ERIC-PCR: A Molecular Typing Tool for Genotyping Multi Drug Resistant Pseudomonas aeruginosa Isolated from the Pus Samples

Pseudomonas aeruginosa is a typical and major human pathogen, it causes numerous risky opportunistic infections like cystic fibrosis, wound infection, urinary tract infection, ear infections, endocarditis, nosocomial infections and bacteremia, etc. Furthermore, due to their phenotypic variations; identification maybe sometimes harder and consequently which delays their early diagnosis and treatment in infected patients.  Hence, a quick and accurate method for the identification of P. aeruginosa would be more helpful in their early diagnosis. The present work was aimed to use species-specific primer for the rapid and precise identification of P. aeruginosa strains. Multiple drug-resistant strains were selected and their genetic variability was studied using ERIC-PCR. The results showed that out of ten P. aeruginosa isolates, eight were found to be unique and genetically diverse. Keywords: Pseudomonas aeruginosa, ERIC-PCR, MDR. Wound infection, Pus sample

Aerosolized adenovirus-vectored vaccine as an alternative vaccine delivery method

Conventional parenteral injection of vaccines is limited in its ability to induce locally-produced immune responses in the respiratory tract, and has logistical disadvantages in widespread vaccine administration. Recent studies suggest that intranasal delivery or vaccination in the respiratory tract with recombinant viral vectors can enhance immunogenicity and protection against respiratory diseases such as influenza and tuberculosis, and can offer more broad-based generalized protection by eliciting durable mucosal immune responses. Controlled aerosolization is a method to minimize vaccine particle size and ensure delivery to the lower respiratory tract. Here, we characterize the dynamics of aerosolization and show the effects of vaccine concentration on particle size, vector viability, and the actual delivered dose of an aerosolized adenoviral vector. In addition, we demonstrate that aerosol delivery of a recombinant adenoviral vaccine encoding H1N1 hemagglutinin is immunogenic and protects ferrets against homologous viral challenge. Overall, aerosol delivery offers comparable protection to intramuscular injection, and represents an attractive vaccine delivery method for broad-based immunization campaigns

Fine mapping of stay-green QTLs on sorghum chromosome SBI-10L ‒ An approach from genome to phenome

Sorghum is the fifth most important C4 cereal crop grown globally in arid and semi-arid climatic conditions. Drought is the major cause for loss of productivity worldwide. Delayed senescence of plants leads to adaptation to drought stress conditions by staying-green and giving high yields. In order to identify and dissect the stay-green genomic regions we have developed a high resolution fine mapping population from introgression line cross RSG04008-6 (stay-green) × J2614-11 (shoot fly resistant). Nearly 1894 F2 genotypes were screened with 8 SSR in order to identify double recombinants for both the parents in the sorghum chromosome-10 long arm (SBI-10L). The selected F2:4 recombinants were GBSed to increase the marker density between flanking markers Xgap001- Xtxp141 for stay-green QTLs on SBI-10L and a high resolution linkage map was developed using GBS SNPs. Out of 182 only 152 recombinants were replicated thrice in field for staygreen screening for two seasons (Summer 2013 and 2014). Fine mapping of the per cent Green Leaf Area (%GLA) traits identified 33 QTLs and 19QTLs were clustered into 7groups where 8 genes were identified. These were AP2/ERF transcription factor family (Sb10g025053), Ankyrin-repeat protein (Sb10g025310), WD40 repeat protein (Sb10g025320), NBSLRR Protein (Sb10g025283), Calcium dependant protein kinase (Sb10g030150), LEA2 protein (Sb10g029570), a putative uncharacterized protein (Sb10g024920) and senescence associated protein (Sb10g030520). With this study SBI-10L staygreen genomic regions were delimited from 15Mb to 8 genes co-localized with GWAS MTAs. Further cloning and expression level studies of the identified candidate genes will improve the development of drought tolerant genotypes

Fine-Mapping of Sorghum Stay-Green QTL on Chromosome10 Revealed Genes Associated with Delayed Senescence

This study was conducted to dissect the genetic basis and to explore the candidate genes underlying one of the important genomic regions on an SBI-10 long arm (L), governing the complex stay-green trait contributing to post-flowering drought-tolerance in sorghum. A fine-mapping population was developed from an introgression line cross—RSG04008-6 (stay-green) J2614-11 (moderately senescent). The fine-mapping population with 1894 F2 was genotyped with eight SSRs and a set of 152 recombinants was identified, advanced to the F4 generation, field evaluated with three replications over 2 seasons, and genotyped with the GBS approach. A high-resolution linkage map was developed for SBI-10L using 260 genotyping by sequencing—Single Nucleotide Polymorphism (GBS–SNPs). Using the best linear unpredicted means (BLUPs) of the percent green leaf area (%GL) traits and the GBS-based SNPs, weidentified seven quantitative trait loci (QTL) clusters and single gene, mostly involved in drought-tolerance, for each QTL cluster, viz., AP2/ERF transcription factor family (Sobic.010G202700), NBS-LRR protein (Sobic.010G205600), ankyrin-repeat protein (Sobic.010G205800), senescence-associated protein (Sobic.010G270300), WD40 (Sobic.010G205900), CPK1 adapter protein (Sobic.010G264400), LEA2 protein (Sobic.010G259200) and an expressed protein (Sobic.010G201100). The target genomic region was thus delimited from 15 Mb to 8 genes co-localized with QTL clusters, and validated using quantitative real-time (qRT)–PCR

Fine genetic mapping of combined shoot fly resistance (SFR) and stay green (STG) traits on sorghum chromosome SBI-10

Sorghum is fifth most important C4 cereal crop used as food, feed, fodder, fuel and a “fail safe” source in semi-arid tropics of the world..

Population Genetics and Structure of a Global Foxtail Millet Germplasm Collection

Foxtail millet is one among the most ancient crops of dryland agriculture. It is the second most important crop among millets, grown for grains or forage. Foxtail millet germplasm resources provide reservoirs of novel alleles and genes for crop improvement that have remained mostly unexplored. We genotyped a set of 190 foxtail millet germplasm accessions (including 155 accessions of the foxtail millet core collection) using genotyping-by-sequencing (GBS) for rapid single nucleotide polymorphisms (SNP) characterization to study population genetics and structure, which enable allele mining through association mapping approaches. After filtering a total 350,000 raw SNPs identified across 190 germplasm accessions for Minor Allele Frequency (MAF), coverage for samples and coverage for sites, we retained 181 accessions with 17,714 high quality SNPs with ≥ 5% MAF. Genetic structure analyses revealed that foxtail millet germplasm accessions are structured along both on the basis of races and geographic origin, and the maximum proportion of variation was due to among individuals within populations. Accessions of race indica were less diverse and are highly differentiated from those of maxima and moharia. Genome-wide linkage disequilibrium (LD) analysis showed on an average LD extends up to ~150 kbp, and varied with individual chromosomes. The utility of these data for performing genome-wide association studies was tested with plant pigmentation and days to flowering, and identified significant marker-trait associations. This SNP data provides a foundation for exploration of foxtail millet diversity and for mining novel alleles and mapping genes for economically important traits

The potential of urinary metabolites for diagnosing multiple sclerosis

A definitive diagnostic test for multiple sclerosis (MS) does not exist; instead physicians use a combination of medical history, magnetic resonance imaging, and cerebrospinal fluid analysis (CSF). Significant effort has been employed to identify biomarkers from CSF to facilitate MS diagnosis; however none of the proposed biomarkers have been successful to date. Urine is a proven source of metabolite biomarkers and has the potential to be a rapid, non-invasive, inexpensive, and efficient diagnostic tool for various human diseases. Nevertheless, urinary metabolites have not been extensively explored as a source of biomarkers for MS. Instead, we demonstrate that urinary metabolites have significant promise for monitoring disease-progression, and response to treatment in MS patients. NMR analysis of urine permitted the identification of metabolites that differentiate experimental autoimmune encephalomyelitis (EAE)-mice (prototypic disease model for MS) from healthy and MS drug-treated EAE mice