Genetic analysis of low survival rate of pups in RR/Sgn inbred mice

Newborn offspring of the inbred mouse RR/Sgn strain have a low survival rate prior to weaning. We hypothesized that this is a consequence of an inferior nurturing ability of RR/Sgn mothers and that RR/Sgn mothers have a tendency to lose their pups. We performed quantitative trait locus (QTL) mapping for inferior nurturing ability and tendency to lose pups in RR/Sgn mothers. The number of pups was adjusted to 6 per dam on the day of delivery, and the number of surviving pups and their total weight (litter weight) were scored at 12 days after birth. Nurturing ability was evaluated by litter weight, and tendency to lose pups was evaluated by scoring whether or not the mothers lost their pups. For litter weight, we identified one significant QTL on chromosome 4 and three suggestive QTLs on chromosomes 7, 9 and 17. The RR/Sgn allele was associated with lower litter weight at all loci. For the tendency to lose pups, we identified three suggestive QTLs on chromosomes 4, 9 and 16. The RR/Sgn allele was associated with an increased tendency to lose pups at all loci. These results supported our hypothesis that the low survival rate phenotype was attributable, at least in part, to a phenotype whereby mothers display inferior nurturing ability and a tendency to lose pups. Thus, it suggests that these two traits share genetic basis.     

Genetic analysis of litter size in mice

We performed quantitative trait locus (QTL) mapping analysis for litter size (total number of pups born and/or number of pups born alive) in 255 backcross mice derived from C57BL/6J and RR/Sgn inbred mice. We identified one significant QTL on chromosome 7 and 4 suggestive QTLs on chromosomes 3, 5, 10 and 13. In addition, two suggestive QTLs were identified on chromosomes 1 and 4 for the number of stillbirth. These results suggested that both litter size and number of stillbirth were heritable traits, although they were controlled by distinct genes. The RR allele was associated with reduced litter size and increased stillbirth at all QTLs. Therefore, RR mothers were observed to have reduced prolificacy in this particular genetic cross.     

Genetic mapping of the QTL affecting body weight in chickens using a F2 family

1. To identify the quantitative trait loci (QTL) affecting growth in chickens, we carried out QTL analysis on chicken growth traits using a population of 227 F2 crosses between a Satsumadori (slow-growing, light-weight Japanese native breed used as a meat chicken) male and a White Plymouth Rock (early-maturing, heavy weight broiler). 2. We chose 78 microsatellite loci from 331 publicly available on 14 linkage groups, with respect to their utility and location. 3. Two QTLs affecting body weight at 13 and 16 weeks were mapped at 220 cM on chromosome 1 (LOD scores, 2.8 and 4.5, respectively, at 13 and 16 weeks), and at 60 cM on chromosome 2 (LOD scores, 6.2 and 8.1, respectively, at 13 and 16 weeks). 4. The closest loci to the QTLs were LEI71 on chromosome 1 and LMU13 and MCW184 on chromosome 2. 5. The sites of the QTLs agreed closely with those already reported. Therefore, it seems likely that QTLs affecting growth of chickens are located at these sites.     

Characterization of Cq3, a quantitative trait locus that controls plasma cholesterol and phospholipid levels in mice

Cq3 was identified in C57BL/6J (B6) x KK-Ay F2 mice as a quantitative trait locus (QTL) that controls plasma cholesterol and phospholipid levels, and normolipidemic B6 allele was associated with increased lipids. Cq3 was statistically significant in F2-a/a, but not in F2-Ay/a; probably because the Cq3 effect was obscured by introduction of the Ay allele, which in itself has a strong hyperlipidemic effect. Because the peak LOD score for Cq3 was identified near D3Mit102 (49.7 cM) on chromosome 3, linkage analyses with microsatellite markers located at 49.7 cM were performed in KK x RR F2, B6 x RR F2, and KK x CF1 F2. However, even a suggestive QTL was not identified in any of the three F2. By testing all pairs of marker loci, I found a significant interaction between Cq3 and the Apoa2 locus, and F2 mice with the Apoa2(KK)/Apoa2(KK); D3Mit102(B6)/D3Mit102(B6) genotype had significantly higher cholesterol levels than did F2 mice with other genotypes. The results showed that the ;round-robin' strategy was not always applicable to the search for QTL genes; probably because specific gene-to-gene interaction limited the validity of the strategy to the utmost extent.     

Quantitative trait locus analysis of plasma cholesterol levels and body weight by controlling the effects of the Apoa2 allele in mice

Colleagues and I previously performed quantitative trait locus (QTL) analysis on plasma total-cholesterol (T-CHO) levels in C57BL/6J (B6) x RR F2 mice. We identified only one significant QTL (Cq6) on chromosome 1 in a region containing the Apoa2 gene locus, a convincing candidate gene for Cq6. Because Cq6 was a highly significant QTL, we considered that the detection of other potential QTLs might be hindered. In the present study, QTL analysis was performed in B6.KK-Apoa2b N(8) x RR F2 mice [B6.KK-Apoa2b N(8) is a partial congenic strain carrying the Apoa2b allele from the KK strain, and RR also has the Apoa2b allele] by controlling of the effects of the Apoa2 allele, for identifying additional QTLs. Although no significant QTLs were identified, 2 suggestive QTLs were found on chromosomes 2 and 3 in place of the effects of the Apoa2 allele. A significant body weight QTL was identified on chromosome 3 (Bwq7, peak LOD score 5.2); its effect on body weight was not significant in previously analyzed B6 x RR F2 mice. Suggestive body weight QTL that had been identified in B6 x RR F2 mice on chromosome 4 (LOD score 3.8) was not identified in B6.KK-Apoa2b N(8) x RR F2 mice. Thus, contrary to expectation, the genetic control of body weight was also altered significantly by controlling of the effects of the Apoa2 allele. The QTL mapping strategy by controlling of the effects of a major QTL facilitated the identification of additional QTLs.     

凉山半细毛羊初生重QTL的研究

选取凉山半细毛羊7个父系半同胞家系,利用绵羊1号染色体上11个微卫星标记,通过聚合酶链式反应（polymerase chain reaction,PCR）对凉山半细毛羊初生重QTL进行了研究。QTL分析的结果表明,在其中一个家系B3K5055中,1号染色体微卫星标记ILSTS004与CSSM004之间检测到影响初生重的QTL,其LOD值为3.338,位于置信区间95%（2.73）和99%（3.49）之间。     

高丹草低氢氰酸含量性状主效QTL PA7-2的精细定位

为了精细定位高丹草低氢氰酸含量性状主效数量性状基因座QTL PA7-2,对进一步开展低氰性状相关候选基因挖掘、功能解析及分子标记辅助育种等研究提供理论依据。本试验在前期研究工作基础上,从高丹草（散穗高粱Scattered ear Sorghum bicolor×红壳苏丹草Red shell Sorghum sudanense）F₂代1 200个分离群体单株中筛选出121个QIRs（Quantitative trait locus（QTL）isogenic recombinants）植株,选出低氰和高氰极端株套袋自交获得F₃代分离群体,选出QIRs群体植株130个,利用BSA-SSR（Bulked segregation analysis（BSA）and simple sequence repeats）技术构建了长度为230.7 cM、密度为4.81 cM的遗传连锁图谱,通过定位分析明确了QTL PA7-2的位置在38 cM处,位于SSR标记Sobic.8 g1-600和XM00242-400之间。经与高粱基因组比对分析,首次将低氰QTL PA7-2精细定位至高粱8号染色体3.77 Mb（51.415 Mb~55.182 Mb）的物理区间,并发现SSR标记SORBI4G3-600与其紧密连锁。     

Primary genome scan to identify putative quantitative trait loci for feedlot growth rate, feed intake, and feed efficiency of beef cattle

Feed intake and feed efficiency of beef cattle are economically relevant traits. The study was conducted to identify QTL for feed intake and feed efficiency of beef cattle by using genotype information from 100 microsatellite markers and 355 SNP genotyped across 400 progeny of 20 Angus, Charolais, or Alberta Hybrid bulls. Traits analyzed include feedlot ADG, daily DMI, feed-to-gain ratio [F:G, which is the reciprocal of the efficiency of gain (G:F)], and residual feed intake (RFI). A mixed model with sire as random and QTL effects as fixed was used to generate an F-statistic profile across and within families for each trait along each chromosome, followed by empirical permutation tests to determine significance thresholds for QTL detection. Putative QTL for ADG (chromosome-wise P < 0.05) were detected across families on chromosomes 5 (130 cM), 6 (42 cM), 7 (84 cM), 11 (20 cM), 14 (74 cM), 16 (22 cM), 17 (9 cM), 18 (46 cM), 19 (53 cM), and 28 (23 cM). For DMI, putative QTL that exceeded the chromosome-wise P < 0.05 threshold were detected on chromosomes 1 (93 cM), 3 (123 cM), 15 (31 cM), 17 (81 cM), 18 (49 cM), 20 (56 cM), and 26 (69 cM) in the across-family analyses. Putative across-family QTL influencing F:G that exceeded the chromosome-wise P < 0.05 threshold were detected on chromosomes 3 (62 cM), 5 (129 cM), 7 (27 cM), 11 (16 cM), 16 (30 cM), 17 (81 cM), 22 (72 cM), 24 (55 cM), and 28 (24 cM). Putative QTL influencing RFI that exceeded the chromosome-wise P < 0.05 threshold were detected on chromosomes 1 (90 cM), 5 (129 cM), 7 (22 cM), 8 (80 cM), 12 (89 cM), 16 (41 cM), 17 (19 cM), and 26 (48 cM) in the across-family analyses. In addition, a total of 4, 6, 1, and 8 chromosomes showed suggestive evidence (chromosome-wise, P < 0.10) for putative ADG, DMI, F:G, and RFI QTL, respectively. Most of the QTL detected across families were also detected within families, although the locations across families were not necessarily the locations within families, which is likely because of differences among families in marker informativeness for the different linkage groups. The locations and direction of some of the QTL effects reported in this study suggest potentially favorable pleiotropic effects for the underlying genes. Further studies will be required to confirm these QTL in other populations so that they can be fine-mapped for potential applications in marker-assisted selection and management of beef cattle.     

Refined mapping of quantitative trait loci for somatic cell score on BTA02 in the German Holstein

The aim of this study was to more precisely map a previously reported quantitative trait locus (QTL) affecting somatic cell score on Bos taurus autosome 2 by increasing the number of markers fourfold, analysing more families and exploiting within‐population linkage disequilibrium (LD). A granddaughter design of 10 German Holstein grandsire families with 1121 progeny tested sons was used. Twenty‐six markers with an average marker spacing of 3.14 cM were genotyped along 81.6 cM. Linkage analysis (LA) was performed using variance‐component methodology. The incorporation of LD was first done using variance‐component methods followed by regression on marker alleles. LA revealed genome‐wide significance (LOD > 3) at 15 contiguous marker‐intervals, with the maximum test‐statistic between DIK2862 and BMS778 and a 1‐lod drop‐off interval of 38 cM. While the variance‐component methods could not detect any LD, two individual markers with a significant effect (ILSTS098, p < 0.05; BMS778, p < 0.01) were found by regression analysis. Compared with previous results QTL‐localisation was substantially narrowed; further fine‐mapping should focus on the close vicinity of BMS778.     

Bone mineral density QTL at sexual maturity and end of lay

1. An F₂ cross of a broiler male line and a White Leghorn layer line was used to identify quantitative trait loci (QTL) for bone density at the onset of lay and at the end of the laying period. A total of 686 measures of humeral bone density were available for analysis.

2. There was no evidence for epistasis.

3. Genome-wide significant QTL for bone density at the onset of lay were identified on chromosomes 1 (311?cM) and 8 (2?cM) and on chromosomes 1 (311?cM), 3 (57?cM) and 8 (2?cM) with a covariate for the number of yellow follicles (a proxy for the concentration of circulating oestrogen).

4. Evidence for only 4 chromosome-wide suggestive QTL were detected at the end of lay (72 weeks).

5. Analysis of the combined data confirmed two genome-wide suggestive QTL on chromosome 1 (137 and 266?cM) and on chromosomes 8 (2?cM) and 9 (10?cM) in analyses with or without the covariate.

6. Positive QTL alleles came from the broiler line with the exception of 2 suggestive QTL at the onset of lay on chromosomes 3 and 5 in an analysis with the covariate.

7. In general, QTL acted additively, except that dominant effects were identified for three suggestive QTL at the onset of lay on chromosomes 3 (57 and 187?cM) and 5 (9?cM).

8. The significant QTL in this study were at similar locations to QTL identified in a range of crosses in other publications, suggesting that they are prime candidates for the search for genes and mutations that could be used as selection criteria to improve bone strength and decrease fractures in commercial laying hens.     

Identification of candidate markers on bovine chromosome 14 (BTA14) under milk production trait quantitative trait loci in Holstein

The objective of this study was to identify single-nucleotide polymorphisms using a bovine chromosome 14 high-density SNP panel after accounting for the effect of DGAT1. Linkage disequilibrium information and sire heterozygosity were used to select markers for linkage analysis on bovine chromosome 14 for milk production traits in 321 Holstein animals. Results show putative milk peaks at 42 and 61 cM, both at p<0.10, a fat yield peak at 42 and 63 cM, both at p<0.05; a protein yield peak at 42 (p<0.01) and 84 cM (p<0.05); fat per cent peaks at 3 (p<0.01) and 29 cM (p<0.05), and a protein per cent peak at 4 cM (p<0.05). Once quantitative trait loci positions were established, allele substitution effects for all markers were evaluated using the same statistical model. Overlaying information between quantitative trait loci (QTL) and allele effect analysis enabled the identification (p<0.01) of 20 SNPs under the milk yield QTL, 2 under both of the fat yield peaks, 8 and 9 under the protein yield peaks, 2 and 6 for the fat per cent peaks and 5 for the protein per cent peak. One SNP in particular, ss61514555:A>C, showed association with 3 of the 5 traits: milk (p=1.59E-04), fat (p=6.88E-05) and protein yields (p=5.76E-05). Overall, combining information from linkage disequilibrium, sire heterozygosity and genetic knowledge of traits enabled the characterization of additional markers with significant associations with milk production traits.     

Identification and fine mapping of quantitative trait loci for backfat on bovine chromosomes 2, 5, 6, 19, 21, and 23 in a commercial line of Bos taurus

Backfat thickness is one of the major quantitative traits that affects carcass quality in beef cattle. In this study, we identified and fine-mapped QTL for backfat EBV on bovine chromosomes 2, 5, 6, 19, 21, and 23 using an identical-by-descent haplotype-sharing analysis in a commercial line of Bos taurus. Eleven haplotypes were found to have significant associations with backfat EBV at the comparison-wise P-value threshold, and one at the chromosome-wise P-value threshold on bovine chromosomes 5, 6, 19, 21, and 23. On average, the 12 significant haplotypes had an effect of 0.62 SD on backfat EBV, ranging from 0.38 SD to 1.33 SD. The 12 significant haplotypes spanned nine chromosomal regions, one on chromosome 5 (65.4 to 70.0 cM), three on 6 (8.2 to 11.8 cM, 63.6 to 68.1 cM, and 81.5 to 83.0 cM), three on 19 (4.8 to 15.9 cM, 39.4 to 46.5 cM, and 65.7 to 99.5 cM), one on 21 (46.1 to 53.1 cM), and one on 23 (45.1 to 50.9 cM). Among the nine chromosomal regions, six were new QTL regions and three showed remarkable agreement with QTL regions that were previously reported. Eight of the nine QTL regions were localized to less than or close to 10 cM in genetic distance. The results provide a useful reference for further positional candidate gene research and marker-assisted selection for backfat.     

Investigations in the character of QTL affecting negatively correlated milk traits

The primary aim of this study was to investigate the quantitative trait loci (QTL) on BTA6 that affect negatively correlated milk traits, using bivariate covariance component analysis of milk yield and fat (or protein) content, protein yield and fat content, and fat yield and protein content. A set of five different genetic models was adapted to differentiate trait‐specific QTL in close linkage from pleiotropy. Using a grand‐daughter design consisting of five half‐sib families from the German Holstein population and 298 sons genotyped for 16 microsatellite markers on BTA6, we found significant trait‐specific QTL for fat content and protein yield, 24 cM apart. Markers BM1329 and FBN12 bracketed the QTL for fat content, and the region between TGLA37 and FBN13 most likely harbours a QTL for protein yield. The analysis based on the close linkage model fully confirmed this result. Despite the pure QTL findings confirming results from the literature, distinguishing pleiotropic and closely linked QTL for competitive traits is a new aspect. Our multivariate analysis results did not suggest a pleiotropic QTL for the investigated negatively correlated traits. The QTL‐based trait correlations were discussed as an important aspect of modelling that needs to be considered in the future.     

Quantitative trait loci analysis for growth and carcass traits in a half-sib family of purebred Japanese Black (Wagyu) cattle

We used a half-sib family of purebred Japanese Black (Wagyu) cattle to locate economically important quantitative trait loci. The family was composed of 348 fattened steers, 236 of which were genotyped for 342 microsatellite markers spanning 2,664 cM of 29 bovine autosomes. The genome scan revealed evidence of 15 significant QTL (<5% chromosome-wise level) affecting growth and carcass traits. Of the 15 QTL, six QTL were significant at the 5% experiment-wise level and were located in bovine chromosomes (BTA) 4, 5, and 14. We analyzed these three chromosomes in more detail in the 348 steers, with an average marker interval of 1.2 cM. The second scan revealed that the same haplotype of the BTA 4 region (52 to 67 cM) positively affected LM area and marbling. We confirmed the QTL for carcass yield estimate on BTA 5 in the region of 45 to 54 cM. Five growth-related QTL located on BTA 14, including slaughter and carcass weights, were positively affected by the same region of the haplotype of BTA 14 (29-51 cM). These data should provide a useful reference for further marker-assisted selection in the family and positional cloning research. The research indicates that progeny design with moderate genotyping efforts is a powerful method for detecting QTL in a purebred half-sib family.     

Interval mapping of growth quantitative trait loci in Japanese Black beef cattle using microsatellite DNA markers and half-sib regression analysis

A confirmatory scan for the regions of bovine chromosome 1 segregating the quantitative trait loci (QTL) influencing birthweight, weaning weight, yearling weight, and preweaning and postweaning average daily gains was performed by genotyping half‐sib progeny of four Japanese Black sires using microsatellite DNA markers. Data were analyzed by generating an F‐statistic every 1 cM on a linkage map by the regression of phenotype on the probabilities of inheriting an allele from the sire after adjusting for the fixed effects of sire, sex, parity and season of birth as well as age as a covariate. Permutation tests at chromosome‐wide significance thresholds were carried out over 10 000 iterations. A significant QTL for birthweight at 114 cM was detected in the sire 2 family. This identification of a birthweight QTL in Japanese Black cattle may be useful for the implementation of marker‐assisted selection.     

Introgressing multiple QTL in breeding programmes of limited size.

The ability to enrich a breed with favourable alleles from multiple unlinked quantitative trait loci (QTL) of a donor breed through marker-assisted introgression (MAI) in a population of limited size was evaluated by considering the effects of the proportion selected, the size of the marker intervals, the number of introgressed QTL and the uncertainty of QTL position. Informative flanking markers were used to select progeny with the largest expected number of donor QTL alleles over five generations of backcrossing and five generations of intercrossing. In the backcrossing phase, with 5% selected and 20 cM marker intervals for three QTL, there were sufficient backcross progeny that were heterozygous for all markers, and QTL frequencies dropped below 0.5 only because of double recombinants. For higher fractions selected, longer marker intervals, and more QTL, frequency reductions from 0.5 were greater and increased with additional generations of backcrossing. However, even with 20% selected, three QTL, and marker intervals of 5 or 20 cM, mean QTL frequencies in generation 5 were 0.35 and 0.30, sufficient to allow subsequent selection of QTL in the intercrossing phase. After five generations of intercrossing, over 90% of individuals were homozygous for all QTL, and 85% when five QTL were introgressed. The higher the proportions selected, the longer the marker intervals, and larger numbers of introgressed QTL increased the number of intercrossing generations required to achieve fixation of QTL. Location of the QTL in the marked intervals did not affect QTL frequencies or the proportion of QTL lost at the end of the introgression programme. In conclusion, introgressing multiple QTL can be accomplished in a MAI programme of limited size without requiring that all individuals selected during the backcrossing phase to be carriers of favourable alleles at all QTL.     

红三叶遗传图谱构建及抗白粉病基因QTL定位

本研究以感(岷山红三叶)、抗(澳大利亚红三叶品种♀Sensation×Renegade♂杂交新品系“甘农PR1”)白粉病红三叶材料为父母本杂交并种植成苗经人工接菌后筛选出抗、感白粉病的F₁群体为作图群体,利用AFLP标记构建红三叶高密度遗传图谱,并利用区间作图法对抗白粉病QTL进行了定位分析,可以为红三叶抗白粉病基因克隆和转基因等分子辅助育种奠定基础。结果表明,149个AFLP标记构建得到的遗传图谱包含7个连锁群(LG1,LG2,LG3,LG4,LG5,LG6和LG7),遗传图谱的总距离为640.5 cM。其中,LG1连锁群的遗传距离(140.6 cM)和标记间平均距离(9.4 cM)均最大;LG4连锁群的遗传距离(55.2 cM)和标记间平均距离(1.8 cM)最小。应用区间作图法对红三叶抗白粉病基因进行QTL分析定位,共检测到5个抗白粉病相关QTL位点(qrp-1,qrp-2,qrp-3,qrp-4和qrp-5),其中qrp-1、qrp-2、qrp-3和qrp-4位于LG4连锁群上,qrp-5位于LG5连锁群上。5个QTL位点对抗白粉病的贡献率为29%~90%,qrp-1对红三叶白粉病抗性的贡献率最大(90%),为主效QTL。     

Identification of quantitative trait loci affecting female reproductive traits in a multigeneration Meishan-White composite swine population.

A multigeneration crossbred Meishan-White composite resource population was used to identify quantitative trait loci (QTL) for age at first estrus (AP) and the components of litter size: ovulation rate (OR; number of ova released in an estrous period) and uterine capacity (UC). The population was established by reciprocally mating Meishan (ME) and White composite (WC) pigs. Resultant F1 females were mated to either ME or WC boars to produce backcross progeny (BC) of either 3/4 WC 1/4 ME or 1/4 WC 3/4 ME. To produce the next generation (F3), 3/4 WC 1/4 ME animals were mated to 1/4 WC 3/4 ME animals yielding half-blood (1/2 WC 1/2 ME) progeny. A final generation (F4) was produced by inter se mating F3 animals. Measurements for AP and OR were recorded on 101 BC, 389 F3, and 110 F4 gilts, and UC data were from 101 BC and 110 F4 first parity litters. A genomic scan was conducted with markers (n = 157) spaced approximately 20 cM apart. All parental, F1, BC, and F4 animals but only 84 F3 animals were genotyped and included in this study. The QTL analysis fitted a QTL at 1-cM intervals throughout the genome, and QTL effects were tested using approximate genome-wide significance levels. For OR, a significant (E[false positive] < .05) QTL was detected on chromosome 8, suggestive (E[false positive] < 1.0) QTL were detected on chromosomes 3 and 10, and two additional regions were detected that may possess a QTL (E[false positive] < 2.0) on chromosomes 9 and 15. Two regions possessed suggestive evidence for QTL affecting AP on chromosomes 1 and 10, and one suggestive region on chromosome 8 was identified for UC. Further analyses of other populations of swine are necessary to determine the extent of allelic variation at the identified QTL.     

Mapping the quantitative trait loci (QTL) for body shape and conformation measurements on BTA1 in Japanese Black cattle

The detection and mapping of segregating quantitative trait loci (QTL) that influence withers height, hip height, hip width, body length, chest width, chest depth, shoulder width, lumbar width, thurl width, pin bone width, rump length, cannon circumference, chest girth, abdominal width and abdominal girth at weaning was conducted on chromosomal regions of bovine chromosome one. The QTL analysis was performed by genotyping half‐sib progeny of five Japanese Black sires using microsatellite DNA markers. Probability coefficients of inheriting allele 1 or 2 from the sire at specific chromosomal locations were computed. The phenotypic data of progeny were regressed on these probability coefficients in a within‐common‐parent regression analysis using a linear model that included fixed effects of sex, parity and season of birth, as well as age as a covariate. F‐statistics were calculated every 1 cM on a linkage map. Permutation tests of 10 000 iterations were conducted to obtain chromosome‐wide significance thresholds. A significant QTL for chest width was detected at 91 cM in family 3. The detection of this QTL boosts the prospects of implementing marker‐assisted selection for body conformation traits in Japanese Black beef cattle.