Experimental population design for estimation of dominant molecular marker effect on egg-production traits

A potential limitation of the use of a dominant molecular marker system such as DNA fingerprinting (DFP) is the inability to distinguish homozygous from heterozygous allele state in an individual, and a resulting inaccuracy in estimating effects of the marker alleles. The objective of this study was to accurately estimate the effect of DFP markers on egg-production traits. A BC1 population was produced from two distinct layer lines. Four DFP bands, each originating predominantly in one of the two parental lines, were evaluated for linkage with egg-production quantitative trait loci in the BC1 population. The egg-production traits consisted of eight early period and seven late period measurements. Eight marker-trait linkages were identified out of 60 total statistical tests. By utilizing information on frequency of DFP bands in two parental lines, selecting F1 sires with DFP bands present, and backcrossing to the line lacking these bands, the population design allowed definitive identification of the DFP zygosity in the BC1 resource population hens. In this manner, accurate estimates of marker allele effects on egg-production traits were obtained from the dominant marker system of DNA fingerprinting.     

Genetic characterization of highly inbred chicken lines by two DNA methods: DNA fingerprinting and polymerase chain reaction using arbitrary primers

Thirteen highly inbred chicken lines were analysed at the DNA level by DNA fingerprinting (DFP) and by polymerase chain reaction (PCR) using random primers. In general, the DFP patterns of individuals within a line were identical. The DFP band-sharing (BS) values among lines from different breeds (Leghorn, Fayoumi, Spanish) ranged from 0.10 to 0.20. The DFP BS values among Leghorn lines from different genetic backgrounds ranged from 0.42 to 0.79. The DFP BS values among lines selected for different major histocompatibility complex serotypes from a common genetic background ranged from 0.70 to 0.95. Some randomly amplified polymorphic DNA (RAPD) PCR products were specific to a single line, some to all lines from the same genetic base, and some to all lines from the same breed. The RAPD-PCR band-sharing values ranged from 0.66 to 0.99 for all between-line comparisons. Thus, the ability to detect biodiversity at the DNA level was greater in this study for DFP than for RAPD-PCR. The possible origin of line-specific bands, relative advantages of detecting biodiversity by using different molecular screening techniques and uses of highly inbred chicken lines in molecular analysis are discussed.     

Genetic markers linked to quantitative traits in poultry

This study utilized DNA fingerprints and crosses of two genetically distinct lines of layer-type chickens to identify genetic markers linked to quantitative trait loci (QTL). In phase I, back-cross (BC¹) hens were separately ranked for each of eight traits and then blood pools were produced in groups along each phenotypic distribution. The DNA was isolated from the blood pools and used in a gradient analysis to screen for DNA fingerprint bands that exhibited intensity gradients associated with the phenotypic traits. To identify linkage of bands with QTL and to estimate band effects, F₂ progeny were produced in phase II from the phase I BC, population. A single-trait animal model was used for analysis of associations of all individual DNA fingerprint bands of sires and their progeny phenotypic performance. Twenty fingerprint bands, only two of which had shown trait-associated gradients in phase I, were identified by the animal model analysis of the progeny test as QTL linked (P≤005) to specific traits of growth, reproduction and egg quality. These 20 bands warrant further study as potentially valuable molecular markers for QTL.     

多个SPF鸡品系的RAPD分析

目的用RAPD方法分析三个品系SPF鸡的遗传变异,以指导SPF鸡群的育种工作.方法对三个SPF鸡品系F、M及B系和对照H纯系分别采血样30个,等量混匀为三个混合样,提取DNA,筛选随机引物,优化反应体系后进行PCR扩增,计算品系内平均相似系数,根据品系间遗传距离进行聚类分析并绘制树状聚类图.结果60个随机引物中有13个产生清晰稳定的多态条带,B系品系内平均相似系数为0.903,F系和M系分别为0.673和0.755,H系为0.968;聚类分析的结果是每个品系的三个样品先聚为一类,然后F系与M系、B系与对照H系分别聚为一类.结论F系和M系的纯合程度均较低,而B系较高;F系与M系的遗传距离较近,而与B系之间距离较远,三者亲缘关系以F系与M系较近,这与各个品系的引种背景相一致.     

Prediction of Heterosis from DNA Fingerprints in Chickens

To assess the value of DNA fingerprints for the prediction of heterosis in chickens, retrospective analyses of data from three crossbreeding experiments and DNA fingerprints (DFP) of parental strains were conducted using two minisatellite and one middle-repetitive DNA probes. DFP bands were assessed on pooled DNA samples of 10-15 individuals per parental genetic group. The number of DFP bands evaluated in the experiments ranged from 81 to 139. The probes varied in their predictive value, but predictability of heterosis generally increased with multiple probes. Highly significant correlations (0.68-0.87) between band sharing ratios (SH) and heterosis were found in 25 crosses of White Leghorns in the first egg production cycle for age at sexual maturity, egg production, and mature body weight: traits with heterosis of 10% or more of the means. Regressions of SH explained 78.4% of the variation in heterosis in age at sexual maturity, 60.2% in egg production and 46.4% in mature body weight. For ``broiler' traits with heterosis of <1%, none of the correlations, based on 13 crosses, were significant. It was concluded that multilocus probe DFP of pooled DNA samples show promise as predictors of heterosis.     

Genome inheritance in populations derived from hexaploid/tetraploid and tetraploid/hexaploid wheat crosses

Hexaploid/tetraploid and tetraploid/hexaploid wheat hybrids were established using the hexaploid (Triticum aestivum L.) bread wheat LRC2010-150 and the tetraploid durum wheat (T. turgidum spp. durum) WID802. Thirty F₂ progeny from each cross were characterised using Diversity Arrays Technology (DArTseq?) markers to determine whether there are differences between the crosses in the proportion of A, B and D genomic material inherited from each parent. Inheritance of the A and B genome from the tetraploid durum parent varied from 32 to 63% among the 60 lines assessed, and results indicated significant differences between the two F₂ populations in the mean overall proportion of chromosomes A and B inherited from each parent. Significant differences were also observed between the crosses in the proportion of chromosomal segments on 2B, 3A, 3B and 4A inherited from the tetraploid parent. The F₂ populations also showed significant differences in the average retention of D chromosomes per line with the tetraploid/hexaploid cross retaining a mean of 2.83 chromosomes while the reciprocal cross retained a mean of 1.8 chromosomes per line. A strong negative correlation was observed in individual lines from both populations between the proportion of the A and B genome inherited from the tetraploid durum parent and the retention of the D genome. The implication of these results for the design of efficient crossing strategies between hexaploid and tetraploid wheats is discussed.     

GENETIC VARIABILITY WITHIN A POPULATION AND BETWEEN DIPLOID/HAPLOID TISSUE OF MACROCYSTIS PYRIFERA (PHAEOPHYCEAE)1

Multi-locus DNA fingerprints using an M13 probe were obtained for eight individuals of giant kelp Macrocystis pyrifera (L.) C. Ag. collected from Monterey Bay, California. For each individual, DNA was extracted from a diploid blade and from ca. 10⁹ haploid spores that were released from four to Jive sporophylls. Viable or swimming spores from one individual were pooled and referred to as a spore group. A total of 34 bands (4–19 kb) was detected in DNA fingerprints from the eight blades and eight spore groups, with individual blade or spore groups exhibiting 7–18 bands (mean = 12.6). One band (4.5 kb) was present in all 16 samples. Eight bands were detected in 11–14 of the 16 samples. Similarity indices were calculated for all pairwise comparisons of fingerprint bands among all possible combinations of blades and spore groups. Mean similarity indices for the eight blades (0.51, SE = 0.032) and spore groups (0.56, SE = 0.031) were significantly lower than for the eight comparisons of the blade and spore groups from a single individual (0.86, SE = 0.052). The data indicate that DNA fingerprints can be used to measure genetic variation within populations of M. pyrifera because variation of DNA fingerprints associated with meiotic products (spores) of a given individual is small relative to variation observed among individuals within the population. Additionally, fingerprint variation between diploid vegetative tissue and haploid meiotic products may be a measure of genetic change due to recombination or DNA turnover mechanisms.     

A new approach for obtaining rapid uniformity in rice (Oryza sativa L.) via a 3x x 2x cross

A triploid (2n = 3x = 36) rice plant was obtained by screening a twin seedling population in which each seed germinated to two or three sprouts that were then crossed with diploid plants. One diploid plant was chosen among the various F(1) progenies and developed into an F (2) population via self-pollination. Compared with the control variety Shanyou 63, this F (2) population had a stable agronomical performance in field trials, as confirmed by the F-test. The stability of the F (2) population was further substantiated by molecular analysis with simple sequence repeat markers. Specifically, of 160 markers assayed, 37 (covering all 12 chromosomes) were polymorphic between the parental lines. Testing the F (1) hybrid individually with these markers showed that each PCR product had only a single band instead of two bands from each parent. The bands were identical to either maternal (23 markers) or paternal (eight markers) bands or distinct from both parents (six markers). The amplified bands of all 60 randomly selected F (2) plants were uniform and identical to those of the F (1) hybrid. These results suggest that the F (1) plant is a non-segregating hybrid and that a stable F (2) population was obtained. This novel system provides an efficient means for shortening the cycle of hybrid rice seed production.     

Hybrid rice (Oryza sativa L.): identification and parentage determination by RAPD fingerprinting

Summary DNA from three families of rice plants selected in Northern China (each comprising the male sterile, the restorer, the hybrid F1 and the maintainer lines) has been extracted and amplified by PCR with different random DNA primers (RAPD analysis). Then, DNA has been analysed by agarose gel electrophoresis and DNA bands scored as present or absent. The generated matrices are reproducible and amenable for identification of each single plant line. Thus, RAPD fingerprinting of the inbred parental lines and of the resulting hybrid is proposed as a convenient tool for the identification, protection and parentage determination of plant hybrids. Furthermore, by offering a molecular tool to verify the degree of dissimilarity between the parental lines, the RAPD analysis may also be used to search for new parental combinations.     

Detection of QTL for immune response to sheep red blood cells in laying hens

The aim of this study is to detect quantitative trait loci (QTL) involved in the regulation of the primary and the secondary immune response to sheep red blood cells (SRBC) in a resource population using microsatellite DNA markers. The F2 resource population originates from a cross of two divergently selected lines for either high (H line) or low (L line) primary antibody response to SRBC. The F2 population consisted of six half-sib families, three families per each of reciprocal crosses. Total antibody titres to SRBC were determined by agglutination in serum from all birds. F2, F1 and F0 generations were genotyped for 170 microsatellite markers, using a whole-genome scan approach. The half-sib and the line-cross analyses were performed to determine QTL regions associated with regulation of the immune response. In the half-sib analysis, four QTL for SRBC primary response have been identified: on GGA3, GGA5, GGA16 and GGA23. No QTL was identified for SRBC secondary response under the half-sib model. In the line-cross analysis, three QTL were identified on GGA10, GGA16 and GGA27 for SRBC primary response and five QTL were identified on GGA6, GGA9, GGA15, GGA16 and GGA27 for SRBC secondary response. Subsequently, the family contribution of individual families to the QTL was analysed. The family with the largest contribution was genotyped with additional microsatellite markers in the QTL region on GGA5. The extended half-sib analysis with additional genotype information results in narrowing down the QTL region on GGA5.     

Genetic variation and relationship of alfalfa populations and their progenies based on RAPD markers

The aim of investigation was to evaluate genetic variation and relationship among alfalfa populations and their offspring, with minimal cost, by using DNA marker analysis. RAPD analysis was performed on bulked DNA samples of five alfalfa parental populations and their progenies: 20 F1 populations from reciprocal diallel crosses and five S1 populations from self-pollination. Twenty primers generated 217 bands, ranging in size from 300 to 6000 bp, with the average number of bands per primer of 10.85 and polymorphism information content of 0.246. Percentage of polymorphic loci, effective number of alleles, expected heterozygosity and Shannon’s information index were used to estimate genetic variation. Higher diversity was observed in F1 progeny populations, while genetic variation in parental populations and S1 progenies remained similar. The genetic relatedness of alfalfa populations was analysed by UPGMA and Bayesian model-based clustering approach. In both types of analysis selfpollinated progenies were grouped. Furthermore, the hybrid offspring where Zuzana, and RSI 20 were maternal parents were placed in separate groups. The results indicate that use of RAPD markers on bulked DNA samples can be fast and cost-effective way for differentiation of alfalfa parental populations and their offspring, as well as for evaluation of their genetic relationships.     

DNA fingerprints of chickens selected for high and low body weight for 31 generations

Two lines of White Plymouth Rock chickens that have been divergently selected for 8-week body weight for 31 generations were compared for patterns of DNA fingerprints (DFP). Digestion of DNA with HinfI and hybridization to Jeffreys' minisatellite probe 33.6 resulted in DFPs that were relatively similar within lines (bandsharing = 0.50) and less similar between lines (bandsharing = 0.22). Analyses of scorable DFP bands produced by mixing DNA from individuals within lines indicated that 48% were line-specific. Causes for the differences in DFP patterns between lines and for occurrence of line-specific bands for the two lines divergently selected for body weight are discussed.     

Assessment of genomic variability through DNA fingerprinting within and between chicken lines divergently selected for residual food consumption

DNA fingerprints obtained by multi-locus probes have been shown to be convenient tools to assess genetic variability and genetic distances in animal populations. This method has been applied in the present study to two Rhode Island Red lines of chickens divergently selected for residual food consumption (RFC) for 16 generations (R-: low RFC; R+: high RFC). Within each line, individuals were sampled from each tail of the phenotypic distribution of RFC for males and for females. The three probes used were two minisatellite probes, R18-1 and 33-6, and the endogenous avian retroviral element probe. Altogether, 101 bands were scored. Band-sharing levels obtained from analysis of pooled DNA were extremely high within-Line (> 0.9) and still high between the lines (0.82 to 0.86). When band frequencies were compared, correlation coefficients were lower between lines than between extreme groups within lines regardless of the performance level. Although the selected lines differed widely in the selected trait RFC and a few correlated traits, it was concluded that they are probably different by only a low proportion of their genome.     

Inheritance of resistance to Bacillus thuringiensis subsp. kurstaki in Trichoplusia ni

The genetic inheritance of resistance to a commercial formulation of Bacillus thuringiensis subsp. kurstaki was examined in a Trichoplusia ni colony initiated from a resistant population present in a commercial vegetable greenhouse in British Columbia, Canada. Progeny of F(1) reciprocal crosses and backcrosses between F(1) larvae and resistant (P(R)) and susceptible (P(S)) populations were assayed at different B. thuringiensis subsp. kurstaki concentrations. The responses of progeny of reciprocal F(1) crosses were identical, indicating that the resistant trait was autosomal. The 50% lethal concentration for the F(1) larvae was slightly higher than that for P(S), suggesting that resistance is partially recessive. The responses of both backcross progeny (F(1) x P(R), F(1) x P(S)) did not correspond to predictions from a single-locus model. The inclusion of a nonhomozygous resistant parental line in the monogenic model significantly increased the correspondence between the expected and observed results for the F(1) x P(R) backcross but decreased the correspondence with the F(1) x P(S) backcross results. This finding suggests that resistance to B. thuringiensis subsp. kurstaki in this T. ni population is due to more than one gene.     

Genetic architecture of growth and body composition in unique chicken populations

A resource population was established by crossing one modern broiler sire from a commercial broiler breeder male line with dams from two unrelated highly inbred lines; F1 birds were intercrossed to produce two F2 populations. A variety of phe notypic measurements related to growth, muscling, internal organs, and skeleton were recorded for the F2 populations and contemporary pure inbred and broiler birds. Based on the means and phenotypic distributions of the F2 populations com pared to their parental lines, the effective number of genes affecting each trait and heterosis were estimated and discussed relative to the known genetic selection history for each trait. The results suggest that a high number of genes with small epistatic effects are involved in determining the phenotype for traits that broilers were traditionally selected for, and a lower number of genes with major effects are involved in determining the phenotype for traits related to fitness. The estimated number of genes and the phenotypic distributions of the different traits suggest that a quantitative trait loci (QTL) search might be more effectively applied for traits with a low number of involved genes and a high phenotypic distribution among the F2 birds than for traits that show a lower phenotypic distribution and a high number of genes.     

Inheritance of female flight in Lymantria dispar (Lepidoptera: Lymantriidae)

A clinal female flight polymorphism exists in the gypsy moth, Lymantria dispar L., where female flight diminishes from east to west across Eurasia. A Russian population where females are capable of sustained ascending flight and a North American population with females incapable of flight were crossed: parentals, reciprocal F(1) hybrids, double reciprocal F(2) hybrids, and all possible backcrosses to both the parental lines were compared. Heritabilities were estimated using a threshold model, female offspring on female parent regressions, and joint-scaling analyses. Heritability of female flight capability measured using a free flight test was at least 0.60, and variation in wing size, muscle strength, and flight behaviors contributed to the flight polymorphism. Relative wing size varied continuously and had a heritability of 0.70. Environmental variation accounted for >90% of the variation in female preflight weight and relative flight muscle strength, as estimated by an inverted female's ability to right herself. Preflight walking behavior and early deposition of eggs were each inherited through a single gene with two co-dominant alleles. There was no evidence for sex-linkage or maternal effects in female flight capability or associated traits. Continued vigilance to exclude and eradicate introductions of strains capable of female flight in North America is warranted even in areas where no females fly, because some of the alleles needed for full flight capability may not be present in the North American populations, and some flight capability is maintained in the hybrids that could increase the rate of spread of L. dispar.     

The effect of Lr19-translocation on in vitro androgenesis and inheritance of leaf-rust resistance in DH3 lines and F2 hybrids of common wheat

Leaf-rust resistance and androgenesis were studied in the anther cultures of Triticum aestivum L., which included Saratovskaya 29 cultivar, the isogenic line Ps29, and three F1 hybrids (L503/S55, L504/S58, ATS7/L1063) with 7DS-7DL-7Ae#1L translocation of Lr19 gene (Lr19 translocation) from Agropyron elongatum (Host) P.B. The Lr19 translocation was shown to affect the induction of embryogenesis and green plant regeneration. The frequencies of Lr19 translocation differed in F2 hybrids obtained by traditional hybridization and in sets of DH lines obtained in F1 anther cultures derived from the same combinations of T. aestivum parental forms. The number of leaf-rust resistant genotypes tended to decrease. The frequency of Lr19 translocation in the set of DH3 lines derived from F1 L504/S58 was significantly lower than in other sets of DH3 lines and F2 hybrid populations.     

RFLP Variations of Common Wheat Doubled Haploid Progenies from Wheat×Maize Crosses

Wheat ( Triticum aestivum L. ) and maize ( Zea mays L. ) crosses (the chromosome elimination system) can be used to produce frequently a large number of doubled haploid (DH) wheat lines by embryo rescue and doubling treatment. The resulting DH lines are genetically homogeneous. Significant RFLP variations were detected in common wheat DH progenies from wheat and maize crosses by using wheat rDNA clone pta71 and two maize DNA clones (MR13 and MRSO) homologous to wheat genome as probes. The results revealed that the copy number and restriction fragment length of rDNA in some wheat DH progenies was changed, and also that deletion was detected in several DH plants when probed with MR13 and MR5O. In particular, the RFLP pattern of DH line No. 18 was greatly changed using MR13 as a probe. In this line, three new bands, 40.0 kb, 2.5 kb and 2.0 kb emerged while a 4.3 kb intense band from the parental common wheat genome disappeared. This change may be related to a quite large DNA rearrangement within the wheat genomic DNA or an insertion by alien maize DNA fragment.     

小麦与多年生黑麦草原生质体的电融合及杂种愈伤组织的形成

多年生黑麦草(Lolium perenne L.)悬浮培养细胞来源的原生质体和小麦(Triticumaestivum L.)含叶绿体的悬浮培养细胞来源的原生质体间,用直流方波脉冲进行电融合,获得了体细胞杂种愈伤组织。小麦的原生质体经过碘乙酰胺失活。愈伤组织的形态和颜色被用作识别预期杂种的标记。为了对杂种愈伤组织进行同工酶分析,观察了亲本的9种同工酶谱,其中3种在亲本间表现出差异(ADH、GOT 和 SDH)。酒精脱氢酶(ADH)的分析结果表明,有6个细胞系表现出杂种带。这些细胞系经过其他两种同工酶分析和 rDNA 探针杂交试验表明,一个细胞系表现出基本完全的亲本基因组间的组合,其余5个细胞系是部分杂种。     

Contributions of heterosis and epistasis to hybrid fitness

Early-generation hybrid fitness is difficult to interpret because heterosis can obscure the effects of hybrid breakdown. We used controlled reciprocal crosses and common garden experiments to distinguish between effects of heterosis and nuclear and cytonuclear epistasis among morphotypes and advanced-generation hybrid derivative populations in the Piriqueta caroliniana (Turneraceae) plant complex. Seed germination, growth, and sexual reproduction of first-generation hybrids, inbred parental lines, and outbred parental lines were compared under field conditions. Average vegetative performance was greater for hybrids than for inbred lines, and first-season growth was similar for hybrids and outbred parental lines. Hybrid survival surpassed that of inbred lines and was equal to or greater than outbred lines' survival, and more F(1) than parental plants reproduced. Reductions in hybrid fitness due to Dobzhansky-Muller incompatibilities (epistasis among divergent genetic elements) were expressed as differences in vegetative growth, survival, and reproduction between plants from reciprocal crosses for both F(1) and backcross hybrid generations. Comparing performance of hybrids against parental genotypes from intra- and interpopulation crosses allowed a more robust prediction of F(1) hybrids' success and more accurate interpretations of the genetic architecture of F(1) hybrid vigor.