Genetic regulation of glucoraphanin accumulation in Beneforté® broccoli

Diets rich in broccoli (Brassica oleracea var italica) have been associated with maintenance of cardiovascular health and reduction in risk of cancer. These health benefits have been attributed to glucoraphanin that specifically accumulates in broccoli. The development of broccoli with enhanced concentrations of glucoraphanin may deliver greater health benefits. Three high-glucoraphanin F1 broccoli hybrids were developed in independent programmes through genome introgression from the wild species Brassica villosa. Glucoraphanin and other metabolites were quantified in experimental field trials. Global SNP analyses quantified the differential extent of B. villosa introgression The high-glucoraphanin broccoli hybrids contained 2.5–3 times the glucoraphanin content of standard hybrids due to enhanced sulphate assimilation and modifications in sulphur partitioning between sulphur-containing metabolites. All of the high-glucoraphanin hybrids possessed an introgressed B. villosa segment which contained a B. villosa Myb28 allele. Myb28 expression was increased in all of the high-glucoraphanin hybrids. Two high-glucoraphanin hybrids have been commercialised as Beneforte broccoli. The study illustrates the translation of research on glucosinolate genetics from Arabidopsis to broccoli, the use of wild Brassica species to develop cultivars with potential consumer benefits, and the development of cultivars with contrasting concentrations of glucoraphanin for use in blinded human intervention studie

Las funciones de tipo como marcador del discurso en el lenguaje juvenil de Buenos Aires: un estudio descriptivo

El objetivo de este trabajo es analizar cómo los jóvenes de Buenos Aires aplican el marcador del discurso tipo en las conversaciones coloquiales para poder dar respuesta a la siguiente pregunta: ¿Qué funciones desempeña tipo como marcador de discurso en las conversaciones coloquiales entre jóvenes porteños, partiendo de los dos hipótesis de que 1)tipo está pasando por un proceso de gramaticalización y que 2)hay seis funciones de tipo como marcador del discurso; atenuante, aproximador, ejemplificador, intensificador, marca de cita directa y retardador.Es un estudio basado en el corpus COLAba (Corpus Oral de Lenguaje Adolescente de Buenos Aires). El corpus COLAba es un corpus de habla informal, transcrita y acoplada al sonido, accesible en Internet para la investigación

Jordtrøtthet i eple - forskjell mellom økologiske og konvensjonelle hager?

Formålet med forprosjektet VitalEPLEJORD var å studere potensielle årsaker til jordtrøtthet i epleproduksjonen, et fenomen som kan oppstå dersom det ikke praktiseres vekstskifte. Jordprøver fra 10 bruk (fem med økologisk og fem med konvensjonell drift) på Øst- og Vestlandet ble undersøkt med tanke på jordkvalitet, nematoder og Phytophthora. Innen hver av de tre kategoriene ble det gjort interessante funn. Både lite fruktbar jord og skadegjørere kan føre til dårlig etablering, svak tilvekst, avlingsreduksjon og at trær i verste fall dør. I de jordkjemiske analysene ble det vist høyere innhold av organisk materiale, totalt nitrogen og plantetilgjengelig fosfor i økologisk enn konvensjonell drift, samt mer stabilt innhold av plantetilgjengelig magnesium, mer stabil pH og mer stabil basemetningsgrad. Det ble identifisert en rekke nematodearter, inkludert flere planteparasittære. Økologisk drift så ut til å kunne øke bakteriekonsumerende nematoder, minske rotkonsumerende nematoder og øke altetere. Innen Phytophthora ble det funnet fire arter der alle er rapportert som alvorlige skadegjørere på treaktige vekster både i Norge og andre land. Phytophthora forekom ved begge driftsformer. Antallet jordprøver var totalt 50 (5 samleprøver/bruk, hver samleprøve kom fra separate rader). Testomfanget var for lite til å dra konklusjoner om årsak til jordtrøtthet, men det har gitt verdifull kunnskap til å bygge videre på.publishedVersio

Jordtrøtthet i eple - forskjell mellom økologiske og konvensjonelle hager?

Formålet med forprosjektet VitalEPLEJORD var å studere potensielle årsaker til jordtrøtthet i epleproduksjonen, et fenomen som kan oppstå dersom det ikke praktiseres vekstskifte. Jordprøver fra 10 bruk (fem med økologisk og fem med konvensjonell drift) på Øst- og Vestlandet ble undersøkt med tanke på jordkvalitet, nematoder og Phytophthora. Innen hver av de tre kategoriene ble det gjort interessante funn. Både lite fruktbar jord og skadegjørere kan føre til dårlig etablering, svak tilvekst, avlingsreduksjon og at trær i verste fall dør. I de jordkjemiske analysene ble det vist høyere innhold av organisk materiale, totalt nitrogen og plantetilgjengelig fosfor i økologisk enn konvensjonell drift, samt mer stabilt innhold av plantetilgjengelig magnesium, mer stabil pH og mer stabil basemetningsgrad. Det ble identifisert en rekke nematodearter, inkludert flere planteparasittære. Økologisk drift så ut til å kunne øke bakteriekonsumerende nematoder, minske rotkonsumerende nematoder og øke altetere. Innen Phytophthora ble det funnet fire arter der alle er rapportert som alvorlige skadegjørere på treaktige vekster både i Norge og andre land. Phytophthora forekom ved begge driftsformer. Antallet jordprøver var totalt 50 (5 samleprøver/bruk, hver samleprøve kom fra separate rader). Testomfanget var for lite til å dra konklusjoner om årsak til jordtrøtthet, men det har gitt verdifull kunnskap til å bygge videre på

Effects of fou8/fry1 mutation on sulfur metabolism: Is decreased internal sulfate the trigger of sulfate starvation response?

The fou8 loss of function allele of adenosine bisphosphate phosphatase FIERY1 results in numerous phenotypes including the increased enzymatic oxygenation of fatty acids and increased jasmonate synthesis. Here we show that the mutation causes also profound alterations of sulfur metabolism. The fou8 mutants possess lower levels of sulfated secondary compounds, glucosinolates, and accumulate the desulfo-precursors similar to previously described mutants in adenosine 5′phosphosulfate kinase. Transcript levels of genes involved in sulfate assimilation differ in fou8 compared to wild type Col-0 plants and are similar to plants subjected to sulfate deficiency. Indeed, independent microarray analyses of various alleles of mutants in FIERY1 showed similar patterns of gene expression as in sulfate deficient plants. This was not caused by alterations in signalling, as the fou8 mutants contained significantly lower levels of sulfate and glutathione and, consequently, of total elemental sulfur. Analysis of mutants with altered levels of sulfate and glutathione confirmed the correlation of sulfate deficiency-like gene expression pattern with low internal sulfate but not low glutathione. The changes in sulfur metabolism in fou8 correlated with massive increases in 3′-phosphoadenosine 5′-phosphate levels. The analysis of fou8 thus revealed that sulfate starvation response is triggered by a decrease in internal sulfate as opposed to external sulfate availability and that the presence of desulfo-glucosinolates does not induce the glucosinolate synthesis network. However, as well as resolving these important questions on the regulation of sulfate assimilation in plants, fou8 has also opened an array of new questions on the links between jasmonate synthesis and sulfur metabolism

Apramycin efficacy against carbapenem- and aminoglycoside-resistant Escherichia coli and Klebsiella pneumoniae in murine bloodstream infection models

Background The aminoglycoside apramycin has been proposed as a drug candidate for the treatment of critical Gram-negative systemic infections. However, the potential of apramycin in the treatment of drug-resistant bloodstream infections (BSIs) has not yet been assessed. Methods The resistance gene annotations of 40 888 blood-culture isolates were analysed. In vitro profiling of apramycin comprised cell-free translation assays, broth microdilution, and frequency of resistance determination. The efficacy of apramycin was studied in a mouse peritonitis model for a total of nine Escherichia coli and Klebsiella pneumoniae isolates. Results Genotypic aminoglycoside resistance was identified in 87.8% of all 6973 carbapenem-resistant Enterobacterales blood-culture isolates, colistin resistance was shown in 46.4% and apramycin in 2.1%. Apramycin activity against methylated ribosomes was &gt; 100-fold higher than that for other aminoglycosides. Frequencies of resistance were &lt; 10-9 at 8 × minimum inhibitory concentration (MIC). Tentative epidemiological cut-offs (TECOFFs) were determined as 8 µg/mL for E. coli and 4 µg/mL for K. pneumoniae. A single dose of 5 to 13 mg/kg resulted in a 1-log colony-forming unit (CFU) reduction in the blood and peritoneum. Two doses of 80 mg/kg resulted in an exposure that resembles the AUC observed for a single 30 mg/kg dose in humans and led to complete eradication of carbapenem- and aminoglycoside-resistant bacteraemia. Conclusion Encouraging coverage and potent in vivo efficacy against a selection of highly drug-resistant Enterobacterales isolates in the mouse peritonitis model warrants the conduct of clinical studies to validate apramycin as a drug candidate for the prophylaxis and treatment of BSI.Niels Frimodt-Møller, Diarmaid Hughes, Carina Vingsbo Lundberg and Sven N. Hobbie share senior authorship</p

Comparison of mRNA levels of key genes of sulfur metabolism.

<p>Col-0, <i>fou8</i>, <i>cad2</i>, <i>rax1</i>, and <i>sultr1;2</i> plants were grown for 2.5 weeks on MS-agarose plates. Relative mRNA levels of <i>ATPS4</i> and <i>APR1</i> in <b>A</b> leaves and <b>B</b> roots and of <i>LS2</i> (At5g48850) and <i>LS5</i> (At5g26220) in <b>C</b> leaves and <b>D</b> roots and genes of glucosinolate synthesis in <b>E</b> leaves were determined. The qRT-PCR reactions were performed in triplicate, the values in Col-0 were set to 1 for all genes. Results are presented as means ± SE from six biological replicates from plants grown in two independent experiments. Different letters mark values significantly different at P<0.05 or in <b>E</b> asterisks show values significantly (P<0.05) different from Col-0.</p

Accumulation of PAP and PAPS in <i>fou8</i> related genotypes.

<p>The various Arabidopsis lines were grown for 5 weeks in controlled environment room. Leaves were harvested and the levels of <b>A</b> PAP and <b>B</b> PAPS were determined by HPLC. Results are presented as means ± SD from four individual plants, grown in two independent experiments. Asterisks represents levels under detection limit, different letters mark values significantly different at P<0.05.</p

Mineral content of Col-0 and <i>fou8</i>.

<p>Col-0 and <i>fou8</i> plants were grown for 30 days in soil in controlled environment room. Whole rosettes were harvested and the mineral levels were determined by X-ray fluorescence spectrophotometry as % of dry weight. Results from one of two independent experiments are presented as means ± SD from three individual plants. Values substantially different between the two genotypes (P<0.05) are marked by asterisks.</p

Sulfate uptake and flux through sulfate assimilation in <i>fou8</i> and related mutants.

<p>WT Col-0 and mutants <i>fou8</i>, <i>fou2</i>, and <i>aos</i> were grown for 3 weeks on MS-phytagel vertical plates in controlled environment room. The seedlings were incubated for four hours with their roots submerged in nutrient solution adjusted to sulfate concentration of 0.2 mM and supplemented with 6.7 μCi [³⁵S]sulfate. Shoot and root material was harvested separately, and the flux was determined as incorporation of ³⁵S from [³⁵S] sulfate to thiols and proteins. <b>A</b> sulfate uptake, <b>B</b> Percentage of ³⁵S transported to leaves from the [³⁵S]sulfate taken up, <b>C</b> relative flux through the sulfate assimilation in the leaves calculated as % of incorporation in thiols and proteins from total [³⁵S]sulfate taken up. Results are presented as means ± SE from six independent pools of 8 seedlings grown in two independent experiments. Values marked with an asterisk show significant (P≤0.05) difference from Col-0.</p