Whole genome sequencing uncovers the structural and transcriptomic landscape of hexaploid wheat/Am. muticum introgression lines

Wheat is a globally vital crop, but its limited genetic variation creates a challenge for breeders aiming to maintain or accelerate agricultural improvements over time. Introducing novel genes and alleles from wheat's wild relatives into the wheat breeding pool via introgression lines is an important component of overcoming this low variation but is limited by poor genomic resolution and limited understanding of the genomic impact of introgression breeding. By sequencing 17 hexaploid wheat/Ambylopyrum muticum introgression lines and the parent lines, we have precisely pinpointed the borders of introgressed segments. We report a genome assembly and annotation of Am. muticum that has facilitated the identification of Am. muticum resistance genes commonly introgressed in lines resistant to stripe rust. Our analysis has identified an abundance of structural disruption and homoeologous pairing across the introgression lines, likely caused by the suppressed Ph1 locus. mRNAseq analysis of six of these introgression lines revealed that introgressed genes tend to be downregulated, shifting the expression balance of triads towards suppression of the introgressed region, with no discernible compensation in the expression of the homoeologous copies. This analysis explores the genomic impact of introgression breeding and provides an affordable way for breeders to better characterise introgression lines and more effectively deploy wild relative variation.

Protein contain in Cyprinus carpio (Ham.) of experimental and control fishes due to administration of ovaprim, ovatide and pituitary gland extract at Fish Seed Production Centre, Paithan, Aurangabad, Maharashtra State, India

Protein contain is a cornerstone mechanism and form part and parcel of nutritive value in particular rely on season, food intake, breeding and spawning capacity. The protein contain in the fish body constitute as a firsthand source of energy for physiological functions incorporating reproduction. Present investigation display that variations in the protein is observed from the mature fishes of Indian major carps such as Cyprinus carpio comprises 20 to 50cm selected for the estimation of protein study considered as experimental and control fishes upon administration of PGE, ovaprim and ovatide synthetic hormonal doses to the male and female sex ratio separately. Protein in the liver of female major carps may be because of vitellogenin, a lipoprotein which is synthesized in the liver and is transported to the ovary (Wallace, 1985). Moreover, gonadal protein enhanced with maturation amongst these fishes. Protein estimation represents the variations throughout the study period amongst the muscle, gonad and liver. After hormonal injections are given then fishes are kept in the breeding pool at the Fish Seed Production Centres (FSPC) located near Paithan barrage. The protein is estimated upon satisfactory administration of hormonal doses Pituitary Gland Extract and by other two synthetic hormonal doses viz. Ovaprim and Ovatide. Study is conducted during 2013 to 2014.

37 Viability of sheep skin fibroblasts after vitrification

Both tissue and cell cryopreservation can be applied for biodiversity conservation. The proper preservation of tissues and cells from a wide range of animals of different species is of paramount importance because these cell samples could be used to reintroduce lost genes back into the breeding pool by somatic cell cloning. The aim of this work was to investigate the effect of vitrification on viability of vitrified sheep fibroblasts for conservation of biodiversity so that it might be used in the future to provide nuclear donors. Skin samples collected from 10 adult sheep were cut into small pieces (1×1mm), placed into culture Petri dishes containing DMEM supplemented with 20% (vol/vol) fetal bovine serum, and covered with coverslips followed by incubation at 5% CO2, 95% RH, and 37°C. During culture, fibroblasts left skin samples and proliferated. Culture medium was changed every 4 days. After 21 to 22 days of incubation, a fibroblast monolayer was observed, culture medium was removed, and cells were incubated for 7 to 10min in the presence of Dulbecco’s PBS+0.25% trypsin. Dissociated fibroblasts were washed with DMEM by centrifugation at 300×g for 10min. For vitrification, fibroblast samples were then diluted at a concentration of 2×106cellsmL−1 in DMEM+ 20% ethylene glycol, 20% dimethylsulfoxide, and 0.5molL−1 of sucrose. The fibroblasts were then exposed to 50 and 100% vitrification solution (VS) at 37°C for 5min and 30s, respectively. Fibroblasts after saturation in VS were transferred and placed into 0.25-mL plastic straws. Straws were sealed with modelling clay and plunged into LN. Viability of frozen-thawed fibroblast samples was detected using the Trypan Blue staining method (frozen-thawed: 53.0±2.6%; control (fresh): 98.5±1.2%). The values obtained are expressed as mean±standard error of the mean. Statistical analysis was done using Student’s t-test. Results indicated that there was a significant difference in viability between fresh and cryopreserved fibroblasts. Importantly, our data suggest that the use of vitrification reduced the toxic elements contained in the cryopreservation solution while maintaining a similar ability to produce viable fibroblasts after cryopreservation. Although further work on the viability of sheep skin fibroblast with the vitrification method is needed, these data suggest that with vitrification a faster cooling rate and high level of cryoprotectants are able to minimize ice crystal formation and should be further evaluated as a routine mechanism for cryopreserving sheep fibroblasts.

Chrysonilia sitophila: Growth on Different Culture Media and Sensitivity to Antifungals

The species Chrysonilia sitophila has a cosmopolitan spread, living on dead or rotting plants, but it is also isolated from home and commercial spaces insulation. The aim of our study was to determining a growth pattern and assessing the susceptibility to different antifungals. The strain was isolated from the water of a trout breeding pool where high mortality due to Saprolegnia infection and various Gram negative and Gram positive bacteria occurred. On all used culture media (except for MacConkey), rapid growth is observed (tendency to spread over the entire surface of the plate in 36-48 hours). The isolated strain has been shown to be sensitive to ketoconazole, econazole, and miconazole; less sensitive to amphotericin B, but resistant to metronidazole, nystatin, griseofulvin and flucytosine. Chrysonilia sitophila can be grown easily, so it can also establish an antifungal sensitivity profile. Additional efforts should be made to study the pattern of susceptibility to different antifungals.

POPULATION GENETIC ANALYSIS OF BRAZILIAN PEACH BREEDING GERMPLASM

ABSTRACT Peach has great economic and social importance in Brazil. Diverse sources of germplasm were used to introduce desirable traits in the Brazilian peach breeding pool, composed mainly by local selections and accessions selected from populations developed by the national breeding programs, adapted to subtropical climate, with low chill requirement, as well as accessions introduced from several countries. In this research, we used SSR markers, selected by their high level of polymorphism, to access genetic diversity and population structure of a set composed by 204 peach selected genotypes, based on contrasting phenotypes for valuable traits in peach breeding. A total of 80 alleles were obtained, giving an average of eight alleles per locus. In general, the average value of observed heterozygosity (0.46) was lower than the expected heterozygosity (0.63). STRUCTURE analysis assigned 162 accessions splitted into two subpopulations based mainly on their flesh type: melting (96) and non-melting (66) flesh cultivars. The remaining accessions (42) could not be assigned under the 80% membership coefficient criteria. Genetic variability was greater in melting subpopulation compared to non-melting. Additionally, 55% of the alleles present in the breeding varieties were also present in the founder varieties, indicating that founding clones are well represented in current peach cultivars and advanced selections developed. Overall, this study gives a first insight of the peach genetic variability available and evidence for population differentiation (structure) in this peach panel to be exploited and provides the basis for genome-wide association studies.

Population viability analysis on a native Danish cattle breed

SummaryMany domestic breeds face challenges concerning genetic variability, because of their small population sizes along with a high risk of inbreeding. Therefore, it is important to obtain knowledge on their extinction risk, along with the possible benefits of certain breeding strategies. Since many domestic breeds face the same problems, results from such studies can be applied across breeds and species. Here a Population Viability Analysis (PVA) was implemented to simulate the future probability of extinction for a population of the endangered Danish Jutland cattle (Bos taurus), based on the software Vortex. A PVA evaluates the extinction risk of a population by including threats and demographic values. According to the results from the PVA the population will go extinct after 122 years with the current management. Four scenarios were created to investigate which changes in the breeding scheme would have the largest effect on the survival probabilities, including Scenario 1: More females in the breeding pool, scenario 2: More males in the breeding pool, scenario 3: Increased carrying capacity, and scenario 4: Supplementing males to the population through artificial insemination using semen from bulls used in the populations in past generations. All scenarios showed a positive effect on the population's probability of survival, and with a combination of the different scenarios, the population size seems to be stabilized.

Individual fluctuations in toxin levels affect breeding site fidelity in a chemically defended amphibian

Behaviours that influence habitat selection strongly determine species movement patterns. One component of animal behaviour that largely influences movement patterns and habitat choice is site fidelity. California newts (family Salamandridae) demonstrate remarkable site fidelity, typically homing to the same pool of a stream each breeding season. Individuals often occupy a specific pool throughout the breeding season, but some males shift among breeding pools, altering their set of potential mates, competitors, and predators. In this study, we measured dermal concentrations of the chemical defence compound tetrodotoxin (TTX) in recaptured male California newts ( Taricha torosa ) over five breeding seasons to evaluate whether relative TTX concentrations are associated with breeding site fidelity in the field. Our five years of field sampling indicates that TTX concentrations of individuals and group means fluctuate tremendously, implying that TTX is not a stable phenotypic trait. Despite such fluctuations, we found that an individual's relative TTX concentration explains fidelity to a breeding pool and suggests that newts may be able to assess both their own concentrations of TTX and that of conspecifics to make decisions about remaining in or abandoning a breeding pool. These results provide us a novel dimension to chemical defence phenotypes in nature and their ecological consequences, potentially requiring a re-evaluation of the coevolutionary dynamics of predation pressure on toxin-laden organisms.