一株可裂解耐黏菌素大肠杆菌噬菌体的分离鉴定及全基因组分析

为研究耐黏菌素大肠杆菌的噬菌体治疗方法,本研究以耐黏菌素大肠杆菌为宿主菌,采用双层琼脂平板法从山东烟台某养鸡场污水样中分离纯化得到裂解性噬菌体SDYTW1-F1-2-2。通过透射电镜观察,最佳感染复数,一步生长曲线,温度稳定性,酸碱度稳定性等生物学特性测定,并对其进行全基因组测序分析。结果表明：1）噬菌体SDYTW1-F1-2-2噬菌斑呈现透亮,外周无晕圈,形态学观察显示其具有可伸缩性尾鞘。2）生物学特性分析表明,噬菌体最佳感染复数为0.1,潜伏期为50 min,爆发期为40 min,爆发量为（331±9）PFU/cell。在温度4～40℃,pH为4～12的环境下,噬菌体活性较为稳定。3）通过噬菌体全基因组测序及核酸类型鉴定显示,该噬菌体基因组全长为74 752 bp,双链DNA,GC含量42.1%。在噬菌体基因组中共鉴定出121个编码蛋白（CDS）,包括结构组成、DNA代谢与复制和包装以及细菌裂解相关的功能基因,发现一个tRNA基因,且未检测到如stx-1等致病性基因和mcr-1等耐药基因相关的基因。综上,噬菌体SDYTW1-F1-2-2裂解谱较宽,具有良好的酸碱性、热稳定性以及安全...     

Genomewide association for a dominant pigmentation gene in sheep

Most published genomewide association studies (GWAS) in sheep have investigated recessively inherited monogenic traits. The objective here was to assess the feasibility of performing GWAS for a dominant trait for which the genetic basis was already known. A total of 42 Manchega and Rasa Aragonesa sheep that segregate solid black or white coat pigmentation were genotyped using the SNP50 BeadChip. Previous analysis in Manchegas demonstrated a complete association between the pigmentation trait and alleles of the MC1R gene, setting an a priori expectation for GWAS. Multiple methods were used to identify and quantify the strength of population substructure between black and white animals, before allelic association testing was performed for 49 034 SNPs. Following correction for substructure, GWAS identified the most strongly associated SNP (s26449) was also the closest to the MC1R gene. The finding was strongly supported by the permutation tree‐based random forest (RF) analysis. Importantly, GWAS identified unlinked SNP with only slightly lower p‐values than for s26449. Random forest analysis indicated these were false positives, suggesting interpretation based on both approaches was beneficial. The results indicate that a combined analytical approach can be successful in studies where a modest number of animals are available and substantial population stratification exists.     

Genomewide association study of immune traits in chicken F2 resource population

Immune traits play pivotal roles in animal immune capacity development and disease resistance. Single nucleotide polymorphisms (SNPs) are common forms of genetic variations among individuals, which are thought to account for the majority of inherited phenotypic variations. In this study, we performed genomewide association, using the Illumina 60K SNP BeadChip studies to detect molecular markers and candidate genes associated with immune traits in an F2 population. Sixteen immune traits were measured. We identified 85 significant SNPs (p < 2.98 × 10^?6) with 5% as the genomewide significance threshold, 380 SNPs of suggestive significance (p < 5.96 × 10^?5) from simple model (general linear model, GLM) and 15 SNPs of suggestive significance (p < 5.96 × 10^?5) from the compressed mixed linear model (MLM), which were also found in GLM (six significant SNPs and seven suggestive SNPs). Three significant SNPs (GGaluGA151406, Gga_rs14554319 and Gga_rs13593979) and candidate genes (LYRM4 and KTN1) were found to be associated with avian influenza antibody titres, and the first two SNPs are from the results of two‐model analysis. For the immune organs, through the analysis of GLM, 19 SNPs were found to be significantly associated with the thymus weight, 61 SNPs were significantly associated with the bursa of Fabricius weight, six of which were located within a 34‐Mb region (125 846 474–159 649 698 bp) on chicken chromosome 1 (GGA1). A candidate region relevant to haematological traits from GLM was found in GGA4 and 9 loci were located on it. Three loci (GGaluGA348521, Gga_rs16098446 and GGaluGA348518) within 179 kb (16 286 868–16 466 134 bp) on GGAZ from GLM provided evidence that this genomic segment may be relevant to red blood cell volume distribution width (RDW). Our study provides a list of significant SNPs and candidate genes that will be valuable information for unveiling the underlying molecular mechanism of immune regulation.     

Genetic analysis of optic nerve head coloboma in the Nova Scotia Duck Tolling Retriever identifies discordance with the NHEJ1 intronic deletion (collie eye anomaly mutation)

Collie eye anomaly (CEA) encompasses a spectrum of different ophthalmic phenotypes from clinically inconsequential choroidal hypoplasia to blindness from coloboma of the optic nerve head (ONH). A previous study found a 7.8‐kb deletion in intron 4 of the NHEJ1 gene to be associated with CEA. A genetic test based on this association is recommended for many breeds, including the Nova Scotia Duck Tolling Retriever (NSDTR). Collection of ONH coloboma‐affected NSDTR showed lack of concordance of the NHEJ1 intronic deletion with ONH coloboma. Using genomewide single nucleotide polymorphism (SNP) genotyping in 7 ONH coloboma‐affected NSDTR cases and 47 unaffected NSDTR controls with no ophthalmic signs, one SNP, located on chromosome 7, demonstrated genomewide significance. However, high genomic inflation may have confounded the results. Therefore, the genomewide association study was repeated using EMMAX to control for population structure in the cohort of 7 cases and 47 controls. However, no regions of the genome were significantly associated with ONH coloboma. These results failed to document significant association with the CEA locus. Due to the complex genetic etiology of ONH coloboma, the NHEJ1 intronic deletion test results should be carefully considered when making breeding decisions. If the goal is to select for visually competent dogs, our data suggest that eye examinations of puppies would be more effective as a guide in selection of breeding pairs than relying solely on currently available genetic tests.     

A genomewide association mapping study using ultrasound‐scanned information identifies potential genomic regions and candidate genes affecting carcass traits in Nellore cattle

The aim of this study was to identify candidate genes and genomic regions associated with ultrasound‐derived measurements of the rib‐eye area (REA), backfat thickness (BFT) and rumpfat thickness (RFT) in Nellore cattle. Data from 640 Nellore steers and young bulls with genotypes for 290 863 single nucleotide polymorphisms (SNPs) were used for genomewide association mapping. Significant SNP associations were explored to find possible candidate genes related to physiological processes. Several of the significant markers detected were mapped onto functional candidate genes including ARFGAP3, CLSTN2 and DPYD for REA; OSBPL3 and SUDS3 for BFT; and RARRES1 and VEPH1 for RFT. The physiological pathway related to lipid metabolism (CLSTN2, OSBPL3, RARRES1 and VEPH1) was identified. The significant markers within previously reported QTLs reinforce the importance of the genomic regions, and the other loci offer candidate genes that have not been related to carcass traits in previous investigations.     

Identification of natural variants affecting chlorophyll content dynamics during rice seedling development

Seedling vigour is an important characteristic in relation to crop growth and yield. Traits such as photosynthetic capacity and chlorophyll content contribute significantly to seedling establishment at the early growth stage in various crop species, including rice. A diverse panel of 227 rice varieties from several countries was evaluated to determine chlorophyll contents at multiple time points during the seedling stage using a soil–plant analysis development (SPAD) meter, a non‐destructive portable device. Using new statistical approaches, several chromosomal regions associated with variations in chlorophyll content in the third leaf at 13, 16 and 19 days after imbibition were detected. A single nucleotide polymorphism (SNP) cluster on the end of chromosome 11 was significantly associated with the onset of leaf senescence. This region was enriched with genes related to cell death and the stress response. We have identified rice germplasm showing delayed‐senescence phenotypes, these could be suitable donors and genetic resources for breeding, and the use of significant SNP markers associated with these traits could enhance the efficiency of their selection in breeding programmes.     

Unravelling the genetic loci for growth and carcass traits in Chinese Bamaxiang pigs based on a 1.4 million SNP array

Bamaxiang pig is from Guangxi province in China, characterized by its small body size and two‐end black coat colour. It is an important indigenous breed for local pork market and excellent animal model for biomedical research. In this study, we performed genomewide association studies (GWAS) on 43 growth and carcass traits in 315 purebred Bamaxiang pigs based on a 1.4 million SNP array. We observed considerable phenotypic variability in the growth and carcass traits in the Bamaxiang pigs. The corresponding SNP based heritability varied greatly across the 43 traits and ranged from 9.0% to 88%. Through a conditional GWAS, we identified 53 significant associations for 35 traits at p value threshold of 10^?6. Among which, 26 associations on chromosome 3, 7, 14 and X passed a genomewide significance threshold of 5 × 10^?8. The most remarkable loci were at around 30.6 Mb on chromosome 7, which had growth stage‐dependent effects on body lengths and cannon circumferences and showed large effects on multiple carcass traits. We discussed HMGA1 NUDT3, EIF2AK1, TMEM132C and AFF2 that near the lead SNP of significant loci as plausible candidate genes for corresponding traits. We also showed that including phenotypic covariate in GWAS can help to reveal additional significant loci for the target traits. The results provide insight into the genetic architecture of growth and carcass traits in Bamaxiang pigs.     

An efficient empirical Bayes method for genomewide association studies

Linear mixed model (LMM) is one of the most popular methods for genomewide association studies (GWAS). Numerous forms of LMM have been developed; however, there are two major issues in GWAS that have not been fully addressed before. The two issues are (i) the genomic background noise and (ii) low statistical power after Bonferroni correction. We proposed an empirical Bayes (EB) method by assigning each marker effect a normal prior distribution, resulting in shrinkage estimates of marker effects. We found that such a shrinkage approach can selectively shrink marker effects and reduce the noise level to zero for majority of non‐associated markers. In the meantime, the EB method allows us to use an ‘effective number of tests’ to perform Bonferroni correction for multiple tests. Simulation studies for both human and pig data showed that EB method can significantly increase statistical power compared with the widely used exact GWAS methods, such as GEMMA and FaST‐LMM‐Select. Real data analyses in human breast cancer identified improved detection signals for markers previously known to be associated with breast cancer. We therefore believe that EB method is a valuable tool for identifying the genetic basis of complex traits.     

Genome‐wide association analysis reveals flowering‐related genes regulating rachis length in rice

Rachis length is correlated with panicle size in rice. Unveiling the genetic basis of rachis length is important for understanding the genetic regulation of panicle size. In this study, we performed a genomewide association study of rachis length using 529 rice accessions from two environments. In total, 20 loci were identified for rachis length and distributed across the 12 chromosomes except for chromosomes 2 and 5. Thirteen of the 20 loci were not linked to the cloned panicle size genes. Six flowering‐related genes, including the CCT domain‐containing genes Ghd7, Ghd7.1, Hd1, OsCCT1 and OsCO3 as well as Ehd1, were associated with rachis length in this study. These findings suggest that the network of these flowering‐related genes probably participates in the regulation of rachis length and thus affects panicle length and yield. Interestingly, haplotype analysis showed that OsCCT1 is a putative candidate gene which plays a key role in the variation of rachis length. Together, the genetic loci identified in this study could be used for rice breeding by marker‐assisted selection to improve grain yield.     

Genome wide association study on beef production traits in Marchigiana cattle breed

A genomewide association study was carried out on a sample of Marchigiana breed cattle to detect markers significantly associated with carcass and meat traits. Four hundred and nine young bulls from 117 commercial herds were genotyped by Illumina 50K BeadChip assay. Eight growth and carcass traits (average daily gain, carcass weight, dressing percentage, body weight, skin weight, shank circumference, head weight and carcass conformation) and two meat quality traits (pH at slaughter and pH 24 h after slaughter) were measured. Data were analysed with a linear mixed model that included fixed effects of herd, slaughter date, fixed covariables of age at slaughter and SNP genotype, and random effects of herd and animal. A permutation test was performed to correct SNP genotype significance level for multiple testing. A total of 96 SNPs were significantly associated at genomewide level with one or more of the considered traits. Gene search was performed on genomic regions identified on the basis of significant SNP position and level of linkage disequilibrium. Interesting loci affecting lipid metabolism (SOAT1), bone (BMP4) and muscle (MYOF) biology were highlighted. These results may be useful to better understand the genetic architecture of growth and body composition in cattle.