Recovery of Cembratrien-Diols from Waste Tobacco (Nicotiana tabacum L.) Flowers by Microwave-Assisted Deep Eutectic Solvent Extraction: Optimization, Separation, and In Vitro Bioactivity.

Deep eutectic solvents (DESs) are novel solvents with physicochemical properties similar to those of ionic liquids, and they have attracted extensive attention for the extraction of bioactive compounds from different plant materials in the context of green chemistry and sustainable development. In this study, seven DESs with different polarities were explored as green extraction solvents for cembratrien-diols (CBT-diols) from waste tobacco flowers. The best solvent, DES-3 (choline chloride: lactic acid (1:3)), which outperformed conventional solvents (methanol, ethanol, and ethyl acetate), was selected and further optimized for microwave-assisted DES extraction using the response surface methodology. The maximum yield of CBT-diols (6.23 ± 0.15 mg/g) was achieved using a microwave power of 425 W, microwave time of 32 min, solid/liquid ratio of 20 mg/mL, and microwave temperature of 40 °C. Additionally, the isolated CBT-diols exhibited strong antimicrobial activity against Salmonella, Staphylococcus aureus, Escherichia coli, Bacillus subtilis, and Pseudomonas aeruginosa and antitumor activity in the human liver cancer HepG2 and SMMC-7721 cell lines. This study highlights the feasibility of recovering CBT-diols from tobacco flower waste using DESs and provides opportunities for potential waste management using green technologies.

Bacterial dynamic of flue-cured tobacco leaf surface caused by change of environmental conditions.

Microorganisms present on the surface of tobacco leaves play a significant role in shaping the composition of the tobacco microbial ecosystem, which undergoes continuous changes throughout the curing process. In the present study, a total of four distinct tobacco curing periods were selected for sampling, namely the fresh, yellowing, leaf-drying, and stem-drying stages. The bacterial 16S rRNA gene sequences of the collected samples were subsequently analyzed to identify operational taxonomic units (OTUs). The findings indicated that the complete dataset of leaf microbial samples was clustered, resulting in the identification of 1,783 operational taxonomic units (OTUs). Furthermore, the analysis of diversity revealed a pattern of initially increasing and subsequently decreasing community diversity. Redundancy Analysis (RDA) and weighted gene correlation networks for analysis (WGCNA) were employed in conjunction with environmental factors to assign OTUs to 22 modules for functional analysis. Additionally, a classification model utilizing the random forest algorithm was utilized to identify seven marker microorganisms (Escherichia coli, Faecalibacterium prausnitzii, Faecalibacterium, Escherichia-Shigella, Peptostreptococcaceae, Peptostreptococcales-Tissierellales, and Proteobacteria) that exhibited discriminative characteristics across different time periods. This study aimed to investigate the dynamic changes in the bacterial community throughout the curing process and their impact on the community's function. Additionally, certain bacteria were identified as potential markers for detecting changes in the curing stage. These findings offer a novel opportunity to accurately regulate the curing environment, thereby enhancing the overall quality of tobacco leaf curing.

The application of plant growth-promoting rhizobacteria enhances the tolerance of tobacco seedling to salt stress.

The application of plant growth-promoting rhizobacteria (PGPR) is a novel and an efficient strategy for improving soil degradation and productivity. However, the effect of PGPR on tobacco (Nicotiana tabacum L.) seedling growth under salt stress remains unclear. Here, microcosm experiments were designed to verify the effects of Bacillus cereus TC012 (BC), Bacillus methylotrophicus TC023 (BM), and Bacillus amyloliquefacien TC037 (BA) on tobacco grown in salt-affected soil. The results showed that BC, BM, and BA treatments significantly increased the height of tobacco plants by 38.65%, 91.94%, and 90.66%, respectively. Furthermore, the growth of various components of tobacco plant, such as stem girth, seedling biomass, carotenoid, and chlorophyll were stimulated in salt-affected soils. The changes in the salinity of the tobacco plant mostly relies on the improvement of proline, soluble protein, soluble sugar content, plant protective enzymatic activity, and K+/Na+ ratios. Increases in indole-3-acetic acid, zeatin riboside and gibberellic acid also promoted tobacco growth. Additionally, inoculation with PGPR enhanced the enzymatic activity of laccase, urease, neutral protease, acid phosphatase, and sucrase in soil samples and had positive effects on the physicochemical properties. The soil bacterial communities significantly improved after inoculation with PGPR. In particular, the relative abundance of Pseudomonas and Bacillus significantly increased. Overall, PGPR inoculation has great potential to alleviate salt damage in tobacco plants and may have far reaching benefits to the agricultural community.

Waste seaweed compost and rhizosphere bacteria Pseudomonas koreensis promote tomato seedlings growth by benefiting properties, enzyme activities and rhizosphere bacterial community in coastal saline soil of Yellow River Delta, China.

This study investigated the effects of waste seaweed compost and rhizosphere bacteria Pseudomonas koreensis HCH2-3 on the tomato seedlings growth in coastal saline soils and chemical properties, enzyme activities, microbial communities of rhizosphere soil. Microcosmic experiment showed that the seaweed compost and rhizosphere bacteria (SC + HCH2-3) significantly alleviated the negative effects of salinity on the growth of tomato seedlings. SC + HCH2-3 amendment significantly increased the plant height and root fresh biomass of tomato seedling by 105.59% and 55.60% in the coastal saline soils, respectively. The soil properties and enzyme activities were also dramatically increased, indicating that the nutrient status of coastal saline soil was improved by SC + HCH2-3 amendment. In addition, Proteobacteria, Actinobacteriota and Firmicutes were the dominant phyla in the rhizosphere soil after adding seaweed compost and rhizosphere bacteria P. koreensis HCH2-3. The relative abundances of Massilia, Azospira, Pseudomonas and Bacillus increased in treatment SC + HCH2-3. Especially, the beneficial bacteria genera, such as Pseudomonas, Bacillus and Azospira, were significantly correlated with the increases of contents of total nitrogen, nitrate nitrogen and ammonium nitrogen in tomato rhizosphere soil samples. Consequently, adding waste seaweed compost and rhizosphere bacteria P. koreensis HCH2-3 into coastal saline soil was suggested as an effective method to relieve salt stress of tomato plants.

Combinations of waste seaweed liquid fertilizer and biochar on tomato (Solanum lycopersicum L.) seedling growth in an acid-affected soil of Jiaodong Peninsula, China.

Biochar application is an effective strategy for improving soil degradation and productivity. However, the effects of the combination of biochar and other fertilizers to improve seedling growth in abiotic stress-affected soils remains unknown. We investigate the effect of biochar derived from reed straw (RBC) and waste seaweed liquid fertilizer (SLF) on tomato (Solanum lycopersicum L.) seedling growth in an acid-affected soil of Jiaodong Peninsula, China. The results revealed RBC, SLF, and the combination of RBC with SLF (RBC+SLF) significantly elevated the dry weight of tomatoes by 23.33 %, 29.93 %, and 63.66 %, respectively. The malondialdehyde content in the tomato seedling roots, stems, and leaves was significantly lower in the RBC+SLF treatment, which might be related to the enhanced contents of proline, soluble sugar, and soluble protein. The synthesis and accumulation of zeatin riboside, indole-3-acetic acid, and gibberellic acid 3 in tomato under RBC+SLF amendment may be attributed to the enhanced plant growth. Moreover, RBC, SLF, and RBC+SLF improved the soil status (including ammonium nitrogen, nitrate nitrogen, laccase, and urease) in the acid-affected soil. Biochar and waste seaweed liquid fertilizer significantly increased the relative abundance of Pseudomonas and Azospira (beneficial bacteria) in tomato rhizosphere. The microbial amino acid metabolism was associated with changes in soil properties and enzyme activities. Consequently, biochar and waste seaweed liquid fertilizer are viable soil conditioners for acid-affected soil.

Chemical Compositions, Extraction Optimizations, and In Vitro Bioactivities of Flavonoids from Perilla Leaves (Perillae folium) by Microwave-Assisted Natural Deep Eutectic Solvents.

Natural deep eutectic solvents (NADESs) have been gradually applied to green extraction of active ingredients. In this study, microwave-assisted NADESs were applied to the extraction of flavonoid compounds from perilla leaves. Through comparative experiments, NADES-3 (choline chloride and malic acid at a molar ratio of 1:1) was found to have the highest extraction efficiency of total flavonoids, including apigenin 7-O-caffeoylglucoside, scutellarein 7-O-diglucuronide, luteolin 7-O-diglucuronide, and scutellarein 7-O-glucuronide by HPLC-MS. The following optimal extraction parameters were obtained based on response surface design: water content in NADES of 23%, extraction power of 410 W, extraction time of 31 min, and solid-liquid ratio of 75 mg/mL, leading to the extraction yield of total flavonoids of 72.54 mg/g. Additionally, the strong antimicrobial and antiallergic activity, inhibition of nitrosation, and antioxidant activity of total flavonoids by using NADESs were confirmed. This new extraction method provides a reference for the further exploration of NADES systems and may be widely used for the green extraction of natural active ingredients.

Pseudomonas Inoculation Stimulates Endophytic Azospira Population and Induces Systemic Resistance to Bacterial Wilt.

Bacterial communities in the rhizosphere play an important role in sustaining plant growth and the health of diverse soils. Recent studies have demonstrated that microbial keystone taxa in the rhizosphere microbial community are extremely critical for the suppression of diseases. However, the mechanisms involved in disease suppression by keystone species remain unclear. The present study assessed the effects of three Pseudomonas strains, which were identified as keystone species in our previous study, on the growth performance and root-associated bacterial community of tobacco plants. A high relative abundance of Ralstonia was found in the non-inoculated group, while a large Azospira population was observed in all groups inoculated with the three Pseudomonas strains. Correspondingly, the activities of the defense-related enzymes and the expression levels of the defense signaling marker genes of the plant were increased after inoculation with the Pseudomonas strains. Moreover, the correlation analyses showed that the relative abundance of Azospira, the activity of superoxide dismutase, catalase, and polyphenol oxidase, and the expression of H1N1, ACC Oxidase, and PR1 a/c had a significantly negative (p<0.05) relationship with the abundance of Ralstonia. This further revealed that the keystone species, such as Pseudomonas spp., can suppress bacterial wilt disease by enhancing the systemic resistance of tobacco plants.

Combined Use of Deep Eutectic Solvents, Macroporous Resins, and Preparative Liquid Chromatography for the Isolation and Purification of Flavonoids and 20-Hydroxyecdysone from Chenopodium quinoa Willd.

: Deep eutectic solvents (DESs) were used in combination with macroporous resins to isolate and purify flavonoids and 20-hydroxyecdysone from Chenopodium quinoa Willd by preparative high-performance liquid chromatography (HPLC). The extraction performances of six DESs and the adsorption/desorption performances of five resins (AB-8, D101, HPD 400, HPD 600, and NKA-9) were investigated using the total flavonoid and 20-hydroxyecdysone extraction yields as the evaluation criteria, and the best-performing DES (choline chloride/urea, DES-6) and macroporous resin (D101) were further employed for phytochemical extraction and DES removal, respectively. The purified extract was subjected to preparative HPLC, and the five collected fractions were purified in a successive round of preparative HPLC to isolate three flavonoids and 20-hydroxyecdysone, which were identified by spectroscopic techniques. The use of a DES in this study significantly facilitated the preparative-scale isolation and purification of polar phytochemicals from complex plant systems.

Porous organic cage incorporated monoliths for solid-phase extraction coupled with liquid chromatography-mass spectrometry for identification of ecdysteroids from Chenopodium quinoa Willd.

Here, a porous organic cage (POC)-incorporated polymeric monolith was fabricated in a syringe through the introduction of the POC into poly(ethylene glycol dimethacrylate) monolith in a one-step traditional free-radical polymerization proceess. The resulting monolithic phases were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FT-IR), powder X-ray diffraction (PXRD), nitrogen adsorption/desorption experiments and thermogravimetric analysis (TGA), which confirmed the successful incorporation of the POC in the monolithic matrix. The functionality of the POC-incorporated poly(EDMA) monolith facilitated for the solid phase extraction (SPE) of 20-hydroxecdysone (an ecdysteroid) from Chenopodium quinoa Willd. extract coupled with UPLC-QqQ-MS/MS, exhibiting satisfactory accuracy (93-106%), precision (< 6.5%) and reusability. In addition, UPLC-Q-Exactive-Orbitrap-MS/MS analysis of the quinoa sample after SPE by POC-incorporated monolith provided the identification of 20-hydroxecdysone and three other ecdysteroids. These results demonstrate the potential of POC-incorporated monoliths for the SPE of ecdysteroids from complex plant systems.

Environmentally-Friendly Extraction of Flavonoids from Cyclocarya paliurus (Batal.) Iljinskaja Leaves with Deep Eutectic Solvents and Evaluation of Their Antioxidant Activities.

Deep eutectic solvents (DESs) are commonly employed as environmentally-friendly solvents in numerous chemical applications owing to their unique physicochemical properties. In this study, a novel and environmentally-friendly extraction method based on ultrasound assisted-deep eutectic solvent extraction (UAE-DES) was investigated for the extraction of flavonoids from Cyclocarya paliurus (Batal.) Iljinskaja (C. paliurus) leaves, and the antioxidant activities of these flavonoids were evaluated. Nine different DES systems based on either two or three components were tested, and the choline chloride/1,4⁻butanediol system (1:5 molar ratio) was selected as the optimal system for maximizing the flavonoid extraction yields. Other extraction conditions required to achieve the maximum flavonoid extraction yields from the leaves of C. paliurus were as follows: DES water content (v/v), 30%; extraction time, 30 min; temperature, 60 °C; and solid-liquid ratio, 20 mg/mL. Liquid chromatography-mass spectrometry allowed the detection of five flavonoids in the extract, namely kaempferol-7-O-α-l-rhamnoside, kaempferol, quercetin, quercetin-3-O-ß-d-glucuronide, and kaempferol-3-O-ß-d-glucuronide. In vitro antioxidant tests revealed that the flavonoid-containing extract exhibited strong DPPH and ABTS radical-scavenging abilities. Results indicate that UAE-DES is a suitable approach for the selective extraction of flavonoids from C. paliurus leaves, and DESs can be employed as sustainable extraction media for other bioactive compounds.