreeding aggression, indicating a role for estrogens. Here, we show that acute (1 day) fadrozole treatment decreases nonbreeding territoriality, suggesting relatively rapid estrogen effects. In spring, fadrozole decreases brain aromatase activity, but acute and chronic fadrozole treatments do not significantly decrease aggression, although trends for some behaviors approach significance. In gonadally intact birds, fadrozole may be less effective at reducing aggression in the spring. This might occur because fadrozole causes a large increase in plasma testosterone in intact breeding males. Alternatively, estradiol may be more important for territoriality in winter than spring. We hypothesize that sex steroids regulate male aggression in spring and winter, but the endocrine mechanisms vary seasonally.

Aggression↗

Growth dynamics of lipizzan horses and their comparison to other horse breeds.

A typical body format of Slovenian Lipizzan horse was investigated. The study included 6 foals (5 colts and 1 filly) at the Lipica stud farm. They were measured from birth to twenty-seven months and again at forty-four months of age. Measurements included body length, chest circumference, withers height and body mass. All those measurements were statistically evaluated and compared to some other horse breeds to determine the similarity of the growth dynamics of those horse breeds. It was concluded that the parameters of the head and neck reached their full growth at the age of 27 months and also exceeded the values of their parents. The same results were obtained with the length and heights of the body (body length, withers height, etc.) and with the parameters of the front and hind legs. However the girth width, hip width and some other parameters had not yet reached their full growth at 27 months. We observed different growth dynamics with the body weight. The parameter continued to grow after 27 months of age. The same growth dynamics also seem to apply to other investigated horse breeds although the actual measurements were slightly different.

Animals↗

Predation by sparrowhawks decreases with increased breeding density in a songbird, the great tit.

Predators may regulate prey populations if predation rate increases with prey density. Alternatively, if space-limited (e.g. territorial) predators become 'satiated' when prey exceed a certain density, increased prey abundance may lead to reduced predation rate. These alternatives have been difficult to test experimentally for mobile prey in the wild. We present such a test, manipulating the density of great tits (Parus major) by adding nest boxes in territories of sparrowhawks (Accipiter nisus). Predation rate was measured for young tits after they left the nests. Although the great tit is an important prey, there was no evidence for regulation during the breeding season: the rate of hawk predation declined with increasing density of tits. This result was not confounded by changes in breeding density of alternative prey species (other songbirds). Hawk predation can therefore favour dense breeding in a territorial (solitary) bird, and conspecific attraction and aggregation reported in several territorial species may partly result from predation pressure. This result also has potential implications for conservation work.

Animals↗

Diagnosis and PrP genotype target of scrapie in clinically healthy sheep of Massese breed in the framework of a scrapie eradication programme.

The application of a selective culling programme in two scrapie affected flocks of Massese breed sheep is described. The genetic susceptibility of this breed and the sensitivity of different diagnostic methods in the pre-clinical diagnosis of scrapie were also investigated. Overall, 2,068 clinically healthy sheep underwent PrP genotyping, providing the basis for selective culling. The prevalence of scrapie infection was investigated in susceptible sheep by two independent diagnostic methods. All the sheep older than 18 months (n = 620) were tested by Prionics Check Western rapid test on the obex, with a prevalence of infection of 3.9%. Furthermore, 385 sheep underwent immunohistochemistry (IHC) on retropharyngeal lymph node (RPLN), with a prevalence of infection of 5.2%. Overall, 32 sheep were diagnosed with pre-clinical scrapie. Of these, 31 were positive by Western blot on the spleen, 29 by IHC on the RPLN and tonsil, 28 by IHC on the obex, 24 by rapid test, and only 18 by IHC on the third eyelid. All the scrapie positive sheep were of the ARQ/ARQ, ARQ/AHQ or ARQ/VRQ genotypes. No significant differences in scrapie prevalence were observed among these genotypes. The estimated risk of the three targeted alleles was also similar, suggesting that in this breed the VRQ allele was not at higher risk for scrapie, compared to the ARQ and AHQ alleles.

Alleles↗

The breeding system of wild red ruffed lemurs (Varecia rubra): a preliminary report.

Captive studies have shown that ruffed lemurs (Varecia) have an unusual suite of reproductive traits combined with extremely high maternal reproductive costs. These traits include the bearing of litters, nesting of altricial young, and absentee parenting. To characterize the breeding system of this enigmatic lemur, reproductive traits must be contextualized in the wild. Here, I provide a preliminary report of mating and infant care in one community of wild red ruffed lemurs (Varecia rubra). Observations span a 15-month period covering two birth seasons and one mating season on the Masoala Peninsula, Madagascar. Factors that are not possible to replicate in captivity are reported, such as mating pattern, natality and mortality rates, the location of nests within the home range, and the structuring of infant care within a natural community. V. rubra at Andranobe have a fission-fusion, multifemale-multimale grouping pattern and a polygamous mating system. They do not mate monogamously or live strictly in family-based groups as suggested by previous workers. During the first 2 months of life, nests and infant stashing localities are situated within each mother's respective core area, and inhabitants of each core area within the communal home range provide care for young. As part of their absentee parenting system, infants are left in concealed, protected, and supportive spots high in the canopy, while mothers travel distantly. This practice is termed 'infant stashing'. Alloparenting appears to be an integral part of V. rubra's overall reproductive strategy in the wild, as it was performed by all age-sex classes. Among the alloparental behaviors observed were infant guarding, co-stashing, infant transport, and allonursing. Alloparenting and absentee parenting may mitigate high maternal reproductive costs. Furthermore, V. rubra may have a breeding system in which genetic partners (i.e., mating partners) do not always correspond to infant care-providers. Combined with recently available information on the behavioral ecology of wild ruffed lemurs, this preliminary report suggests directions for in-depth studies on Varecia's breeding system.

Animals↗

Genetic polymorphism of beta-lactoglobulin gene in native Turkish sheep breeds.

The genetic polymorphism of the beta-lactoglobulin gene was investigated in three native Turkish sheep breeds. The study was carried out on 108 sheep (29 Kivircik, 38 Gökçeada, and 41 Sakiz) by means of PCR-RFLP methods. Two genetic variants (A and B) and three genotypes (AA, AB, and BB) of beta-lactoglobulin have been identified. The gene frequencies of beta-LG A and B were 0.7759 and 0.2241 in Kivircik, 0.7632 and 0.2368 in Gökçeada, and 0.9756 and 0.0244 in Sakiz breeds, respectively. The populations were in Hardy-Weinberg equilibrium in all samples from the three breeds.

Animals↗

Mercury and other contaminants in common loons breeding in Atlantic Canada.

Common loons (Gavia immer) were studied to assess the biomagnification of persistent contaminants in lake ecosystems in Atlantic Canada. Forty-two breeding adults and 20 juvenile loons were captured in August, 1995-1997 on lakes in four areas of southern New Brunswick (Lepreau and Fundy National Park) and Nova Scotia (Halifax and Kejimkujik National Park). Blood samples were collected for analysis of mercury, methylmercury, lead, and selenium. Plasma samples from adult loons were analyzed for PCB congeners and organochlorine pesticides. Secondary flight feathers were collected from adult loons for mercury analysis. Kejimkujik loons had significantly higher geometric mean concentrations of mercury in blood (adults: 5.5 microg/g, wet wt) and feathers (adults: 15 microg/g, fresh wt), and higher levels of PCBs, DDE, oxychlordane, trans-nonachlor, mirex and hexachlorobenzene than loons at Halifax or Lepreau. Blood selenium levels were lowest in Kejimkujik loons. Selenium and mercury concentrations were negatively related in blood of adults across all sites. Adult males had higher concentrations of mercury in blood and feathers and PCBs, DDE, and oxychlordane in plasma, and lower levels of selenium in blood, than adult females. Juvenile loons had lower blood mercury and selenium concentrations than adults. Several contaminants showed significant inter-lake variation within the study sites. Mean blood mercury concentrations in Kejimkujik loons are the highest found in breeding common loons across North America. Of adult loons at Kejimkujik, 92% had blood mercury levels > 4 microg/g (wet wt), which have been associated with impaired reproduction, elevated corticosterone levels, asymmetry in plumage development, and altered breeding behavior in loons there and elsewhere.

Animals↗

Efficiency of DNA repair mechanisms of domestic dog primary fibroblasts isolated from small and large breeds of different ages in response to double stranded breaks (DSB).

Aging is associated with increased genomic instability, a phenomenon largely driven by the accumulation of DNA damage over time, and large species of mammals seem to have more robust DNA repair systems associated with longer lives. Among DNA lesions, double-strand breaks (DSBs) are particularly deleterious and have been implicated in age-related functional decline and disease. In this study, we investigated how age and body mass affect the efficiency of DSB repair (DSBr) in primary fibroblast cells isolated from domestic dogs, a species that exhibits significant intraspecies variation in lifespan and body mass. Primary fibroblast cells were isolated from puppies and senior dogs of both large and small breeds. Cells were treated with 100&#xa0;&#xb5;M etoposide to induce DSBs and subsequently analyzed at two post-treatment recovery intervals (2 and 24&#xa0;h) to correlated with the two pathways associated with DSBr, the fast, non-homologous end joining (NHEJ) and the much slower, homologous recombination (HR). Cells were stained for &#x3b3;-H2AX foci and images were collected using confocal microscopy. We found that mean fluorescence per cell was higher in older dogs of both size classes in the 2&#xa0;h recovery, indicating higher amounts of DNA damage but suggesting similar efficiencies through the NHEJ repair mechanism in older dogs despite size class. We also show that mean fluorescence per cell was higher in the older large breed dogs in the 24&#xa0;h recovery, suggesting that the slower phase associated with HR seems to be deficient in cells from older, larger breeds of dogs. These findings support the broader theory that aging is associated with impaired genomic maintenance and establish domestic dogs as a valuable model for studying the cellular mechanisms of age-related genomic instability.

Animals↗

Response of pineal beta-adrenoceptors in different breeds of sheep to immunization against selected steroids.

Active immunization of Merino and Wiltshire Horn X Merino cross-bred sheep against a range of gonadal steroids has revealed that pineal beta-adrenoceptors in both breeds are sensitive to hormonal modification by androgens, but only in the less seasonal, non-shedding Merino do these receptors appear to be sensitive to regulation by estrogens. Neither breed showed sensitivity of its pineal beta-adrenoceptors to active immunization against the pineal hormone melatonin under either normal or reversed photoperiod treatment. These results (a) suggest that pineal related seasonal differences between the breeds (i.e. wool shedding and/or reproductive function) may reside in differential sensitivity of the pineal gland to regulation by specific circulating steroid hormones, and (b) indicate an absence of end-product regulation of beta-adrenoceptor-mediated pineal melatonin biosynthesis in both seasonally shedding and non-shedding sheep.

Androgens↗

Plasma gonadal steroid levels in wild starlings (Sturnus vulgaris) during the annual cycle and in relation to the stages of breeding.

Blood samples were collected from male and female free-living starlings in every month during the year and at all stages of the breeding cycle. The samples from females were assayed for oestradiol-17 beta, oestrone, and progesterone, and the samples from males were assayed for testosterone and 5 alpha-dihydrotestosterone. The testicular weights were recorded. All hormones other than oestrone showed a pronounced unimodal cycle. In females, oestradiol was highest in April, corresponding to the period of greatest nest-building activity. It decreased during the later stages of the breeding cycle and maintained low levels until the autumn when levels began to increase. Progesterone was highest in laying birds and reached a peak in May. During the rest of the year it remained at a lower but fairly constant level. Oestrone was normally only detectable in laying birds. In males, testicular weight and plasma concentrations of testosterone and 5 alpha-dihydrotestosterone were all highest during April. Plasma testosterone and 5 alpha-dihydrotestosterone were highest in nest-builders. They decreased during the later stages of breeding, remained low during the summer and increased slightly in the autumn. Testicular weight was high in nest-builders, but peaked during laying, by which time testosterone and 5 alpha-dihydrotestosterone had decreased significantly. Testicular weight decreased during incubation and feeding the young and remained low until the following year.

Animals↗