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Abstract Cadmium (Cd) toxicity significantly threatens agricultural productivity and food safety. Developing effective strategies to enhance plant tolerance to Cd stress is essential. This study investigates the synergistic effects of biochar (BC) and gibberellic acid (GA3) on mitigating Cd toxicity in maize (Zea mays), focusing on their impact on oxidative stress markers and antioxidant enzyme activities. Soil samples were collected from the Cholistan Institute of Desert Studies (CIDS) and analyzed for trace metal ions and other properties. Biochar was produced from fruit and vegetable waste, washed, washed, deashed, and mixed with 10 ppm GA3. FH-1036 hybrid maize seeds were sterilized and planted in pots containing soil with varying Cd levels (0, 8, and 16 mg Cd/kg soil). Twelve treatments were established, including control, GA3, BC, and their combinations under different Cd stress levels. Plants were irrigated to maintain 60% field capacity and harvested at the V10 growth stage. Hydrogen peroxide (H2O2) contents and activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) were measured in roots, stems, and leaves. Statistical analysis was performed using OriginPro 2021, with ANOVA and Fisher’s LSD test used to determine significant differences. GA3 and BC treatments significantly reduced H2O2 levels in maize roots, stems and leaves under Cd stress. The combined treatment of GA3 + BC showed the most significant reduction in H2O2 levels across all plant parts, reducing root H2O2 by 50%, stem H2O2 by 55%, and leaf H2O2 by 53% under severe Cd stress (16 mg Cd/kg). SOD activity increased under non-stress conditions but decreased under Cd stress, with the highest activity observed in the combined treatment. POD activity followed a similar pattern, with GA3 + BC treatment resulting in the most significant increases under non-stress conditions and the least reductions under Cd stress. CAT activity showed substantial increases with GA3 + BC treatment, particularly under severe Cd stress, with a notable rise over the control. APX activity also exhibited enhancements with GA3 and BC treatments, especially in the combined treatment under various Cd stress levels. This study highlights the potential of combined BC and GA3 treatments in improving Cd stress tolerance in maize. Future research should focus on field trials and the long-term impacts of these treatments on crop productivity and soil health.

Abstract The plant extract-mediated method is eco-friendly, simple, safe, and low-cost, using biomolecules as a reducing agent to separate nanoparticles. Lanthanum (La) is a rare earth metal that positively affects plant growth and agriculture. Citrus limon is a leading citrus fruit with many varieties. Conventional vegetative propagation methods depend on season, availability of plant material and are time-consuming. It is the main reason for limiting the acceptance of new varieties. So, In-vitro propagation of the lemon method is practiced overcoming all these problems. Lanthanum oxide nanoparticles (La2O3-NPs) were synthesized using plant extract of C. aurantium. Ultraviolet (UV)-Visible Spectroscopy, Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM), Fourier Transform Infrared (FTIR) spectroscopy, and Thermal Gravimetric Analysis (TGA) were used to characterize the synthesized La2O3-NPs. Fabricated La2O3-NPs were oval and spherical, with an average size of 51.1 nm. UV-visible absorption spectra of La2O3-NPs were shown at a sharp single peak at 342 nm and FTIR showed stretching frequency at 455 cm−1-516 cm−1. In the TGA outcome, mass loss was 9.1%. In vitro experiments demonstrated that La2O3-NPs significantly enhanced the germination and growth of C. limon seeds, achieving an 83% germination rate at 5 mg/L concentration, with uncoated seeds showing root initiation at 10 days and shoot formation at 15 days. Furthermore, La2O3-NPs effectively stimulated callus induction and maturation, with optimal responses observed in media containing MS and 2 mg/L 2,4-D, resulting in a maximum callus frequency of 100% from leaves and 87.5% from shoots at 5 mg/L concentration. These findings underscore the potential of La2O3-NPs to improve seed germination rates, seedling vigor, and callogenesis efficiency, suggesting their promising integration into agricultural practices for sustainable crop production, especially in suboptimal growing conditions. Future research is recommended to explore the mechanisms and broader applications of La2O3-NPs across various plant species and environments.

Abstract Matricaria chamomilla (Asteraceae), commonly known as chamomile can tolerate freezing temperatures and grows in many soil types. This plant is found on all continents and has significant medicinal value. There are more than 120 chemicals detected in chamomile flowers, with the majority found in the essential oil. In this study, M. chamomilla was given the NaCl stress of 0 mM, 1 mM, 100 mM, and 150 mM concentrations This study was the first to assess the efficacy of German chamomile upon exposure to salt stress hence plant particles that had been dried and powdered were analyzed using, phytochemical tests, Fourier Transform Infrared and UV–Vis spectroscopy, thin layer chromatography, fluorescence recovery after photobleaching assay, antibacterial and antioxidant activity. The characterization and results of these activities show amazing results which enhance their antibacterial property with an increased zone of inhibition when the samples of salt stress of the above-given concentrations were compared to the control samples. More graph analysis indicates an effective impact of salt stress on the phytoconstituents of M. chamomilla. Other than that, there was a clear flower induction upon salt stress, as a variety of compounds are regarded as essential to the biological functions of chamomile flowers according to the phytoconstituent screening which can be further used in the cosmetic industry, pharmaceutical industry, and all other fields as well for various application as a nano-drug or bio-drug. Due to this, this plant became essential for plant biotechnology research.

Abstract Dragon fruit (Selenicereus undatus), known for its captivating appearance and remarkable nutritional profile, has garnered considerable attention in recent years. Despite its popularity, there's a dearth of research on optimal conditions for seed germination and early growth stages such as seedling shoot length, which are crucial for optimal crop yield. This study aims to bridge this gap by evaluating various growing media's performance on dragon fruit germination and early growth stages. Dragon fruit seeds were obtained from local markets in Pakistan and evaluated in five different growing media: cocopeat, peat moss, sand, vermiculite, and compost. Germination parameters were observed for 45 days, including seed germination percentage, mean germination time, and mean daily germination percentage, among others while early growth was monitored for 240 days. Statistical analysis was conducted using ANOVA and Tukey’s HSD test. Significant differences were found among the growing media regarding germination percentage, mean germination time, and mean daily germination. Vermiculite exhibited the highest germination rate (93.33%), while compost showed the least (70%). Peat moss and sand media facilitated rapid germination, while compost showed slower rates. Stem length was significantly influenced by the growth media, with compost supporting the longest stems. Vermiculite emerged as the most effective medium for dragon fruit seed germination, while compost showed slower but steady growth. These findings provide valuable insights for optimizing dragon fruit cultivation, aiding commercial growers and enthusiasts in achieving higher yields and quality. Further research could explore additional factors influencing dragon fruit growth and development.

Abstract Cadmium (Cd), being a heavy metal, tends to accumulate in soils primarily through industrial activities, agricultural practices, and atmospheric deposition. Maize, being a staple crop for many regions, is particularly vulnerable to Cd contamination, leading to compromised growth, reduced yields, and potential health risks for consumers. Biochar (BC), a carbon-rich material derived from the pyrolysis of organic matter has been shown to improve soil structure, nutrient retention and microbial activity. The choice of biochar as an ameliorative agent stems from its well-documented capacity to enhance soil quality and mitigate heavy metal stress. The study aims to contribute to the understanding of the efficacy of biochar in combination with GA3, a plant growth regulator known for its role in promoting various physiological processes, in mitigating the adverse effects of Cd stress. The detailed investigation into morpho-physiological attributes and biochemical responses under controlled laboratory conditions provides valuable insights into the potential benefits of these interventions. The experimental design consisted of three replicates in a complete randomized design (CRD), wherein soil, each containing 10 kg was subjected to varying concentrations of cadmium (0, 8 and 16 mg/kg) and biochar (0.75% w/w base). Twelve different treatment combinations were applied, involving the cultivation of 36 maize plants in soil contaminated with Cd (T1: Control (No Cd stress; T2: Mild Cd stress (8 mg Cd/kg soil); T3: Severe Cd stress (16 mg Cd/kg soil); T4: 10 ppm GA3 (No Cd stress); T5: 10 ppm GA3 + Mild Cd stress; T6: 10 ppm GA3 + Severe Cd stress; T7: 0.75% Biochar (No Cd stress); T8: 0.75% Biochar + Mild Cd stress; T9: 0.75% Biochar + Severe Cd stress; T10: 10 ppm GA3 + 0.75% Biochar (No Cd stress); T11: 10 ppm GA3 + 0.75% Biochar + Mild Cd stress; T12: 10 ppm GA3 + 0.75% Biochar + Severe Cd stress). The combined application of GA3 and BC significantly enhanced multiple parameters including germination (27.83%), root length (59.53%), shoot length (20.49%), leaf protein (121.53%), root protein (99.93%), shoot protein (33.65%), leaf phenolics (47.90%), root phenolics (25.82%), shoot phenolics (25.85%), leaf chlorophyll a (57.03%), leaf chlorophyll b (23.19%), total chlorophyll (43.77%), leaf malondialdehyde (125.07%), root malondialdehyde (78.03%) and shoot malondialdehyde (131.16%) across various Cd levels compared to the control group. The synergistic effect of GA3 and BC manifested in optimal leaf protein and malondialdehyde levels indicating induced tolerance and mitigation of Cd detrimental impact on plant growth. The enriched soils showed resistance to heavy metal toxicity emphasizing the potential of BC and GA3 as viable strategy for enhancing maize growth. The application of biochar and gibberellic acid emerges as an effective means to mitigate cadmium-induced stress in maize, presenting a promising avenue for sustainable agricultural practices.

BackgroundZea mays (maize) is a globally significant cereal crop with diverse applications in food, feed, and industrial products. However, maize cultivation is often challenged by environmental stressors such as heavy metal toxicity and drought stress (DS). Heavy metals like cadmium (Cd) and lead (Pb) can accumulate in soil through industrial activities and improper waste disposal, posing significant threats to plant growth and development. Drought stress further exacerbates these challenges by limiting water availability and affecting physiological processes in plants. This study explores the impact of Cd and Pb toxicity, as well as DS, on maize growth and development, and investigates the potential mitigating effects of various treatments, including gibberellic acid (GA3), biochar (BC), rhizobacteria (RB), and their combinations.MethodsThe experiment involved maize plants subjected to different stress conditions: cadmium (Cd) at concentrations of 0, 6, and 12 ppm, lead (Pb) at 0 and 400 ppm, and drought stress (DS). Treatments included the application of 10 ppm GA3, 0.75% BC, a combined treatment of 10 ppm GA3 and 0.75% BC, rhizobacteria (RB), and a combined treatment of 0.5% BC and RB. The study measured germination rates, shoot and root lengths, and biochemical parameters such as shoot and root protein, phenolics, and chlorophyll contents under these conditions.ResultsIn the absence of Cd stress (0 Cd), the application of 10 ppm GA3 and 0.75% BC significantly enhanced germination rates by 72% and 76%, respectively, compared to the control, with the combined treatment exhibiting the highest enhancement of 86%. Under Cd stress (6 ppm Cd), GA3 and BC individually improved germination by 54% and 57%, respectively, with the combined treatment showing the largest increase of 63%. Drought stress influenced germination, with notable improvements observed with the application of 0.5% BC (50% increase) and RB (49% increase). Similar trends were observed in shoot and root lengths, where the combined treatment of GA3 and BC resulted in the most significant improvements. The treatments positively influenced shoot and root protein, phenolics, and chlorophyll contents, particularly under stress conditions.ConclusionThese findings highlight the potential of combined treatments, such as the application of GA3 and BC or BC with RB, in alleviating the adverse effects of heavy metals (Cd and Pb) and drought stress in maize cultivation. The combined treatments not only improved germination rates but also significantly enhanced shoot and root growth, as well as important biochemical parameters under stress conditions. This suggests that GA3 and BC, alone or in combination with RB, can play a crucial role in enhancing maize resilience to environmental stressors. The study highlights the importance of exploring sustainable agricultural practices to mitigate the impacts of heavy metal toxicity and drought stress. Future research should focus on long-term field trials to validate these findings and further investigate the mechanistic pathways involved in stress mitigation by these amendments, as well as their economic feasibility and environmental impact on a larger scale to ensure their practical applicability in real-world agricultural settings.

Abstract Background This ethnobotanical study in Dunyapur, District Lodhran, Pakistan, focuses on traditional medicinal knowledge, exploring 41 plants across 28 families. The research involves 496 informants from diverse backgrounds, including farmers, herbalists, housewives, teachers, and shopkeepers. The prevalence of herbs (68%) aligns with their accessibility and rapid regrowth, shaping the local medicinal landscape. The study investigates socio-demographic features, emphasizing the importance of considering the community's diverse perspectives. Methods The research employs quantitative ethnobotanical data analysis, introducing various indices like PPV, FUV, FIV, RFC, UV, and RI. The analysis of plant growth habits underscores the dominance of herbs, and the method of preparation evaluation identifies decoction as the most common (23%). Leaves (27%) are the most utilized plant part, and Resedaceae stands out with the highest FUV (0.38). FIV highlights the ecological and cultural significance of Poaceae, Boraginaceae, Fabaceae, and Solanaceae. Results The RFC values range from 0.016 to 0.032, with Cucumis melo having the highest value (0.032), indicating its frequent citation and cultural significance. The study reveals specific plants like Melia azedarach, Peganum harmala and Salvadora oleoides with high PR values for skin issues, reflecting their widespread acceptance and effectiveness. Oligomeris linifolia emerges with the highest UV (0.38), emphasizing its greater significance in local traditional practices. Leptadenia pyrotechnica records the highest RI (9.85), underlining its exceptional importance in the community's traditional pharmacopeia. Conclusion The findings offer a holistic understanding of ethnobotanical knowledge in Dunyapur, emphasizing the role of local contexts and ecological factors in shaping traditional plant uses. The study contributes valuable insights into the diverse practices within the community, laying the foundation for sustainable integration of traditional knowledge into broader healthcare frameworks.

Abstract Salinity poses significant challenges to agricultural productivity, impacting crops’ growth, morphology and biochemical parameters. A pot experiment of three months was conducted between February to April 2023 in the Department of Botany, The Islamia University of Bahawalpur. Four brinjal (eggplant) varieties: ICS-BR-1351, HBR-313-D, HBR-314-E, and HBR-334-D were selected and assessed for the effects of salinity on various growth and biochemical attributes. The experiment was completely randomized in design with three replicates each. This study revealed that increased salinity significantly reduced the shoot length, root length, and leaf number across all varieties, with maximum adverse effects observed at a 300mM NaCl concentration. Among the tested varieties, ICS-BR-1351 demonstrated superior performance in most growth parameters, suggesting potential salt tolerance. Biochemically, salinity decreased chlorophyll content across all varieties, with the sharpest decline observed at the highest salt concentration. V4 (HBR-334-D) showed a 57% decrease in chlorophyll followed by V3 (HBR-314-E) at 56%, V2 (HBR-313-D) at 54%, and V1 (ICS-BR-1351) at 33% decrease at maximum salt levels as compared to control. Conversely, carotenoid content increased up to -42.11% in V3 followed by V2 at -81.48%, V4 at -94.11%, and − 233% in V1 at 300mM NaCl stress as compared to respective controls. V3 (HBR-314-E) has the maximum value for carotenoids while V1 has the lowest value for carotenoids as compared to the other three brinjal varieties. In addition to pigments, the study indicated a salinity-induced decrease in total proteins and total soluble sugar, whereas total amino acids and flavonoids increased. Total proteins showed a decrease in V2 (49.46%) followed by V3 (36.44%), V4 (53.42%), and V1 (53.79%) at maximum salt concentration as compared to plants treated with tap water only. Whereas, total soluble sugars showed a decrease of 52.07% in V3, 41.53% in V2, 19.49% in V1, and 18.99% in V4 at the highest salt level. While discussing total amino acid, plants showed a -9.64% increase in V1 as compared to V4 (-31.10%), V2 (-36.62%), and V3 (-22.61%) with high salt levels in comparison with controls. Plant flavonoid content increased in V3 (-15.61%), V2 (-19.03%), V4 (-18.27%) and V1 (-27.85%) at 300mM salt concentration. Notably, salinity elevated the content of anthocyanin, lycopene, malondialdehyde (MDA), and hydrogen peroxide (H2O2) across all varieties. Antioxidant enzymes like peroxidase, catalase, and superoxide dismutase also increased under salt stress, suggesting an adaptive response to combat oxidative damage. However, V3 (HBR-314-E) has shown an increase in anthocyanin at -80.00%, lycopene at -24.81%, MDA at -168.04%, hydrogen peroxide at -24.22%, POD at -10.71%, CAT as-36.63 and SOD as -99.14% at 300mM NaCl stress as compared to control and other varieties. The enhanced accumulation of antioxidants and other protective compounds suggests an adaptive mechanism in brinjal to combat salt-induced oxidative stress. The salt tolerance of different brinjal varieties was assessed by principal component analysis (PCA), and the order of salt tolerance was V1 (ICS-BR-1351) > V4 (HBR-334-D), > V2 (HBR-313-D) > V3 (HBR-314-E). Among the varieties studied, ICS-BR-1351 demonstrated resilience against saline conditions, potentially offering a promising candidate for saline-prone agricultural areas.

Abstract Potato (Solanum tuberosum) is the third crucial global crop facing threats from Alternaria solani, a necrotrophic fungal pathogen causing early blight disease. Beyond crop impact, it leads to substantial production reduction and economic losses worldwide. This study introduces a green synthesis method for producing Ferric Oxide nanoparticles (FNPs) using dried Guava (Psidium guajava) leaves. Guava leaf extract acts as a reducing agent, with iron (III) chloride hexahydrate (FeCl3·6H2O) as the oxidizing agent. This study employed various characterization techniques for Ferric Oxide nanoparticles (FNPs). Fourier Transform Infrared Spectroscopy (FTIR) revealed peaks at 877 cm−1, 1180 cm−1, 1630 cm−1, 1833 cm−1, 2344 cm−1, and 3614 cm−1, associated with Maghemite vibrations, polyphenol compounds, and amino acids. UV–Vis spectroscopy exhibited a characteristic absorbance peak at 252 nm for FNPs. Scanning Electron Microscope (SEM) images illustrated particle sizes of 29-41 nm, and Energy Dispersive Spectroscopy (EDS) indicated elemental composition. X-ray diffraction (XRD) confirmed crystalline FNPs with peaks at 26.78, 30.64, 36.06, 38.21, 43.64, 53.52, 57.42, 63.14 and 78.32. Disease resistance assays demonstrated FNPs’ effectiveness against A. solani, reducing disease incidence and severity. In the leaf detach assay, concentrations of 15, 10 and 5 mg/L showed a dose-dependent reduction in disease severity and incidence. The Greenhouse Assay confirmed FNPs’ concentration-dependent effect on disease incidence and severity. The study also explored FNPs’ potential as biocontrol agents showing no adverse effects on overall plant development. Additionally, the study highlighted the agronomic potential of FNPs in enhancing plant growth and development emphasizing their role as micronutrients in biofortification. The findings suggest the promising application of FNPs in plant protection and biofortification strategies.

Abstract Osmotic stress is a significant concern in agricultural crop production as it can harm crop growth, development, and productivity. Agriculture crops are particularly vulnerable to osmotic stress due to their reliance on water availability for various physiological processes. Organic amendments like activated carbon biochar and growth hormone gibberellic acid (GA3) can play a vital role. However, the time needed is to modify the established amendment to achieve better results. That’s why the current study used potassium-enriched biochar (KBC = 0.75%) with and without GA3 (15 mg/L) as amendments under no osmotic stress and osmotic stress in wheat. Results showed that GA3 + KBC caused significant enhancement in germination (9.44%), shoot length (29.30%), root length (21.85%), shoot fresh weight (13.56%), shoot dry weight (68.38), root fresh weight (32.68%) and root dry weight (28.79%) of wheat over control under osmotic stress (OS). A significant enhancement in chlorophyll a, chlorophyll b and total chlorophyll, while the decline in electrolyte leakage of wheat, also validated the effectiveness of GA3 + KBC over control in OS. In conclusion, GA3 + KBC is the most effective among all applied treatments for improving wheat growth attributes under no osmotic and osmotic stress. Further research is needed at the field level, focusing on various cereal crops, to establish GA3 + KBC as the optimal treatment for effectively mitigating the impacts of osmotic stress.

Abstract Soil salinization is a prevalent form of land degradation particularly in water-deficient regions threatening agricultural sustainability. Present desalinization methods demand excessive water use. Biochar has been recognized as a potential remedy for saline soils and Gibberellic acids (GA3) are known to mediate various biochemical processes aiding in stress mitigation. This study was undertaken at The Islamia University of Bahawalpur during winter 2022–23 to explore the combined effect of biochar and GA3 on wheat (Triticum aestivum L.) in saline conditions. Employing a fully randomized design wheat seeds in 24 pots were subjected to two salinity levels with three replications across eight treatments: T1 to T8 ranging from controls with different soil electrical conductivities (ECs) to treatments involving combinations of GA3, biochar and varying soil ECs. These treatments included T1 (control with soil EC of 2.43dS/m), T2 (salinity stress with soil EC of 5.11dS/m), T3 (10 ppm GA3 with soil EC of 2.43dS/m), T4 (10 ppm GA3 with soil EC of 5.11dS/m), T5 (0.75% Biochar with soil EC of 2.43dS/m), T6 (0.75% Biochar with soil EC of 5.11dS/m), T7 (10 ppm GA3 combined with 0.75% biochar at soil EC of 2.43dS/m) and T8 (10 ppm GA3 plus 0.75% biochar at soil EC of 5.11dS/m). The results indicated that the combined applications of GA3 and biochar significantly enhanced plant growth in saline conditions viz. germination rate by 73%, shoot length of 15.54 cm, root length of 4.96 cm, plant height of 16.89 cm, shoot fresh weight 43.18 g, shoot dry weight 11.57 g, root fresh weight 24.26 g, root dry weight 9.31 g, plant water content 60.77%, photosynthetic rate 18.58(CO2 m−2 s−1) carotenoid 3.03 g, chlorophyll a 1.01 g, chlorophyll b 0.69 g, total chlorophyll contents by 1.9 g as compared to the control. The findings suggest that the combined application of these agents offers a sustainable and effective strategy for cultivating wheat in saline soils. The synergy between biochar and GA3 presents a promising avenue for sustainable wheat cultivation in saline conditions. This combined approach not only improves plant growth but also offers an innovative, water-efficient solution for enhancing agricultural productivity in saline-affected regions.

Abstract Drought and cadmium (Cd) stress are both major issues that significantly affect the growth and development of wheat plants. Both drought stress and Cd toxicity disrupt physiological processes i.e., nutrient uptake, cell expansion, and enzymatic reactions resulting in poor crop growth. To overcome these issues, the use of activated carbon and gibberellic acid (GA3) are considered valuable amendments. However, the current study aimed to add value using GA3-enriched biochar (GA3-BC). That’s why, a lab experiment was conducted on wheat to assess the effectiveness of GA3-BC against Cd and drought stress. For GA3 enrichment in biochar, 10 µg GA3/g biochar was mixed. There were 3 levels of GA3-BC i.e., 0, 0.6 (GA3-BC1), and 0.9% (GA3-BC). All levels were applied in 3 replicates under no stress (0Cd + no drought), drought stress (DS), and 6 mg Cd/ kg soil (6Cd). Results showed that GA3-BC2 caused a significant improvement in shoot length (44.99%), root length (99.73%), seedling length (60.13%) and shoot fresh weight (63.59%) over control at 6Cd + drought stress. A significant improvement in chlorophyll a, chlorophyll b, and total chlorophyll while a decrease in electrolyte leakage and regulation of antioxidants i.e., lipid peroxidation, SOD, CAT, APx, GR, GPx, GST, and DPHH also signified the effectiveness of GA3-BC2 compared to control at 6Cd + drought stress. In conclusion, GA3-BC2 is an efficacious amendment for simultaneously alleviating drought and Cd stress in wheat. More investigations are recommended at the field level on different cereal crops cultivated in different soil textures to declare GA3-BC2 as the best treatment for mitigation of drought stress and Cd toxicity.

The genus Trichoderma holds economic significance due to its widespread distribution and diverse applications, including biological control, enzyme production, and various biotechnological uses. The accurate identification of Trichoderma species is crucial given their close association with human activities. Despite previous efforts in classification, a comprehensive analysis combining morphological and molecular approaches is necessary. This study focuses on the isolation of four Trichoderma species from industrial wastewater in Pakistan, expanding on the known diversity in the region; isolation involved collecting samples from industrial wastewater effluents at specific sites in Punjab, Pakistan. Trichoderma strains were cultured and purified on solid media, with subsequent biomass production for bisorptional activity. Morphological characterization included colony features and microscopic examinations. DNA extraction, polymerase chain reaction (PCR), and sequencing of the internal transcribed spacer (ITS) region were conducted for molecular analysis. Phylogenetic analysis was performed using the Maximum Likelihood Algorithm. The study identified three Trichoderma species, viz. T. citrinoviride, T. erinaceum, and T. longibrachiatum. Each species was characterized morphologically and supported by molecular–phylogenetic analysis. Illustrations of microscopic features and a phylogenetic tree based on the ITS-nrDNA region were recorded. T. citrinoviride and T. longibrachiatum, isolated from steel mill and tanneries wastewater, respectively, were differentiated based on morphological characteristics such as phialides and conidia. The combination of morphological and molecular techniques enhances the accuracy of species identification. The study highlights the significance of Trichoderma in industrial wastewater environments and underscores the need for continued research in this area. Future research should focus on exploring the ecological roles and potential applications of the newly identified Trichoderma species. Additionally, further investigations into the biotechnological potential of these species, including enzyme production and bioremediation capabilities, would contribute to their practical applications.

The production of citric acid, a vital agricultural commodity utilized across various industries such as food, beverages, pharmaceuticals, agriculture, detergents, and cosmetics, predominantly relies on microbial fermentation, with Aspergillus niger accounting for approximately 90% of global production. In this study, we aimed to optimize the key factors influencing citric acid production, with a focus on strains, fermentation techniques, and carbon sources, particularly sugarcane molasses. A. niger, sourced from the Botany department/Biotechnology laboratories at Govt. College of Science, Lahore, was employed for citric acid production. The process involved inoculum preparation through spore collection from 3 to 5 days of cultured PDA slants. The fermentation medium, comprising cane molasses with a 15% sugar concentration, was meticulously prepared and optimized for various factors, including magnesium sulfate, potassium ferrocyanide, time of addition of potassium ferrocyanide, ammonium oxalate, and calcium chloride. Our optimization results shed light on the significant impact of different factors on citric acid production. For instance, the addition of 0.4 g/L magnesium sulfate led to a maximum yield of 75%, while 2 g/L potassium ferrocyanide, added at 24 h, achieved a yield of 78%. Remarkably, ammonium oxalate, at a concentration of 10 g/L, resulted in a notable 77% yield. Conversely, the addition of calcium chloride exhibited negligible effects on citric acid production, with the control group yielding more at 78%. Our study underscores the potential for optimizing factors to enhance citric acid production by A. niger in submerged fermentation. These findings highlight the pivotal role of magnesium sulfate, potassium ferrocyanide, and ammonium oxalate in augmenting citric acid yields while emphasizing the minimal impact of calcium chloride. Ultimately, these insights contribute to advancing our understanding of microbial citric acid biosynthesis, providing valuable implications for industrial applications and future research endeavors.

Abstract Background Cardiovascular diseases (CVDs) are the major cause of deaths all over the world. The high level of blood cholesterol and oxidative stress are major risk factors for heart diseases. The phytotherapeutics have attracted attention as potential agents for preventing and treating oxidative stress associated diseases. The objective of present study was to evaluate the synergetic cardio-protective and antilipidemic potential of medicinal plants viz. Coriandrum sativum, Piper nigrum and Cactus grandiflorus. Cardio-protective and anti-lipidemic potential of herbal mixture was evaluated against salbutamol induced cardiotoxicity in rabbits. For this purpose, rabbits were divided into six groups as normal control, salbutamol control, curative and standard drug curative. Results Salbutamol significantly (p

Background: Traditional and ethnic foods, rooted in centuries-old practices, have long been revered for their potential health benefits. In contemporary society, the sustainability and application of these foods for basic health care are garnering increased attention, offering promising prospects for holistic well-being. Methods: In our research, we aimed to document the medicinal plants utilized by the indigenous community residing within the study area. Between 2021 and 2022, interviews were conducted with 457 participants, with 100 of them being identified as key informants. The snowball sampling technique was utilized to identify respondents. A group discussion was also held concerning the conservation and challenges associated with medicinal plants and traditional wisdom. The medicinal plants were assessed by applying various quantitative metrics, including Use Value (UV), Fidelity Level (FL), Informant Consensus Factor (ICF), Relative Frequency Citation (RFC) and Relative Popularity Level (RPL). Results: The study identified 40 medicinal plant species from 22 different families that are employed to treat 33 unique human ailments. It was observed that perennial herbs accounted for 78% of these species in use. The leaves were shown to be the primary plant part in use, accounting for 35% and decoction was determined as the most prevalent preparation method. Some plants, such as Polygonum plebejum with the highest UV and RI values, and Centaurium pulchelum known for its use in treating jaundice with the highest RFC, were highlighted. In contrast to the findings presented in earlier literature, our study revealed a 14% concurrence rate, accompanied by a 6% disparity rate, and notably, 69% of the usages were newly documented. Conclusion: This research serves as the inaugural quantitative investigation into ethnomedicinal practices within the study area, underscoring the importance of indigenous herbal treatments. The sustainability of traditional and ethnic foods offers promising prospects for the future of health care in human society. These foods, rooted in age-old wisdom, not only preserve cultural heritage but also present holistic health benefits. Embracing them can lead to more natural, accessible, and effective health solutions, bridging the gap between traditional wisdom and modern healthcare needs.

Saffron, the "golden spice" derived from Crocus sativus L., is renowned for its richness in secondary metabolites such as crocin and safranal, contributing to its unique properties. Facing challenges like decreasing global production, optimizing cultivation techniques becomes imperative for enhanced yields. Although the impact of factors like planting density, planting depth, spacing, and corm size on saffron growth has been studied, the interaction between corm size and planting depth remains underexplored. This study systematically investigates the interactive effects of corm size and planting depth on saffron growth and yield, providing evidence-based guidelines for optimizing cultivation. A factorial experiment, employing a completely randomized design, was conducted to assess the influence of corm size (05-10g, 10.1-15g, 15.1-20g) and planting depth (10cm, 15cm, 20cm) on saffron yield. Uniform-sized corms were obtained, and a suitable soil mixture was prepared for cultivation. Morphological and agronomic parameters were measured, and statistical analyses were performed using ANOVA and Tukey's HSD test. The study revealed that planting depth significantly affected saffron emergence. The corms sown under 15cm depth showed 100% emergence regardless of corm size (either 05-10g, 10.1-15g, 15.1-20g) followed by 10cm depth corms. Corm dry weight exhibited a complex interaction, where larger corms benefited from deeper planting, while intermediate-sized corms thrived at shallower depths. Similar patterns were observed in shoot fresh weight and dry weight. Specifically, the largest corm size (t3, 15.1-20g) produced the greatest fresh-weight biomass at the deepest planting depth of 20cm (T3), while intermediate-sized corms (t2, 10.1-15g) were superior at the shallowest 10cm depth (T1). The total plant biomass demonstrated that larger corms excelled in deeper planting, while intermediate-sized corms were optimal at moderate depths. This research highlights the intricate interplay between corm size and planting depth in influencing saffron growth. Larger corms generally promote higher biomass, but the interaction with planting depth is crucial. Understanding these dynamics can aid farmers in tailoring cultivation practices for optimal saffron yields. The study emphasizes the need for a coordinated approach to corm selection and depth placement, providing valuable insights for sustainable saffron production and economic growth.

To create weed management techniques capable of improving agroecosystem sustainability, it is essential to comprehend the different weed flora composition and diversification dynamics in conservation agriculture production systems. In North Western Himalaya from 2018 to 2020, evaluation of various tillage techniques with and without residue incorporation and weed management techniques on diverse weed flora composition was performed. Fifteen treatment amalgamations of five tillages viz. CT (conventional tillage) in maize-CT in wheat, CT-ZT (ZT, zero tillage), ZT-ZT, ZT-ZTR (ZTR, zero tillage in amalgamation with remainder integration) & ZTR-ZTR and three weed management practices viz. H (suggested herbicide in maize)-H (suggested herbicide in wheat), IWM-IWM (IWM, integrated weed supervision) & HW-HW; (HW, hand weeding) were calculated in strip plot design. Significant differences (p = 0.05) of weed density and total weed count were recorded during summer (maize) and winter (wheat) seasons. Among tillage treatments, lowest weed population and total weed population were recorded in conservation tillage (ZTR-ZTR) in summer and conservative tillage followed by zero tillage in winter season (CT-ZT), respectively. IWM-IWM and H-H had a least weed population and total weed population in summer and winter seasons, respectively. Highest important value index during summer and winter seasons was recorded in Echinochloa colona (55.90%) and Avena ludoviciana (76.99%), respectively. Higher Shannon Wiener index and Evenness were recorded for CT+HW-CT+HW (1.83, 0.91) and ZT+IWM-ZTR+IWM (1.73, 0.86) which indicate higher weed species diversity during summer and winter seasons, respectively. ZTR-ZTR led to less diverse weed diversity compared to intensive conventional tillage (CT-CT) practices. However, higher Simpson’s diversity was recorded for CT+HW-CT+HW, i.e. 0.83 and 0.82 for maize and wheat, respectively, compared to the other conventional tillage treatments. This study helps in understanding the crop-weed relation such that the implementation of sound and economic weeds management strategies. Further, long-term experiments are required in order to interpret the complicated relationships of diverse weed species under different tillage and weed management strategies for sustainable and adoptive conservation agriculture production system for North Western Himalayas.

The current study aimed to explore the traditional knowledge and practices of indigenous communities in Bahawalpur and adjacent regions, Pakistan, to treat a variety of diseases with a particular focus on the usage of ethnomedicinal wild plants. The objective of the study was to collect and evaluate local and indigenous knowledge regarding the quantity, variety, use and management of resources by the people. Data were gathered using Rapid Appraisal Approach (RAA), which involved in-person interviews. The data were analyzed using descriptive statistics methods as well as common ethnobotanical analytical techniques viz. use value (UV), relative frequency of citation (RFC), informant consensus factor (ICF), fidelity level (FL), relative importance (RI), frequency index (FI), family use value (FUV), family importance value (FIV), popular therapeutic use value (POPUT), plant part value (PPV), preference ranking (PR), cultural significance index (CSI), rank order priority (ROP) and Jaccard index (JI). A total of 158 plant species from 49 families were identified. The perennial herbs made up 72% of the documented species of therapeutic plants. There were 21 different species in each of the two major plant families i.e., Fabaceae and Poaceae. The maximum ICF were calculated for gonorrhea and paralysis (1.0) and maximum POPUT was calculated for skin disorders (0.088) and cough (0.077), respectively. The maximum UV was found for Rumex crispus (0.57) and RI for Leucaena leucocephala (4.38). Heliotropium crispum had the highest FL value (83%) and is used to treat kidney diseases. Alhagi maurorum had the greatest RFC (0.009) and FI (0.93). Arecaceae (0.45) and Fabaceae (13.39%) had the greatest FUV while Poaceae and Fabaceae had the highest FIV (13.29 for each). Leaves were the most frequently used plant part (35%). Alhagi maurorum (4.0) and Rumex crispus (32.57) had the highest CSI and ROP, respectively. The maximum JI (28.31) was calculated for Cholistan desert. These results showed the local population’s reliance on herbal remedies to treat prevalent illnesses. To demonstrate the viability of species, it′s crucial to emphasize both the use and conservation of these species. Adopting innovative applications, enhancing their value, and prioritizing the protection of multipurpose wild plants in inhabited environments is vital.

Lysine is an essential amino acid that is not biologically manufactured in the body. Different chemical methods for lysine production are expensive and give low yields. The present study was conducted with the purpose to evaluate the biochemical production of lysine by different carbon sources using bacterial isolates. Three carbon sources namely glucose, sucrose, and fructose were used to evaluate the biochemical production of lysine by Escherichia coli and Klebsiella spp. isolates. Optimum incubation periods were between 48-96 hours. An extensive amount of lysine was produced by all of these isolates in L6 fermentation medium. Maximum lysine was produced by Klebsiella isolate K1 6.48 g/L after 96 hours of incubation by using glucose as carbon source followed by 6.0 g/L by Klebsiella isolates K3 after 72 hours of incubation when sucrose was used as a carbon source at 37 °C. Highest amount of lysine was produced at 96 hours by Klebsiella isolates in addition to E. coli. From all three carbon sources using Klebsiella isolates and E. coli, glucose showed better lysine production.

The threat of invasion is growing globally and endangers biodiversity. Exotic invasive plants are putting a harm to the vegetation of Pakistan’s Pothwar region, which is a biodiversity hotspot. In the current study, the effects of Broussonetia papyrifera, Parthenium hysterophorus, Xanthium strumarium, and Lantana camara on the local flora in the Pothwar area were examined. Two categorical groups (invaded and non-invaded) were used in a dichotomized experimental design to collect data. Using the software PRIMER-7 and IBM SPSS v. 21, different diversity indices including Margalef’s index of species richness, Shannon index of diversity, and Simpson index of dominance were measured and compared between invaded and control plots. In comparison to the experimental plots, the control plots had an average of more individuals and diversity. On a multivariate scale, non-metric multidimensional scaling (nMDS) and analysis of similarity (ANOSIM) revealed higher species richness in control plots. The invasion effect of L. camara was the highest, followed by X. strumarium, P. hysterophorus, and B. papyrifera.

Within integrated pest management programs, biological control of unwanted plants has remarkable capacity to provide viable, effective, and economic control of weeds. When using bio-herbicides, crop production and quality improve with virtually no damage to the ecosystem. Bioherbicides are target-specific, destroy only selected weeds that have been sprayed for and do not cause harm to non-target plants. Bio-herbicides can be quickly incorporated into weed control programs, which can reduce chemical herbicide dependence. We are also raising the chance of environmental pollution by pesticides. There are only a few bio-herbicides available on commercial bases although work began earlier in the 1940s. Sources of commercialized bioherbicides include Phytophthora palmivora (Devine), Collectotrichum gleosporiodes (Collego), Colletotrichum gloeosporioides (Binomial) and Streptomyces viridochromogenes (Bialaphos and Glufosinate). Virulence for pathogens and their environmental requirement are major constraints for bioherbicide development. Specific bio-herbicides should be useful in finding position in irrigated fields, wildlife while thriving weeds with pests or resistant weed control.

Throughout the tropical and subtropical climates, the genus Citrus can be found. The current study was conducted to extract the Citrus sinensis peel oil and evaluate its antibacterial, antifungal and antiparasitic potential. Petroleum ether was used to extract the C. sinensis peel oil through a Soxhlet apparatus. The antimicrobial and antifungal potential was determined via agar well diffusion method and minimum inhibitory concentrations (MIC) were calculated (test bacterial strains: Staphylococcus aureus, Escherichia coli and Streptococcus agalactiae; test fungal strains: Aspergillus flavus, Aspergillus niger, Altrnaria alternata). Antiparasitic activity against Leishmaniatropica was determined following standard protocol using amphotericin-B as positive and Dimethyl Sulfoxide (DMSO) as a negative control and the percentage inhibition was calculated. The oil extracted was brownish yellow with a tangy smell, water-insoluble, density (0.778 g/cm3) and specific gravity (0.843 g/cm). In antibacterial activity, the diameter of the zone of inhibition was maximum against E. coli (14 mm) and minimum for S. agalactiae (10 mm). While in antifungal activity diameter of the zone of inhibition was maximum against A. flavus (12.5 mm) and minimum for A. alternata (8.6 mm). S. agalactiae exhibited the minimum MIC value (6 mg/mL) and in fungal strains A. alternata exhibited the minimum value (2 mm). Citrus sinensis peel oil displayed antileishmanial efficiency of 60% at 50 μg/mL concentration after 48 h of incubation. C. sinensis peel oil demonstrated antimicrobial capabilities, implying that it could be used as a natural preservative in food or as an effective treatment against a variety of pathogenic organisms. Industries should extract oil from the waste of citrus fruits which will be beneficial from an economic point of view.

The current study was designed to assess the comparative morphology of eight olive cultivars with different geographical origins and diverse genetic backgrounds, introduced to a new climatic zone. The morphological parameters of eight (five exotic and three domestic) olive cultivars (Bari Zaitoon-1, Bari Zaitoon-2, Favolosa (FS-17), Koroneiki, Balkasar, Ottobratica, Leccino, and Arbequina) were compared at the experimental area of the Department of Botany, The Islamia University of Bahawalpur, Pakistan (29°24′0″ North, 71°41′0″ East, 401–421 feet above sea level). Plant height, number of leaves/15 cm shoot, leaf size characteristics (leaf length, leaf width, leaf area, and length/width ratio), leaf shape characteristics (margin, leaf axil, base, and apex angles), leaf pigments (Chlorophyll a, Chlorophyll b, total chlorophyll contents, and carotenoids), phyllotaxy, and leaf color and venation were recorded. The highest plant height (28 cm) was obtained by Bari Zaitoon-2 followed by Bari Zaitoon-1 (24 cm), both of which are domestic cultivar of Pakistan, while the shortest height (5 cm) was obtained by Koroneiki. Leccino displayed the highest average number of leaves (17.8) on main shoot, followed by BARI-2 (16.4) and the lowest score was from Balkasar (10.4). Leaf area ranged from 5.66 cm2 (Bari Zaitoon-1) to 3.08 cm2 (Koroneiki). The longest leaf length (5.74 cm) was found in Bari Zaitoon-1 and the shortest (4.04 cm) in Koroneiki, while the broadest leaves were found in Leccino (1.54 cm) and the narrowest (1.12 cm) in Koroneiki. Bari Zaitoon-2 led in leaf length to width ratio (4.058) followed by Bari Zaitoon-1 (3.772) with small lanceolate leaves hardly reaching the value of 4, with the lowest value illustrated by Leccino. The total chloroplast pigments were highest in FS-17 followed by Bari Zaitoon-1 and Bari Zaitoon-2, while the lowest was in Arbequina. Chlorophyll a was highest in Bari Zaitoon-1 followed by FS-17 and Balkasar, with the lowest rate in Arbequina. Chlorophyll b content of FS-17 was the highest whereas the Chlorophyll b and total chlorophyll contents in Arbequina were the lowest of all the cultivars. The highest value of total carotenoids was found in Balkasar followed by FS-17 with the lowest value in Arbequina. The phyllotaxy was categorized into three types, i.e., alternate, opposite, and whorled. The combination of two or more types was usually observed on the same branch. The whorl of four leaves was also present in rare cases. Leaf venation was both pinnate and reticulate. The leaf base of most (four) of the olive cultivars, i.e., Arbequina, Balkasar, Leccino and FS-17, were cuneate having acute, rounded, apiculate, and cuspidate leaf tips, respectively. The findings revealed remarkable variations in olive morphology, especially in the leaves and a successful record of the preliminary data of olive cultivars from the study area was made. The present research demonstrated that local olive cultivars have unique characteristics that differentiate them from imported cultivars. Thus, local cultivars provide novel genetic resources that should be conserved.

Aim of the study: Heavy doses of synthetic weed control chemicals have caused herbicide resistance in weeds. Natural compounds produced by living organisms constitute a wide field for ecologically safe herbicides. The current investigation was directed to test allelopathic potential of hexane extracts of selected plants against common weeds of agriculture. Material and methods: Allelopathic potential of Carica papaya, Rhazya stricta, Lantana camara and Pinus roxburghii hexane extracts against weeds viz. Euphorbia helioscopia, Rumex dentatus, Phalaris minor, Avena fatua and Chenopodium album was determined at 100%, 75% and 50% concentration on soil, filter paper and agar. Parameters for assessing allelopathic potential were the germination (%), plumule and radicle size (cm). Data analysis was performed using the software STATISTIX 9. Results and conclusions: Based on the findings, it was determined that R. stricta, C. papaya, L. camara and P. roxburghii hexane extract possesses possible suppression effects among which L. camara had the most conspicuous inhibition effects on selected weeds. The inhibitory effects of germination and growth were establishing in order R. stricta > L. camara > C. papaya > P. Roxburghii. Field analysis to assess the phytotoxic ability of these species to be used as herbicide is recommended.

Green nanotechnology has emerged as a viable option for the production of nanoparticles. The purpose of the current investigation was to synthesize silver nanoparticles (AgNPs) using Eucalyptus camaldulensis and Terminalia arjuna extracts, as well as their combinations, as green reducing and capping agents. The parameters (concentration of silver nitrate solution and plant extract, time, pH, and temperature) were optimized for maximal yields, regulated size, and stability of silver nanoparticles. The ultraviolet–visible spectrophotometer (UV-Vis) and the surface plasmon resonance band (SPR) were used to validate the synthesis of AgNPs. The size, shape, and stability of nanoparticles were assessed using a zeta analyzer and a scanning electron microscope (SEM). The biomolecules responsible for the reduction of silver ion (Ag+) and the stability of silver nanoparticles generated with the plant extracts were identified using Fourier-transform infrared spectroscopy (FTIR). The agar-well diffusion method was used to test the antimicrobial activity of biosynthesized nanoparticles against Bacillus subtilis, Staphylococcus aureus, Pasteurella multocida, and Escherichia coli. When 1 mM of silver nitrate (AgNO3) was added to plant extracts and incubated for 60 min at 75°C in a neutral medium, maximum nanoparticles were produced. Biosynthesized silver nanoparticles were stable, spherical, and monodispersed according to zeta potential and scanning electron microscopy. Silver nanoparticles synthesized with combination 2 and T. arjuna showed the highest zone of inhibition (16 mm) against B. subtilis while combination 3 showed the largest zone of inhibition against S. aureus (17 ± 0.8). It was concluded that greenly produced silver nanoparticles showed good antibacterial activity while causing negligible cytotoxicity.

To meet the current challenges concerning the removal of dyes from wastewater, an environmentally friendly and efficient treatment technology is urgently needed. The recalcitrant, noxious, carcinogenic and mutagenic compound dyes are a threat to ecology and its removal from textile wastewater is challenge in the current world. Herein, biochar-mediated zirconium ferrite nanocomposites (BC-ZrFe2O5 NCs) were fabricated with wheat straw-derived biochar and applied for the adsorptive elimination of Tartrazine dye from textile wastewater. The optical and structural properties of synthesized BC-ZrFe2O5 NCs were characterized via UV/Vis spectroscopy, Fourier transform Infra-red (FTIR), X-Ray diffraction (XRD), Energy dispersive R-Ray (EDX) and Scanning electron microscopy (SEM). The batch modes experiments were executed to explore sorption capacity of BC-ZrFe2O5 NCs at varying operative conditions, i.e., pH, temperature, contact time, initial dye concentrations and adsorbent dose. BC-ZrFe2O5 NCs exhibited the highest sorption efficiency among all adsorbents (wheat straw biomass (WSBM), wheat straw biochar (WSBC) and BC-ZrFe2O5 NCs), having an adsorption capacity of (mg g−1) 53.64 ± 0.23, 79.49 ± 0.21 and 89.22 ± 0.31, respectively, for Tartrazine dye at optimum conditions of environmental variables: pH 2, dose rate 0.05 g, temperature 303 K, time of contact 360 min and concentration 100 mg L−1. For the optimization of process variables, response surface methodology (RSM) was employed. In order to study the kinetics and the mechanism of the adsorption process, kinetic and equilibrium mathematical models were used, and results revealed 2nd order kinetics and a multilayer chemisorption mechanism due to complexation of hydroxyl, Fe and Zr with dyes functional groups. The nanocomposites were also recovered in five cycles without significant loss (89 to 63%) in adsorption efficacy. This research work provides insight into the fabrication of nanoadsorbents for the efficient adsorption of Tartrazine dye, which can also be employed for practical engineering applications on an industrial scale as efficient and cost effective materials.

Noble metal nanostructures are being used broadly as catalysts for energy conversion in fuel cells. To overcome the future energy crises, fuel cells are anticipated as clean energy sources because they can be operated at low temperature, their energy conversion is high and their carbon release is almost zero. However, an active and stable electrocatalyst is essential for the electrochemical reactions in fuel cells. Therefore, properties of the nanostructures greatly depend on the shape of the nanostructures. Individual as well as interaction properties are greatly affected by changes in the surface area of the nanostructures. By shape controlled synthesis, properties of the nanostructures could be further enhanced by increasing the surface area or active sites for electrocatalysts. Therefore, an efficient approach is needed for the fabrication of nanostructures to increase their efficiency, activity, or durability in fuel cells by reducing the usage of noble metals. Different types of hollow nanostructures until now have been prepared including nanoboxes, nanocages, nanoshells, nanoframes (NFs), etc. NFs are the hollow unique three-dimensional structure which have no walls—they only contain corners or edges so they have large surface area. In electrocatalytic reactions, the molecules involved in the reaction can easily reach the inner surface of the nanoframes, thus noble metals' utilization efficiency increases. NFs usually have high surface area, greater morphological and compositional stabilities, allowing them to withstand harsh environmental conditions. By considering the current challenges in fabrication of noble metal based alloy NFs as electrocatalysts, this review paper will highlight recent progress, design, and fabrication of noble metal alloy NFs through different strategies—mainly photocatalytic template, electrodeposition, Kirkendall effect, galvanic replacement, chemical/oxidative etching, combination of both and other methods. Then, electrochemical applications of NFs in fuel cells toward formic acid, methanol, ethanol, oxygen reduction reaction as well as bifunctional catalyst will also be highlighted. Finally, we will summarize different challenges in the fabrication of highly proficient nanocatalysts for the fuel cells with low cost, high efficiency and high durability, which are the major issues for the highly commercial use of fuel cells in the future.