RESUMO
MAIN CONCLUSION: Induced mutations in the SC-uORF of the tomato transcription factor gene SlbZIP1 by the CRISPR/Cas9 system led to the high accumulation of sugar and amino acid contents in tomato fruits. Tomato (Solanum lycopersicum) is one of the most popular and consumed vegetable crops in the world. Among important traits for tomato improvement such as yield, biotic and abiotic resistances, appearance, post-harvest shelf life and fruit quality, the last one seems to face more challenges because of its genetic and biochemical complexities. In this study, a dual-gRNAs CRISPR/Cas9 system was developed to induce targeted mutations in uORF regions of the SlbZIP1, a gene involved in the sucrose-induced repression of translation (SIRT) mechanism. Different induced mutations in the SlbZIP1-uORF region were identified at the T0 generation, stably transferred to the offspring, and no mutation was found at potential off-target sites. The induced mutations in the SlbZIP1-uORF region affected the transcription of SlbZIP1 and related genes in sugar and amino acid biosynthesis. Fruit component analysis showed significant increases in soluble solid, sugar and total amino acid contents in all SlbZIP1-uORF mutant lines. The accumulation of sour-tasting amino acids, including aspartic and glutamic acids, raised from 77 to 144%, while the accumulation of sweet-tasting amino acids such as alanine, glycine, proline, serine, and threonine increased from 14 to 107% in the mutant plants. Importantly, the potential SlbZIP1-uORF mutant lines with desirable fruit traits and no impaired effect on plant phenotype, growth and development were identified under the growth chamber condition. Our result indicates the potential utility of the CRISPR/Cas9 system for fruit quality improvement in tomato and other important crops.
Assuntos
Solanum lycopersicum , Fatores de Transcrição , Fatores de Transcrição/genética , Aminoácidos/metabolismo , Açúcares/metabolismo , Solanum lycopersicum/genética , Sistemas CRISPR-Cas , Frutas/genética , Frutas/metabolismoRESUMO
Contamination status, spatial variability, and exposure risk of triclosan (TCS) and triclocarban (TCC) in indoor dusts from different micro-environments were evaluated for the first time in Vietnam as well as in Southeast Asian region. TCS and TCC were measured in 89 dust samples collected from bedrooms, living rooms, and kitchens of private houses in four northern cities including Hanoi, Bac Ninh, Hung Yen, and Nam Dinh, by means of liquid chromatography-tandem mass spectrometry. Concentrations of TCS and TCC ranged from <5 to 1090 (median 33.2) and from <3 to 531 (median 19.3) ng g-1, respectively. Concentrations of TCS and TCC in the kitchen and bedroom dusts were markedly higher than levels found in the living room samples, probably due to their applications in kitchen utensils, household cleaning reagents, and personal care products. A strong positive correlation between TCS and TCC concentrations was detected in the whole dataset (R2 = 0.810, p < 0.001). For samples in which both TCS and TCC were quantified, TCS/TCC ratios ranged from 0.3 to 12 with a median value of 1.8, and did not show big differences between micro-environments. Human exposures to TCS and TCC through dust ingestion were estimated for various age groups with 95% CI daily intake doses ranging from (0.032-0.070) to (0.340-0.740) and from (0.017-0.033) to (0.175-0.345) ng kg-bw-1 d-1 for adults and infants respectively. Although our derived values were much lower than reference doses, more comprehensive risk assessment considering multiple exposure pathways of TCS and TCC is needed.