Computation of identity by descent probabilities conditional on DNA markers via a Monte Carlo Markov Chain method

The accurate estimation of the probability of identity by descent (IBD) at loci or genome positions of interest is paramount to the genetic study of quantitative and disease resistance traits. We present a Monte Carlo Markov Chain method to compute IBD probabilities between individuals conditional on DNA markers and on pedigree information. The IBDs can be obtained in a completely general pedigree at any genome position of interest, and all marker and pedigree information available is used. The method can be split into two steps at each iteration. First, phases are sampled using current genotypic configurations of relatives and second, crossover events are simulated conditional on phases. Internal track is kept of all founder origins and crossovers such that the IBD probabilities averaged over replicates are rapidly obtained. We illustrate the method with some examples. First, we show that all pedigree information should be used to obtain line origin probabilities in F2 crosses. Second, the distribution of genetic relationships between half and full sibs is analysed in both simulated data and in real data from an F2 cross in pigs.     

Prediction of identity by descent probabilities from marker-haplotypes

The prediction of identity by descent (IBD) probabilities is essential for all methods that map quantitative trait loci (QTL). The IBD probabilities may be predicted from marker genotypes and/or pedigree information. Here, a method is presented that predicts IBD probabilities at a given chromosomal location given data on a haplotype of markers spanning that position. The method is based on a simplification of the coalescence process, and assumes that the number of generations since the base population and effective population size is known, although effective size may be estimated from the data. The probability that two gametes are IBD at a particular locus increases as the number of markers surrounding the locus with identical alleles increases. This effect is more pronounced when effective population size is high. Hence as effective population size increases, the IBD probabilities become more sensitive to the marker data which should favour finer scale mapping of the QTL. The IBD probability prediction method was developed for the situation where the pedigree of the animals was unknown (i.e. all information came from the marker genotypes), and the situation where, say T, generations of unknown pedigree are followed by some generations where pedigree and marker genotypes are known.     

Estimation of coefficient of coancestry using molecular markers in maize

Summary The coefficient of coancestry (f_AB) between individuals A and B is the classical measure of genetic relationship. f_AB is determined from pedigree records and is the probability that random alleles at the same locus in A and B are copies of the same ancestral allele or identical by descent (ibd). Recently, the proportion of molecular marker variants shared between A and B (S_AB) has been used to measure genetic relationship. But S_AB is an upwardly-biased estimator of f_AB, especially between distantly-related lines. f_AB, S_AB, and adjusted (to remove bias) estimates of molecular marker similarity (f _AB ^M ) were compared. RFLP banding patterns at 46 probe-restriction enzyme combinations were obtained for 23 maize inbred lines derived from the Iowa Stiff Stalk Synthetic (BSSS) maize (Zea mays L.) population, and for 4 non-BSSS lines. f _AB ^M was estimated as , where _A (or _B) was the average proportion of RFLP variants shared between inbred A (or inbred B) and the non-BSSS lines. The average f_AB among 253 pairwise combinations of BSSS lines was 0.212, whereas the average S_AB was 0.397. The average f _AB ^M was 0.162, indicating that the upward bias in S_AB was effectively removed. S_AB and f_AB were significantly different ( = 0.05) in 76.3% of the comparisons, whereas 24.9% of the f _AB ^M values differed significantly from f_AB. The latter result suggests that selection and/or drift were present during inbred line development and that f_AB may not be an accurate measure of the true proportion of ibd alleles between two lines. Cluster analyses based on S _AB ^M and f _AB ^M grouped lines according to pedigree, although several exceptions were noted. The presence of shared molecular marker variants between unrelated lines must be considered when setting S_AB-based minimum distances for varietal protection. Under simplified conditions, more than 250 molecular marker loci are necessary to obtain sufficiently precise estimates of coefficient of coancestry using molecular markers.A contribution from Limagrain Genetics, a Group Limagrain company     

Identification of genetic factors for alachlor tolerance in maize by molecular markers analysis

Genetic factors controlling tolerance to the herbicide Alachlor in maize were localised by means of two different strategies. In the first approach, backcross (BC) plants, derived from pollen which had been subjected to selective pressure for resistance to the herbicide, were analysed for segregation distortion at 47 RFLP loci and compared to BC plants obtained from non-selected pollen. Preferential transmission of five chromosomal regions where putative QTLs (Quantitative Trait Loci) are localised was revealed in the BC plants from selected pollen. A second approach was based on a classical linkage analysis for segregation of the same set of RFLPs and factors controlling the trait, in a BC population of 210 individuals, by means of regression analysis. This study detected seven significant loci in four genomic regions. Overall, two loci revealed both segregation distortion and association with the expression of the trait, indicating linkage to genes expressed in both gametophytic and sporophytic phase. Three chromosomal regions appeared to carry factors involved in plant tolerance to Alachlor which are not expressed in pollen. Conversely, three loci were linked to factors selectable in pollen, but did not reveal significant association with tolerance in the plant in the segregating populations.     

Marker‐based estimates of relatedness and inbreeding coefficients: an assessment of current methods

Inbreeding (F) of and relatedness (r) between individuals are now routinely calculated from marker data in studies in the fields of quantitative genetics, conservation genetics, forensics, evolution and ecology. Although definable in terms of either correlation coefficient or probability of identity by descent (IBD) relative to a reference, they are better interpreted as correlations in marker‐based analyses because the reference in practice is frequently the current sample or population whose F and r are being estimated. In such situations, negative estimates have a biological meaning, a substantial proportion of the estimates are expected to be negative, and the average estimates are close to zero for r and equivalent to F_IS for F. I show that although current r estimators were developed from the IBD‐based concept of relatedness, some of them conform to the correlation‐based concept of relatedness and some do not. The latter estimators can be modified, however, so that they estimate r as a correlation coefficient. I also show that F and r estimates can be misleading and become biased and marker dependent when a sample containing a high proportion of highly inbred and/or closely related individuals is used as reference. In analyses depending on the comparison between r (or F) estimates and a priori values expected under ideal conditions (e.g. for identifying genealogical relationship), the estimators should be used with caution.     

A score test for the linkage analysis of qualitative and quantitative traits based on identity by descent data from sib-pairs

We propose a general likelihood-based approach to the linkage analysis of qualitative and quantitative traits using identity by descent (IBD) data from sib-pairs. We consider the likelihood of IBD data conditional on phenotypes and test the null hypothesis of no linkage between a marker locus and a gene influencing the trait using a score test in the recombination fraction theta between the two loci. This method unifies the linkage analysis of qualitative and quantitative traits into a single inferential framework, yielding a simple and intuitive test statistic. Conditioning on phenotypes avoids unrealistic random sampling assumptions and allows sib-pairs from differing ascertainment mechanisms to be incorporated into a single likelihood analysis. In particular, it allows the selection of sib-pairs based on their trait values and the analysis of only those pairs having the most informative phenotypes. The score test is based on the full likelihood, i.e. the likelihood based on all phenotype data rather than just differences of sib-pair phenotypes. Considering only phenotype differences, as in Haseman and Elston (1972) and Kruglyak and Lander (1995), may result in important losses in power. The linkage score test is derived under general genetic models for the trait, which may include multiple unlinked genes. Population genetic assumptions, such as random mating or linkage equilibrium at the trait loci, are not required. This score test is thus particularly promising for the analysis of complex human traits. The score statistic readily extends to accommodate incomplete IBD data at the test locus, by using the hidden Markov model implemented in the programs MAPMAKER/SIBS and GENEHUNTER (Kruglyak and Lander, 1995; Kruglyak et al., 1996). Preliminary simulation studies indicate that the linkage score test generally matches or outperforms the Haseman-Elston test, the largest gains in power being for selected samples of sib-pairs with extreme phenotypes.     

Multivariate analysis of polypeptide synthesis in field-grown maize inbreds and hybrids

Summary A leaf disc method is described to permit the localized incorporation of ³⁵S-l-methionine into polypeptides synthesized in individual leaves of maize plants grown in the field. The method of incorporation employs minimal external manipulation of the intact leaf, is simple, repeatable, and may be used at any plant age after leaf emergence. Incorporation (cpm/g protein) in 12 leaves per plant was compared among three inbred (Oh43, W23, M14) and three F₁ hybrid (Oh43/M14, W23/M14, Oh43/W23) genotypes. The incorporation was 40% higher (hybrid versus inbred) in 9 of the 12 leaves studied. Samples from leaf 07 (7th leaf numbered from base of plant) for four inbreds (Oh43, M14, B73, Mol 7) and two pairs of reciprocal F₁ hybrids (Oh43/M14, M14/Oh43; B73/Mo17, Mo17/B73) were labelled in situ using the leaf disc method. Each cultivar was sampled at three different ages in each of 1985, 1986, and 1987. High-resolution, two-dimensional isoelectric focusing sodium-dodecyl-sulfate polyacrylamide gel electrophoresis and fluorography were used to display the polypeptides synthesized in the samples. Multivariate methods — Principal Coordinate Analysis, Cluster Analysis, and Standard Deviation Distance — were used to analyze variation and to identify trends in the variation for year, genotype, and age sampled. Our analyses disclose a hierarchy to polypeptide synthesis variation in maize leaves: differences in polypeptide synthesis are greater for year-to-year comparisons than differences due to sample age, which in turn are greater than differences for inbred versus hybrid comparisons.     

Parental contribution and coefficient of coancestry among maize inbreds: pedigree, RFLP, and SSR data

The genetic relationship between inbreds i and j can be estimated from pedigree or from molecular marker data. The objectives of this study were to: (1) determine whether pedigree, restriction fragment length polymorphism (RFLP), and simple sequence repeat (SSR) data give similar estimates of parental contribution and coefficient of coancestry (f _ij ) among a set of maize (Zea mays L.) inbreds, and (2) compare the usefulness of RFLP and SSR markers for estimating genetic relationship. We studied 13 maize inbreds with known pedigrees. The inbreds were genotyped using 124 RFLP and 195 SSR markers. For each type of marker, parental contributions were estimated from marker similarity among an inbred and both of its parents, and were subsequently used to estimate f _ij . Estimates of parental contribution differed significantly (α<0.05) between pedigree data and either type of marker, but not between the marker systems. The RFLP estimates of parental contribution failed to sum to 1.0, reflecting a higher frequency of non-parental bands with RFLP than with SSR markers. The f _ij estimated from pedigree, RFLP, and SSR data were highly correlated (r=0.87–0.97), although significant differences were found among the three sets of f _ij estimates. We concluded that pedigree and marker data often lead to different estimates of parental contribution and f _ij , and that SSR markers are superior to RFLP markers for estimating genetic relationship. A relevant question is whether or not the inbreds previously genotyped with an older marker system (e.g., RFLP) need to be re-analyzed with a newer marker system (e.g., SSR) for the purpose of estimating genetic relationship. Such re-analysis seems unnecessary if data for the same type of marker are available for a given inbred and both of its parents. Received: 2 June 1999 / Accepted: 30 July 1999     

Genes encoding oleosins in maize kernel of inbreds Mo17 and B73

We have investigated all three oleosin genes which are expressed in the kernel of maize (Zea mays L., Mo17). Oleosin genes, ole16, ole17, and ole18, encode OLE16, OLE17, and OLE18, respectively, in proportional amounts of approximately 2:1:1 in isolated oil bodies. None of the three genes has an intron or a sequence encoding an N-terminal signal peptide. The three genes are expressed coordinately during seed maturation, and their encoded oleosins are present in similar proportional amounts in oil bodies isolated from the embryonic axis, scutellum, and aleurone layer. OLE16 represents one oleosin isoform, whereas OLE17 and OLE18 are close members of another oleosin isoform. ole16 and ole18 have been mapped to single loci on chromosomes 2 (near b1 gene) and 5S (near phya2), respectively. We predict that ole17 is located on chromosome 1 (near phya1), in a chromosomal segment duplicated on chromosome 5.     

Genetic relationships and clonal identity in a collection of commercially relevant poplar cultivars assessed by AFLP and SSR

A collection of 66 poplar commercial clones widely cultivated in Italy, China and in other countries of southern Europe and belonging to various poplar species and hybrids, have been fingerprinted using both amplified fragment length polymorphism (AFLP) and simple sequence repeats (SSR) techniques. Three AFLP primer combinations and six SSRs unambiguously genotyped the analysed poplar collection, with the exception of three groups of six, four and two individuals, which turned out to be indistinguishable even if they met the standards currently applied for distinctness, uniformity and stability (DUS) testing when registered. High levels of variation were detected with both molecular techniques; a total of 201 AFLP bands were amplified of which 96% turned out to be polymorphic and up to 15 SSR alleles were identified at a single locus, with a mean of 9.3 alleles per locus in the case of Populus × canadensis. The probability of matching fortuitously any two genotypes at all the SSR loci in the case of P. × canadensis was less then 7.5×10^–9. The AFLP-derived dendrogram and principal coordinate analysis (PCOORDA) clustered the clones with respect to their taxonomic classification, and allowed their genetic interrelationships to be established. Correct identification of poplar varieties is essential for ensuring the effective correspondence between the real and the declared identity of a clone, to avoid commercial frauds, and to establish breeding programmes. Molecular markers may play a major role to satisfy all these needs.     

Genetic improvement of anther culture response in maize: relationships with molecular, Mendelian and agronomic traits

 Two cycles of androgenetic in vitro doubled haploid (DH) plant production and intermating were implemented in an experimental synthetic population of maize. In vitro traits, including androgenetic embryo production, the regeneration potential and the frequency of spontaneous chromosome doubling, were studied. The success of the regenerated plants to self pollinate was also observed. Impressive genetic progress is reported for all the steps of the androgenetic process including seed set. Differential genetic progress is recorded according to the trait measured. Using a set of Mendelian and molecular markers that mapped to the different maize chromosomes, we were able to characterize the variation in the genetic variability in the population. Progress in the in vitro response was not found to be associated with any noticeable decline in global genetic variability. In addition, the QTL chromosomic regions tested, which were involved in androgenetic response, were not found to be subjected to a strong variation during the breeding experiment. Some phenotypical and morphological traits were also evaluated, and these showed that there was no depreciation effect in the agronomic value of the population. DH plant production and intermating the regenerated plants may be considered for the introduction and use of androgenesis in material which responds poorly. Received: 3 October 1997 / Accepted: 25 November 1997     

Prediction of maize hybrid performance using similarity in state and similarity by descent information

We evaluated the efficiency of the best linear unbiased predictor (BLUP) and the influence of the use of similarity in state (SIS) and similarity by descent (SBD) in the prediction of untested maize hybrids. Nine inbred lines of maize were crossed using a randomized complete diallel method. These materials were genotyped with 48 microsatellite markers (SSR) associated with the QTL regions for grain yield. Estimates of four coefficients of SIS and four coefficients of SBD were used to construct the additive genetic and dominance matrices, which were later used in combination with the BLUP for predicting genotypic values and specific combining ability (SCA) in unanalyzed hybrids under simulated unbalance. The values of correlations between the genotypic values predicted and the means observed, depending on the degree of unbalance, ranged from 0.48 to 0.99 for SIS and 0.40 to 0.99 using information from SBD. The results obtained for the SCA ranged from 0.26 to 0.98 using the SIS and 0.001 to 0.990 using the SBD information. It was also observed that the predictions using SBD showed less biased than SIS predictions demonstrating that the predictions obtained by these coefficients (SBD) were closer to the observed value, but were less efficient in the ranking of genotypes. Although the SIS showed a bias due to overestimation of relatedness, this type of coefficient may be used where low values are detected in the SBD in the group of parents because of its greater efficiency in ranking the candidates hybrids.     

Comparison of identity by descent and identity by state for detecting genetic regions under selection in a sorghum pedigree breeding program

Seventy sorghum inbred lines which formed part of the Queensland Department of Primary Industries (QDPI) sorghum breeding program were screened with 104 previously mapped RFLP markers. The lines were related by pedigree and consisted of ancestral source lines, intermediate lines and recent releases from the program. We compared the effect of defining marker alleles using either identity by state (IBS) or identity by descent (IBD) on our capacity to trace markers through the pedigree and detect evidence of selection for particular alleles. Allelic identities defined using IBD were much more sensitive for detecting non-Mendelian segregation in this pedigree. Only one marker allele showed significant evidence of selection when IBS was used compared with ten regions with particular allelic identities when IBD was used. Regions under selection were compared with the location of QTLs for agronomic traits known to be under selection in the breeding program. Only two of the ten regions were associated with known QTLs that matched with knowledge of the agronomic characteristics of the ancestral lines. Some of the other regions were hypothesised to be associated with genes for particular traits based on the properties of the ancestral source lines.     

Genomic inbreeding and population structure of northern pike (Esox lucius) in Xinjiang,China

Northern pike (Esox lucius) was widely distributed in the high latitudes of the northern hemisphere. In China, northern pike was originally distributed only in the upper reaches of the Irtysh River in Xinjiang and has appeared in many water bodies outside the Irtysh River Basin in Northern Xinjiang. A total of four populations were collected from north to south in Xinjiang, including Irtysh River (RIR), Ulungu Lake (LUL), a small lake nearby Ulungu River (LJD), and Bosten Lake (LBO). We estimated population genomic parameters, performed gene flow analysis, and estimated the effective population size of each population. The proportion of individuals with high inbreeding coefficient (F ≥ 0.0625) accounted for 36.4% (44/121) of all sequenced individuals, approximately 4.5% (1/22) in LUL, 25.9% (7/27) in LBO, 42.9% (18/42) in RIR, and 60% (18/30) in LJD. RIR had the highest mean of genomic relatedness (coancestry coefficient = 0.025 ± 0.040, IBD = 0.036 ± 0.078). Gene flow results showed that the population spreading was from RIR into two branches, one was LBO, and the other continued to split into LUL and LJD, and migration signal from LBO to LUL was detected. Our results suggested that the extinction risk of northern pike was very low in Xinjiang of China, and the controlled capture fishery of northern pike could be developed reasonably.     

Mapping quantitative trait loci controlling silking date in a diallel cross among four lines of maize

 We describe and apply an interval mapping method for quantitative trait locus (QTL) detection using F₃ and testcross progenies derived from F₂ populations obtained from a diallel cross among four elite lines of maize. Linear model-based procedures were used for the test and estimation of putative QTL effects together with genetic interactions including epistasis. We mapped QTL associated with silking date and explored their genetic effects. Ten QTL were detected, and these explained more than 40% of the phenotypic variance. Most of these QTL had consistent and stable effects among genetic backgrounds and did not show significant epistasis. QTL-by-environment interaction was important for four QTL and was essentially due to changes in magnitude of allelic effects. These results show the efficiency of our method in several genetic situations as well as the power of the diallel design in detecting QTL simultaneously over several populations. Received: 2 September 1996 / Accepted: 20 December 1996     

Comparative analysis of genetic similarity among maize inbred lines detected by RFLPs, RAPDs, SSRs, and AFLPs

 DNA-based fingerprinting technologies have proven useful in genetic similarity studies. RFLP is still most commonly used in the estimation of genetic diversity in plant species, but the recently developed PCR-based marker techniques, RAPDs, SSRs and AFLPs, are playing an increasingly important role in these investigations. Using a set of 33 maize inbred lines we report on a comparison of techniques to evaluate their informativeness and applicability for the study of genetic diversity. The four assays differed in the amount of polymorphism detected. The information content, measured by the expected heterozygosity and the average number of alleles, was higher for SSRs, while the lowest level of polymorphism was obtained with AFLPs. However, AFLPs were the most efficient marker system because of their capacity to reveal several bands in a single amplification. In fact, the assay efficiency index was more than ten-fold higher for AFLPs compared to the other methods. Except for RAPDs, the genetic similarity trees were highly correlated. SSR and AFLP technologies can replace RFLP marker in genetic similarity studies because of their comparable accuracy in genotyping inbred lines selected by pedigree. Bootstrap analysis revealed that, in the set of lines analysed, the number of markers used was sufficient for a reliable estimation of genetic similarity and for a meaningful comparison of marker technologies. Received: 11 April 1998 / Accepted: 19 May 1998     

Prediction of multi-locus inbreeding coefficients and relation to linkage disequilibrium in random mating populations

An algorithm to predict the level of identity by descent simultaneously at multiple loci is presented, which can in principle be extended to any number of loci. The model assumes a random mating population, with random association of haplotypes. The relationship is shown between coefficients of multi-locus identity or non-identity by descent and moments of multi-locus linkage disequilibrium. Thus, these moments can be computed from the multilocus identity or, using algorithms derived previously to predict the disequilibria moments, vice-versa. The results can be applied to predict multi-locus identity in, for example, gene mapping.     

Gene frequencies and admixture estimates in the state of Puebla, Mexico

Three hundred ninety-three individuals from the Universidad Autónoma de Puebla and its University Hospital were studied to determine the distribution of ABO, MN, Rh-Hr, Duffy, and Diego blood groups; red cell hemoglobin and glucose-6-phosphate dehydrogenase variants; and serum haptoglobins, albumins, and factor Bf types. With the results we estimated that the proportions of black, indian, and white genes are 10.7%, 56.3%, and 33.0%, respectively, in a trihybrid model. Reasons are given as to why the black ancestry may be artifically high, and it is pointed out that independent confirmation with other markers is needed before the figure can be accepted as a true value.     

Genetic diversity of maize inbred lines within and among heterotic groups revealed by RFLPs

Summary The objectives of this study were (1) to investigate genetic diversity for RFLPs in a set of important maize inbreds commonly used in Italian breeding programs, (2) to compare genetic similarities between unrelated lines from the same and different heterotic groups, and (3) to examine the potential of RFLPs for assigning maize inbreds to heterotic groups. Forty inbreds were analyzed for RFLPs with two restriction enzymes (EcoRI and HindIII) and 82 DNA clones uniformly distributed over the maize genome. Seventy clone-enzyme combinations gave single-banded RFLP patterns, and 79 gave multiple-banded RFLP patterns. The average number of RFLP patterns detected per clone-enzyme combination across all inbreds was 5.8. RFLP data revealed a wide range of genetic diversity within the two heterotic groups assayed, Iowa Stiff Stalk Synthetic (BSSS) and Lancaster Sure Crop (LSC). Genetic similarity (GS) between lines was estimated from binary RFLP data according to the method of Nei and Li (1979). The mean GS for line combinations of type BSSS × LSC (0.498) was substantially smaller than for unrelated line combinations or type BSSS × BSSS (0.584) but almost as great as for un-related line combinations of type LSC × LSC (0.506). Principal coordinate and cluster analyses based on GS values resulted in the separate groupings of lines, which is consistent with known pedigree information. A comparison between both methods for multivariate analyses of RFLP data is presented.