15 research outputs found

    Functional characterization of genes controlling the production of prenylflavonoids in hop (Humulus lupulus L.)

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    Identification of Cuscuta Campestris Yuncker in UAE: Study of Bar Code Loci- Rbcl, Matk and Trnh-Psba in the UAE and Egyptian Cultivars and In the Respective Host Plants Basil and Jute

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    Cuscuta campestris is a stem holoparasite. We observed Cuscuta parasite on basil host plant Ocimum basilicum, in Al Mohadub Umm Al Quwain, UAE. The parasite was pale green in color, twined around the host in anti-clock wise direction, with white flowers that had green ovaries at maturity. Based on the morphology and floral structures, we identified the parasite as C. campestris Yuncker. To authenticate the species, three “Bar-code loci†viz, rbcL, matK and inter-spacer region trnH-psbA were studied. A portion of rbcL locus and the trnH-psbA non-coding spacer region seem to be intact, revealed by PCR amplification and sequencing, while three sets of primers failed to amplify the maturase K locus. Although the stem and floral structures were light green in color, RuBisCo protein could not be detected in polyacrylamide gels, indicating its total dependency on the host at that stage of development. To validate thus obtained results, frozen samples of C. campestris were collected from Egypt and the three bar code loci (rbcL, matK and trnH-psbA) were amplified with the same set of primers; the PCR products were sequenced. There was 100% similarity with respect to the sequenced loci (rbcl and trnH-psbA) between the two cultivars of C. campestris Yuncker. Sequences were deposited in Genbank with accession numbers KXO15762 (C. campestris, UAE) and KXO15761 (C. campestris, Egypt). C. campestris is being reported for the first time from Al Mohadub Umm Al Quwain, UAE. There is no difference in both the candidate bar code gene loci rbcL and trn-H psbA between the UAE and Egyptian cultivars of Cuscuta campestris and the region is conserved

    Metagenomic Analysis of the Outdoor Dust Microbiomes: A Case Study from Abu Dhabi, UAE

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    Outdoor dust covers a shattered range of microbial agents from land over transportation, human microbial flora, which includes pathogen and commensals, and airborne from the environment. Dust aerosols are rich in bacterial communities that have a major impact on human health and living environments. In this study, outdoor samples from roadside barricades, safety walls, and fences (18 samples) were collected from Abu Dhabi, UAE and bacterial diversity was assessed through a 16S rRNA amplicon next generation sequencing approach. Clean data from HiSeq produced 1,099,892 total reads pairs for 18 samples. For all samples, taxonomic classifications were assigned to the OTUs (operational taxonomic units) representative sequence using the Ribosomal Database Project database. Analysis such as alpha diversity, beta diversity, differential species analysis, and species relative abundance were performed in the clustering of samples and a functional profile heat map was obtained from the OTUs by using bioinformatics tools. A total of 2814 OTUs were identified from those samples with a coverage of more than 99%. In the phylum, all 18 samples had most of the bacterial groups such as Actinobacteria, Proteobacteria, Firmicutes, and Bacteroidetes. Twelve samples had Propionibacteria acnes and were mainly found in RD16 and RD3. Major bacteria species such as Propionibacteria acnes, Bacillus persicus, and Staphylococcus captis were found in all samples. Most of the samples had Streptococcus mitis, Staphylococcus capitis. and Nafulsella turpanensis and Enhydrobacter aerosaccus was part of the normal microbes of the skin. Salinimicrobium sp., Bacillus alkalisediminis, and Bacillus persicus are halophilic bacteria found in sediments. The heat map clustered the samples and species in vertical and horizontal classification, which represents the relationship between the samples and bacterial diversity. The heat map for the functional profile had high properties of amino acids, carbohydrate, and cofactor and vitamin metabolisms of all bacterial species from all samples. Taken together, our analyses are very relevant from the perspective of out-door air quality, airborne diseases, and epidemics, with broader implications for health safety and monitoring

    A Methodological Review of Tools That Assess Dust Microbiomes, Metatranscriptomes and the Particulate Chemistry of Indoor Dust

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    Indoor house dust is a blend of organic and inorganic materials, upon which diverse microbial communities such as viruses, bacteria and fungi reside. Adequate moisture in the indoor environment helps microbial communities multiply fast. The outdoor air and materials that are brought into the buildings by airflow, sandstorms, animals pets and house occupants endow the indoor dust particles with extra features that impact human health. Assessment of the health effects of indoor dust particles, the type of indoor microbial inoculants and the secreted enzymes by indoor insects as allergens merit detailed investigation. Here, we discuss the applications of next generation sequencing (NGS) technology which is used to assess microbial diversity and abundance of the indoor dust environments. Likewise, the applications of NGS are discussed to monitor the gene expression profiles of indoor human occupants or their surrogate cellular models when exposed to aqueous solution of collected indoor dust samples. We also highlight the detection methods of dust allergens and analytical procedures that quantify the chemical nature of indoor particulate matter with a potential impact on human health. Our review is thus unique in advocating the applications of interdisciplinary approaches that comprehensively assess the health effects due to bad air quality in built environments

    Metal composition and contamination assessment of urban roadway dusts on the Abu Dhabi-Liwa Highway, UAE

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    The metal composition of road-deposited dust along the Abu Dhabi-Liwa Highway was investigated to provide insight into the contamination profile and levels of road dust. The average concentrations of metals decreased in the order Al (28668 ± 4631 mg/kg)\u3e Fe (21461 ± 2594 mg/kg) \u3e Mn (711.8 ± 76.3 mg/kg) \u3e Zn (210.6 ± 51.6 mg/kg) \u3e Cu (94.9 ± 15.8 mg/kg) \u3e Pb (83.6 ± 5.3 mg/kg) \u3e Cd (75.1 ± 1.6 mg/kg) \u3e Co (62.6 ± 6.4 mg/kg) \u3e As (4.7 ± 2.9 mg/kg) \u3e Ni (0.10 ± 0.19 mg/kg) \u3e Cr (0.08 ± 0.06 mg/kg). The spatial variations of metals suggest different sources and contributing factors for these metals, with most dust metals having mixed traffic and non-traffic origins. The contamination factor (CF) and enrichment factor (EF) showed identically the same order, Cd\u3e Pb\u3e As\u3e Zn\u3e Co\u3e Cu\u3e Mn\u3e Ni\u3e Cr, whereas the geoaccumulation index (Igeo) follows a slightly different ranking, Cd\u3e Pb\u3e Zn\u3e Co\u3e As\u3e Cu\u3e Mn\u3e Ni\u3e Cr. Based on EF and CF levels, roadway dusts are enriched in all metals, except for Ni and Cr. Similarly, the average Igeo values show differing rates of pollution for all metals except for Mn, Ni, and Cr. All pollution indicators suggest extreme pollution with Cd. The pollution loading index values showed sites 1–10 are generally polluted, while sampling sites from 11 to 19 are unpolluted with decreasing pollution loadings. Dusts collected from both sides of highway were higher in metal content than those obtained from the central reservation area. This may be due to the prevailing southeast wind direction, resuspension of road dust, and farmlands, among others. Soils bordering the highway showed high metal contents with potential consequences on the agricultural products

    Housekeeping gene selection for real time-PCR normalization in female hop (Humulus lupulus L) tissues

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    The variability of transcript accumulation of six genes encoding chloropyll-a/b (Chla/b) binding protein, Nicotinamide Adenine Dinucleotide Hydride (NADH) dehydrogenase, Histone H3 (H3); DEAD-box RNA helicase 1 (DRH1) and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and of the 7SL component of the. signal recognition particle (7SL-RNA), was estimated in leaves and female inflorescences of the two hop cultivars, White Golding and Admiral at different developmental stages by RT-PCR. The value of these genes as internal, normalization controls in gene transcript accumulation studies was assessed. The combination of the three housekeeping genes DRH1, GAPDH and 7SL-RNA as internal standards for hop, provided reliable results in the quantitative analysis of the transcript, accumulation of Hua Enhancer 1 transcription factor (HEN1) homologue in hop tissues and is recommended for future studies of gene expression in female hop tissues

    RecQ helicases function in development, DNA repair, and gene targeting in Physcomitrella patens

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    RecQ DNA helicases are genome surveillance proteins found in all kingdoms of life. They are characterized best in humans, as mutations in RecQ genes lead to developmental abnormalities and diseases. To better understand RecQ functions in plants we concentrated on Arabidopsis thaliana and Physcomitrella patens, the model species predominantly used for studies on DNA repair and gene targeting. Phylogenetic analysis of the six P. patens RecQ genes revealed their orthologs in humans and plants. Because Arabidopsis and P. patens differ in their RecQ4 and RecQ6 genes, reporter and deletion moss mutants were generated and gene functions studied in reciprocal cross-species and cross-kingdom approaches. Both proteins can be found in meristematic moss tissues, although at low levels and with distinct expression patterns. PpRecQ4 is involved in embryogenesis and in subsequent development as demonstrated by sterility of ΔPpRecQ4 mutants and by morphological aberrations. Additionally, ΔPpRecQ4 displays an increased sensitivity to DNA damages and an increased rate of gene targeting. Therefore, we conclude that PpRecQ4 acts as a repressor of recombination. In contrast, PpRecQ6 is not obviously important for moss development or DNA repair but does function as a potent enhancer of gene targeting

    Development of allelic discrimination assay to detect Mediterranean G6PD mutation and its linked inheritance with normal vision/colorblindness loci for 4 generations among Egyptian and Emirati families

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    G6PD deficiency c563T is the most common inherent blood disease among the Mediterranean populations and its molecular diagnosis is critical as the enzyme assay fails for heterozygous individuals. The purpose of the study is to estimate the ubiquity of the heterozygous G6PD Med (c563T) variants among Egyptians and UAE nationals living in Dubai. We validated two molecular methods, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and qPCR allelic discrimination assay for detection of G6PD Med variants. Among 100 screened individuals, G6PD c563T variants are 30% of whom 15 % are carriers. Sanger sequencing validated the qPCR discrimination assays. In search of a phenotypic marker to detect G6PD heterozygous variants, inheritance of G6PD locus and red-green color vision genes is studied in 1 Egyptian and 2 Emirati families. Among the 3 families, G6PD is polymorphic, displaying 4 phenotypes: in phenotype-1, person is normal, in phenotype-2 the person has no G6PD deficiency but with deuteranopia/deuteranomaly, in phenotype-3 the person is G6PD Med variant with deuteranopia/deuteranomaly and in phenotype 4 the person is G6PD Med variant has normal vision. Based on the molecular analysis of G6PD and Ishihara vision test it can be concluded that the two mutations at the two loci arose independent of each other without any interaction (epistatic effect) between them. Following the pedigree analysis of the two genes for 4 generations it is presumed that it is infeasible to use “deuteranopia /deuteranomaly” as a phenotypic marker to detect G6PD c563T heterozygous individuals among the Egyptian populations

    Cloning and molecular analysis of HlbZip1 and HlbZip2 transcription factors putatively involved in the regulation of the lupulin metabolome in hop (Humulus lupulus L.)

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    Hop (Humulus lupulus L.), the essential source of beer flavor is of interest from a medicinal perspective in view of its high content in health-beneficial terpenophenolics including prenylflavonoids. The dissection of biosynthetic pathway(s) of these compounds in lupulin glands, as well as its regulation by transcription factors (TFs), is important for efficient biotech no logical manipulation of the hop metabolome. TFs of the bZIP class were preselected from the hop transcriptome using a cDNA-AFLP approach and cloned from a cDNA library based on glandular tissue-enriched hop cones. The cloned TFs HlbZIP1A and HlbZIP2 have predicted molecular masses of 27.4 and 34.2 kDa, respectively, and both are similar to the group A3 bZIP TFs according to the composition of characteristic domains. While HlbZIP1A is rather neutral (pl 6.42), HlbZIP2 is strongly basic (pl 8,51). A truncated variant of HlbZIP1 (HlbZIP1B), which is strongly basic but lacks the leucine zipper domain, has also been cloned from hop. Similar to the previously cloned HIMyb3 from hop, both bZIP TFs show a highly specific expression in lupulin glands, although low expression was observed also in other tissues including roots and immature pollen. Comparative functional analyses of HbZip1A, HlbZip2, and subvariants of HIMyb3 were performed in a transient expression system using Nicotiana benthamiana leaf coinfiltration with Agrobacterium tumefaciens strains bearing hop TFs and selected promoters fused to the GUS reference gene. both hop bZIP TFs and HIMyb3 mainly activated the promoters of chalcone synthase chs_H1 and the,newly cloned O-methyl transferase 1 genes, while the response of the valerophenone synthase promoter to the cloned hop TFs was very low. These analyses also showed that the cloned bZIP TFs are not strictly G-box-specific. HPLC analysis of secondary metabolites in infiltrated Petunia hybrida showed that both hop bZIP TFs interfere with the accumulation and the composition of flavonol glycosides, phenolic acids, and anthocyanins, suggesting the possibility of coregulating flavonoid biosynthetic pathways in hop glandular tissue
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