Applied Biological Chemistry

The adsorption of phenanthrene onto activated carbons produced from Vitisvinifera leaf litter (a waste plant biomass) was investigated in this study. Zinc chloride (ZnCl₂) and phosphoric acid (H₃PO₄) were utilised as activating agents in producing the activated carbons. The characterisation of the activated carbons was achieved with Fourier transform infrared spectroscopy (for surface functional groups), scanning electron microscopy (for surface morphology) and Brunauer–Emmett–Teller (BET) (for surface area determination). The adsorption of phenanthrene onto the activated carbons was optimised in terms of solution pH, adsorbent dosage, initial concentration of adsorbate solution and contact time. Experimental results showed that H₃PO₄ modified activated carbon gave better yield (up to 58.40%) relative to ZnCl₂ modified activated carbon (only up to 47.08%). Meanwhile, surface characterisation showed that ZnCl₂ modification resulted in higher BET surface area (up to 616.60 m²/g) and total pore volume (up to 0.289 cm³/g) relative to BET surface area of up to 295.49 m²/g and total pore volume of up to 0.185 cm³/g obtained from H₃PO₄ modified activated carbons. Adsorption equilibrium data fitted well into Freundlich isotherm model relative to other applied isotherm models, with maximum K_f value of 1.27 for ZnCl₂ modified activated carbon and 1.16 K_f value for H₃PO₄ modified activated carbon. The maximum adsorption capacity for ZnCl₂ and H₃PO₄ activated carbons for the removal of phenanthrene were 94.12 and 89.13 mg/g, respectively. Kinetic studies revealed that dynamic equilibrium was reached at 80 min contact time. Experimental data fitted best into the Elovich kinetic model relative to other kinetic models, based on the correlation coefficient (R²) values obtained from kinetic studies. Chemisorption was deduced as a major phenanthrene removal pathway from aqueous solution and the physicochemical characteristics of the adsorbents have major influence on phenanthrene removal efficiencies.

Agsatache rugosa (Korean mint), belongs to the mint family and it has various medicinal properties. In addition, it has several valuable compounds such as monoterpenes and phenylpropanoid compounds. Amongst these, two compounds viz., rosmarinic acid (RA), and tilianin are well-known natural compounds that have numerous pharmacological properties. The phenylpropanoid biosynthetic gene expression under stress conditions and the subsequent accumulation of phenylpropanoid content has not been extensively studied in Korean mint. Here, we investigated the effect of light-emitting diodes (LEDs) on the expression levels of phenylpropanoid biosynthetic pathway genes and the accumulation of phenylpropanoid compounds such as RA and tilianin in A. rugosa. Real-time PCR analysis showed that the phenylpropanoid pathway genes responded to the LED lights. The transcript levels of downstream genes (C4H, CHS, CHI, and RAS) were comparatively higher than those of upstream genes (PAL, TAT, and HPPR). In addition, HPLC analysis showed that the content of RA and tilianin were significantly higher in plants cultivated under white light than those grown under red, blue, green, and orange lights. The RA and tilianin content were the highest in the plantlets after three weeks of exposure to white light. These results suggested that white LED lights significantly enhanced the accumulation of phenylpropanoid compounds in A. rugosa.

Herbicide resistance is one of the main crop traits that improve farming methods and crop productivity. CRISPR-Cas9 can be applied to the development of herbicide-resistant crops based on a target site resistance mechanism, by editing genes encoding herbicide binding proteins. The sgRNAs capable of editing the target genes of herbicides, pds (phytoene desaturase), ALS (acetolactate synthase), and EPSPS (5-Enolpyruvylshikimate-3-phosphate synthase), were designed to use with the CRISPR-Cas9 system in tomato (Solanum lycopersicum cv. Micro-Tom). The efficiency of the sgRNAs was tested using Agrobacterium mediated transient expression in the tomato cotyledons. One sgRNA designed for editing the target site of PDS had no significant editing efficiency. However, three different sgRNAs designed for editing the target site of ALS had significant efficiency, and one of them, ALS2-P sgRNA, showed over 0.8% average efficiency in the cotyledon genome. The maximum efficiency of ALS2-P sgRNA was around 1.3%. An sgRNA for editing the target site of EPSPS had around 0.4% editing efficiency on average. The sgRNA efficiency testing provided confidence that editing of the target sites could be achieved in the transformation process. We confirmed that 19 independent transgenic tomatoes were successfully edited by ALS2_P or ALS1_W sgRNAs and two of them had three base deletion mutations, which are expected to have altered herbicide resistance. In this study, we demonstrated the usefulness of performing an sgRNA efficiency test before crop transformation, and confirmed that the CRISPR-Cas9 system is a valuable tool for breeding herbicide-resistant crops.

This study aimed to comprehensively analyze primary and secondary metabolites of three different-colored (white, pale green, and green) pak choi cultivars (Brassica rapa subsp. chinensis) using gas chromatography attached with time-of-flight mass spectrometry (GC-TOFMS) and high-performance liquid chromatography (HPLC). In total, 53 primary metabolites were identified and subjected to partial least-squares discriminant analysis. The result revealed a significant difference in the primary and secondary metabolites between the three pak choi cultivars. In addition, 49 hydrophilic metabolites were detected in different cultivars. Total phenolic and glucosinolate contents were highest in the pale green and green cultivars, respectively, whereas total carotenoid and chlorophyll contents were highest in the white cultivar. Superoxide dismutase activity, 2,2-diphenyl-1-picrylhydraz scavenging, and reducing power were slightly increased in the white, pale green, and green cultivars, respectively. In addition, a negative correlation between pigments and phenylpropanoids was discovered by metabolite correlation analysis. This approach will provide useful information for the development of strategies to enhance the biosynthesis of phenolics, glucosinolates, carotenoids, and chlorophyll, and to improve antioxidant activity in pak choi cultivars. In addition, this study supports the use of HPLC and GC-TOFMS-based metabolite profiling to explore differences in pak choi cultivars.