Identify known and novel candidate genes associated with backfat thickness in Duroc pigs by large-scale genome-wide association analysis

Backfat thickness (BFT) is complex and economically important traits in the pig industry, since it reflects fat deposition and can be used to measure the carcass lean meat percentage in pigs. In this study, all 6,550 pigs were genotyped using the Geneseek Porcine 50K SNP Chip to identify SNPs related to BFT and to search for candidate genes through genome-wide association analysis in two Duroc populations. In total, 80 SNPs, including 39 significant and 41 suggestive SNPs, and 6 QTLs were identified significantly associated with the BFT. In addition, 9 candidate genes, including a proven major gene MC4R, 3 important candidate genes (RYR1, HMGA1 and NUDT3) which were previously described as related to BFT, and 5 novel candidate genes (SIRT2, NKAIN2, AMH, SORCS1 and SORCS3) were found based on their potential functional roles in BFT. The functions of candidate genes and gene set enrichment analysis indicate that most important pathways are related to energy homeostasis and adipogenesis. Finally, our data suggests that most of the candidate genes can be directly used for genetic improvement through molecular markers, except that the MC4R gene has an antagonistic effect on growth rate and carcass lean meat percentage in breeding. Our results will advance our understanding of the complex genetic architecture of BFT traits, and laid the foundation for additional genetic studies to increase carcass lean meat percentage of pig through marker-assisted selection and/or genomic selection.

Identification and Genetic Analysis of Major Gene ST3GAL4 Related to Serum Alkaline Phosphatase in Chickens

The full text of this preprint has been withdrawn by the authors while they make corrections to the work. Therefore, the authors do not wish this work to be cited as a reference. Questions should be directed to the corresponding author.

Detection of Novel Variations Related to Litter Size in BMP15 Gene of Luzhong Mutton Sheep (Ovis aries)

Litter size is an important economic trait in the mutton sheep industry. BMP15 is one of the key candidate genes for litter size in sheep. In this study, the entire ORF region of BMP15 was sequenced in 154 Luzhong mutton ewes, and the novel variations were determined. The association between polymorphism in BMP15 and litter size was analyzed using a general linear model. Six out of a total of thirteen variations were identified to be novel. Association analysis indicated that four (SNPs ENSOART00000010201.1:c.352+342C>A, c.352+1232T>C, c.352+1165A>G and c.353-2036T>A) were significantly associated with litter size. The joint analysis among three major genes (BMP15, BMPR1B and GDF9) exhibited significant interaction effects in three combinations (FecB and c.352+1232T>C of BMP15; FecB and c.352+1165A>G of BMP15; c.352+342C>A of BMP15 and ENSOART00000014382.1:c.994G>A of GDF9). For the SNPs c.352+1232T>C and c.352+342C>A, the global distribution of allele frequencies showed that the highest variation frequency occurs in Western Europe. In conclusion, the results demonstrated that BMP15 is a major gene for litter size in Luzhong mutton sheep and candidate SNPs associated with litter size were identified.

Microbial ecology of sulfur biogeochemical cycling at a mesothermic hot spring atop Northern Himalayas, India

Graphical AbstractSulfur Related Prokaryotes (SRP) residing in hot spring present good opportunity for exploring the limitless possibilities of integral ecosystem processes. Metagenomic analysis further expand the phylogenetic breadth of these extraordinary sulfur metabolizing microorganisms, as well a their complex metabolic networks and syntrophic interactions in environmental biosystems. Through this study, we explored and expanded the microbial genetic repertoire with focus on sulfur cycling genes through metagenomic analysis of sulfur (S) contaminated hot spring, located at the Northern Himalayas. The analysis revealed rich diversity of microbial consortia with established roles in S cycling such as Pseudomonas, Thioalkalivibrio, Desulfovibrio and Desulfobulbaceae (Proteobacteria). The major gene families inferred to be abundant across microbial mat, sediment and water were assigned to Proteobacteria as reflected from the RPKs (reads per kilobase) categorized into translation and ribosomal structure and biogenesis. Analysis of sequence similarity showed conserved pattern of both dsrAB genes (n=178) retrieved from all metagenomes while other sulfur disproportionation proteins were diverged due to different structural and chemical substrates. The diversity of sulfur oxidizing bacteria (SOB) and sulfate reducing bacteria (SRB) with conserved (r)dsrAB suggests for it to be an important adaptation for microbial fitness at this site. Here, we confirm that (i) SRBs belongs to δ-Proteobacteria occurring independent LGT of dsr genes to different and few novel lineages (ii) also, the oxidative and reductive dsr evolutionary time scale phylogeny, proved that the earliest (not first) dsrAB proteins belong to anaerobic Thiobacillus with other (rdsr) oxidizers. Further, the structural prediction of unassigned DsrAB proteins confirmed their relatedness with species of Desulfovibrio (TM score= 0.86; 0.98; 0.96) and Archaeoglobus fulgidus (TM score= 0.97; 0.98). We proposed that the genetic repertoire might provide the basis of studying time scale evolution and horizontal gene transfer of these genes in biogeochemical S cycling and the complementary genes could be implemented in biotechnology and bioremediation applications.

Heritability and Genomic Architecture of Episodic Exercise-Induced Collapse in Border Collies

An episodic nervous system disorder triggered by strenuous exercise, termed border collie collapse (BCC), exists in border collies and related breeds. The genetic basis of BCC is unknown but is believed to be a complex genetic disorder. Our goal was to estimate the heritability (h2SNP) of BCC, define its underlying genetic architecture, and identify associated genomic loci using dense whole-genome single-nucleotide polymorphism (SNP) genotyping data. Genotype data were obtained for ~440,000 SNPs from 343 border collies (168 BCC cases and 175 controls). h2SNP was calculated to be 49–61% depending on the estimated BCC prevalence. A total of 2407 SNPs across the genome accounted for nearly all the h2SNP of BCC, with an estimated 2003 SNPs of small effect, 349 SNPs of moderate effect, and 56 SNPs of large effect. Genome-wide association analyses identified significantly associated loci on chromosomes 1, 6, 11, 20, and 28, which accounted for ~5% of the total BCC h2SNP. We conclude that BCC is a moderately- to highly- heritable complex polygenetic disease resulting from contributions from hundreds to thousands of genetic variants with variable effect sizes. Understanding how much the BCC phenotype is determined by genetics and whether major gene mutations are likely to exist inform veterinarians and working/stock dog communities of the true nature of this condition.

Quantitative Disease Resistance to White Pine Blister Rust at Southwestern White Pine’s (Pinus strobiformis) Northern Range

White pine blister rust, caused by the non-native, invasive fungal pathogen Cronartium ribicola, is a significant cause of mortality in white pines (Pinus subgenus Strobus) in North America. Along with climate-driven range contraction, mortality from blister rust can seriously impact the abundance and distribution of the nine white pine species native to the United States and Canada. Very little evaluation of this disease in southwestern white pine (Pinus strobiformis) has been previously undertaken, but genetic resistance to the disease has been documented, including major gene resistance (MGR) conferred by a dominant R gene. Data is emerging suggesting that the species also has quantitative disease resistance (QR). Our results suggest QR occurs at low frequency, with perhaps 10% of trees having a moderate level (> 35% survival). We assessed progeny arrays from 40 P. strobiformis families (1873 seedlings), originating from three populations, inoculated with C. ribicola. Subsequently, the seedlings were assessed for signs, symptoms and resulting impact in a common garden trial over a 7.5-year period to determine the types and frequency of resistance in a portion of this species’ range. There was a high incidence of both stem symptoms and mortality in the P. strobiformis families tested, and families ranged in survival from 0 to 84.6%. Three families had > 70% survival, representing perhaps the highest documented QR to date in a North American white pine species. Approximately 29.1% of the 441 surviving seedlings showed no stem symptoms, and of the approximately 70.8% of seedlings surviving with infections only few (24 of 316) had infections of moderate to high severity. QR traits associated with improved survival were primarily related to lower severity of infection, a reduced number of stem symptoms, and an increased number of bark reactions. Despite the high overall susceptibility, the presence of QR appears to be at a frequency and level useful to forest managers involved in restoration and reforestation efforts.

Active DNA demethylation of developmental cis-regulatory regions predates vertebrate origins

DNA methylation (5-methylcytosine; 5mC) is a repressive gene-regulatory mark required for vertebrate embryogenesis. Genomic 5mC is tightly regulated through the coordinated action of DNA methyltransferases, which deposit 5mC, and TET enzymes, which participate in its active removal through the formation of 5-hydroxymethylcytosine (5hmC). TET enzymes are essential for mammalian gastrulation and activation of vertebrate developmental enhancers, however, to date, a clear picture of 5hmC function, abundance, and genomic distribution in non-vertebrate lineages is lacking. By employing base-resolution 5mC and 5hmC quantification during sea urchin and lancelet embryogenesis, we shed light on the roles of non-vertebrate 5hmC and TET enzymes. We find that these invertebrate deuterostomes employ TET enzymes for targeted demethylation of regulatory regions associated with developmental genes and show that the complement of identified 5hmC-regulated genes is conserved to vertebrates. This work thus demonstrates that active 5mC removal from regulatory regions is a common feature of deuterostome embryogenesis suggestive of unexpected deep conservation of a major gene-regulatory module.

Comparative association mapping reveals conservation of major gene resistance to white pine blister rust in southwestern white pine (Pinus strobiformis) and limber pine (P. flexilis)

All native North American white pines are highly susceptible to white pine blister rust (WPBR) caused by Cronartium ribicola. Understanding genomic diversity and molecular mechanisms underlying genetic resistance to WPBR remains one of the great challenges in improvement of white pines. To compare major gene resistance (MGR) present in two species, southwestern white pine (Pinus strobiformis) Cr3 and limber pine (P. flexilis) Cr4, we performed association analyses of Cr3-controlled resistant traits using SNP assays designed with Cr4-linked polymorphic genes. We found that ~ 70% of P. flexilis SNPs were transferable to P. strobiformis. Furthermore, several Cr4-linked SNPs were significantly associated with the Cr3-controlled traits in P. strobiformis families. The most significantly associated SNP (M326511_1126R) almost co-localized with Cr4 on the Pinus consensus linkage group 8 (LG-8), suggesting that Cr3 and Cr4 might be the same R locus, or have localizations very close to each other in the syntenic region of the P. strobiformis and P. flexilis genomes. M326511_1126R was identified as a non-synonymous SNP, causing amino acid change (Val376Ile) in a putative pectin acetylesterase (PAE), with coding sequences identical between the two species. Moreover, top Cr3-associated SNPs were further developed as TaqMan genotyping assays, suggesting their usefulness as marker-assisted selection (MAS) tools to distinguish genotypes between quantitative resistance (QR) and MGR. This work demonstrates the successful transferability of SNP markers between two closely related white pine species in the hybrid zone, and the possibility for deployment of MAS tools to facilitate long-term WPBR management in P. strobiformis breeding and conservation.

Molecular Detection of blaTEM, blaCTX-M and blaSHV Genes in Extended Spectrum β-Lactamase (ESBL) Escherichia Coli from Clinical Samples

BackgroundExtended spectrum β-lactamases are the group of beta-lactamase enzymes which confer resistance to the oxyimino-cephalosporins and monobactams. Emergence of ESBL producing genes which possess a serious threat for the treatment of infections both in community and hospitals since it is found to be increasing trends of multidrug resistance. This study was focused to find out the ESBLs producing genes.MethodsThis was a cross-sectional study conducted over a period of 2 years (September 2018 to April 2020) at microbiology laboratory of Nepal Mediciti Hospital. Clinical samples were processed in microbiology laboratory and culture isolates were indetified and characterized by standard microbiological techniques. following standard procedures.Antibiotic susceptibility testing was performed by modified Kirby-Bauer disc diffusion method as recommended by Clinical and Laboratory Standard Institute. Extended spectrum beta-lactamases were phenotypically confirmed by combined disc method.ESBL producing genes i.e. blaTEM,blaCTX-M and blaSHV were confirmed by PCR.ResultsOf the 1449 total E.coli isolates, 323/1449(22.29%) isolates were multi -drug resistance.Among total MDR Escherichia coli isolates, 215/323(66.56%) isolates were ESBL producers. The maximum number of ESBL Escherichia coli was isolated from urine 194(90.23%), followed by sputum 12(5.58%), swab 5 (2.32%), pus 2 (0.93%) and blood 2 (0.93%).Antibiotic susceptibility pattern of ESBL E.coli producers showed highest sensitivity towards tigecycline (100%) followed by polymyxin b, colistin and meropenem.Out of 215 phenotypically confirmed ESBL E.coli,only 186(86.51%) isolates were found to positive by PCR.The last 29(13.49%) were negative for any of the resistant genes.Among the ESBL genotypes,most common was blaTEM 118(63.4%) followed by blaCTX-M 68(36.6%).ConclusionThe emergence of MDR and ESBL producing E.coli isolates with high antibiotic resistant rates to commonly used antibiotics and increased predominance of major gene types blaTEM is a serious concern to the clinicians as well as microbiologist. This study forwarded a real massage to all the clinicians for the emergence of XDR and PDR resistant bacteria and preservation of antibiotics for their proper use in near future, if past experience with MDR and ESBLs is any indicator.

Exploring Aromatic Medicinal Compounds for the Treatment of Amyotrophic Lateral Sclerosis

Purpose: Amyotrophic lateral sclerosis (ALS) is a lethal neurodegenerative condition, in which motor neurons start to degenerate due to the accumulation of protein aggregates in the neuron cytoplasm. The formation of aggregates causes neurotoxicity, facilitated by the N-terminal domain (NTD) of the transactive response DNA-binding protein-43 (TDP-43). Therapies used to treat ALS manage secondary symptoms, but do not stop the activity of the rogue NTD domain of TDP-43. Therefore, new drug candidates should be designed to deal efficiently with this disease by inhibiting the domains involved in the development of ALS. This study determined the chemical affinity of aromatic medicinal compounds with NTD. Screening of 1323 medicinal compounds was conducted with PYRX 0.9 software against NTD. Compounds obtained from this analysis were further used to predict absorption, distribution, metabolism, excretion, and toxic (ADMET) properties and their effect on major gene targets of ALS. Results: From 1300 + compounds, acetovanillone showed binding affinity for NTD and had good ADMET and drug likeness attributes. This compound reduced the expression of CXCL2, NOP56, and SOD1 genes implicated in ALS pathogenesis. Conclusion: These results concluded that acetovanillone is a candidate drug for in vitro and clinical studies into the exploitation of drugs within ALS therapeutics.