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High-Energy Emulsified Clove Essential Oil Nanoemulsion as a Natural Herbicidal Product: Germination Suppression and Seed Structure Alteration in Echinochloa crus-galli
(2026-03-01)
Sikhao, Potjana
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Somala, Naphat
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Manichart, Nutcha
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Dimak, Jantra
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Poonpaiboonpipat, Thanatsan
Clove (Syzygium aromaticum (L.) Merr. & L.M. Perry) essential oil (EO)-based nanoemulsions may have a promising future in eco-friendly herbicide development. Clove EO was found to have a high eugenol content of 87.27%. Organic-solvent-free nanoemulsions using clove EO as a bioactive ingredient were fabricated using ultrasonication and microfluidization emulsification methods. Fourier-transform infrared spectroscopy confirmed that both emulsification methods did not affect the EO components. The droplet size of optimized nanoemulsions was determined using dynamic light scattering. The smallest size of 66.9 nm was obtained by microfluidization at 20,000 psi and eight passes. Additionally, the smallest droplet size for a sonicated nanoemulsion was 103.9 nm, obtained by ultrasonication at 20% for 6 min. Transmission electron microscopy confirmed the droplet sizes of both optimized nanoemulsions. In a storage test, both optimized nanoemulsions were stored at 4 °C for at least four weeks. Finally, both nanoemulsions were evaluated on pre-emergence herbicidal activities against Echinochloa crus-galli. The results showed that both nanoemulsions inhibited E. crus-galli germination and seedling growth, and additionally, inhibited seed imbibition and α-amylase activity. Micro-morphological and ultrastructural analysis was observed using a scanning electron microscope and an energy dispersive X-ray spectrometer (SEM-EDS). SEM-EDS micrographs of the treated seeds showed that the seed structure was damaged, especially the endosperm, leading to the inhibition of seed germination.
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Ultrasonic emulsification of Cananga odorata nanoemulsion formulation for enhancement of herbicidal potential
(2025-12-01)
Laosinwattana, Chamroon
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Somala, Naphat
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Dimak, Jantra
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Teerarak, Montinee
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Chotsaeng, Nawasit
A nanoemulsion was fabricated from Cananga odorata essential oil (EO) and stabilized by incorporation of Tween 80 using ultrasonication. The major constituents of the EO were benzyl benzoate, linalool, and phenylmethyl ester. Differing sonication amplitude (20–60%) and time (2–10 min) were assessed for effects on nanoemulsion droplet size and polydispersity index (PI). The smallest droplet size of 43.98 nm (PI 0.222) was obtained using 40% amplitude for 8 min; this nanoemulsion was evaluated for its droplet characteristics and pre-emergence herbicidal activities on Amaranthus tricolor. FT-IR confirmed ultrasonic emulsification to not affect the EO components. Regarding stability, storage at 4 °C was determined appropriate, with droplet size changing slightly after five weeks. Assays of herbicidal potential showed the coarse emulsion and nanoemulsion to both reduce A. tricolor germination and growth, with the nanoemulsion being more effective at a given concentration and the difference in effectivity correlating to droplet size. Remarkably, treatment with 250 ppm nanoemulsion and coarse emulsion respectively resulted in 100% and 63.75% germination inhibition. Both emulsions decreased seed imbibition and alpha-amylase activity. The highest relative electrolyte leakage was achieved in seed treated with the nanoemulsion. Therefore, this ultrasonic-based nanoemulsion may have utility as bioherbicide alternative.
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Differential Biochemical Responses of Resistant and Susceptible Genotypes of Chili to Pepper Yellow Leaf Curl Thailand Virus
(2025-09-01)
Mueangkhong, Manthana
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Suwor, Patcharaporn
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Techawongstien, Suchila
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Teerarak, Montinee
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Tsai, Wen Shi
Chili (Capsicum annuum L.) production is threatened by the pepper yellow leaf curl virus (PepLCV), transmitted by whiteflies, leading to reduced yields. This study investigated the biochemical changes in two chili genotypes, PEP6 (tolerant to PepLCV) and Homsuphan (susceptible to PepLCV), following inoculation with the Thailand strain of PepLCV (known as Pepper Yellow Leaf Curl Thailand Virus, PepYLCTHV). Inoculation was performed using whitefly transmission (WF) and graft transmission (GT) methods, and disease severity was evaluated using a standardized index. The level of total phenolic compounds and the activities of peroxidase (POD) and polyphenol oxidase (PPO) enzymes were analyzed in virus-infected plants and compared with those in uninoculated controls. Both chili genotypes exhibited a more rapid increase in disease severity when inoculated with WF than with GT. In PEP6, disease severity was lower than Homsuphan in both WF and GT inoculations. Disease severity in WT-inoculated PEP6 plants increased gradually, reaching 100% by day 36, whereas Homsuphan plants had a more rapid progression, attaining 100% by day 21. The GT method led to slower disease severity progression in both genotypes, reaching 80–85% by day 36. In PEP6 plants, total phenolic compound content increased significantly following WF, indicating an active defense response, whereas levels remained stable in GT plants. Phenolic content in the tolerant genotype Homsuphan remained stable across all conditions. Notably, peroxidase (POD) activity was elevated in GT plants of both genotypes, which correlated with reduced disease severity. Polyphenol oxidase (PPO) activity was lowest in control plants, but WT and GT increased the PPO level. Among the treatments, GT induced the highest PPO activity, which was associated with the lowest disease severity. These findings suggest that GT may enhance disease resistance by modulating phenolic compound accumulation and increasing POD and PPO activity.
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Siamese neem tree as a natural preservative: Chemical profile, antioxidant properties, and antibacterial efficacy against foodborne pathogens and spoilage bacteria
(2025-03-01)
Puttongsiri, Tongchai
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Manichart, Nutcha
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Teerarak, Montinee
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Sikhao, Potjana
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Somala, Naphat
The objective of the present study was to investigate the effect of various ethanol in water solvent systems (0, 25, 50, 75 and 100 % v/v) on the extraction crude yield, total phenolic content, total flavonoid content, and chemical profile of Azadirachta indica (Siamese neem tree) leaf extracts. In addition, the antioxidant and antimicrobial properties were investigated using in vitro assays. Crude yield was highest for the water extract (5.78 g/100 g DW) and decreased with increased ethanol. Total phenolic and flavonoid contents were highest in the 75 % ethanol extract (26.95 mg GAE/g crude and 23.73 mg QE/g crude, respectively). GC-MS analysis of the 75 % ethanol extract revealed 33 bioactive compounds, accounting for 96.33 % of the total area. Significant components included benzene, 1,4-bis(phenylmethyl)- (15.05 %), and naphthalene, 1,6-dimethyl- (10.98 %); also present were sterols, fatty acids, hydrocarbons, aromatic, and heterocyclic compounds, indicating a complex chemical profile. In antioxidant assays, the 75 % ethanol extract revealed the highest DPPH radical scavenging activity (IC<inf>50</inf>: 143.03 mg/L), reducing power (IC<inf>50</inf>: 378.69 mg/L), and lipid peroxidation (IC<inf>50</inf>: 526.25 mg/L). However, the absolute ethanol extract exhibited both the highest metal chelating activity (IC<inf>50</inf>: 382.60 mg/L) and the strongest antimicrobial activity, inhibiting all 11 tested foodborne pathogenic and spoilage bacteria. The lowest minimum inhibitory concentration (MIC) of 25 mg/mL was observed for Staphylococcus aureus, Aeromonas hydrophila, Pseudomonas fluorescens, Latilactobacillus sakei, Streptococcus sp., and Lactococcus cremoris, with inhibition zone diameters of 27.21, 28.79, 28.19, 17.60, 15.09 and 11.20 mm, respectively. Similarly, the lowest minimum bactericidal concentration (MBC) of 25 mg/mL was obtained for S. aureus and L. sakei, indicating potent antibacterial effects against these species. Future studies should include direct comparative analyses of neem extracts and synthetic preservatives to evaluate further their relative efficacy and applicability in real-world food systems.
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Formulation and evaluation of nanoemulsions from Jasminum officinale essential oil for controlling postharvest browning and maintaining quality in jasmine (Jasminum sambac) flowers
(2025-01-01)
Yeamsuriyotai, Kittiya
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Pradabkun, Natthamon
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Manichart, Nutcha
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Yonsawad, Nipaporn
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Khamchatra, Na Monrug
The jasmine (Jasminum sambac (L.) Aiton) flower has delicate petals, resulting in rapid browning after harvest. The aim of this study was to search for an innovative postharvest treatment for delaying browning of jasmine petals using plant essential oils. J. officinale L. f. var. grandiflorum (L.) essential oil was found to reduce peroxidase activity in jasmine flower by 44.21% in the in vitro condition. The antioxidant activities and chemical composition of J. officinale essential oil were subsequently characterized. The essential oil exhibited the ability to scavenge 2,2-diphenyl-1-picrylhydrazil (DPPH) radicals with a 50% inhibition (EC<inf>50</inf>) value of 6.72 ± 0.89 mg/mL, a chelating effect with EC<inf>50</inf> value of 7.42 ± 1.59 mg/mL, and reducing power with EC<inf>0.5</inf> value of 14.89 ± 0.73 mg/mL. GC-MS analysis detected 29 compounds in the oil, with benzyl alcohol (20.68%) and benzyl acetate (19.87%) predominating. As plant essential oils have restricted water solubility, an oil-in-water emulsion was formulated using a spontaneous emulsification method. The resulting J. officinale essential oil naonoemulsion (JEN) had an oil droplet size of 70.2 ± 0.39 nm and a narrow polydispersity index. In vivo testing confirmed the inhibitory effects of JEN on jasmine flower browning and relevant enzyme activities. Jasmine flowers were soaked in various concentrations of JEN for 5 min, packed in polyethylene plastic bags, and stored in a refrigerator at 10 ± 3°C with relative humidity 66 ± 5%. Flowers treated with 1 and 2 mg/mL JEN showed effective delay of petal browning and maintained good quality with minimum flower opening index, high freshness score, and high color retention index. JEN treatment also reduced phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), and peroxidase (POD) activities, indicating postponement of the browning process. In addition, scanning electron microscopy micrographs of treated flower epidermis cells revealed delayed cell wall collapse, indicating retention of intact cells. Taken together, these results support JEN as a potential preventative of enzymatic browning and hence petal browning in jasmine flower.
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Bioassay-Guided Extraction and Isolation of Natural Herbicides from Dried Zanthoxylum limonella Alston Fruits
(2025-01-01)
Chotpatiwetchkul, Warot
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Charoenying, Patchanee
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Teerarak, Montinee
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Meesin, Jatuporn
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Jongkon, Nathjanan
Weeds are problematic plant species around the world. Various strategies exist for controlling weeds, but chemical treatment remains the preferred method, particularly when using natural substances. In this research, a crude aqueous-methanol extract from dried Zanthoxylum limonella fruits was acid-base partitioned into four fractions: neutral extract (NE), acid extract (AE), basic extract (BE), and aqueous extract (AQ). These fractions were further separated into seventeen subfractions: NEF1 to NEF7, AEF1 to AEF5, and BEF1 to BEF5, which were then tested for herbicidal activity against the growth of Chinese amaranth (Amaranthus tricolor) and barnyard grass (Echinochloa crus-galli). Active subfractions were isolated via column chromatography and identified using spectroscopic methods, yielding seven active compounds: xanthoxyline (1), tambulin (2), atanine (3), prudomestin (4), skimmianine (5), p-methoxybenzoic acid (6), and methyl caffeate (7). Compounds 2–7 had not been previously reported in Z. limonella. Xanthoxyline (1) was identified as the most potent botanical herbicide, fully inhibiting seed germination of Chinese amaranth and barnyard grass. This compound also decreased seed imbibition and α-amylase activity in both species. Molecular docking studies on the α-amylase enzyme (PDB ID: 1BG9) revealed that the aromatic, hydroxy, and carbonyl groups of xanthoxyline (1) interact with the enzyme's active sites.
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Impact of Nigrospora oryzae-Derived Natural Products on Photosynthesis and Oxidative Stress in Eichhornia crassipes
(2025-01-01)
Laosinwattana, Chamroon
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Manichart, Nutcha
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Wichittrakarn, Pattharin
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Yoneyama, Kaori
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Teerarak, Montinee
Interest in natural herbicides has been growing due to government policies restricting synthetic herbicide use in many countries. In that regard, this study investigates the potential of Nigrospora oryzae extract as a natural herbicide against the aquatic invasive weed Eichhornia crassipes. A stable formulation was developed with a droplet size of 36.44 ± 0.36 nm and a zeta potential of -62.59 mV. Pot-based experiments revealed the N. oryzae extract induced 38.33% phytotoxicity within 24 hours, increasing to 84.72% by 28 days post-treatment. Scanning electron microscopy demonstrated morphoanatomical changes in epidermal tissue and stroma of E. crassipes, such as erosion of epicuticular waxes and degeneration of epidermis cells. The treatment significantly reduced the photosynthetic pigment content while increasing hydrogen peroxide (46.26%), malondialdehyde (17.49%), and proline (19.16%) levels, causing cellular electrolyte leakage. Activities of superoxide dismutase, catalase, ascorbate peroxidase, and guaiacol peroxidase were significantly elevated (p<0.05), indicating oxidative damage. These findings demonstrate that N. oryzae extract can disrupt growth and photosynthesis and induce oxidative stress in E. crassipes, suggesting its potential as a source of natural herbicide for industrial application.
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Influence of ginger essential oil nanoemulsion delivery system on antioxidant activity and postharvest Davallia frond vase life
(2025-01-01)
Kowwilaisang, Piangpassorn
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Teerarak, Montinee
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Laosinwattana, Chamroon
Plant essential oils have limited water solubility, for which problem oil-in-water emulsions provide a good solution. The aim of this work was to prepare a ginger essential oil (Zingiber officinale Rosc.; GEO) coarse emulsion, microemulsion, and nanoemulsion all with the same formula, determine their characteristics, and compare their antioxidant activities and utility as holding solutions for Davallia fronds. A coarse emulsion was firstly formed by a magnetic stirrer of GEO with a 1:2 by weight mixture of Tween 20 and Span 20 at weight ratio of 1:8, respectively. The coarse emulsion droplets (810.0 nm) were then broken by sonication and high-pressure homogenization to create the microemulsion (426.1 nm) and nanoemulsion (76.4 nm), respectively. The three emulsions exhibited ζ-potential values more negative than -30 mV, indicating them to be stable. All three emulsions demonstrated DPPH• and ABTS•+ free radical scavenging capacities significantly higher than those of GEO in ethanol. Meanwhile, the nanoemulsion significantly improved the Fe<sup>2+</sup> chelating effect Finally, in a bioefficacy experiment with Davallia fronds, both the microemulsion at concentrations of 5 and 10 mg/mL and the coarse emulsion at 10 mg/mL were found to extend frond vase life. The nanoemulsion demonstrated superior frond longevity at low concentration (5 mg/mL), but at higher concentration (10 mg/mL), vase life and leaf chlorophyll were not improved and malondialdehyde formation increased. Based on the current investigation, emulsification significantly enhances the antioxidant activities of GEO. Vase solutions containing high concentrations of coarse emulsion (10 μg/mL), microemulsion (5 and 10μg/mL), and low concentration of nanoemulsion (5μg/mL) increased Davallia frond longevity from 6.3 days to up to 11.9 days. Our findings suggest that there is an optimal concentration range for using essential oil emulsions in different delivery systems as a preservation solution in cut fronds.
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Characterization of Diaporthe fungal extract composition and phytotoxicity on the aquatic noxious weed Eichhornia crassipes: inhibitory effects on photosynthetic machinery and membrane integrity
(2025-01-01)
Somala, Naphat
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Manichart, Nutcha
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Thongbang, Muanfan
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Wichittrakarn, Pattharin
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Laosinwattana, Chamroon
Fungal extract-based natural herbicides are a promising avenue for the development of sustainable weed management. The study investigates the herbicidal activity of fungal extracts against the prominent aquatic invasive weed Eichhornia crassipes (water hyacinth). The ethanol crude fraction derived from Diaporthe sp. strain EC010 demonstrated the highest phytotoxicity. Chemical characterization using gas chromatography-mass spectrometry revealed 2(3H)-furanone, dihydro-4-hydroxy- (22.81%), linoelaidic acid (6.87%), and hordenine (6.62%) as major constituents. Phytotoxicity was evaluated by wrapping bioassay under greenhouse conditions. Observable foliar damage, such as necrosis, chlorosis, and depigmentation, occurred within 1 day after treatment (DAT). The rapid onset of visible damage within 1 DAT and the detailed analysis of tissue damage are particularly noteworthy. Lesion progression reflected the phytotoxicity of the extract, increasing to 77.33% visible phytotoxicity at 14 DAT for the highest concentration (8.0% w/v). Microscopic analysis revealed disintegration and complete deformation of epidermal and parenchymal tissues, and treated plants featured extensive aerenchyma spaces. Furthermore, the Diaporthe sp. extract decreased chlorophyll a, b, and carotenoid concentrations while increasing electrolyte leakage and malondialdehyde, indicative of weed deterioration. The current work offers valuable insights for sustainable and eco-friendly strategies in the management of water hyacinth populations in a lentic ecosystem. The findings suggest that Diaporthe sp. extract could serve as a natural herbicide, offering an environmentally friendly alternative to synthetic chemicals in managing water hyacinth.
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The effect of natural herbicide from Fusarium equiseti crude extract on the aquatic weed water hyacinth (Eichornia crassipes (Mart.) Solms)
(2024-12-01)
Laosinwattana, Chamroon
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Manichart, Nutcha
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Thongbang, Muanfan
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Wichittrakarn, Pattharin
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Somala, Naphat
In this study, Fusarium equiseti was isolated from the weed plant Tridax procumbens in an agricultural field and a crude extract produced with 75% ethanol for use as active ingredient material in natural herbicides. The herbicidal effect of F. equiseti extract was tested on water hyacinth (Eichornia crassipes), an invasive aquatic weed, by leaf disk assay at concentrations of 0.05%, 0.1%, and 0.2% w/v crude extract. Dose-dependent visual toxicity symptoms were evident after three days, namely chlorosis, yellow leaves surrounded by dark brown edges. Photosynthetic pigments (chlorophyll a, b, and carotenoids) and membrane integrity (as electrolyte leakage and malondialdehyde content) were evaluated following the leaf disk test. 3 days after treatment, photosynthetic pigment contents showed dose-dependent decreases, while both measures of membrane integrity showed dose-dependent increases with increasing extract concentration. In addition, a cytogenetic assay was conducted on Allium cepa L. root, in which mitotic index reduction and depigmentation were evident as early as 24 h after herbicide application. Finally, anatomical analysis of treated E. crassipes leaves revealed degradation or damage of the ground tissue. All told, our results support the F. equiseti crude-based natural herbicide cloud as a sustainable alternative in agriculture.

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