Search Results (183)

Mismanagement of rural wastewater can lead to environmental contamination with the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D). Fungi with bioremediating potential constitute a sustainable alternative to decontaminate such wastewater before its reuse. This study evaluated the ability of Aspergillus oryzae pellets to remove 2,4-D from natural and sterile rural wastewater (i.e., with/without native microbiota). The pellets were produced by incubating conidial solutions of A. oryzae strains RCA2, RCA4, RCA5, and RCA10 in synthetic wastewater for 21 days at 25 °C. The wastewater samples were characterized physicochemically and microbiologically upon arrival at the laboratory. Afterwards, they were supplemented with 1, 2.5, or 5 mmol L⁻¹ of 2,4-D and inoculated with the pellets. Physicochemical characterization was repeated throughout the experiment. Herbicide removal and the presence of 2,4-D degradation intermediate, 2,4-dichlorophenol (2,4-DCP), were assessed through high-pressure liquid chromatography with UV/Vis detection (HPLC-UV) and mass spectrometry. At the beginning of the assay, the macro- and micronutrient content in the samples were suitable to sustain fungal growth. By the end, pH had increased and sodium and nitrate levels decreased in comparison with the control. RCA2, RCA4, and RCA10 removed over 80% of 2,4-D after 7 days of incubation, at the three herbicide concentrations tested. Moreover, wet fungal biomass had increased by the end of the assay. These findings demonstrate that RCA2, RCA4, and RCA10 can grow, form pellets, and remove 2,4-D in natural rural wastewater, which makes them potential candidates for bioremediation strategies aimed at improving the quality of water set to be reused. Full article

Agrobacterium-mediated transformation systems are extensively applied in japonica rice varieties. However, the adaptability of local rice varieties to existing transformation systems remains limited, owing to their complex genotypes, posing a substantial challenge to transformation. In this study, four local rice varieties were selected to optimize the effects of different culture media on callus induction, browning resistance, contamination resistance, callus tolerance, differentiation, regeneration, and root development, and then two varieties were selected to improve plant architecture and tiller development by CRISPR/Cas9-mediated gene editing, based on constructive transformation systems. The goal was to enhance the transformation efficiency of local varieties and innovate germplasms. The results demonstrated that japonica rice varieties XG293 and WD68 exhibited higher induction rates under the treatment of 2 mg/L 2,4-D (2,4-Dichlorophenoxyacetic acid) + 1 mg/L NAA (Naphthaleneacetic acid), whereas indica rice varieties H128 and E33 performed the best under 3 mg/L 2,4-D + 1 mg/L NAA. Severe browning in H128 was effectively mitigated by a carbon source of 20 g/L maltose supplemented with 40 mg/L ascorbic acid. Contamination after Agrobacterium infection was controlled by 300 mg/L Tmt (Timentin). Under a treatment of 200 µM/L acetosyringone +10 min infection duration, XG293 and WD68 exhibited higher callus tolerance, differentiation rates, and GUS staining rates, achieving transformation efficiencies of 43.24% and 52.38%, respectively. In contrast, H128 and E33 performed better under the treatment of 200 µM/L Acetosyringone + 5 min, with transformation efficiencies of 40.00% and 40.74%, respectively. The mutants after OsCCD7 gene editing in WD68 and H128 displayed a dwarfness of plant height, a significant increase in tiller numbers, and compact architecture. These findings demonstrate that an optimized combination of plant growth regulators and infection durations effectively improves transformation efficiency for local varieties, and the OsCCD7 gene regulates plant architecture and tiller development with variable effects, depending on the rice complex genotypes. This study provides a theoretical basis for the efficient transformation of local rice varieties and germplasm innovation. Full article

Melastoma dodecandrum is primarily propagated through stem cuttings, which limits genetic variation and constrains breeding efforts. To overcome this limitation and facilitate molecular breeding, the establishment of a reliable and efficient regeneration system is essential. This study investigated the effects of plant growth regulators (PGRs) and culture media on the in vitro regeneration system of M. dodecandrum. The highest rate of callus induction (96.67%) was achieved when sterile leaf explants were cultured on Murashige and Skoog (MS) basal medium supplemented with 2.00 mg·L⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.50 mg·L⁻¹ 6-benzylaminopurine (6-BA). For callus differentiation, the optimal formulation of MS + 2.0 mg·L⁻¹ 6-BA + 0.5 mg·L⁻¹ naphthylacetic acid (NAA) resulted in a differentiation frequency of 83.33%. The optimal PGR combinations for shoot proliferation were 1.5 mg·L⁻¹ 6-BA + 0.1 mg·L⁻¹ NAA and 0.5 mg·L⁻¹ 6-BA + 0.2 mg·L⁻¹ NAA. The optimal rooting media were MS medium supplemented with 0.1, 0.2, or 0.5 mg·L⁻¹ indole-3-butyric acid (IBA) or 1/2MS medium supplemented with 0.1 mg·L⁻¹ IBA. Additionally, this study investigated the dynamic changes in endogenous hormones during the regeneration process. The levels and ratios of hormones, including gibberellin (GA₃), abscisic acid (ABA), indole-3-acetic acid (IAA), and zeatin (ZT), collectively regulated the regeneration process. Elevated levels of ABA and GA₃ may promote callus initiation as well as the growth and development of adventitious roots during the early induction stage. Reduced levels of ABA and IAA favored callus differentiation into shoots, whereas elevated GA₃ levels facilitated proliferation of adventitious shoots. Throughout the regeneration process, fluctuations in ZT levels remained relatively stable. This study successfully established an in vitro regeneration system for M. dodecandrum using leaf explants, providing theoretical guidance and technical support for further molecular breeding efforts, genetic transformation, and industrial development. Full article

An efficient in vitro propagation protocol for Eulophia bicallosa was developed using asymbiotic seed germination and protocorm proliferation. The effect of light on seed germination and development was evaluated on Vacin and Went (VW) medium under five conditions: darkness, white, green, red, and blue light for 24 weeks. Blue and red light significantly accelerated seed development, allowing progression to stage 5 within 24 weeks. For protocorm proliferation, six semi-solid culture media were tested. Half-strength Murashige and Skoog (½MS) medium yielded the best results after 8 weeks, producing the highest numbers of shoots (1.0), leaves (1.1), and roots (4.2) per protocorm, with 100% survival. The effects of organic additives were also evaluated using coconut water and potato extract. A combination of 200 mL L⁻¹ coconut water and 50 g L⁻¹ potato extract enhanced shoot formation (1.7 shoots), while 150 mL L⁻¹ coconut water with 50 g L⁻¹ potato extract increased both leaf (1.9) and root (8.8) numbers. The effects of cytokinins (benzyladenine (BA), kinetin (6-furfurylaminopurine), and thidiazuron (TDZ)) and auxins (indole-3-acetic acid (IAA), α-naphthalene acetic acid (NAA), indole-3-butyric acid (IBA), and 2,4-dichlorophenoxyacetic acid (2,4-D)) were investigated using ½MS medium supplemented with each plant growth regulator individually at concentrations of 0, 0.1, 0.5, 1.0, and 2.0 mg L⁻¹. Among the cytokinins, 0.1 mg L⁻¹ BA produced the highest survival rate (96%), while 1.0 mg L⁻¹ BA induced the greatest shoot formation (93%, 2.3 shoots). Among the auxins, 0.1 mg L⁻¹ IAA resulted in the highest survival (96%), and 1.0 mg L⁻¹ IAA significantly enhanced root induction (4.2 roots per protocorm). Acclimatization in pots containing a 1:1:1 (v/v) mixture of pumice, sand, and soil resulted in 100% survival. This protocol provides a reliable and effective approach for the mass propagation and ex situ conservation of E. bicallosa. Full article

The common gardening herbicides and fertilizers are crucial for weed control and plant growth, yet they may have potentially harmful impacts on neurological health. This review explored the possible effects of these chemicals on neurodegenerative disorders, especially Alzheimer’s disease (AD) and Parkinson’s disease (PD). The mode of action of several frequently used gardening chemicals (paraquat, glyphosate, 2,4-dichlorophenoxyacetic acid: 2,4-D, and ammonium chloride) in AD and PD has been highlighted. The mechanisms involved are glutamate excitotoxicity, dopaminergic pathway disruption, oxidative stress, mitochondrial dysfunction, neuroinflammation, synaptic dysfunction, and gut–brain-axis dysregulation, crucial in the pathophysiology of AD and PD. Although the links between these substances and neurodegenerative conditions remained to be seen, growing evidence indicated their detrimental effects on brain health. This highlights the need for further research to understand their long-term consequences and develop effective interventions to mitigate the adverse effects of commonly used chemicals on human health and the environment. Full article

Bombax ceiba is an important medicinal and ornamental tree widely distributed in tropical and subtropical areas. However, its seeds lose viability rapidly after harvest, which has created hurdles in large-scale propagation. Here, we describe the development of a rapid and efficient de novo organogenesis system for Bombax ceiba, incorporating both indirect and direct regeneration pathways. The optimal basal medium used throughout the protocol was ½ MS supplemented with 30 g/L glucose, with all cultures maintained at 26–28 °C. For the indirect pathway, callus was induced from both ends of each hypocotyl on basal medium supplemented with 0.2 mg·L⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg·L⁻¹ 6-Benzylaminopurine (6-BA) under dark conditions. The induced calluses were subsequently differentiated into adventitious shoots on basal media containing 0.5 mg·L⁻¹ Indole-3-butyric acid (IBA), 0.15 mg·L⁻¹ Kinetin (KIN), and 1 mg·L⁻¹ 6-BA under a 16 h photoperiod, resulting in a callus induction rate of 140% and a differentiation rate of 51%. For the direct regeneration pathway, shoot buds cultured on medium with 0.5 mg·L⁻¹ IBA and 1 mg·L⁻¹ 6-BA achieved a 100% sprouting rate with a regeneration coefficient of approximately 3.2. The regenerated adventitious shoots rooted successfully on medium supplemented with 0.5 mg·L⁻¹ Naphthylacetic acid (NAA) and were acclimatized under greenhouse conditions to produce viable plantlets. This regeneration system efficiently utilizes sterile seedling explants, is not limited by seasonal or environmental factors, and significantly improves the propagation efficiency of Bombax ceiba. These optimized micropropagation methods also provide a robust platform for future genetic transformation studies using hypocotyls and shoot buds as explants. Full article

This paper presents a study on the optimization of 2,4-Dichlorophenoxyacetic (2,4-D) acid removal from synthetic wastewater by batch Fenton-Ozonation. The aim of this study is to evaluate the potential of the catalytic system Fe-L27 coupled to ozonation in the presence and absence of H₂O₂ as an effective and affordable technique for the treatment of organic pollutants in water. Fenton-like catalysts for the removal of 2,4-D in aqueous solutions were elaborated using catalysts synthesized by the wet impregnation method. The ACs and prepared catalysts were characterized by nitrogen adsorption–desorption isotherms at 77 K, TGA, XPS, SEM, and TEM. Their efficiency as Fenton-like catalysts was studied. In a first step, a response surface modeling method was employed in order to find the optimal parameters of the Fenton process, and then the optimal O₃/H₂O₂ ratio was established at laboratory scale. Finally, the investigated advanced oxidation processes were carried out at pilot scale. The results show that Fenton-like catalysts obtained by the direct impregnation method enhance the degradation rate and mineralization of 2,4-D. Full article

This study evaluated the histopathological impact of three commonly used pesticides—pirimiphos-methyl, propamocarb hydrochloride, and 2,4-dichlorophenoxyacetic acid (2,4-D)—on the kidneys of common carp (Cyprinus carpio Linnaeus, 1758) after 96-h acute exposure. The histopathological analysis demonstrated that all three tested pesticides induced structural changes. The histopathological changes were assessed using a semi-quantitative scoring system and categorised into circulatory, degenerative, proliferative, and inflammatory alterations. While circulatory alterations were absent in all treatments, clear and statistically significant degenerative, proliferative, and inflammatory responses were recorded, which escalated with increasing pesticide concentrations. Additionally, various statistical analyses were conducted to evaluate the lesions in kidney structure and function. Before the statistical analysis, normality and variance homogeneity were assessed using the Shapiro–Wilk and Levene’s tests, respectively. Due to non-normal data distribution, non-parametric methods were applied. Hence, the non-parametric statistical methods showed distinct group-level differences in the kidney damage indices. The Kruskal–Wallis test revealed significant differences across treatments (p < 0.001), and Mann–Whitney U tests identified specific pairwise differences. The degenerative and proliferative lesions were most prominent in fish exposed to 2,4-D at 100 µg/L (IK = 34), followed by pirimiphos-methyl and propamocarb hydrochloride. Inflammatory changes were mainly observed in the pirimiphos-methyl groups. The histopathological lesions were concentration-dependent, with 2,4-D causing irreversible renal damage at higher concentrations. These findings highlight the nephrotoxic risks posed by common pesticides and validate that the use of histopathological indices, combined with robust non-parametric testing, provides a reliable approach to evaluating organ-specific pesticide toxicity. These biomarkers offer sensitive early warning indicators of environmental risk, reinforcing the suitability of common carp as a model species for ecotoxicological assessment. Full article

Six activated carbons from tomato (Solanum lycopersicum L.) stems (TS-AC) were synthesized by carbonization and chemical activation using potassium hydroxide (KOH) and sodium hydroxide (NaOH) at temperatures of 550, 650, and 750 °C. These TS-ACs were then evaluated as adsorbents to remove 2,4-dichlorophenoxyacetic acid (2,4-D) and 2-methyl-4-chlorophenoxyacetic acid (MCPA) from aqueous solutions. The adsorption kinetics of both herbicides followed the pseudo-second-order model, closely correlating with the mesopore volume of the TS-AC. The Langmuir isotherm accurately described the adsorption process for both 2,4-D and MCPA. The porous structure of TS-AC, characterized by micropore volume and specific surface area, significantly influenced the maximum adsorption capacities. The adsorption of both herbicides was pH dependent, but ionic strength had no significant effect. Regeneration testing, conducted over three cycles, showed less than a 15% reduction in herbicide adsorption capacity. This study demonstrates that agricultural waste, specifically tomato stems, can be effectively valorized by using simple activation techniques in TS-AC that are efficient adsorbents to remove organic pollutants, such as herbicides, from aqueous media. Full article

Amphibians face a multitude of threats and therefore make a prime example of the current biodiversity crisis. Multiple amphibian stressors in anthropogenic landscapes include infectious diseases and agrochemicals. Synergic effects between these stressors may increase the negative impact of infections on amphibian health. In a 56-day trial, we assessed the impact of co-exposure to the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) and the pathogenic fungus Batrachochytrium salamandrivorans (Bsal) on infection parameters (infection intensity and disease severity) and health (body condition and telomere length) in Italian crested newts (Triturus carnifex). Twenty days post-inoculation with Bsal, newts were either exposed to 2,4-D for 12 days or not exposed (control). Most newts developed high infection loads that steadily increased towards the end of the trial. While body condition was negatively correlated with pathogen burden, only one out of 23 animals died. Telomere length remained unaffected by the pesticide and the pathogen. The 2,4-D treatment did not exacerbate Bsal infection. Most newts survived almost two months with significant pathogen loads; thus, even in a pesticide-infested environment, T. carnifex could be an important long-term Bsal reservoir for co-occurring species on the Italian peninsula, a urodele diversity hotspot. Full article

Traditionally, medicinal plants have served as the main resource for treating various human health conditions. Prionosciadium dissectum is a plant used in traditional medicine in the southern region of Jalisco, Mexico, to treat inflammatory respiratory problems. However, this species has not undergone pharmacological or biotechnological studies that validate these popular uses. The aim of this study was to induce calluses on P. dissectum leaves and then evaluate the antioxidant, anti-inflammatory, and antiproliferative activity of their extracts. The best callus induction was obtained using Murashige and Skoog (MS) culture medium with 1 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 1 mg/L kinetin (KIN). Extracts of hexane, dichloromethane, and methanol were obtained from the dry biomass, and the highest yield was obtained with methanol. The total phenolic content and antioxidant activity of the methanolic extracts were quantified. The methanolic extract showed 26.5 ± 0.4 mg equivalents of gallic acid/g extract, while, for antioxidant activity, it demonstrated IC₅₀ values of 49.4 ± 0.2 and 10.0 ± 0.0 μg/mL for 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ((2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) (ABTS), respectively. Regarding anti-inflammatory potential, the extracts did not significantly affect cell viability in RAW 264.7 macrophages. In contrast, it was clear that all extracts significantly decreased nitric oxide (NO) production at concentrations of 5–40 µg/mL. Additionally, extracts evaluated in human cancer cell lines only had a significant inhibitory effect at 100 µg/mL after 48 h, mainly with dichloromethane extract. This first biotechnological study indicates that P. dissectum cell cultures may produce compounds that favor the biological activities evaluated; however, it is necessary to carry out more in-depth evaluations of its extracts. This study is the basis for future research to enable the sustainable use of this valuable resource. Full article

Extensive utilization of the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) has resulted in contamination of the aquatic environment; this situation requires effective treatment technology. Ultraviolet-based advanced oxidation processes (UV-AOPs) are widely employed for the removal of organic contaminants from water. This study’s aim was to compare the degradation of the pesticide 2,4-D in UV-C-activated peroxide and peroxydisulfate systems. UV-C irradiation alone exhibited a negligible effect on pesticide degradation, whereas the addition of oxidants significantly enhanced the degradation efficiency relative to 2,4-D. Complete pesticide removal was achieved after 15 min of UV/H₂O₂ treatment, while twice as much time was required with the UV/S₂O₈²⁻ process. COD decreased by 74% and 28% for UV-C-activated peroxide and peroxydisulfate, respectively. Both investigated systems demonstrated good performance for 2,4-D dechlorination. Pesticide degradation rates increased with increasing dosages of the applied oxidants. Acidic conditions were more favorable for degradation of 2,4-D, compared to neutral and basic conditions, for both systems studied. The degradation efficiency relative to 2,4-D decreased in the presence of HA, Cl⁻ and HCO₃⁻ in water matrices. The predominant radical for the UV-C-activated peroxydisulfate was determined to be a sulfate radical. These findings are of fundamental and practical significance in understanding UV-C-activated 2,4-D degradation, paving the way for the selection of preferred processes for the optimal removal of pesticides from various aqueous matrices. Full article

Rice (Oryza sativa L.) is a staple food for billions of people globally. Rice cultivar RD43 has been recognized for its health benefits but has faced declining productivity due to climate change. Plant tissue culture serves as a powerful tool for studying and improving rice cultivars, yet a standardized protocol for rice cv. RD43 is lacking. This study aims to establish an efficient plant tissue culture protocol for rice cv. RD43 by evaluating concentrations of plant growth regulators for callus induction, proliferation, and regeneration. Callus induction was most effective with 4.0 mg/L of 2,4-dichlorophenoxyacetic acid (2,4-D), while callus proliferation was effective with 2.0 mg/L of 2,4-D. Furthermore, 2.0 mg/L of 6-benzyladenine (BA) yielded the highest plant regeneration, achieving a 50% regeneration rate and producing 9.60 shoots per callus. These findings lay the groundwork for a robust tissue culture protocol for rice cv. RD43 as a means for advanced breeding studies and contributing to global food security amid climate change challenges. Full article

Propagation of hinoki cypress (Japanese cypress, Chamaecyparis obtusa, Cupressaceae) through adventitious bud multiplication was performed using leaf-segment explants from cutting plants of selected adult trees. Explants were successfully surface-sterilized (>90% asepsis) by agitating them in 2.5% (w/v available chlorine) sodium hypochlorite solution for 15 min and then rinsed with sterile distilled water. Explants approximately 2 cm long were cultured on plates containing medium supplemented with 6-benzylaminopurine (BAP) and 2,4-dichlorophenoxyacetic acid (2,4-D), 20 g/L sucrose, and 7 g/L agar. The cultures were kept at 25 ± 1 °C under a 16-h photoperiod with a photon flux density of approximately 65 µmol m⁻² s⁻¹. The optimal adventitious bud multiplication (31.5 buds per explant) was obtained on a medium supplemented with 10 µM BAP in combination with 1 µM 2,4-D. Proliferated adventitious buds were elongated better on medium supplemented with 1 µM trans-zeatin. The best rooting result (86%) was achieved on a rooting medium supplemented with 1 µM 3-indolebutyric acid in combination with 0.1 µM 1-naphthaleneacetic acid. However, rooting response varied according to genotypes. Clones related to the cultivar ‘Nangouhi’ (Na18, Na14 x Isa, Na14-14, Isa x Na14, and NaS) were easier to root than those derived from the cultivar ‘ShizuokaKenZairai’ (SKZ5 and SKZ8). Regenerated plantlets did not show morphological abnormalities and showed a high survival rate after acclimatization (>90%). Full article

In vitro anther culture is a technique used to produce haploid plants when regenerating varieties with specific traits. To generate haploid plants with preferred characteristics, an anther culture technique was established for Lilium longiflorum “Show Up”. Morphological characteristics were recorded, including the flower bud length and anther color corresponding to different stages of microspore development. The effects of different flower bud lengths, various concentrations of exogenous plant growth regulators (PGRs), low-temperature pretreatment at 4 °C, and incubation under dark conditions on the induction of callus formation were studied. When the flower buds were 2.2–2.4 cm in length and the microspores were in the mononuclear development phase, callus induction reached the highest rate (15.6%). Callus was not induced when the PGRs 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin (KT) were added separately to the growth medium, but the highest callus induction rate occurred when anthers were cultured on the medium containing 2,4-D (0.75–1.0 mg/L) and KT (4 mg/L). The low-temperature pretreatment significantly enhanced the induction rate of anthers, but prolonged low-temperature pretreatment reduced the induction rate. The optimal period of cultivation in darkness was 6 d. After 15 days of cultivation, the number of swollen anthers was recorded, and these were transferred onto the differentiation medium Murashige and Skoog (MS) + 1-naphthaleneacetic acid (NAA) (2.0 mg/L), sucrose (30 g/L), and agar (7 g/L) at pH 5.8, whereon 100% differentiation was recorded. Overall, 14 regenerated lines were obtained by in vitro anther culture. Chromosome ploidy was determined by counting chromosomes in the root tips of ten regenerated plants, and four were found to be haploids. This study lays the foundation for anther culture in lilies to shorten the breeding cycle, improve selection efficiency, facilitate efficient genetic transformation, and enable the effective production of both haploid and double-haploid plants. Full article