RESUMO
Mineral elements in barley (Hordeum vulgare) play an important physiological role in global human health. In this study, quantitative trait loci (QTLs) for concentration of nine mineral elements in barley grain and grass powder were detected in a population of 193 recombinant inbred lines of the barley cross Ziguangmangluoerling x Schooner and the parents. We observed large genetic variation contributing to element concentrations in both grains and grass powder. The mean K, Ca, and Fe concentrations in grass powder were 6.67, 12.00, and 4.58 times that of regenerating barley grains. In grains, 17 QTLs that accounted for 6.36-64.08% of the phenotypic variation in Zn, Mg, Ca, K, Na, Mn, Fe, and P concentrations were identified. In grass powder, seven QTLs were identified; these accounted for 6.03-21.86% of the variation in Ca, Zn, Mg, K, Fe, and Cu concentrations. These QTLs affecting elements in grain and grass powder are so far unreported in barley. To our knowledge, QTLs with pleiotropic effects for three elements were also identified for the first time in barley. The qK1/qMg1/qCa1 region between markers Bmag0211 and GBMS0014 on chromosome 1H was shown to have large additive effects for Mg, Ca, and K concentrations in grains. These additive effects indicated that the high element (Mg, Ca, Zn, Mn, and K) alleles were contributed by Ziguangmangluoerling. These results will further our understanding of the genetic basis of mineral elements and help us develop markers linked with mineral elements for marker-assisted selection breeding in barley.
Assuntos
Hordeum/genética , Minerais/análise , Locos de Características Quantitativas , DNA de Plantas/genética , Grão Comestível/genética , Variação Genética , Melhoramento Vegetal , Seleção GenéticaRESUMO
An F3 population consisting of 117 F2:3 families derived from a cross between two varieties of rice, Gongmi No. 3 and Diantun 502, with a large difference in their resistant starch and total alkaloid content, was used for quantitative trait locus (QTL) mapping. Two QTLs of resistant starch for rice (qRS7-1, qRS7-2) were identified in a linkage group on chromosome 7, which could explain phenotypic variance from 7.6 to 17.3%, due to additive effects for resistant starch from Gongmi No. 3 or over-dominance effects for qRS7-2 of the marker interval (RM3404-RM478) on chromosome 7 from Gongmi No. 3, accounting for 13.8-17.3% of the phenotypic variance. Two QTLs of total alkaloids for brown rice (qALb7-1, qALb7-2) were identified in the same linkage group, which could explain phenotypic variance from 7.7 and 19.3%, respectively, due to dominance or over-dominance effects for total alkaloids on chromosome 7 from Diantun 502. To our knowledge, these are the first QTLs to be identified, which are related to resistant starch and total alkaloid content in rice. These results are beneficial for understanding the genetic basis of, as well as for developing markers linked with, resistant starch and total alkaloids of functional components for marker-assisted selection breeding in rice.
Assuntos
Cromossomos de Plantas/genética , Oryza/genética , Locos de Características Quantitativas , Alcaloides/genética , Alcaloides/metabolismo , Mapeamento Cromossômico , Cruzamentos Genéticos , Genes de Plantas , Estudos de Associação Genética , Ligação Genética , Loci Gênicos , Marcadores Genéticos , Oryza/metabolismo , Fenótipo , Melhoramento Vegetal , Amido/genética , Amido/metabolismoRESUMO
Black disease is an acute disease of sheep and cattle. The pathogen is the obligate anaerobe, Clostridium novyi. Due to difficulties of anaerobic culturing in the country or disaster sites, a simple, rapid, and sensitive method is required. In this study, an electrochemical method, the cyclic voltammetry method, basing on loop-mediated isothermal amplification (LAMP), electrochemical ion bonding (positive dye, methylene blue), was introduced. DNA extracted from C. novyi specimens was amplified through the LAMP reaction. Then the products combined were with methylene blue, which lead to a reduction in the oxidation peak current (ipA) and the reduction peak current (ipC) of the cyclic voltammetry. The changes of ipA/ipC were real-time measured by special designed electrode, so the DNA was quantitatively detected. The results displayed that this electrochemical detection of C. novyi could be completed in 1-2 h with the lowest bacterial concentration of 10(2) colony forming units/mL, and high accuracy (96.5%), sensitivity (96%), and specificity (97%) compared to polymerase chain reation. The cyclic voltammetry method was a simple and fast method, with high sensitivity and high specificity, and has great potential to be a usable molecular tool for fast diagnosis of Black disease.
Assuntos
Doenças dos Animais/genética , Clostridium/genética , DNA Bacteriano/genética , Carneiro Doméstico/microbiologia , Doenças dos Animais/microbiologia , Animais , Bovinos , Clostridium/patogenicidade , Técnicas de Amplificação de Ácido Nucleico/métodos , Reação em Cadeia da Polimerase/métodosRESUMO
We used a newly developed electrochemical method, real-time resistance measurement, based on loop-mediated isothermal amplification (LAMP), with real-time resistance monitoring and derivative analysis. DNA extracted from specimens was amplified through LAMP reaction. The 2 products of LAMP, DNA and pyrophosphate, both are negative ions; they combine with positive dye (crystal violet) and positive ions (Mg(2+)), which leads to an increase in the resistivity of the reaction liquid. The changes of resistivity were measured in real-time with a specially designed resistance electrode, to detect Clostridium difficile DNA. We found that electrochemical detection of C. difficile could be completed in 0.5-1 h, with a detection limit of 10(2) CFU/mL, with high accuracy (95.0%), sensitivity (91.1%), and specificity (97.3%) compared to PCR methods. C. difficile is commonly associated with antibiotic-induced diarrhea. Due to the difficulty in performing anaerobic culture and cytotoxicity neutralization assays, a simple, rapid, sensitive, and accurate method is preferred. We conclude that real-time resistance measurement is a rapid, sensitive, and stable method for the diagnosis of C. difficile infection that could be applied to gene chips and pocket instruments.
Assuntos
Clostridioides difficile/isolamento & purificação , Infecções por Clostridium/diagnóstico , DNA Bacteriano/isolamento & purificação , Clostridioides difficile/genética , Clostridioides difficile/patogenicidade , Infecções por Clostridium/genética , Fezes/microbiologia , Humanos , Limite de Detecção , Sensibilidade e EspecificidadeRESUMO
Ayu17-449, a novel gene in mice, has been identified as a tumor-suppressor gene in myeloid malignancy; its product catalyzes the conversion of 5-methylcytosine of DNA to 5-hydroxymethylcytosine. However, in vivo, its functional target genes and biological function have remained unclear. Based on the assumption that alterations in the expression of the Ayu17-449 gene affect the expression of other related genes, we screened a microarray of altered gene expression in Ayu17-449(-/-) and Ayu17-449(+/+) mice. We identified 4049 genes with altered expression, including 1296 up-regulated (fold change ≥2) and 2753 down-regulated (fold change ≤0.5) genes in knockout mice compared with control mice. We then used qRT-PCR and RT-PCR to validate the chip data. Gene ontology and pathway analysis were performed on these altered genes. We found that these altered genes are functional genes in the complement and coagulation cascades, metabolism, biosynthesis, transcriptional regulation, proteolysis, and intracellular signaling pathways, such as the peroxisome proliferator-activated-receptor signaling pathway, the TNF-α-NF-κB pathway, the Notch signaling pathway, the MAPK signaling pathway, and the insulin signaling pathway. The results of our genome-wide comprehensive study could be helpful for comprehending the underlying functional mechanisms of the Ayu17-449 gene in mammals.