Number Of Different Alleles Research Articles

Genetic relatedness of sweet cherry (Prunus avium L.) cultivars from Ukraine determined by microsatellite markers

ABSTRACTMicrosatellite (SSR) markers were used to characterise 23 sweet cherry cultivars of Ukrainian, and four cultivars of non-Ukrainian, origin. To assess their genetic diversity and relatedness, 11 pairs of primers were applied to microsatellite loci, resulting in amplification of 66 SSR alleles. The mean value of the number of different alleles, and the polymorphic index content, amount to 7.333 and 0.700, respectively, demonstrating a significant genetic diversity of the investigated sweet cherry cultivars. Four highly polymorphic SSR loci (EMPAS02, EMPAS06, PceGA34, UDP98-412), which belong to the list recommended by the European Cooperative Program for Plant Genetic Resources, can be used as a minimum genetic marker set for identification of the majority of the studied cultivars; however, for successful discrimination of the most similar cultivars, more markers, located on all chromosomes of sweet cherry, appear to be necessary. Application of unweighted variable-group method using averages clustering allowed elucidation of the relatedness among the sweet cherry varieties, and showed that the Ukrainian cultivars combine genetic material of local, western European, and probably Caucasian origin; however, the origin of several cultivars still remains unclear, and should be studied additionally.

Large-scale analysis of variation in the insulin-like growth factor family in humans reveals rare disease links and common polymorphisms

The insulin-like growth factors IGF1 and IGF2 are closely related proteins that are essential for normal growth and development in humans and other species and play critical roles in many physiological and pathophysiological processes. IGF actions are mediated by transmembrane receptors and modulated by IGF-binding proteins. The importance of IGF actions in human physiology is strengthened by the rarity of inactivating mutations in their genes and by the devastating impact caused by such mutations on normal development and somatic growth. Large-scale genome sequencing has the potential to provide new insights into human variation and disease susceptibility. Toward this end, the availability of DNA sequence data from 60,706 people through the Exome Aggregation Consortium has prompted the analyses presented here. Results reveal a broad range of potential missense and other alterations in the coding regions of every IGF family gene, but the vast majority of predicted changes were uncommon. The total number of different alleles detected per gene in the population varied over an ∼15-fold range, from 57 for IGF1 to 872 for IGF2R, although when corrected for protein length the rate ranged from 0.22 to 0.59 changes/codon among the 11 genes evaluated. Previously characterized disease-causing mutations in IGF2, IGF1R, IGF2R, or IGFALS all were found in the general population but with allele frequencies of <1:30,000. A few new highly prevalent amino acid polymorphisms were also identified. Collectively, these data provide a wealth of opportunities to understand the intricacies of IGF signaling and action in both physiological and pathological contexts.

The New Genomics: What Molecular Databases Can Tell Us About Human Population Variation and Endocrine Disease.

Major recent advances in genetics and genomics present unique opportunities for enhancing our understanding of human physiology and disease predisposition. Here I demonstrate how analysis of genomic information can provide new insights into endocrine systems, using the human growth hormone (GH) signaling pathway as an illustrative example. GH is essential for normal postnatal growth in children, and plays important roles in other biological processes throughout life. GH actions are mediated by the GH receptor, primarily via the JAK2 protein tyrosine kinase and the STAT5B transcription factor, and inactivating mutations in this pathway all lead to impaired somatic growth. Variation in GH signaling genes has been evaluated using DNA sequence data from the Exome Aggregation Consortium, a compendium of information from >60,000 individuals. Results reveal many potential missense and other alterations in the coding regions ofGH1,GHR,JAK2, andSTAT5B, with most changes being uncommon. The total number of different alleles per gene varied by ~threefold, from 101 forGH1 to 338 forJAK2. Several known disease-linked mutations inGH1,GHR, andJAK2 were present but infrequent in the population; however, three amino acid changes inGHR were sufficiently prevalent (~4% to 44% of chromosomes) to suggest that they are not disease causing. Collectively, these data provide new opportunities to understand how genetically driven variability in GH signaling and action may modify human physiology and disease.

Molecular epidemiology of Brucella abortus isolated from cattle in Brazil, 2009–2013

The aims of the present study were to genotype Brucella abortus strains isolated from cattle in Brazil between 2009 and 2013, and to analyze their distribution to support the Programa Nacional de Controle e Erradicação de Brucelose e Tuberculose (PNCEBT) (National Brucellosis and Tuberculosis Control and Eradication Program). One hundred forty B. abortus strains isolated from cattle in Brazil between 2009 and 2013 were genotyped using a set of 18 variable number of tandem repeats (VNTR) (MLVA16+HOOF-Print 3 and 4). The multiple locus VNTR analysis (MLVA) composed by eight markers (MLVA8) revealed eight different genotypes among B. abortus strains, including five previously described and three new ones. Analysis of the MLVA16 loci revealed fifty-eight distinct genotypes, from which three were identical, thirty-eight were considered very close, and seventeen were considered distant compared to those previously described and deposited in MLVAbank. Analysis of the HOOF-Prints 3 and 4 revealed the larger number of different alleles among all VNTR assessed, exhibiting maximum resolution when associated with MLVA16 markers. This study also provides insights on the genotypes of B. abortus circulating in Brazil, which certainly contribute for the better understanding of the epidemiology and control of bovine brucellosis in the country. Moreover, our data showed a high genetic diversity among the B. abortus strains isolated between 2009 and 2013, and a close relationship among these strains and Brazilian B. abortus deposited by MLVAbank.

Genetic diversity of the Dwarf honeybee (Apis florea Fabricius, 1787) populations based on microsatellite markers.

Apis florea is one of two species of small, wild honeybee. The present study was conducted to evaluate the genetic diversity of Apis florea honeybee from 48 nests (colonies) using microsatellite markers in the South of Iran. All honeybee samples were analyzed for six microsatellite loci (A88, A107, A7, B124, A113 and A35). The six loci had different numbers of alleles in the sampled colonies ranging from 7 (loci A107) to 3 (loci A7, A35). Gene diversity in Apis florea ranged from 0.491 to 0.595. This range probably reflects the spreading of nests in a large region with a varied climate. Phylogenetic tree showed two distinct clusters including a) Minab region samples and b) Bandar Abbas, Bandar Khamir and Qeshm Island regions. All of these regions are geographically rich, having varied vegetation and climate conditions. Our findings are an important contribution to the methods of studying distribution and conservation of Apis florea.

P114 Validation of high resolution HLA typing using NXTypeTM Ion-Torrent next generation sequencing. The University of Miami experience

Aim HLA typing using next generation sequencing (NGS) takes the Sanger based sequencing HLA typing to a higher dimension. The technology permits multiplexing of several target regions and various samples in a single run. The advantages of this technology include high throughput, improved DNA sequencing accuracy and faster data analysis. Our aim was validate the NXTypeTM Ion-Torrent NGS HLA typing procedure to implement it in our clinical laboratory. Methods 214 samples were analyzed with NXType™ Ion-Torrent NGS and compared to high resolution typing obtained by Sanger sequencing. HLA Class I (full HLA-A, -B, and -C genes) and HLA Class II (Exon 2 thru intron 3 of HLA-DR and HLA-DQ, and Exon 2 thru 3’UTR of HLA-DP) were amplified with NXType™ Class I and Class II primers set, respectively. Amplicons were fragmented with Ion Shear™ Plus Reagents Kit to sizes from 350 to 950bp. Libraries were enriched in OneTouch ES and run in an Ion PGM™. Data was analyzed with HLATypeStream (v1.0.0.86) software. Results We obtained good coverage of the target regions when multiplexing 24 samples per run using a 318™ v2 chip. Of the 1070 alleles analyzed (214 samples), 143 were homozygous and 927 heterozygous. The concordance between Sanger sequencing results and NGS was 99.2%, 99.6%, 99.8%, 100% and 99.8% for loci A, B, C, DRB1 and DQB1, respectively. The number of different alleles identified for loci A, B, C, DRB1 and DQB1 were 40, 69, 36, 42, and 18, respectively. Our results revealed that 269 NMDP codes reported by Sanger sequencing were resolved by NGS, except 3 codes in DRB1 because of mismatches present in exons 1 or 4, which are not targeted by NXType™ Class II primers. We found two new alleles in exons (A ∗ 24, exon 1 and C ∗ 01, exon 2) and variations in intronic regions of 46 alleles. All null alleles A ∗ 68:11N (10), B ∗ 15:01N (11) and C ∗ 04:09N/B ∗ 44:03 (3) seen by Sanger were resolved by NGS. Conclusions The NGS is a reliable and accurate method to determine HLA typing of a considerable number of samples relatively fast, solving most allele ambiguities (98.7%) without the need of alternative tests. We confirmed that NXTypeTM Ion-Torrent NGS is a suitable and quick method for routine use in clinical laboratories to obtain complete ultra-high-resolution HLA typing that ultimately will improve on current HLA matching for transplant.

Genetic diversity of Plasmodium falciparum isolates from uncomplicated malaria cases in Ghana over a decade.

BackgroundGenotyping malaria parasites to assess their diversity in different geographic settings have become necessary for the selection of antigenic epitopes for vaccine development and for antimalarial drug efficacy or resistance investigations. This study describes the genetic diversity of Plasmodium falciparum isolates from uncomplicated malaria cases over a ten year period (2003–2013) in Ghana using the polymorphic antigenic marker, merozoite surface protein 2 (msp2).MethodsArchived filter paper blood blots from children aged nine years and below with uncomplicated malaria collected from nine sites in Ghana were typed for the presence of the markers. A total of 880 samples were genotyped for msp2 for the two major allelic families, FC27 and 3D7, using nested polymerase chain reaction (PCR). The allele frequencies and the multiplicity of infection were determined for the nine sites for five time points over a period of ten years, 2003–2004, 2005–2006, 2007–2008, 2010 and 2012–2013 malaria transmission seasons.ResultsThe number of different alleles detected for the msp2 gene by resolving PCR products on agarose gels was 14. Both of the major allelic families, 3D7 and FC27 were common in all population samples. The highest multiplicity of infection (MOI) was observed in isolates from Begoro (forest zone, rural site): 3.31 for the time point 2007–2008. A significant variation was observed among the sites in the MOIs detected per infection (Fisher's exact test, P < 0.001) for the 2007 isolates and also at each of the three sites with data for three different years, Hohoe, P = 0.03; Navrongo, P < 0.001; Cape Coast, P < 0.001. Overall, there was no significant difference between the MOIs of the three ecological zones over the years (P = 0.37) and between the time points when data from all sites were pooled (P = 0.40).ConclusionsThe diversity and variation between isolates detected using the msp2 gene in Ghanaian isolates were observed to be profound; however, there was homogeneity throughout the three ecological zones studied. This is indicative of gene flow between the parasite populations across the country probably due to human population movements (HPM).Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1692-1) contains supplementary material, which is available to authorized users.

Major histocompatibility complex class II DAB alleles associated with intestinal parasite load in the vulnerable Chinese egret (Egretta eulophotes).

The maintenance of major histocompatibility complex (MHC) polymorphism has been hypothesized to result from many mechanisms such as rare‐allele advantage, heterozygote advantage, and allele counting. In the study reported herein, 224 vulnerable Chinese egrets (Egretta eulophotes) were used to examine these hypotheses as empirical results derived from bird studies are rare. Parasite survey showed that 147 (65.63%) individuals were infected with 1–3 helminths, and 82.31% of these infected individuals carried Ascaridia sp. Using asymmetric polymerase chain reaction technique, 10 DAB1, twelve DAB2, and three DAB3 exon 2 alleles were identified at each single locus. A significant association of the rare allele Egeu‐DAB2*05 (allele frequency: 0.022) with helminth resistance was found for all helminths, as well as for the most abundant morphotype Ascaridia sp. in the separate analyses. Egeu‐DAB2*05 occurred frequently in uninfected individuals, and individuals carrying Egeu‐DAB2*05 had significantly lower helminth morphotypes per individual (HMI) (the number of HMI) and the fecal egg count values. Further, the parasite infection measurements were consistently lower in individuals with an intermediate number of different alleles in the duplicated DAB loci. Significantly, heterozygosity within each DAB locus was not correlated with any parasite infection measurements. These results indicate that the diversity in MHC Egeu‐DAB gene is associated with intestinal parasite load and maintained by pathogen‐driven selection that probably operate through both the rare‐allele advantage and the allele counting strategy, and suggest that Egeu‐DAB2*05 might be a valuable indicator of better resistance to helminth diseases in the vulnerable Chinese egret.

Characterization of Expression Quantitative Trait Loci in Pedigrees from Colombia and Costa Rica Ascertained for Bipolar Disorder.

The observation that variants regulating gene expression (expression quantitative trait loci, eQTL) are at a high frequency among SNPs associated with complex traits has made the genome-wide characterization of gene expression an important tool in genetic mapping studies of such traits. As part of a study to identify genetic loci contributing to bipolar disorder and other quantitative traits in members of 26 pedigrees from Costa Rica and Colombia, we measured gene expression in lymphoblastoid cell lines derived from 786 pedigree members. The study design enabled us to comprehensively reconstruct the genetic regulatory network in these families, provide estimates of heritability, identify eQTL, evaluate missing heritability for the eQTL, and quantify the number of different alleles contributing to any given locus. In the eQTL analysis, we utilize a recently proposed hierarchical multiple testing strategy which controls error rates regarding the discovery of functional variants. Our results elucidate the heritability and regulation of gene expression in this unique Latin American study population and identify a set of regulatory SNPs which may be relevant in future investigations of complex disease in this population. Since our subjects belong to extended families, we are able to compare traditional kinship-based estimates with those from more recent methods that depend only on genotype information.

Population genetic analysis of Theileria parva isolated in cattle and buffaloes in Tanzania using minisatellite and microsatellite markers

A population genetic study of Theileria parva was conducted on 103 cattle and 30 buffalo isolates from Kibaha, Lushoto, Njombe Districts and selected National parks in Tanzania. Bovine blood samples were collected from these study areas and categorized into 5 populations; Buffalo, Cattle which graze close to buffalo, Kibaha, Lushoto and Njombe. Samples were tested by nested PCR for T. parva DNA and positives were compared for genetic diversity to the T. parva Muguga vaccine reference strain, using 3micro and 11 minisatellite markers selected from all 4 chromosomes of the parasite genome. The diversity across populations was determined by the mean number of different alleles, mean number of effective alleles, mean number of private allele and expected heterozygosity. The mean number of allele unique to populations for Cattle close to buffalo, Muguga, Njombe, Kibaha, Lushoto and Buffalo populations were 0.18, 0.24, 0.63, 0.71, 1.63 and 3.37, respectively. The mean number of different alleles ranged from 6.97 (Buffalo) to 0.07 (Muguga). Mean number of effective alleles ranged from 4.49 (Buffalo) to 0.29 (Muguga). The mean expected heterozygosity were 0.07 0.29, 0.45, 0.48, 0.59 and 0.64 for Muguga, cattle close to buffalo, Kibaha, Njombe, Lushoto and Buffalo populations, respectively. The Buffalo and Lushoto isolates possessed a close degree of diversity in terms of mean number of different alleles, effective alleles, private alleles and expected heterozygosity. The study revealed more diversity in buffalo isolates and further studies are recommended to establish if there is sharing of parasites between cattle and buffaloes which may affect the effectiveness of the control methods currently in use.

GENETIC VARIATION OF E. oleifera POPULATION AND POPULATION E. guineensis x E. oleifera ON NUTFAH PLASMA COLLECTION PPKS

The oil palm species of Elaeis oleifera have some superior characters not possessed by E. guineensis species. The cross between E. oleifera x E. guineensis (E. o x E. g.) Is carried out to insert the superior character of E. oleifera into the E. guineensis species and to obtain individuals who possess a superior characteristic blend of both species. This study aims to determine the genetic diversity within and between populations of E. oleifera, as well as relationships between E. oleifera, E. guineensis and its interspecific hybrids. A total of 8 germplasm populations used were E. oleifera originating from Suriname and Brazil, E. guineensis population of Dura Deli and SP540T, and interspecific crosses of hybrid E. guineensis x E. oleifera from Colombia, hybrid E. guineensis x E. oleifera which is presumably from Colombia, as well as hybrid E. guineensis x E. oleifera from Brazil. A total of sixteen Simple Sequence Repeat (SSR) markers were used to analyze 92 individuals from 8 populations. PCoA results show that 8 Elaeis populations cluster in each group. The hybrid population of E. guineensis x E. oleifera suspected of Colombia has a very close kinship relationship with hybrid E. guineensis x E. oleifera from Colombia and clumped in the same quadrant. This suggests that the hybrid is indeed a hybrid of E. guineensis x E. oleifera from Colombia. The population of E. oleifera from Suriname showed the lowest genetic diversity, with the number of different alleles, specific alleles, heterozygosity values, and the percentages of polymorphic loci sequentially were 1.37, 0.18, 0.09, and 37.50%. While the population of interspecific interspecific crossbreeds of the E. oleifera species showed the highest genetic diversity with values ​​of 3.81, 0.43, 0.62 and 100%.

Mitochondrial DNA-based genetic diversity and population structure of Zymoseptoria tritici in Tunisia

A total of 108 isolates of the wheat pathogen Zymoseptoria tritici were collected from four distinct locations of Tunisia and characterized for three mitochondrial DNA sequences to provide insight into the genetic diversity and population structure of the fungus in this country. The number of different alleles averaged over all loci within locations ranged from 3.33 to 5.67, with an average of 4.42 per location. Multilocus analysis identified 27 (25 %) distinct haplotypes among the 108 assessed isolates, revealing a high mtDNA-based clonality within the population. Three of the highlighted haplotypes occurred in all locations and nine of them covered at least two locations. Significant levels of genetic diversity were found for the whole population and within each of the sampled locations, as indicated by the Nei’s gene diversity (0.52), unbiased gene diversity (0.58) and allele richness (4.43) indices. Further analyzes using Bayesian and non-Bayesian statistical models, as well as AMOVA, showed a lack of mtDNA-based genetic structure (GST = 0.06), hence supporting previous reports on Z. tritici in Tunisia performed using nuclear-DNA markers. A high level of gene flow (Nm = 8.27) corroborates the lack of structure and suggests regular cycles of sexual reproduction, leading to allelic pool homogenization via wind-born ascospores in the Tunisian population of Z. tritici.

Maximization of minority classes in core collections designed for association studies

Core collections are nowadays widely employed in diverse studies on plant genetics. The more extensively used method to build core collections (maximization strategy) is based on the selection, from a global collection, of those accessions which maximize the number of different alleles and phenotypic classes (classes’ richness). However, different core collections should be created for different types of studies, and though several years ago most of core collections were developed to make the characterization and use of germplasm collections easier with a smaller sample size, for either conservation or breeding purposes, today, they are widely employed for association studies that are broadly applied in plant genetic improvement. Following the M strategy, some alleles or phenotypic classes often appear in a very low frequency, which may reduce the power of the analysis, avoiding the detection of real associations (false negatives). In this work, we propose and evaluate a new way to build core collections using the maximization strategy in several sequential steps, to maximize the frequency of minority classes, thus increasing the statistical power of the association study.

Can genetic diversity be maintained across multiple mass selection lines of Sydney rock oyster, Saccostrea glomerata despite loss within each?

This study assesses the genetic diversity of four different hatchery bred lines of Sydney rock oyster, Saccosstrea glomerata (SRO) after three, five and seven generations of mass selection using microsatellite DNA markers and mitochondrial cytochrome C oxidase subunit 1 sequences (COX1). This was achieved using seven newly developed microsatellite markers, along with three published loci. Considering the same number of samples in each line, the number of different alleles in each of the four hatchery lines ranged from 49 to 70 alleles from 10 loci, which was approximate 50% less than the 120 alleles found in the samples from the wild population. However, if we pool all our four hatchery lines together, then their total number of alleles was not significantly different from the wild population. Similar to the patterns found for the DNA microsatellite alleles, mtDNA haplotype numbers were considerably lower in each of the hatchery lines than in the samples from the wild population. Again, pooling the hatchery lines gave a total haplotype number not statistically significantly different from that in the samples from the wild. Considering together the DNA microsatellite alleles and the mtDNA haplotypes, we conclude that there was a substantial loss of genetic diversity within all lines separately over generations of mass selection, and that such mass selection was not sustainable in the long term. However, importantly, these data indicate that for oysters, if multiple independent mass selection lines are kept, then their combined genetic diversity may approach that of the ancestral line or wild population even after many generations of mass selection. These results for oysters, finding preservation of diversity among lines yet loss within them, are similar to recent findings for multiple banana shrimp lines; considering both species together it suggests that one relatively simple option suitable for some farms and industry to maintain genetic diversity during mass selection over many generations of selection is to subdivide their breeding nucleus and keep multiple different and independent lines over generations. It remains to be tested if this approach is general across many aquaculture species and circumstances. Statement of relevanceThis paper attempts to falsify the published hypothesis set out by Knibb et al., 2014 that “the use of multiple lines under some circumstances may be a hedge against loss of diversity” using a second independent example, namely Sydney rock oysters.The data of this paper do not refute the generality of the above hypothesis of Knibb et al., 2014 that splitting mass selection lines into multiple sublines preserves genetic variation.

Analysis of population structure and genetic diversity of Egyptian and exotic rice (Oryza sativa L.) genotypes

Understanding the population structure and genetic diversity is a very important goal to improve the economic value of crops. In rice, a loss of genetic diversity in the last few centuries is observed. To address this challenge, a set of 22 lines from three different regions – India (two), and Philippines (six), and Egypt (14) – were used to assess the genetic diversity and the features of population structure. These genotypes were analyzed using 106 SSR alleles that showed a clear polymorphism among the lines. Genetic diversity was estimated based on the number of different alleles, polymorphism information content (PIC), and gene diversity. A total of 106 SSR alleles was identified from the 23 SSR loci and used to study the population structure and carry out a cluster analysis. All SSR loci showed a wide range of the number of different alleles extended from two (one loci) to seven alleles (three loci). Five and eight loci showed high PIC and gene diversity (≥0.70), respectively. The results of population structure are in agreement with cluster analysis results. Both analyses revealed two different subpopulations (G1 and G2) with different genetic properties in number of private alleles, number of different alleles (Na), number of effective alleles (Ne), expected heterozygosity (He), and Shannon's Information Index (SII). Our findings indicate that five SSR loci (RM 111, RM 307, RM 22, RM 19, and RM 271) could be used in breeding programs to enhance the marker-assisted selection through QTL mapping and association studies. A high genetic diversity found between genotypes which can be exploited to improve and produce rice cultivars for important traits (e.g. high agronomic features and tolerance to biotic or/and abiotic stresses).

Examination of Genetic Diversity of Common Bean from the Western Balkans

In this study, genetic diversity of 119 accessions of common bean (Phaseolus vulgaris) from five former Yugoslav republics constituting the western Balkans was assessed by 13 microsatellite markers. This set of markers has proven before to efficiently distinguish between bean genotypes and assign them to either the Andean or the Mesoamerican gene pool of origin. In this study, 118 alleles were detected or 9.1 per locus on average. Four groups (i.e., Slovene, Croatian, Bosnian, and Serbian) showed similarly high levels of genetic diversity as estimated by the number of different alleles, number of effective alleles, Shannon’s information index, and expected heterozygosity. Mildly narrower genetic diversity was identified within a group of Macedonian accessions; however, this germplasm yielded the highest number of private alleles. All five germplasms share a great portion of genetic diversity as indicated by the analysis of molecular variance (AMOVA). On the basis of the scored number of migrants, we concluded that the most intensive gene flow in the region exists in Bosnia and Herzegovina. Cluster analysis based on collected molecular data classified the accessions into two large clusters that corresponded to two gene pools of origin (i.e., Andean and Mesoamerican). We found that Andean genotypes are more prevalent than Mesoamerican in all studied countries, except Macedonia, where the two gene pools are represented evenly. This could indicate that common bean was introduced into the western Balkans mainly from the Mediterranean Basin. Bayesian cluster analysis revealed that in the area studied an additional variation exists which is related to the Andean gene pool. Different scenarios of the origin of this variation are discussed in the article.

Genetic characterization of red-colored heartwood genotypes of Chinese fir using simple sequence repeat (SSR) markers.

The present study investigated the genetic characterization of red-colored heartwood Chinese fir [Cunninghamia lanceolata (Lamb.) Hook.] in Guangxi using 21 simple sequence repeat (SSR) markers and analyzes of the genetic variation (N = 149) in samples obtained from five sites in Guangxi Province, China. The number of different alleles and the Shannon's information index per locus ranged from 3 to 12 and from 0.398 to 2.258 with average values of 6 and 1.211, respectively, indicating moderate levels of genetic diversity within this germplasm collection. The observed and expected heterozygosity ranged from 0.199 to 0.827 and from 0.198 to 0.878 with an average of 0.562 and 0.584, respectively. Although, the mean fixation index was 0.044, indicative of a low level of genetic differentiation among germplasms, analysis of molecular variance revealed considerable differentiation (99%) within the samples. The neighbor-joining dendrogram revealed that the majority of red-colored Chinese fir genotypes were apparently not associated with their geographic origins. Further analysis by STRUCTURE showed that this Guangxi germplasm collection could be divided into three genetic groups comprising 76, 37, and 36 members, respectively; these were classified into mixed groups with no obvious population structure. These results were consistent with those of the cluster analysis. On the whole, our data provide a starting point for the management and conservation of the current Guangxi germplasm collection as well as for their efficient use in Chinese fir-breeding programs.

Swine Leukocyte Antigen (SLA) class I allele typing of Danish swine herds and identification of commonly occurring haplotypes using sequence specific low and high resolution primers

The swine major histocompatibility complex (MHC) genomic region (SLA) is extremely polymorphic comprising high numbers of different alleles, many encoding a distinct MHC class I molecule, which binds and presents endogenous peptides to circulating T cells of the immune system. Upon recognition of such peptide-MHC complexes (pMHC) naïve T cells can become activated and respond to a given pathogen leading to its elimination and the generation of memory cells. Hence SLA plays a crucial role in maintaining overall adaptive immunologic resistance to pathogens.Knowing which SLA alleles that are commonly occurring can be of great importance in regard to future vaccine development and the establishment of immune protection in swine through broad coverage, highly specific, subunit based vaccination against viruses such as swine influenza, porcine reproductive and respiratory syndrome virus, vesicular stomatitis virus, foot-and-mouth-disease virus and others.Here we present the use of low- and high-resolution PCR-based typing methods to identify individual and commonly occurring SLA class I alleles in Danish swine. A total of 101 animals from seven different herds were tested, and by low resolution typing the top four most frequent SLA class I alleles were those of the allele groups SLA-3*04XX, SLA-1*08XX, SLA-2*02XX, and SLA-1*07XX, respectively. Customised high resolution primers were used to identify specific alleles within the above mentioned allele groups as well as within the SLA-2*05XX allele group. Our studies also suggest the most common haplotype in Danish pigs to be Lr-4.0 expressing the SLA-1*04XX, SLA-2*04XX, and SLA-3*04XX allele combination.

House sparrowPasser domesticussurvival is not associated with MHC-I diversity, but possibly with specific MHC-I alleles

The MHC (Major Histocompatibility Complex) plays an important role in the immune system of vertebrates. MHC genes are extremely polymorphic and this variation is considered to be maintained by selection from pathogens. We investigate whether MHC diversity (number of different alleles per individual) affects the survival and recruitment of nestling house sparrows. We hypothesize that individuals with higher MHC diversity can recognize and combat a wider range of pathogens, and therefore are more likely to survive and recruit into the breeding population. Additionally, we hypothesize that specific MHC class I alleles (MHC-I) could be associated with survival and recruitment. We screened MHC-I genotypes in 518 house sparrow chicks hatched on Lundy Island but we found no evidence for a relationship between nestling survival, post-fledging survival or recruitment success with MHC diversity. Then we investigated effects of specific MHC-I alleles in 195 individuals from a single cohort. Twenty-one MHC-I alleles were tested for relationships with nestling survival, post-fledging survival and recruitment, and we detected associations with survival for three different alleles. This pattern was, however, not different to what would be expected from random, so we could not conclude that particular MHC-I alleles are associated with survival in house sparrows on Lundy Island. Nonetheless, one of these alleles (1105) showed both a tendency for a higher probability of surviving in nestlings, and a significant association with survival in fledglings. We envision that allele 1105 could be an interesting candidate gene for testing associations with survival in house sparrows in the future.

Conditional asymmetric linkage disequilibrium (ALD): extending the biallelic r2 measure.

For multiallelic loci, standard measures of linkage disequilibrium provide an incomplete description of the correlation of variation at two loci, especially when there are different numbers of alleles at the two loci. We have developed a complementary pair of conditional asymmetric linkage disequilibrium (ALD) measures. Since these measures do not assume symmetry, they more accurately describe the correlation between two loci and can identify heterogeneity in genetic variation not captured by other symmetric measures. For biallelic loci the ALD are symmetric and equivalent to the correlation coefficient r. The ALD measures are particularly relevant for disease-association studies to identify cases in which an analysis can be stratified by one of more loci. A stratified analysis can aid in detecting primary disease-predisposing genes and additional disease genes in a genetic region. The ALD measures are also informative for detecting selection acting independently on loci in high linkage disequilibrium or on specific amino acids within genes. For SNP data, the ALD statistics provide a measure of linkage disequilibrium on the same scale for comparisons among SNPs, among SNPs and more polymorphic loci, among haplotype blocks of SNPs, and for fine mapping of disease genes. The ALD measures, combined with haplotype-specific homozygosity, will be increasingly useful as next-generation sequencing methods identify additional allelic variation throughout the genome.