Enhancing growth and suppressing root rot in Phaseolus vulgaris using ginger powder derived natural antifungal agent for optimum disease control and crop health

Enhancing growth and suppressing root rot in Phaseolus vulgaris using ginger powder derived natural antifungal agent for optimum disease control and crop health | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Enhancing growth and suppressing root rot in Phaseolus vulgaris using ginger powder derived natural antifungal agent for optimum disease control and crop health Shazia Alam, Asma Hanif, Sumara Shaheen, Min Zhu, Mohammad Mehdizadeh, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7054212/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Chemical fungicides impart pollution, toxicity to non-target organisms and health risks. Utilization of plant derivatives hold potential as a valuable source of bioactive for both disease control and the promotion of plant growth. This study investigated the biological activity of Zingiber officinale (ginger) derived plant extract on Ricinus communis L. (castor), Brassica spp., and Lantana camara L., using different solvents for extraction against major root-roting fungi, Macrophomina phaseolina , Fusarium oxysporum and Aspergillus flavus , using disc diffusion and agar well diffusion methods. At 100% concentration, both fresh and dried ginger extracts displayed total suppression (100%) of M. phaseolina , F. oxysporum , and A. flavus , with no apparent fungal growth. However, there was a little zone of inhibition (< 10%) for both dry, and fresh ginger, against R. solani . In comparison to the disc approach, the agar well method consistently yielded a great zone of inhibition. On the other hand, fungal overgrowth was noted, and no antifungal activity was demonstrated by ethanol-based extracts of Lantana camara, Brassica nigra , and Ricinus communis . When compared to untreated controls, in vivo tests showed that seeds treated with 100% fresh and dried ginger extracts dramatically increased root rot pathogen colonization by over 80% (P < 0.01). A substantial decrease in pathogen colonization was also seen after 100% extracts were soaked into the soil (P < 0.05). When ginger-treated seeds were combined with soil amendments, growth indices, including biomass, shoot length, and root length significantly enhanced. For instance, dry ginger + fudan resulted in 38% increase in shoot length, whereas dry ginger + DAP resulted in a 41% rise in root weight (P < 0.05). When soil amendment with 0.5 fresh ginger powder, the greatest decrease in fungal colonization (up to 95%) was observed. These results demonstrate the potential of ginger extracts, especially at 100% concentration, as efficient, environmentally friendly treatments for Phaseolus vulgaris L. root rot illnesses and enhancing plant development. Further research is recommended to optimize the application methods and explore the underlying mechanisms of ginger extract in promoting plant growth and disease suppression. Root-rotting Phytopathogens Zone of Inhibition Seed treatment Soil drenching Soil amendment Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 1.0 Introduction Microbial invasion from virus to viroid, prokaryotic bacteria, eukaryotic fungi, oomycetes, and nematodes, causes significant crop damage annually during both pre-harvest and post-harvest phases (Jamiołkowska 2020 ; Debebe 2022 ). These plant diseases cause 40 billion dollars in consequential losses globally and are extremely persistent in their attack (Jamiołkowska 2020 ; Singh et al. 2023 ). Microbes related to plants can sustain several detrimental strategies by producing viroids, viruses, bacteria and fungi, and lead to infectious diseases af fecting only the plant kingdom. Fungal disease frquency depends upon the plant health, the coincidence of a susceptible host, a virulent pathogen, and a favorable environment. The fungal disease invasion in plants also depends on the life strategy and the the pathogen population and the host plant change during their life cycle. The idea of biocontrol has sparked a significant political, economic, and technological discussion to create sustainable agriculture at a lower environmental cost (Barratt et al. 2018 ; Sun et al. 2025 ). Recent plant safeguarding tactics must immediately be based on natural resources due to the growing worries about ecological toxicity and environmental pollution brought on by the careless use of chemical formulation (Jamiołkowska 2020 ; Jabeen et al. 2024 ). Phaseolus vulgaris L. (red bean) is a herbaceous annual plant belongs to Leguminosae family known for their enriched macronutrients, micronutrients and antioxidant substances that nutrition for both humans and animals (Fernández-Ruiz et al. 2017 ; Chen et al. 2020 ). Various pathogenic fungi including, Rhizoctonia solani, Macrophomina phaseolina , Fusarium oxysporum , and Aspergillus flavus cause huge economic losses to the bean plants. R. solani is the major root-rotting fungi, as found 91.8% during disease in bean plants (Mayo et al. 2015 ). This pathogen is more functional in disposed soil and at temperatures between 15 and 18°C. R. solani is a necrotrophic pathogen and one of the root and hypocotyl pathogens which help them in plant invasion that leads to alter plant physiology and destroy fruit yield as a consequence economic loss globally. Macrophomina. phaseolina is a soil-borne fungus that affects more than 500 plant species in around 100 families and causes charcoal rot, seedling blight, and root and stem rot (Ghosh et al. 2018 ). This fungus causes disease in soybeans, ground nut cultivars and sorghum under hot weather (30–35°C) and lower soil moisture (below 60%) (Marquez et al. 2021 ). Fusarium wilt, which is origin by Fusarium oxysporum and occurs in all regions, can also effects the productivity of common beans. F.oxysporum can be invaded through wounds, natural openings, or intact roots of bean plants with a preference for the tap root and lateral root interconnections (de-Borba et al. 2017 ). The most general fungus that causes contamination of food and feed and build aflatoxins is Aspergillus flavus (Lahouar et al. 2015 ). It is the source to cause contamination during food storage because it has the capacity to produce aflatoxins and its ability to survive as a pathogen and saprophyte in the food supply before and after harvest (Tai et al. 2020 ). Aspergillus flavus and Aspergillus parasiticus create secondary metabolites which are known for their high mutagenic, carcinogenic, and teratogenic properties (Shi et al. 2023 ). Traditional Chinese utilized Ginger ( Zingiber officinale Roscoe) as a source of spice and medication for more than 200 years. The ginger extract could be used as an additive in food and medical components due to their antifungal and therapeutic properties. Ginger essential oil and ginger extract both have antifungal activities as opposed to a variety of plant pathogens (Peng et al. 2022 ). Xi et al. 2022 ). The findings show that ginger contains phenolic compounds monoterpenoids, sesquiterpenoids, and their derivatives consist of ketones, aldehydes, alcohols, and esters. These chemicals offer a broad range of antimicrobial activity against various microorganisms, making them a promising substitute for synthetic microbial (Gao et al. 2022 ). Utilization of ginger could be an effective biostimulant as the extracted sap from them could be as disease protecting agents in many plant species and maintain the crop heallth of industrial importance. Natural ginger application could be used to enhance the productivity of crop plants and raised the life hood of developing nations and ensure food security and zero hunger. Bioactive qualities found in Ricinus communis L., Brassica nigra (L.) Koch, and Lantana camara were selected in this study were based on optimum accumulation of bioactive qualities. Medicinal potential of Ricinus communis L. (castor) leaf, roots and seed oil was used as therapeutic uses have confirmed the larvicidal efficacy (Carolina et al. 2019 ). Likewise, essential oils (EOs) derived from mustard plants ( Brassica nigra L.), have potent antibacterial qualities that successfully prevent microbial growth and support both natural preservation and therapeutic applications (Mejía-Garibay et al. 2015 ). Shrub Lantana camara L belong to the genus Lantana contains over 150 species gain attention due to its medical potential and phytochemical richness. Lantana camara parts employed as biological control agents, stimulants, antibacterials, anti-inflammatory agents, and to treat rheumatism (Shi et al. 2023 ). These antifungal characteristics highlight the importance of these plants as tests to evaluate their biological activities using standardized extraction techniques for both vitro and vivo assessment. Using plants as fertilizers, those have higher antifungal activity are most reliable in contemporary agricultural production, have performed a fundamental role in enhancing crop yield and quality (Li et al. 2019 ; Akbar et al. 2024 ). Although unnecessary fertilization not only prevents the improvement of crop quality and yield, it also causes serious problems such as nutrient loss through leaching, hardness, and acidification of soil, crop pest annoyance, and groundwater safety issues (Du et al. 2021 ). The objective of the current study was to evaluate the assess the antifungal activity of ginger powder promotes growth and inhibits root-rotting fungi in Phaseolus vulgaris L. Study of such kind could be helpful toimprove plant disease management techniques and offer an alternative to chemical fertilizers which not only increase production but also pose no environmental risks. 2.0 Materials and Methods 2.1 Collection of material and seeds Fresh ginger rhizome and dry powder were collected from Mosamiyat, near University Road Karachi, Pakistan. The leaves of Lantana camara L. and castor ( Ricinus communis L.) were gathered from the University of Karachi and leaves of mustard ( Brassica nigra (L.) Koch) were collected from a local market near Mosamiyat, Karachi, Pakistan. Organic fertilizers, cow dung and goat dung, inorganic fertilizers fudan, and diammonium phosphate (DAP) were collected from a nursery near the University of Karachi. Red bean seeds were collected from New Sabzi Mandi Karachi, Pakistan, for pots experiment. 2.2 Preparation of stock solution of ginger, mustard, castor, and lantana camara L. For the preparation of stock solution 10g of fresh and dry ginger powder were soaked in 100 mL of appropriate organic solvent (n-hexane). The flasks were covered with aluminum foil and left for 72 hours. Fresh leaves of mustard, castor, and Lantana camara dried under sunlight for 24 hours. For the preparation of stock solution, 10g of the tested leaves powder was weighed and added to the flask containing 100 mL of the appropriate organic solvent (ethanol), separately. Flasks were covered with aluminum foil and left for 1 week. The solutions were centrifuged at 3000 rpm for 10 minutes and used for in-vitro studies while residue was discarded (Michiels et al. 2012 ). The crude plant extract was considered as 100% stock concentration. One gram of crude extract diluted in one mL of solvent, for instance, equal 100% (w/v) stock. 2.3 The Isolation of Pathogenic Fungi 2.3.1 Isolation of M. phaseolina by using the wet sieving technique Soil samples were taken from the rhizosphere of infected soybean plants exhibiting classic charcoal rot signs in the field station of the University of Karachi, Pakistan. In order to extract microsclerotia from the soil, the pathogen was separated by using the wet sieving approach, as outlined by Ghaffar (1976). In short, distilled water was used to suspend 100g of air-dried soil, which was then run through a succession of sieves with progressively smaller mesh sizes (60, 100 and 200). To stop bacteria from growing, the material that was left on the last sieve was cultivated on potato dextrose agar (PDA) treated with streptomycin and penicillin. Emerging fungal colonies were preliminarily identified as Macrophomina phaseolina based on morphological traits, including the formation of numerous black microsclerotia, absence of conidia, and fast-growing grey to black mycelia (Schafer & Kiewnick. 2010). 2.3.2 Isolation of R. solani by baiting technique Samples of soil were taken from the rhizosphere of infected chilli ( Capsicum annuum ) plants from agricultural fields of the University of Karachi, Pakistan, that were exhibiting indications of basal stem rot. The soil plating method outlined by Anderson and Papavizas ( 1965 ) was used to recover the fungal isolate. Plates were incubated at 25 ± 1°C for 3–5 days after 10 g of soil was spread out on water agar (WA) supplemented with streptomycin sulphate (100 mg/L). For purification, hyphal tips from newly formed colonies were subcultured onto potato dextrose agar (PDA). Preliminary identification of the fungal isolate as Rhizoctonia solani was based on morphological traits, including right-angled hyphal branching, absence of conidia, and presence of typical multinucleate cells seen under a compound microscope following safranin staining. DAPI staining was used to confirm whether the condition was binucleate or multinucleate. The internal transcribed spacer (ITS) region of rDNA was amplified using the universal primers ITS1 and ITS4 in order to perform molecular identification and verify the identity (White et al. 1990 ). 2.3.3 Soil dilution for the F. oxysporum isolation Soil samples were taken from the rhizosphere of wilted tomato ( Solanum lycopersicum ) plants from agricultural fields of the University of Karachi, Pakistan, that displayed the typical indications of vascular wilt. According to Leslie and Summerell (2006), the pathogen was identified using the soil dilution plate approach. The fusarium- selective medium from Komada, which is known to promote the development of Fusarium spp. while inhibiting the growth of bacterial and other fungal pollutants, was plated onto 100 grams of air dried soil that had been serially diluted in sterile distilled water. Subculturing on potatoes dextrose agar (PDA) allowed for the purification of newly formed fungal colonies. Based on morphological features such as development of many microconidia, three to five septate sickle-shaped macroconidia, and thick walled chlamydospores seen under a compound microscope, Fusarium oxysporum was initially identified (Booth, 1971). On PDA, the colony morphology displayed cottony aerial mycelium and characteristic white to pinkish-purple coloring. 2.3.4 Isolation of Aspergillus flavus In agricultural fields at the University of Karachi, Pakistan, an area known for frequent contamination by Aspergillus , soil samples were taken from the rhizosphere of ground nuts ( Arachis hypogaea ) plants. The dilution plate technique was used for isolation, in accordance with Pitt and Hocking’s (20009) methodology. To promote specific growth, serial dilutions of air-dried soil were made in sterile distilled water and plated on semi-selective media such as Aspergillus flavus and parasiticu s agar (AFPA) and Czapek- Dox agar (CDA). For five to seven days, plates were incubated at 28 ± 1 \(\:℃\) . To examine morphological traits, colonies that resembled Aspergillus flavus were subcultured on PDA. Both macroscopic and microscopic characteristics, such as yellow green conidial head, biseriate conidiophores, and the presence of rough walled conidia organized in chains, were used for the first time identification (Klich, 2002 ). 2.4 In-vitro experiment 2.4.1 Antifungal Activity using Paper Disc Diffusion Method Sterilized Whatman’s paper discs were socked in stock solution of 100 µL of fresh and dried ginger (extracts separately), n-hexane (control for ginger), ethanol (control for castor, mustard, Lantana camara ) and fungicide (carbendazim) for half an hour. Pour the petri plates with Potato Dextrose Agar (PDA) medium containing streptomycin 0.2/L and penicillin 100 units/L. Sterilized Whatman’s paper discs soaked in respective solutions (castor, mustard, Lantana camara , fresh and dry ginger powder) were placed on three sides of petri plates. In the center of PDA poured petri plates, a disc of each fungus ( M. phaseolina, F. oxysporum, R. solani , and A. flavus ) was placed, and each fungus was replicated thrice. At 25–33°C temperature petri plates were incubated for 7 days. After an incubation period, the zone of inhibition was measured by scale in mm units (Silva &Domingues, 2017 ). 2.4.2 Antifungal activity using Agar well method PDA medium having penicillin 100 units/L and streptomycin 0.2/L was poured into petri plates. By using a sterilized 5mm cork borer three wells were made in PDA poured petri plates, and each well was inoculated with each stock solution (castor, mustard, lantana camara , fresh and dry ginger powder), n-hexane (control for ginger), ethanol (control for castor, mustard, Lantana camara ) and fungicide (carbendazim). Discs of fungus (i.e. M. phaseolina, F. oxysporum, R. solani , and A. flavus ) was placed in the middle of PDA-poured petri plates. Each examined fungus was replicated three times. At 25–33°C Petri plates were incubated for 7 days. After an incubation period, the zone of inhibition was measured by scale in mm units (Espinel-Ingroff, et al. 2007 ). The zone of growth inhibition was measured and calculated by the following formula. X= \(\:\frac{Y}{Z}\:\) x 100 X = percentage of growth inhibition Y = Growth of mycelia measured in control plates (mm) Z = Growth of mycelia measured in treated plates (mm) 2.5 In-vivo experiment Fresh and dry ginger powder which showed effective results in the in-vitro studies among all other treatments (castor, mustard, and lantana camara ) was carried to the in-vivo studies on red beans. 2.5.1 Collection of soil and seed treatment Sandy loam soil obtained from a nursery near the University of Karachi, sieved the soil, and the debris was discarded. The soil was transferred into clay pots (600g/) pots. Red bean seeds were sterilized with 1% NaClO, using distilled water, cleaned twice, then allowed to air dry. Tested seeds were treated with 100 and 75% concentrations of both dry and fresh ginger powder separately for about half an hour while untreated seeds were used as control. Five seeds were planted in pots, and there were three replicates for each treatment. Infection percentage and growth parameters were documented after one month of seed germination. 2.5.3 Soil amendment and drenching Each pot had 600 g of soil and was amended with dry and fresh ginger (0.1% and 0.5%) powder. After amendment for one day, five seeds of red beans after surface sterilization with 1% NaClO were planted in each pot. Three replicates were made for each treatment. After one month of seed germination, infection percentage and growth parameters were documented. For soil drenching, 75% extract is made by dilution of 25% of the stock, and 100% extract is undiluted stock. 25 mL prepared concentration of 75% and 100% of both fresh and dry ginger were drenched into clay pots having soil(600g) while untreated soil was used as control. Five seeds were planted in pots and, three replicates for each treatment. Growth parameters and percentage of infection were documented after one month of seed germination. 2.5.5 Growth experiment Pots were filled with 600g of soil. Organic fertilizers (cow dung and goat dung) and inorganic fertilizers (DAP and fudan) amended in soil at 0.1 and 0.5% per pot. Tested bean seeds were treated with extracts of both fresh and dry ginger powder. Another experiment was set up of these fertilizers without treating seeds with ginger. After one day of amendment, 5 seeds were sown in each container having 600g soil, and were watered regularly. Growth parameters such as infection percentage, number of leaves and nodules, and shoot and root length and weight were recorded after 1 month of seed germination. 2.5.6 Colonization % of pathogenic fungi from roots After one month of red bean growth, the plants were pulled out, the roots were cleaned with running water and dried with paper. The taproot of each treatment was cut into 5 small pieces and soaked in 1% of sodium hypochlorite NaClO for three minutes for surface sterilization, and washed with sterilized distilled water three times. After the roots were dried the fragments were put on a PDA-poured petri dish having antibiotics (penicillin and streptomycin) to inhibit the growth of bacteria and incubated at room temperature for one week. Fungi that emerged from the root segment after incubation were examined under the microscope and the percentage of colonization was calculated by the following formula; a = \(\:\:\frac{b}{c}\:\) x 100 a = Colonization percentage b = Number of fungi colonized by root pieces c = Total roots pieces 2.6 Statistical analysis Data was reported as mean ± standard deviation. The zone of inhibition of fungi was expressed in percentage and represented by using a chord diagram. To examine the effects of seed treatment, soil drenching, and soil amendment and the treatments using IBM SPSS statistics 26.0, the principal component analysis was performed. With significance of 0.000, Barlett’s Test of Sphericity confirms that Principal Component Analysis could be carried out on the data subjected to the analysis. The colonization percentage of the root-rotting pathogens was represented using a Heat map with labels. The graphs were plotted using OriginPro 2024b. 3.0 Experimental Results 3.1 In-vitro test Two different methods (disc diffusion and agar-well method) were used to find the effect of extracts against the tested fungi namely, M. phaseolina, R. solani, F. oxysporum , and A. flavus (Fig. 1 -Fig. 5S). Test fungi were suppressed and distinct zones of inhibitions were produced when extract at 100% concentration of both fresh ginger and dry ginger powder was used except for R. solani , which didn’t show the zone of inhibition against fresh ginger extract. Dry ginger powder extract showed a significant zone of inhibition against F. oxysporum in the agar well method. However, fresh ginger extract showed a minimum zone of inhibition against M. phaseolina . The extracts of Lantana camara , mustard, and castor didn’t show a zone of inhibition against the test fungi, however overlapping of test fungi over the extracts was observed. Overall results showed the complete inhibition of M. phaseolina, F. oxysporum , and A.flavus by both ginger extracts but inhibition of the R. solani by both ginger extracts was negligible. Extracts of castor, Lantana camara , and mustard were not effective against tested fungi. Among both of the methods, the Agar well method showed the highest zone of inhibition followed by the paper disc method (Figs. 1 and 2 ). 3.2 In-vivo results Principal component analysis The principal component analysis was carried out on the physical parameters of red bean plants, including root length, root weight, shoot length, shoot weight, no. of leaves, no. of nodules, and dry root and shoot weight in the greenhouse experiment. Three different pot experiments were plotted. The first experiment shows the effect of fertilizer and fertilizer + seed treatment by fresh ginger on the growth of red beans and against root-rotting fungi. The second experiment shows the effect of fertilizer and fertilizer + seed treatment by dry ginger powder on the growth of red beans and against root-rotting fungi. The third pot experiment shows the effect of soil amendment, soil drenching, and seed treatment by fresh ginger and dry ginger powder on the growth of red beans and against root-rotting fungi. It demonstrates how organic amendment and other attributes influence heterogeneity compared to the controlled plants. For the 1st pot experiment, a total of 76.98% combined variance was seen in the data set. Out of which, the 1st principal component gives 51.20% variance while 25.78% was shown by the 2nd principal component. The comparative distance of the treatments from the control on the score plot shows their positive correlation with one another (Supp. Figure 1 ). For the 2nd pot experiment, a total of 67.53% combined variance was seen in the data set. Out of which, the 1st principal component gives 44.30% variance while 23.23% was shown by the 2nd principal component. The comparative distance of the treatments from the control on the score plot shows their positive correlation with one another (Supp. Figure 2 ). For the 3rd pot experiment, a total of 55.18% combined variance was seen in the data set. Out of which, the 1st principal component gives 34.11% variance while 21.28% was shown by the 2nd principal component. The comparative distance of the treatments from the control on the score plot shows their positive correlation with one another (Supp. Figure 3 ). Different methods were used with fresh ginger and dry ginger powder and extract for the management of root pathogenic fungi, and to enhance the growth of red bean plants. Red Bean seeds were treated with 75% (P < 0.05) of fresh ginger increased the number of leaves, shoot length, and shoot weight as compared to control. Similar results were also recorded for dry ginger powder. Seeds treated with 100% concentration of dry ginger powder was found to be best to improve the length and weight of bean plants. Soil 600g drenched with 100% concentration of dry ginger powder significant (P < 0.01) to improve the length and weight of red bean root. Pots drenched with 75% of dry ginger increased (P < 0.05) the shoot length and weight as compared to control. Similar results were also recorded for fresh ginger powder. Soil amended with 0.5% concentration of dry ginger powder was significantly (P < 0.01) enhanced the length and weight of roots of beans. Pots soil amended with 0.5% dry ginger showed an increase the number of leaves, length and weight of shoot followed by 0.1%. Similar results were also recorded in fresh ginger powder (Fig. 3 ). Among the three methods, all were found effective but seed treated at 100% concentration could be used as an inexpensive tool in commercial scale as compare to soil drenching and soil amendment method which is recommended for low scale farming. By using organic fertilizers (goat dung and cow dung) and inorganic fertilizer (DAP) amended @0.1% along with seeds treated at 100% gave maximum increase in growth parameter of shoot and root of beans plant and impressively reduced the infection of root rotting fungi. Shoot length was increased significantly (P < 0.05) when soil was amended with Fudan in combination with seed treatment with dry ginger powder. Shoot weight and root length increased significantly (P < 0.05)when soil was amended with cow dung in addition with seeds treated with dry ginger powder. Similarly, root weight was increased significantly (P < 0.05) when soil was amended with DAP in combination with seed treatment with dry ginger powder. Shoot weight and root length were increased significantly when soil was amended with goat dung in combination with seed treated with fresh ginger. Similarly, root weight increased significantly when soil was amended with cow dung and seed treated with fresh ginger. Shoot length was increased when soil was amended with fudan in combination with seed treatment with fresh ginger (Fig. 3 – 5 ). Colonization percentage of root-rotting fungi ( M.phaseolina, R. solani, F.solani , and F. oxysporum ) Combine effect of fertilizers with seed treatment by fresh and dry ginger. Soil amended with fudan in combination with seeds treated with fresh ginger reduced (P < 0.01) the colonization percentage of root-rotting fungi. Similarly, soil amended with cow dung, DAP, fudan, and goat dung in combination with seeds treated with fresh ginger and dry ginger extracts were more effective against root-rotting fungi. Seed treatment, Soil drenching, and Soil amendment method by fresh ginger and dry ginger Sterilized seeds with 1% NaClO were treated by fresh ginger and dry ginger powder with 75% and 100% concentrations separately for 30 minutes. The seeds of red beans were sown in each pot. Each treatment was replicated three times. The seed not treated acts as control. The colonization percentage of root rot pathogen was reduced (P < 0.01) when seeds were treated with 100% of the extract of both fresh ginger and dry ginger powders. Seeds treated with 100% show maximum inhibition as compared to the control. Both treatments show antifungal activity againstroot-rotting fungi on bean plants. Sterilized seed after 1% NaClO were sowed in the soil, and the soil was drenched at 75% and 100% concentration of both tested ginger extracts. Each treatment was replicated thrice. Untreated seeds were served as control. The colonization percentage of root rot pathogens was reduced (P < 0.05) when soil was drenched with 100% extracts of both fresh ginger and dry ginger powder. Both treatments at different concentrations showed antifungal activity against root rotting fungi and improved the growth of red bean plants. Sterilized seeds with 1% NaClO were sowed in soil at different concentrations (0.5% and 0.1%); each treatment was replicated three times. The seed not treated act as control. The colonization percentage of root rot pathogen significantly decreased when soil was amended with 0.5 and 0.1% of fresh ginger powder. However, maximum inhibition (P < 0.01) of M. phaseolina, F. solani, R. solani , and F. oxysporum were recorded as 0.5% as compared to control. At different concentrations (P < 0.01) both fresh ginger and dry ginger powders showed antifungal activity against root rotting fungi by using soil amendment method experiment. 4.0 Discussion Overuse of chemical fertilizers is a multilayered problem with severe consequences, adversely affecting human health and crop health. Extensive use of agrochemicals, in agriculture is inducing the chemical resistance in the phytopathogens against the commonly used chemical fertilizers in the field (Bódalo et al. 2023 ). The current reserch was designed to use organic material instead of chemicals to control root pathogenic fungi associated with beans. For this purpose, the antifungal effect of ginger, mustard, castor, and Lantana camara was accessed in the in-vitro experiment, and the effective organic material, i.e. , ginger, was carried to the in-vivo experimentation, where its effects on the growth parameters and root rot colonization were observed. One-third of the world’s crop production is lost due to plant pathogens each year. It has been reported that the fungal pathogens cause severe damage among the biotic stresses, followed by bacterial and viral infections (Wani et al. 2023 ). Root rot, stem-blight, and stem-rot are the major diseases of common beans and cause huge economic losses. According to Ali et al. ( 2022 ), ginger is rich source of bioactive substances with potent antifungal and antibacterial properties, including zingerone, shogaol, and gingerol. Present results showed that ginger extract in the in-vitro experiment showed the maximum zone of inhibition against test fungi due to the anti-microbial effect. Our results are also in concordance with Yadav et al. ( 2023 ) that ginger can stop the growth of some of the most prevalent fungal pathogens. Previous research also showed that ginger oil has been estimated to possess antimicrobial effects (Beristain-Bauza et al. 2019 ). The research carried out by Gao et al. ( 2022 ) reported that monoterpenoids, sesquiterpenoids, phenolic compounds and their derivatives, aldehydes, ketones, alcohols, and esters are present in ginger, which gives a broad antimicrobial array against various pathogens and makes it a potential alternative to synthetic fertilizers. The study carried out by Xi et al. ( 2022 ) reported the antifungal potential of ginger extract and ginger essential oil against F. oxysporum . In the present research, castor, mustard, and Lantana camara didn’t show effective results against the pathogenic fungi. However, various researchers have reported their antifungal activities. Carolina et al. ( 2019 ) have reported that ricinine is the main compound found in the castor leaves responsible for the toxic effects against various pathogens. Compound in Ricinus communis L.’s seeds, leaves, and oil are mostly responsible for its remarkable antifungal qualities. Ricinoleic acid, a fatty acid with well-known antibacterial properties, is present in castor oil. Numerous studies demonstrate its effectiveness against harm full fungi that affect both plants and humans, such as Rhizoctonia solani , Aspergilus species, and Fusarium species. Furthermore, Mejia-Garibay et al. (2015) reported that the essential compounds of mustard i.e. , allyl-isothiocyanate (AITC) shows antimicrobial activity, in both vapor and liquid phase against Aspergillus niger, A. ochraceus and Penicillium citrinum. Glucosinolates, which are abundant in Brassica nigra, break down into isothiocyanates, which are strong antifungal substances. Fungal cell membranes and metabolic processes are disrupted by these bioactives. Extract from mustard has demonstrated efficacy against fungi, including species of Alternaria , Fusarium , and Botrytis . Triterpenoids, flavonoids, and essential oils found in Lantana camara L. have broad-spectrum antibacterial properties. M. phaseolina, F. oxysporum , and A. flavus are among the phytopathogenic fungi whose growth has been demonstrated to be inhibited by it’s extracts. The disruption of fungal cell wall synthesis and mitochondrial dysfunction are probably the cause of the antifungal action (Shi, Y et al. 2023 ). Due to the efficient results of ginger in the in-vitro experiment, it was carried to the pot experiment where it successfully improved the physical parameters and suppressed the root-rotting systemic fungi. The research carried out by Ali et al. ( 2022 ) reported that the application of ginger extract enhanced damask rose growth characteristics and it is as a result in the availability of nutrients for enhanced growth. Similarly, Xi et al. ( 2022 ) reported that the constituents of ginger, including 4-hydroxybenzaldehyde and quercetin, inhibit the growth of F. solani completely and suggested their use as a natural fungicide in disease management. Additionally, investigations on plant-microbe interactions have shown that ginger powder may affect the dynamics of microorganisms in the soil, favoring beneficial bacteria while decreasing pathogens (Carolina et al. 2019 ). Increased nutrient absorption, better root system, and eventually higher crop output are the outcomes of the combined impact. Generally, the colonization percentage of M. phaseolina, F. solani and F. oxysporum were more suppressed than R. solani when combined with fertilizers and seeds treated by fresh and dry ginger powder also improved the growth of red plants as compared to individual treatment. Overall result showed that root pathogenic fungi were more suppressed by ginger dry powder along with treatment with fudan and increased the plant growth as compared to other. Among three methods (seed treatment, soil drenching, and soil amendment), all were found effective but seed treated with 100% concentration could be used as an inexpensive tool on commercial scale as compare to soil drenching and soil amendment methods which is recommended for low scale farming. It is suggested from the present research that the application of fresh and dry ginger extracts should be used against root-rotting fungi and for the improvement of the growth of red bean plants on large agricultural fields, which is cheap, easily available and non-toxic to the soil environment. So, it is a useful alternative to fungicides and bactericides. 5.0 Conclusion Based on the study on the application of ginger to enhance the growth of red beans and suppress root-rotting pathogenic fungi, the results suggest that ginger can be an effective organic amendment for improving the growth and health of red beans. The study found that the application of ginger extract improved the seed germination rate, plant growth, and yield of Red beans, while also suppressing the growth of root-rotting pathogenic fungi. This was attributed to the presence of various bioactive compounds in ginger, including gingerol and shogaol, which have been shown to have antifungal and plant growth-promoting properties. The use of organic amendments, such as ginger, can have several benefits for plant growth and soil health. Organic amendments can improve soil fertility, increase microbial activity, and suppress the growth of soil-borne pathogens. This can lead to improved plant growth, yield, and quality, while also reducing the need for synthetic fertilizers and pesticides. Overall, the use of organic amendments like ginger can be an effective strategy for improving the growth and health of Red beans, while also promoting sustainable agriculture practices. An efficient method for promoting Phaseolus vulgaris development and inhibiting root rotting fungal infections, including M. phaseolina, F. oxysporum , and R. solani is the application of powdered ginger. This purpose demonstrates that ginger ( Zingiber officinale) promises a sustainable, natural, and environmentally friendly substitute for chemical fungicides in the treatment of legume diseases. In order to maximize ginger powder’s effectiveness, future studies should concentrate on adjusting its concentration and administration method under various agronomic circumstances. To evaluate the effect of recurrent ginger treatment on soil microbiota in group health, long-term research is also required. Additionally, combining ginger powder with other biological control medications may enhance its ability to inhibit illness. Declarations Ethics approval Not applicable Consent to Participate We confirm that the manuscript has been read and approved by all named authors and that there are no other people who satisfied the criteria for authorship but are not listed. We further confirm that all the authors listed in the manuscript have been approved by all of us. Authors contribution Shazia Alam, Asma Hanif, Sumara Shaheen and Zainul Abideen: Conceptualization, Investigation, Formal analysis, Methodology. Shazia Alam, Min Zhu, Writing – original draft. Supervision, Conceptualization and review & editing Mohammad Mehdizadeh, Min Zhu and Zainul Abideen Funding Not applicable. Competing Interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Data availability statement The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request. References Akbar, R., Manzoor, S., Azad, R., Makai, G., Rahim, J., Sheikh, U.A.A., Ali, A., Aziz, T., Ahmad, H.I., Ahmed, M. and Du, D., 2024. Botanical Pesticides: Role of Ricinus communis in Managing Bactrocera zonata (Tephritidae: Diptera). Insects, 15(12), p.959. Ali, E. F., Al-Yasi, H. M., Issa, A. A., Hessini, K., & Hassan, F. A. (2022). Ginger extract and fulvic acid foliar applications as novel practical approaches to improve the growth and productivity of Damask Rose. Plants , 11 (3), 412. Anderson, J. A., & Papavizas, G. C. (1965). A method for isolating Rhizoctonia solani from soil and plant tissue. Phytopathology , 55 , 1299–1301. Barratt, B. I. P., Moran, V. C., Bigler, F., & Van Lenteren, J. C. (2018). The status of biological control and recommendations for improving uptake for the future. BioControl , 63 , 155-167. Beristain-Bauza, S. D. C., Hernández-Carranza, P., Cid-Pérez, T. S., Ávila-Sosa, R., RuizLópez, I. I., & Ochoa-Velasco, C. E. (2019). Antimicrobial activity of ginger ( Zingiber Officinale ) and its application in food products. Food Reviews International , 35 (5), 407-426. Bódalo, A., Borrego, R., Garrido, C., Bolivar-Anillo, H. J., Cantoral, J. M., Vela-Delgado, M. D., ... & Carbú, M. (2023). In Vitro Studies of Endophytic Bacteria Isolated from Ginger ( Zingiber officinale ) as Potential Plant-Growth-Promoting and Biocontrol Agents against Botrytis cinerea and Colletotrichum acutatum . Plants , 12 (23), 4032.Booth, C. (1971). The Genus Fusarium . Commonwealth Mycological Institute, Kew, UK. Carolina, A., Herliyana, E. N., and Sulastri, H. (2019). Antifungal activity of castor (Ricinus communis) leaves methanolic extract on Aspergillus niger . International Food Research Journal , 26 (2):595-598. Carolina, F. R., Lopez, S., & Morales, M. (2019). Biological activities of Ricinus communis plant parts: A review. Journal of Medicinal Plants Research , 13(11), 255–264. Chen, Y. C., Hsieh, S. L., & Hu, C. Y. (2020). Effects of red-bean tempeh with various strains of Rhizopus on GABA content and cortisol level in zebrafish. Microorganisms , 8 (9), 1330. Chiejina, N. V., and Onaebi, C. N. (2016). Phytochemical constituents and antifungal properties of Chromolaenaodorata and Moringa oleifera Lam on fungal rot of cucumber ( Cucumis sativus L.) fruit. Asian Journal of Plant Sciences , 2 (1): 10-15 de Borba, M. C., Garcés-Fiallos, F. R., & Stadnik, M. J. (2017). Reactions of black bean seedlings and adult plants to infection by Fusarium oxysporum f. sp. phaseoli. Crop protection , 96 , 221-227. Debebe, S., 2022. Post-harvest losses of crops and its determinants in Ethiopia: tobit model analysis. Agriculture & food security, 11(1), pp.1-8. Du, Q. J., Xiao, H. J., Li, J. Q., Zhang, J. X., Zhou, L. Y., & Wang, J. Q. (2021). Effects of different fertilization rates on growth, yield, quality and partial factor productivity of tomato under non-pressure gravity irrigation. PLoS One , 16 (3), e0247578. Espinel-Ingroff, A., Arthington-Skaggs, B., Iqbal, N., Ellis, D., Pfaller, M. A., Messer, S., Rinaldi, M., Fothergill, A., Gibbs, D. L., & Wang, A. (2007). Multicenter evaluation of a new disk agar diffusion method for susceptibility testing of filamentous fungi with voriconazole, posaconazole, itraconazole, amphotericin B, and caspofungin. Journal of clinical microbiology , 45 (6), 1811–1820. Fernández-Ruiz V., Morales P., Ruiz-Rodríguez B.M., TorijaIsasa E. (2017). Nutrients and bioactive compounds in wild fruits through different continents. Wild Plants Mushrooms Nuts Funct. Food Prop. Appl, 17 :263–314. Gao, Y.; Lu, Y.; Zhang, N.; Udenigwe, C.C.; Zhang, Y.; Fu, Y. Preparation, pungency and bioactivity of gingerols from ginger ( Zingiber officinale Roscoe): A review. Crit. Rev. Food Sci. Nutr. 2022, 22 , 1–26. Ghosh T., Biswas M. K., Guin C., Roy P. (2018). A review on characterization, therapeutic approaches and pathogenesis of Macrophomina phaseolina . Plant Cell Biotechnol. Mol. Biol. 19 , 72–84 Ibrahim, E., Ahmad, A. A., Abdo, E. S., Bakr, M. A., Khalil, M. A., Abdallah, Y., ... & Galal, A. A. (2024). Suppression of root rot fungal diseases in common beans ( Phaseolus vulgaris L.) through the application of biologically synthesized silver nanoparticles. Nanomaterials , 14 (8), 710. Jabeen, S., Tariq, M., Abid, R., Hanif, A., Shehzad, K., Rehman, M.W.U., Dawar, S., Dias, D.A., Alarjani, K.M. and Abideen, Z., 2024. Application of powdered Medicago sativa L. enhances eco-physiological output and protect against root rot fungi disease in okra and cowpea. Scientia Horticulturae, 337, p.113458. Jamiołkowska, A. (2020). Natural compounds as elicitors of plant resistance against diseases and new biocontrol strategies. Agronomy , 10 (2), 173. Klich, M. A. (2002). Identification of Common Aspergillus Species . Centraalbureau voor Schimmelcultures (CBS), Utrecht, The Netherlands. Lahouar, A., Crespo-Sempere, A., Marín, S., Saïd, S., & Sanchis, V. (2015). Toxigenic molds in Tunisian and Egyptian sorghum for human consumption. Journal of Stored Products Research , 63 , 57-62. Leslie, J. F., & Summerell, B. A. (2006). The Fusarium Laboratory Manual . Blackwell Publishing. Li, Y., Xue, X., Guo, W., Wang, L., Duan, M., Chen, H., & Chen, F. (2019). Soil moisture and nitrate-nitrogen dynamics and economic yield in the greenhouse cultivation of tomato and cucumber under negative pressure irrigation in the North China Plain. Scientific Reports , 9 (1), 4439. Marquez, N., Giachero, M. L., Declerck, S., & Ducasse, D. A. (2021). Macrophomina phaseolina : General characteristics of pathogenicity and methods of control. Frontiers in Plant Science , 12 , 634397. Mayo, S., Gutierrez, S., Malmierca, M. G., Lorenzana, A., Campelo, M. P., Hermosa, R., & Casquero, P. A. (2015). Influence of Rhizoctonia solani and Trichoderma spp. in growth of bean ( Phaseolus vulgaris L.) and in the induction of plant defense-related genes. Frontiers in plant science , 6 , 685. Mejía-Garibay, B., Palou, E., & Pez-Malo, A. L. (2015). Composition, diffusion, and antifungal activity of black mustard ( Brassica nigra ) essential oil when applied by direct addition or vapor phase contact. Journal of food protection , 78 (4), 843-848. Michiels, J. A., Kevers, C., Pincemail, J., Defraigne, J. O., and Dommes, J. (2012). Extraction conditions can greatly influence antioxidant capacity assays in plant food matrices. Food Chemistry, 130 (4), 986-993. Nash, S.M., and Snyder, W.C. (1962). Quantitative estimations by plate counts of propagules of the bean root rot Fusarium in field soils. Phytopathology, 52 : 567- 572. Peng, H., Hu, H., Xi, K., Zhu, X., Zhou, J., Yin, J., ... & Zhu, Y. (2022). Silicon nanoparticles enhance ginger rhizomes tolerance to postharvest deterioration and resistance to Fusarium solani . Frontiers in Plant Science , 13 , 816143. Pitt, J. I., & Hocking, A. D. (2009). Fungi and Food Spoilage (3rd ed.). Springer. Schafer, W., & Kiewnick, S. (2010). Morphological characteristics and molecular identification of Macrophomina phaseolina . Plant Pathology Journal , 26 (3), 223–229. https://doi.org/10.5423/PPJ.2010.26.3.223 Sheikh, A.H., and Ghaffar, A. (1975). Population study of sclerotia of Macrophomina phaseolina in cotton field. Pakistan Journal of Botany, 7 : 13-17. Shi, H., Li, J., Zhao, Y., Mao, J., Wang, H., & Zhu, J. (2023). Effect of Aspergillus flavus contamination on the fungal community succession, mycotoxin production and storage quality of maize kernels at various temperatures. Food Research International , 174 , 113662. Shi, Y., Liu, W., Wang, C., & Li, H. (2023). Traditional uses, phytochemistry, and pharmacological properties of Lantana camara : A review. Journal of Ethnopharmacology , 301, 115789. Silva, F., and Domingues, F. C. (2017). Antimicrobial activity of coriander oil and its effectiveness as food preservative. Critical reviews in food science and nutrition, 57 (1), 35-47. Singh, B.K., Delgado-Baquerizo, M., Egidi, E., Guirado, E., Leach, J.E., Liu, H. and Trivedi, P., 2023. Climate change impacts on plant pathogens, food security and paths forward. Nature Reviews Microbiology, 21(10), pp.640-656. Sun, X.P., Qiu, H.J., Qiu, C.S., Wu, Z.M., Song, X., Tang, L., Liu, D., Cao, Y. and Wang, F., 2025. Research progress on biological prevention and control in tobacco green production. Cogent Food & Agriculture, 11(1), p.2448595. Tai, B., Chang, J., Liu, Y., & Xing, F. (2020). Recent progress of the effect of environmental factors on Aspergillus flavus growth and aflatoxins production on foods. Food Quality and Safety , 4 (1), 21-28. Wani, S., Nisa, Q., Fayaz, T., Nabi, N., Nabi, A., Lateef, I., ... & Padder, B. A. (2023). An Overview of Major Bean Diseases and Current Scenario of Common Bean Resistance. Diseases in Legume Crops: Next Generation Breeding Approaches for Resistant Legume Crops , 99-123. White, T. J., Bruns, T., Lee, S., & Taylor, J. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In PCR Protocols: A Guide to Methods and Applications (pp. 315–322). Academic Press. Wilhelm, S. (1955). Longevity of the Verticillium wilt fungus in the laboratory and field. Phytopathology, 45 , 180–181. Xi, K. Y., Xiong, S. J., Li, G., Guo, C. Q., Zhou, J., Ma, J. W., ... & Zhu, Y. X. (2022). Antifungal Activity of Ginger Rhizome Extract against Fusarium solani . Horticulturae , 8 (11), 983. Yadav, D., Gaurav, H., Yadav, R., Waris, R., Afzal, K., & Shukla, A. C. (2023). A comprehensive review on soft rot disease management in ginger ( Zingiber officinale ) for enhancing its pharmaceutical and industrial values. Heliyon . Additional Declarations No competing interests reported. Supplementary Files supplementalinformation.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7054212","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":494468020,"identity":"11e17796-a556-430b-8e01-2703d82d8c38","order_by":0,"name":"Shazia Alam","email":"","orcid":"","institution":"Yangzhou University","correspondingAuthor":false,"prefix":"","firstName":"Shazia","middleName":"","lastName":"Alam","suffix":""},{"id":494468021,"identity":"907fd76e-3534-45b0-9d7a-0fb3a1910d24","order_by":1,"name":"Asma Hanif","email":"","orcid":"","institution":"University of Karachi","correspondingAuthor":false,"prefix":"","firstName":"Asma","middleName":"","lastName":"Hanif","suffix":""},{"id":494468022,"identity":"9216426d-2224-4a2f-884e-d0dcfa17c7b3","order_by":2,"name":"Sumara Shaheen","email":"","orcid":"","institution":"University of Karachi","correspondingAuthor":false,"prefix":"","firstName":"Sumara","middleName":"","lastName":"Shaheen","suffix":""},{"id":494468023,"identity":"a89f885a-b7fe-4e51-99a2-e175c3ad650d","order_by":3,"name":"Min Zhu","email":"","orcid":"","institution":"Yangzhou University","correspondingAuthor":false,"prefix":"","firstName":"Min","middleName":"","lastName":"Zhu","suffix":""},{"id":494468024,"identity":"d7a1a52c-6bd6-46ca-a433-ee18410cd645","order_by":4,"name":"Mohammad Mehdizadeh","email":"","orcid":"","institution":"University of Mohaghegh Ardabili","correspondingAuthor":false,"prefix":"","firstName":"Mohammad","middleName":"","lastName":"Mehdizadeh","suffix":""},{"id":494468025,"identity":"9d855d23-6395-465e-8c08-9591a7a826cd","order_by":5,"name":"Zainul Abideen","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA5ElEQVRIiWNgGAWjYHACNoYENiDF3sDA2MAAxsxEauE5QIoWEGKQSCBSi/yM5GcPHpTZyJlLvjF8OIPBRnbDAd7DBvi0MM5IMzdIOJdmbDk7x9hwA0Oa8YYDfMkJ+LQwSySYSSS2HU7ccDvHTPIBA5BxgMf4AF6PSKR/A2r5X7/h5hmQlv+EtfBI5IBsOZBgcIPHTHIDwwGwFrwOk+B5Uw70S7LhhjNpxYYzDJKNZx7mS8brffn29G0Pf5TZyRscP7zxYU+FnWzf8d7DEvi0oAGQ8cw8JGiAAjK0jIJRMApGwbAGANaJStVjSDa0AAAAAElFTkSuQmCC","orcid":"","institution":"University of Karachi","correspondingAuthor":true,"prefix":"","firstName":"Zainul","middleName":"","lastName":"Abideen","suffix":""}],"badges":[],"createdAt":"2025-07-05 16:38:08","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7054212/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7054212/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":88269958,"identity":"77cd46f4-276e-4255-b441-ed4c195b436e","added_by":"auto","created_at":"2025-08-04 16:56:38","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":191545,"visible":true,"origin":"","legend":"\u003cp\u003eThe Chord diagram shows the \u003cem\u003eIn-vitro\u003c/em\u003eanti-fungal activity of various extracts against pathogenic fungi viz., \u003cem\u003eF.oxysporum, R.solani, M.phaseolina, \u003c/em\u003eand \u003cem\u003eA.flavus\u003c/em\u003e using disc diffusion method. It displays the interrelation between data radially around the circle. The band width of the cord shows the significance of the data and the cords are arranged in the descending order of their weight.\u003c/p\u003e","description":"","filename":"image1.png","url":"https://assets-eu.researchsquare.com/files/rs-7054212/v1/acf5e94fbaf9ee77886ea280.png"},{"id":88269959,"identity":"eb1e8990-fd27-4469-ad77-a252e8c892f0","added_by":"auto","created_at":"2025-08-04 16:56:38","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":132389,"visible":true,"origin":"","legend":"\u003cp\u003eThe Chord diagram shows the \u003cem\u003eIn-vitro\u003c/em\u003eanti-fungal activity of various extracts against pathogenic fungi viz., \u003cem\u003eF.oxysporum, R.solani, M.phaseolina, \u003c/em\u003eand \u003cem\u003e\u0026nbsp;A.flavus\u003c/em\u003e using agar-well method. It displays the interrelation between data radially around the circle. The band width of the cord shows the significance of the data and the cords are arranged in the descending order of their weight.\u003c/p\u003e","description":"","filename":"image2.png","url":"https://assets-eu.researchsquare.com/files/rs-7054212/v1/1e76463925a371e3faa56c8c.png"},{"id":88269964,"identity":"f5fc4542-2134-43d3-b8be-9f40b3b48614","added_by":"auto","created_at":"2025-08-04 16:56:38","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":88684,"visible":true,"origin":"","legend":"\u003cp\u003eApplication of fresh and dry ginger on the growth improvement of red beans.\u003c/p\u003e\n\u003cp\u003eWhereas, T1= control, T2=100% S.T with fresh ginger, T3=75% S.T with fresh ginger, T4=100% S.T with dry ginger, T5=75% S.T with dry ginger, T6=100% S.D with fresh ginger, T7=75% S.D with fresh ginger, T8=100% S.D with dry ginger, T9=75% S.D with dry ginger, T10=0.5% S.A with fresh ginger, T11= 0.1% S.A with fresh ginger, T12= 0.5% S.A with dry ginger, T13=0.1% S.A with dry ginger\u003c/p\u003e","description":"","filename":"image3.png","url":"https://assets-eu.researchsquare.com/files/rs-7054212/v1/667aa9dd8807d546ba5d6b43.png"},{"id":88269962,"identity":"89303331-d403-4b53-8f48-76939a08aff7","added_by":"auto","created_at":"2025-08-04 16:56:38","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":68109,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of seed treatment by fresh ginger powder in addition with fertilizers of red bean plant on the growth. Whereas, \u0026nbsp;T1=control, T2= seed treatment 100%, T3= cowdung, T4= goat dung, T5=DAP, T6= fudan, T7= cow dung +seed treatment 100%, T8= goat dung + seed treatment 100%, T9= DAP + seed treatment 100%, T10= fudan + seed treatment 100%\u003c/p\u003e","description":"","filename":"image4.png","url":"https://assets-eu.researchsquare.com/files/rs-7054212/v1/3da434dfec24effd0534f11b.png"},{"id":88270614,"identity":"c2f86ef8-37cb-4a47-933b-ea9330c53d8d","added_by":"auto","created_at":"2025-08-04 17:04:38","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":64197,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of seed treatment by dry ginger powder in addition with fertilizers of red bean plant on the growth. Whereas, T1=control, T2= seed treatment 100%, T3= cow dung, T4= goat dung, T5=DAP, T6= fudan, T7= cowdung +seed treatment 100%, T8= goat dung + seed treatment 100%, T9= DAP + seed treatment 100%, T10= fudan + seed treatment 100%\u003c/p\u003e","description":"","filename":"image5.png","url":"https://assets-eu.researchsquare.com/files/rs-7054212/v1/e1736b14b22a3d6db7ae2519.png"},{"id":88272420,"identity":"a2bfe61d-fc6f-4b1a-b3dd-77512ff1be0e","added_by":"auto","created_at":"2025-08-04 17:20:38","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":196969,"visible":true,"origin":"","legend":"\u003cp\u003eThe heat map with labels shows the colonization percentage of \u003cem\u003eF.solani, F.oxysporum, R.solani, M. phaseolina, \u003c/em\u003eand \u003cem\u003eA. flavus \u003c/em\u003efor seed treatment with fresh ginger, dry ginger, and for soil drenching and soil amendment with fresh and dry ginger\u003cem\u003e. \u003c/em\u003eThe\u003cem\u003e \u003c/em\u003escale indicates the ascending order of the colonization percentages which shifts to the dark shade of red when the colonization percentage increases while the blue color indicates the minimum colonization percentage.\u003c/p\u003e\n\u003cp\u003eWhereas 6A,\u003c/p\u003e\n\u003cp\u003e1=Control, 2=Seed treatment with FG , 3=Cow dung (CD), 4=Goat dung (GD), 5=DAP, 6=Fudan (F), 7=CD+ seed treatment with FG, 8=GD + seed treatment with FG, 9=DAP + seed treatment with FG, 10=F + seed treatment with FG\u003c/p\u003e\n\u003cp\u003eWhereas 6B,\u003c/p\u003e\n\u003cp\u003e1=Control, 2=Seed treatment with DG , 3=Cow dung (CD), 4=Goat dung (GD), 5=DAP, 6=Fudan (F), 7=CD+ seed treatment with DG, 8=GD + seed treatment with DG, 9=DAP + seed treatment with DG, 10=F + seed treatment with DG\u003c/p\u003e\n\u003cp\u003eWhereas 6C,\u003c/p\u003e\n\u003cp\u003e1=Control, 2=100% S.T with FG, 3=75% S.T with FG, 4=100%S.T with DG, 5=75%S.T with DG, 6=100% S.D with FG, 7=75% S.D with FG, 8=100%S.D with DG, 9=75%S.D with DG, 10=0.5 % S.A with FG, 11=0.1 % S.A with FG, 12=0.5 % S.A with DG, and 13=0.1 % S.A with DG.\u003c/p\u003e","description":"","filename":"image6.png","url":"https://assets-eu.researchsquare.com/files/rs-7054212/v1/8dfa0b19d527cadb3c93a5d3.png"},{"id":88273162,"identity":"19fa54ad-97b5-4e6d-8310-cfb1760d4ef6","added_by":"auto","created_at":"2025-08-04 17:28:39","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1785854,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7054212/v1/d4808b1c-a7cd-4fe4-93da-a35c94e2183e.pdf"},{"id":88271335,"identity":"821e5558-e41f-400a-a8ab-94df9f596304","added_by":"auto","created_at":"2025-08-04 17:12:38","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":162240,"visible":true,"origin":"","legend":"","description":"","filename":"supplementalinformation.docx","url":"https://assets-eu.researchsquare.com/files/rs-7054212/v1/8b843fe43714df63f8ce224b.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Enhancing growth and suppressing root rot in Phaseolus vulgaris using ginger powder derived natural antifungal agent for optimum disease control and crop health","fulltext":[{"header":"1.0 Introduction","content":"\u003cp\u003eMicrobial invasion from virus to viroid, prokaryotic bacteria, eukaryotic fungi, oomycetes, and nematodes, causes significant crop damage annually during both pre-harvest and post-harvest phases (Jamiołkowska \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Debebe \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). These plant diseases cause 40\u0026nbsp;billion dollars in consequential losses globally and are extremely persistent in their attack (Jamiołkowska \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Singh et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Microbes related to plants can sustain several detrimental strategies by producing viroids, viruses, bacteria and fungi, and lead to infectious diseases af fecting only the plant kingdom. Fungal disease frquency depends upon the plant health, the coincidence of a susceptible host, a virulent pathogen, and a favorable environment. The fungal disease invasion in plants also depends on the life strategy and the the pathogen population and the host plant change during their life cycle. The idea of biocontrol has sparked a significant political, economic, and technological discussion to create sustainable agriculture at a lower environmental cost (Barratt et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Sun et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Recent plant safeguarding tactics must immediately be based on natural resources due to the growing worries about ecological toxicity and environmental pollution brought on by the careless use of chemical formulation (Jamiołkowska \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Jabeen et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cem\u003ePhaseolus vulgaris\u003c/em\u003e L. (red bean) is a herbaceous annual plant belongs to Leguminosae family known for their enriched macronutrients, micronutrients and antioxidant substances that nutrition for both humans and animals (Fern\u0026aacute;ndez-Ruiz et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Chen et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Various pathogenic fungi including, \u003cem\u003eRhizoctonia solani, Macrophomina phaseolina\u003c/em\u003e, \u003cem\u003eFusarium oxysporum\u003c/em\u003e, and \u003cem\u003eAspergillus flavus\u003c/em\u003e cause huge economic losses to the bean plants. \u003cem\u003eR. solani\u003c/em\u003e is the major root-rotting fungi, as found 91.8% during disease in bean plants (Mayo et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). This pathogen is more functional in disposed soil and at temperatures between 15 and 18\u0026deg;C. \u003cem\u003eR. solani\u003c/em\u003e is a necrotrophic pathogen and one of the root and hypocotyl pathogens which help them in plant invasion that leads to alter plant physiology and destroy fruit yield as a consequence economic loss globally. \u003cem\u003eMacrophomina. phaseolina\u003c/em\u003e is a soil-borne fungus that affects more than 500 plant species in around 100 families and causes charcoal rot, seedling blight, and root and stem rot (Ghosh et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). This fungus causes disease in soybeans, ground nut cultivars and sorghum under hot weather (30\u0026ndash;35\u0026deg;C) and lower soil moisture (below 60%) (Marquez et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). \u003cem\u003eFusarium\u003c/em\u003e wilt, which is origin by \u003cem\u003eFusarium oxysporum\u003c/em\u003e and occurs in all regions, can also effects the productivity of common beans. \u003cem\u003eF.oxysporum\u003c/em\u003e can be invaded through wounds, natural openings, or intact roots of bean plants with a preference for the tap root and lateral root interconnections (de-Borba et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The most general fungus that causes contamination of food and feed and build aflatoxins is \u003cem\u003eAspergillus flavus\u003c/em\u003e (Lahouar et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). It is the source to cause contamination during food storage because it has the capacity to produce aflatoxins and its ability to survive as a pathogen and saprophyte in the food supply before and after harvest (Tai et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). \u003cem\u003eAspergillus flavus\u003c/em\u003e and \u003cem\u003eAspergillus parasiticus\u003c/em\u003e create secondary metabolites which are known for their high mutagenic, carcinogenic, and teratogenic properties (Shi et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eTraditional Chinese utilized Ginger (\u003cem\u003eZingiber officinale\u003c/em\u003e Roscoe) as a source of spice and medication for more than 200 years. The ginger extract could be used as an additive in food and medical components due to their antifungal and therapeutic properties. Ginger essential oil and ginger extract both have antifungal activities as opposed to a variety of plant pathogens (Peng et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Xi et al. \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The findings show that ginger contains phenolic compounds monoterpenoids, sesquiterpenoids, and their derivatives consist of ketones, aldehydes, alcohols, and esters. These chemicals offer a broad range of antimicrobial activity against various microorganisms, making them a promising substitute for synthetic microbial (Gao et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Utilization of ginger could be an effective biostimulant as the extracted sap from them could be as disease protecting agents in many plant species and maintain the crop heallth of industrial importance. Natural ginger application could be used to enhance the productivity of crop plants and raised the life hood of developing nations and ensure food security and zero hunger.\u003c/p\u003e\u003cp\u003eBioactive qualities found in \u003cem\u003eRicinus communis\u003c/em\u003e L., \u003cem\u003eBrassica nigra\u003c/em\u003e (L.) Koch, and \u003cem\u003eLantana camara\u003c/em\u003e were selected in this study were based on optimum accumulation of bioactive qualities. Medicinal potential of \u003cem\u003eRicinus communis\u003c/em\u003e L. (castor) leaf, roots and seed oil was used as therapeutic uses have confirmed the larvicidal efficacy (Carolina et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Likewise, essential oils (EOs) derived from mustard plants (\u003cem\u003eBrassica nigra\u003c/em\u003e L.), have potent antibacterial qualities that successfully prevent microbial growth and support both natural preservation and therapeutic applications (Mej\u0026iacute;a-Garibay et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Shrub \u003cem\u003eLantana camara\u003c/em\u003e L belong to the genus Lantana contains over 150 species gain attention due to its medical potential and phytochemical richness. \u003cem\u003eLantana camara\u003c/em\u003e parts employed as biological control agents, stimulants, antibacterials, anti-inflammatory agents, and to treat rheumatism (Shi et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). These antifungal characteristics highlight the importance of these plants as tests to evaluate their biological activities using standardized extraction techniques for both \u003cem\u003evitro\u003c/em\u003e and \u003cem\u003evivo\u003c/em\u003e assessment. Using plants as fertilizers, those have higher antifungal activity are most reliable in contemporary agricultural production, have performed a fundamental role in enhancing crop yield and quality (Li et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Akbar et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Although unnecessary fertilization not only prevents the improvement of crop quality and yield, it also causes serious problems such as nutrient loss through leaching, hardness, and acidification of soil, crop pest annoyance, and groundwater safety issues (Du et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The objective of the current study was to evaluate the assess the antifungal activity of ginger powder promotes growth and inhibits root-rotting fungi in \u003cem\u003ePhaseolus vulgaris\u003c/em\u003e L. Study of such kind could be helpful toimprove plant disease management techniques and offer an alternative to chemical fertilizers which not only increase production but also pose no environmental risks.\u003c/p\u003e"},{"header":"2.0 Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003e2.1 Collection of material and seeds\u003c/h2\u003e\u003cp\u003eFresh ginger rhizome and dry powder were collected from Mosamiyat, near University Road Karachi, Pakistan. The leaves of \u003cem\u003eLantana camara\u003c/em\u003e L. and castor (\u003cem\u003eRicinus communis\u003c/em\u003e L.) were gathered from the University of Karachi and leaves of mustard (\u003cem\u003eBrassica nigra\u003c/em\u003e (L.) Koch) were collected from a local market near Mosamiyat, Karachi, Pakistan. Organic fertilizers, cow dung and goat dung, inorganic fertilizers fudan, and diammonium phosphate (DAP) were collected from a nursery near the University of Karachi. Red bean seeds were collected from New Sabzi Mandi Karachi, Pakistan, for pots experiment.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\u003ch2\u003e\u003cb\u003e2.2 Preparation of stock solution of ginger, mustard, castor, and\u003c/b\u003e \u003cb\u003elantana camara\u003c/b\u003e L.\u003c/h2\u003e\u003cp\u003eFor the preparation of stock solution 10g of fresh and dry ginger powder were soaked in 100 mL of appropriate organic solvent (n-hexane). The flasks were covered with aluminum foil and left for 72 hours. Fresh leaves of mustard, castor, and \u003cem\u003eLantana camara\u003c/em\u003e dried under sunlight for 24 hours. For the preparation of stock solution, 10g of the tested leaves powder was weighed and added to the flask containing 100 mL of the appropriate organic solvent (ethanol), separately. Flasks were covered with aluminum foil and left for 1 week. The solutions were centrifuged at 3000 rpm for 10 minutes and used for \u003cem\u003ein-vitro\u003c/em\u003e studies while residue was discarded (Michiels et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). The crude plant extract was considered as 100% stock concentration. One gram of crude extract diluted in one mL of solvent, for instance, equal 100% (w/v) stock.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\u003ch2\u003e2.3 The Isolation of Pathogenic Fungi\u003c/h2\u003e\u003cdiv id=\"Sec6\" class=\"Section3\"\u003e\u003ch2\u003e2.3.1 Isolation of \u003cem\u003eM. phaseolina\u003c/em\u003e by using the wet sieving technique\u003c/h2\u003e\u003cp\u003eSoil samples were taken from the rhizosphere of infected soybean plants exhibiting classic charcoal rot signs in the field station of the University of Karachi, Pakistan. In order to extract microsclerotia from the soil, the pathogen was separated by using the wet sieving approach, as outlined by Ghaffar (1976). In short, distilled water was used to suspend 100g of air-dried soil, which was then run through a succession of sieves with progressively smaller mesh sizes (60, 100 and 200). To stop bacteria from growing, the material that was left on the last sieve was cultivated on potato dextrose agar (PDA) treated with streptomycin and penicillin. Emerging fungal colonies were preliminarily identified as \u003cem\u003eMacrophomina phaseolina\u003c/em\u003e based on morphological traits, including the formation of numerous black microsclerotia, absence of conidia, and fast-growing grey to black mycelia (Schafer \u0026amp; Kiewnick. 2010).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec7\" class=\"Section3\"\u003e\u003ch2\u003e2.3.2 Isolation of \u003cem\u003eR. solani\u003c/em\u003e by baiting technique\u003c/h2\u003e\u003cp\u003eSamples of soil were taken from the rhizosphere of infected chilli (\u003cem\u003eCapsicum annuum\u003c/em\u003e) plants from agricultural fields of the University of Karachi, Pakistan, that were exhibiting indications of basal stem rot. The soil plating method outlined by Anderson and Papavizas (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e1965\u003c/span\u003e) was used to recover the fungal isolate. Plates were incubated at 25\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C for 3\u0026ndash;5 days after 10 g of soil was spread out on water agar (WA) supplemented with streptomycin sulphate (100 mg/L). For purification, hyphal tips from newly formed colonies were subcultured onto potato dextrose agar (PDA).\u003c/p\u003e\u003cp\u003ePreliminary identification of the fungal isolate as \u003cem\u003eRhizoctonia solani\u003c/em\u003e was based on morphological traits, including right-angled hyphal branching, absence of conidia, and presence of typical multinucleate cells seen under a compound microscope following safranin staining. DAPI staining was used to confirm whether the condition was binucleate or multinucleate. The internal transcribed spacer (ITS) region of rDNA was amplified using the universal primers ITS1 and ITS4 in order to perform molecular identification and verify the identity (White et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e1990\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec8\" class=\"Section3\"\u003e\u003ch2\u003e2.3.3 Soil dilution for the \u003cem\u003eF. oxysporum\u003c/em\u003e isolation\u003c/h2\u003e\u003cp\u003eSoil samples were taken from the rhizosphere of wilted tomato (\u003cem\u003eSolanum lycopersicum\u003c/em\u003e) plants from agricultural fields of the University of Karachi, Pakistan, that displayed the typical indications of vascular wilt. According to Leslie and Summerell (2006), the pathogen was identified using the soil dilution plate approach. The fusarium- selective medium from Komada, which is known to promote the development of \u003cem\u003eFusarium\u003c/em\u003e spp. while inhibiting the growth of bacterial and other fungal pollutants, was plated onto 100 grams of air dried soil that had been serially diluted in sterile distilled water. Subculturing on potatoes dextrose agar (PDA) allowed for the purification of newly formed fungal colonies. Based on morphological features such as development of many microconidia, three to five septate sickle-shaped macroconidia, and thick walled chlamydospores seen under a compound microscope, \u003cem\u003eFusarium oxysporum\u003c/em\u003e was initially identified (Booth, 1971). On PDA, the colony morphology displayed cottony aerial mycelium and characteristic white to pinkish-purple coloring.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec9\" class=\"Section3\"\u003e\u003ch2\u003e2.3.4 Isolation of \u003cem\u003eAspergillus flavus\u003c/em\u003e\u003c/h2\u003e\u003cp\u003eIn agricultural fields at the University of Karachi, Pakistan, an area known for frequent contamination by \u003cem\u003eAspergillus\u003c/em\u003e, soil samples were taken from the rhizosphere of ground nuts (\u003cem\u003eArachis hypogaea\u003c/em\u003e) plants. The dilution plate technique was used for isolation, in accordance with Pitt and Hocking\u0026rsquo;s (20009) methodology. To promote specific growth, serial dilutions of air-dried soil were made in sterile distilled water and plated on semi-selective media such as \u003cem\u003eAspergillus flavus\u003c/em\u003e and \u003cem\u003eparasiticu\u003c/em\u003es agar (AFPA) and Czapek- Dox agar (CDA). For five to seven days, plates were incubated at 28\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:℃\\)\u003c/span\u003e\u003c/span\u003e. To examine morphological traits, colonies that resembled \u003cem\u003eAspergillus flavus\u003c/em\u003e were subcultured on PDA. Both macroscopic and microscopic characteristics, such as yellow green conidial head, biseriate conidiophores, and the presence of rough walled conidia organized in chains, were used for the first time identification (Klich, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2002\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\u003ch2\u003e2.4 \u003cem\u003eIn-vitro\u003c/em\u003e experiment\u003c/h2\u003e\u003cdiv id=\"Sec11\" class=\"Section3\"\u003e\u003ch2\u003e2.4.1 Antifungal Activity using Paper Disc Diffusion Method\u003c/h2\u003e\u003cp\u003eSterilized Whatman\u0026rsquo;s paper discs were socked in stock solution of 100 \u0026micro;L of fresh and dried ginger (extracts separately), n-hexane (control for ginger), ethanol (control for castor, mustard, \u003cem\u003eLantana camara\u003c/em\u003e) and fungicide (carbendazim) for half an hour. Pour the petri plates with Potato Dextrose Agar (PDA) medium containing streptomycin 0.2/L and penicillin 100 units/L. Sterilized Whatman\u0026rsquo;s paper discs soaked in respective solutions (castor, mustard, \u003cem\u003eLantana camara\u003c/em\u003e, fresh and dry ginger powder) were placed on three sides of petri plates. In the center of PDA poured petri plates, a disc of each fungus (\u003cem\u003eM. phaseolina, F. oxysporum, R. solani\u003c/em\u003e, and \u003cem\u003eA. flavus\u003c/em\u003e) was placed, and each fungus was replicated thrice. At 25\u0026ndash;33\u0026deg;C temperature petri plates were incubated for 7 days. After an incubation period, the zone of inhibition was measured by scale in mm units (Silva \u0026amp;Domingues, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section3\"\u003e\u003ch2\u003e2.4.2 Antifungal activity using Agar well method\u003c/h2\u003e\u003cp\u003ePDA medium having penicillin 100 units/L and streptomycin 0.2/L was poured into petri plates. By using a sterilized 5mm cork borer three wells were made in PDA poured petri plates, and each well was inoculated with each stock solution (castor, mustard, \u003cem\u003elantana camara\u003c/em\u003e, fresh and dry ginger powder), n-hexane (control for ginger), ethanol (control for castor, mustard, \u003cem\u003eLantana camara\u003c/em\u003e) and fungicide (carbendazim). Discs of fungus (i.e. \u003cem\u003eM. phaseolina, F. oxysporum, R. solani\u003c/em\u003e, and \u003cem\u003eA. flavus\u003c/em\u003e) was placed in the middle of PDA-poured petri plates. Each examined fungus was replicated three times. At 25\u0026ndash;33\u0026deg;C Petri plates were incubated for 7 days. After an incubation period, the zone of inhibition was measured by scale in mm units (Espinel-Ingroff, et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2007\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe zone of growth inhibition was measured and calculated by the following formula.\u003c/p\u003e\u003cp\u003eX= \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\frac{Y}{Z}\\:\\)\u003c/span\u003e\u003c/span\u003ex 100\u003c/p\u003e\u003cp\u003eX\u0026thinsp;=\u0026thinsp;percentage of growth inhibition\u003c/p\u003e\u003cp\u003eY\u0026thinsp;=\u0026thinsp;Growth of mycelia measured in control plates (mm)\u003c/p\u003e\u003cp\u003eZ\u0026thinsp;=\u0026thinsp;Growth of mycelia measured in treated plates (mm)\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\u003ch2\u003e2.5 \u003cem\u003eIn-vivo\u003c/em\u003e experiment\u003c/h2\u003e\u003cp\u003eFresh and dry ginger powder which showed effective results in the \u003cem\u003ein-vitro\u003c/em\u003e studies among all other treatments (castor, mustard, and \u003cem\u003elantana camara\u003c/em\u003e) was carried to the in-vivo studies on red beans.\u003c/p\u003e\u003cdiv id=\"Sec14\" class=\"Section3\"\u003e\u003ch2\u003e2.5.1 Collection of soil and seed treatment\u003c/h2\u003e\u003cp\u003eSandy loam soil obtained from a nursery near the University of Karachi, sieved the soil, and the debris was discarded. The soil was transferred into clay pots (600g/) pots. Red bean seeds were sterilized with 1% NaClO, using distilled water, cleaned twice, then allowed to air dry. Tested seeds were treated with 100 and 75% concentrations of both dry and fresh ginger powder separately for about half an hour while untreated seeds were used as control. Five seeds were planted in pots, and there were three replicates for each treatment. Infection percentage and growth parameters were documented after one month of seed germination.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec15\" class=\"Section3\"\u003e\u003ch2\u003e2.5.3 Soil amendment and drenching\u003c/h2\u003e\u003cp\u003eEach pot had 600 g of soil and was amended with dry and fresh ginger (0.1% and 0.5%) powder. After amendment for one day, five seeds of red beans after surface sterilization with 1% NaClO were planted in each pot. Three replicates were made for each treatment. After one month of seed germination, infection percentage and growth parameters were documented. For soil drenching, 75% extract is made by dilution of 25% of the stock, and 100% extract is undiluted stock. 25 mL prepared concentration of 75% and 100% of both fresh and dry ginger were drenched into clay pots having soil(600g) while untreated soil was used as control. Five seeds were planted in pots and, three replicates for each treatment. Growth parameters and percentage of infection were documented after one month of seed germination.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec16\" class=\"Section3\"\u003e\u003ch2\u003e2.5.5 Growth experiment\u003c/h2\u003e\u003cp\u003ePots were filled with 600g of soil. Organic fertilizers (cow dung and goat dung) and inorganic fertilizers (DAP and fudan) amended in soil at 0.1 and 0.5% per pot. Tested bean seeds were treated with extracts of both fresh and dry ginger powder. Another experiment was set up of these fertilizers without treating seeds with ginger. After one day of amendment, 5 seeds were sown in each container having 600g soil, and were watered regularly. Growth parameters such as infection percentage, number of leaves and nodules, and shoot and root length and weight were recorded after 1 month of seed germination.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec17\" class=\"Section3\"\u003e\u003ch2\u003e2.5.6 Colonization % of pathogenic fungi from roots\u003c/h2\u003e\u003cp\u003eAfter one month of red bean growth, the plants were pulled out, the roots were cleaned with running water and dried with paper. The taproot of each treatment was cut into 5 small pieces and soaked in 1% of sodium hypochlorite NaClO for three minutes for surface sterilization, and washed with sterilized distilled water three times. After the roots were dried the fragments were put on a PDA-poured petri dish having antibiotics (penicillin and streptomycin) to inhibit the growth of bacteria and incubated at room temperature for one week. Fungi that emerged from the root segment after incubation were examined under the microscope and the percentage of colonization was calculated by the following formula; a =\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\:\\frac{b}{c}\\:\\)\u003c/span\u003e\u003c/span\u003ex 100\u003c/p\u003e\u003cp\u003ea\u0026thinsp;=\u0026thinsp;Colonization percentage\u003c/p\u003e\u003cp\u003eb\u0026thinsp;=\u0026thinsp;Number of fungi colonized by root pieces\u003c/p\u003e\u003cp\u003ec\u0026thinsp;=\u0026thinsp;Total roots pieces\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\u003ch2\u003e2.6 Statistical analysis\u003c/h2\u003e\u003cp\u003eData was reported as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation. The zone of inhibition of fungi was expressed in percentage and represented by using a chord diagram. To examine the effects of seed treatment, soil drenching, and soil amendment and the treatments using IBM SPSS statistics 26.0, the principal component analysis was performed. With significance of 0.000, Barlett\u0026rsquo;s Test of Sphericity confirms that Principal Component Analysis could be carried out on the data subjected to the analysis. The colonization percentage of the root-rotting pathogens was represented using a Heat map with labels. The graphs were plotted using OriginPro 2024b.\u003c/p\u003e\u003c/div\u003e"},{"header":"3.0 Experimental Results","content":"\u003cdiv id=\"Sec20\" class=\"Section2\"\u003e\n \u003ch2\u003e3.1 \u003cem\u003eIn-vitro\u003c/em\u003e test\u003c/h2\u003e\n \u003cp\u003eTwo different methods (disc diffusion and agar-well method) were used to find the effect of extracts against the tested fungi namely, \u003cem\u003eM. phaseolina, R. solani, F. oxysporum\u003c/em\u003e, and \u003cem\u003eA. flavus\u003c/em\u003e (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e-Fig. 5S). Test fungi were suppressed and distinct zones of inhibitions were produced when extract at 100% concentration of both fresh ginger and dry ginger powder was used except for \u003cem\u003eR. solani\u003c/em\u003e, which didn\u0026rsquo;t show the zone of inhibition against fresh ginger extract. Dry ginger powder extract showed a significant zone of inhibition against \u003cem\u003eF. oxysporum\u003c/em\u003e in the agar well method. However, fresh ginger extract showed a minimum zone of inhibition against \u003cem\u003eM. phaseolina\u003c/em\u003e. The extracts of \u003cem\u003eLantana camara\u003c/em\u003e, mustard, and castor didn\u0026rsquo;t show a zone of inhibition against the test fungi, however overlapping of test fungi over the extracts was observed.\u003c/p\u003e\n \u003cp\u003eOverall results showed the complete inhibition of \u003cem\u003eM. phaseolina, F. oxysporum\u003c/em\u003e, and \u003cem\u003eA.flavus\u003c/em\u003e by both ginger extracts but inhibition of the \u003cem\u003eR. solani\u003c/em\u003e by both ginger extracts was negligible. Extracts of castor, \u003cem\u003eLantana camara\u003c/em\u003e, and mustard were not effective against tested fungi. Among both of the methods, the Agar well method showed the highest zone of inhibition followed by the paper disc method (Figs. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e and \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec21\" class=\"Section2\"\u003e\n \u003ch2\u003e3.2 \u003cem\u003eIn-vivo\u003c/em\u003e results\u003c/h2\u003e\n \u003cp\u003e\u003cstrong\u003ePrincipal component analysis\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThe principal component analysis was carried out on the physical parameters of red bean plants, including root length, root weight, shoot length, shoot weight, no. of leaves, no. of nodules, and dry root and shoot weight in the greenhouse experiment. Three different pot experiments were plotted. The first experiment shows the effect of fertilizer and fertilizer\u0026thinsp;+\u0026thinsp;seed treatment by fresh ginger on the growth of red beans and against root-rotting fungi. The second experiment shows the effect of fertilizer and fertilizer\u0026thinsp;+\u0026thinsp;seed treatment by dry ginger powder on the growth of red beans and against root-rotting fungi. The third pot experiment shows the effect of soil amendment, soil drenching, and seed treatment by fresh ginger and dry ginger powder on the growth of red beans and against root-rotting fungi. It demonstrates how organic amendment and other attributes influence heterogeneity compared to the controlled plants.\u003c/p\u003e\n \u003cp\u003eFor the 1st pot experiment, a total of 76.98% combined variance was seen in the data set. Out of which, the 1st principal component gives 51.20% variance while 25.78% was shown by the 2nd principal component. The comparative distance of the treatments from the control on the score plot shows their positive correlation with one another (Supp. Figure \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). For the 2nd pot experiment, a total of 67.53% combined variance was seen in the data set. Out of which, the 1st principal component gives 44.30% variance while 23.23% was shown by the 2nd principal component. The comparative distance of the treatments from the control on the score plot shows their positive correlation with one another (Supp. Figure \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). For the 3rd pot experiment, a total of 55.18% combined variance was seen in the data set. Out of which, the 1st principal component gives 34.11% variance while 21.28% was shown by the 2nd principal component. The comparative distance of the treatments from the control on the score plot shows their positive correlation with one another (Supp. Figure \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e). Different methods were used with fresh ginger and dry ginger powder and extract for the management of root pathogenic fungi, and to enhance the growth of red bean plants.\u003c/p\u003e\n \u003cp\u003eRed Bean seeds were treated with 75% (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) of fresh ginger increased the number of leaves, shoot length, and shoot weight as compared to control. Similar results were also recorded for dry ginger powder. Seeds treated with 100% concentration of dry ginger powder was found to be best to improve the length and weight of bean plants. Soil 600g drenched with 100% concentration of dry ginger powder significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) to improve the length and weight of red bean root. Pots drenched with 75% of dry ginger increased (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) the shoot length and weight as compared to control. Similar results were also recorded for fresh ginger powder. Soil amended with 0.5% concentration of dry ginger powder was significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) enhanced the length and weight of roots of beans. Pots soil amended with 0.5% dry ginger showed an increase the number of leaves, length and weight of shoot followed by 0.1%. Similar results were also recorded in fresh ginger powder (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e). Among the three methods, all were found effective but seed treated at 100% concentration could be used as an inexpensive tool in commercial scale as compare to soil drenching and soil amendment method which is recommended for low scale farming.\u003c/p\u003e\n \u003cp\u003eBy using organic fertilizers (goat dung and cow dung) and inorganic fertilizer (DAP) amended @0.1% along with seeds treated at 100% gave maximum increase in growth parameter of shoot and root of beans plant and impressively reduced the infection of root rotting fungi. Shoot length was increased significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) when soil was amended with Fudan in combination with seed treatment with dry ginger powder. Shoot weight and root length increased significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05)when soil was amended with cow dung in addition with seeds treated with dry ginger powder. Similarly, root weight was increased significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) when soil was amended with DAP in combination with seed treatment with dry ginger powder. Shoot weight and root length were increased significantly when soil was amended with goat dung in combination with seed treated with fresh ginger. Similarly, root weight increased significantly when soil was amended with cow dung and seed treated with fresh ginger. Shoot length was increased when soil was amended with fudan in combination with seed treatment with fresh ginger (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e\u0026ndash;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eColonization percentage of root-rotting fungi (\u003c/strong\u003e\u003cstrong\u003eM.phaseolina, R. solani, F.solani\u003c/strong\u003e, \u003cstrong\u003eand\u003c/strong\u003e \u003cstrong\u003eF. oxysporum\u003c/strong\u003e\u003cstrong\u003e)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eCombine effect of fertilizers with seed treatment by fresh and dry ginger.\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eSoil amended with fudan in combination with seeds treated with fresh ginger reduced (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) the colonization percentage of root-rotting fungi. Similarly, soil amended with cow dung, DAP, fudan, and goat dung in combination with seeds treated with fresh ginger and dry ginger extracts were more effective against root-rotting fungi.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eSeed treatment, Soil drenching, and Soil amendment method by fresh ginger and dry ginger\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eSterilized seeds with 1% NaClO were treated by fresh ginger and dry ginger powder with 75% and 100% concentrations separately for 30 minutes. The seeds of red beans were sown in each pot. Each treatment was replicated three times. The seed not treated acts as control. The colonization percentage of root rot pathogen was reduced (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) when seeds were treated with 100% of the extract of both fresh ginger and dry ginger powders. Seeds treated with 100% show maximum inhibition as compared to the control. Both treatments show antifungal activity againstroot-rotting fungi on bean plants. Sterilized seed after 1% NaClO were sowed in the soil, and the soil was drenched at 75% and 100% concentration of both tested ginger extracts. Each treatment was replicated thrice. Untreated seeds were served as control. The colonization percentage of root rot pathogens was reduced (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) when soil was drenched with 100% extracts of both fresh ginger and dry ginger powder. Both treatments at different concentrations showed antifungal activity against root rotting fungi and improved the growth of red bean plants. Sterilized seeds with 1% NaClO were sowed in soil at different concentrations (0.5% and 0.1%); each treatment was replicated three times. The seed not treated act as control. The colonization percentage of root rot pathogen significantly decreased when soil was amended with 0.5 and 0.1% of fresh ginger powder. However, maximum inhibition (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) of \u003cem\u003eM. phaseolina, F. solani, R. solani\u003c/em\u003e, and \u003cem\u003eF. oxysporum\u003c/em\u003e were recorded as 0.5% as compared to control. At different concentrations (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) both fresh ginger and dry ginger powders showed antifungal activity against root rotting fungi by using soil amendment method experiment.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"4.0 Discussion","content":"\u003cp\u003eOveruse of chemical fertilizers is a multilayered problem with severe consequences, adversely affecting human health and crop health. Extensive use of agrochemicals, in agriculture is inducing the chemical resistance in the phytopathogens against the commonly used chemical fertilizers in the field (B\u0026oacute;dalo et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). The current reserch was designed to use organic material instead of chemicals to control root pathogenic fungi associated with beans. For this purpose, the antifungal effect of ginger, mustard, castor, and \u003cem\u003eLantana camara\u003c/em\u003e was accessed in the \u003cem\u003ein-vitro\u003c/em\u003e experiment, and the effective organic material, \u003cem\u003ei.e.\u003c/em\u003e, ginger, was carried to the \u003cem\u003ein-vivo\u003c/em\u003e experimentation, where its effects on the growth parameters and root rot colonization were observed. One-third of the world\u0026rsquo;s crop production is lost due to plant pathogens each year. It has been reported that the fungal pathogens cause severe damage among the biotic stresses, followed by bacterial and viral infections (Wani et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Root rot, stem-blight, and stem-rot are the major diseases of common beans and cause huge economic losses.\u003c/p\u003e\u003cp\u003eAccording to Ali et al. (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), ginger is rich source of bioactive substances with potent antifungal and antibacterial properties, including zingerone, shogaol, and gingerol. Present results showed that ginger extract in the \u003cem\u003ein-vitro\u003c/em\u003e experiment showed the maximum zone of inhibition against test fungi due to the anti-microbial effect. Our results are also in concordance with Yadav et al. (\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) that ginger can stop the growth of some of the most prevalent fungal pathogens. Previous research also showed that ginger oil has been estimated to possess antimicrobial effects (Beristain-Bauza et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). The research carried out by Gao et al. (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) reported that monoterpenoids, sesquiterpenoids, phenolic compounds and their derivatives, aldehydes, ketones, alcohols, and esters are present in ginger, which gives a broad antimicrobial array against various pathogens and makes it a potential alternative to synthetic fertilizers. The study carried out by Xi et al. (\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) reported the antifungal potential of ginger extract and ginger essential oil against \u003cem\u003eF. oxysporum\u003c/em\u003e.\u003c/p\u003e\u003cp\u003eIn the present research, castor, mustard, and \u003cem\u003eLantana camara\u003c/em\u003e didn\u0026rsquo;t show effective results against the pathogenic fungi. However, various researchers have reported their antifungal activities. Carolina et al. (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) have reported that ricinine is the main compound found in the castor leaves responsible for the toxic effects against various pathogens. Compound in \u003cem\u003eRicinus communis\u003c/em\u003e L.\u0026rsquo;s seeds, leaves, and oil are mostly responsible for its remarkable antifungal qualities. Ricinoleic acid, a fatty acid with well-known antibacterial properties, is present in castor oil. Numerous studies demonstrate its effectiveness against harm full fungi that affect both plants and humans, such as \u003cem\u003eRhizoctonia solani\u003c/em\u003e, \u003cem\u003eAspergilus\u003c/em\u003e species, and \u003cem\u003eFusarium\u003c/em\u003e species. Furthermore, Mejia-Garibay et al. (2015) reported that the essential compounds of mustard \u003cem\u003ei.e.\u003c/em\u003e, allyl-isothiocyanate (AITC) shows antimicrobial activity, in both vapor and liquid phase against \u003cem\u003eAspergillus niger, A. ochraceus\u003c/em\u003e and \u003cem\u003ePenicillium citrinum.\u003c/em\u003e Glucosinolates, which are abundant in Brassica nigra, break down into isothiocyanates, which are strong antifungal substances. Fungal cell membranes and metabolic processes are disrupted by these bioactives. Extract from mustard has demonstrated efficacy against fungi, including species of \u003cem\u003eAlternaria\u003c/em\u003e, \u003cem\u003eFusarium\u003c/em\u003e, and \u003cem\u003eBotrytis\u003c/em\u003e. Triterpenoids, flavonoids, and essential oils found in \u003cem\u003eLantana camara\u003c/em\u003e L. have broad-spectrum antibacterial properties. \u003cem\u003eM. phaseolina, F. oxysporum\u003c/em\u003e, and \u003cem\u003eA. flavus\u003c/em\u003e are among the phytopathogenic fungi whose growth has been demonstrated to be inhibited by it\u0026rsquo;s extracts. The disruption of fungal cell wall synthesis and mitochondrial dysfunction are probably the cause of the antifungal action (Shi, Y et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Due to the efficient results of ginger in the \u003cem\u003ein-vitro\u003c/em\u003e experiment, it was carried to the pot experiment where it successfully improved the physical parameters and suppressed the root-rotting systemic fungi. The research carried out by Ali et al. (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) reported that the application of ginger extract enhanced damask rose growth characteristics and it is as a result in the availability of nutrients for enhanced growth. Similarly, Xi et al. (\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) reported that the constituents of ginger, including 4-hydroxybenzaldehyde and quercetin, inhibit the growth of \u003cem\u003eF. solani\u003c/em\u003e completely and suggested their use as a natural fungicide in disease management. Additionally, investigations on plant-microbe interactions have shown that ginger powder may affect the dynamics of microorganisms in the soil, favoring beneficial bacteria while decreasing pathogens (Carolina et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Increased nutrient absorption, better root system, and eventually higher crop output are the outcomes of the combined impact.\u003c/p\u003e\u003cp\u003eGenerally, the colonization percentage of \u003cem\u003eM. phaseolina, F. solani\u003c/em\u003e and \u003cem\u003eF. oxysporum\u003c/em\u003e were more suppressed than \u003cem\u003eR. solani\u003c/em\u003e when combined with fertilizers and seeds treated by fresh and dry ginger powder also improved the growth of red plants as compared to individual treatment. Overall result showed that root pathogenic fungi were more suppressed by ginger dry powder along with treatment with fudan and increased the plant growth as compared to other. Among three methods (seed treatment, soil drenching, and soil amendment), all were found effective but seed treated with 100% concentration could be used as an inexpensive tool on commercial scale as compare to soil drenching and soil amendment methods which is recommended for low scale farming. It is suggested from the present research that the application of fresh and dry ginger extracts should be used against root-rotting fungi and for the improvement of the growth of red bean plants on large agricultural fields, which is cheap, easily available and non-toxic to the soil environment. So, it is a useful alternative to fungicides and bactericides.\u003c/p\u003e"},{"header":"5.0 Conclusion","content":"\u003cp\u003eBased on the study on the application of ginger to enhance the growth of red beans and suppress root-rotting pathogenic fungi, the results suggest that ginger can be an effective organic amendment for improving the growth and health of red beans. The study found that the application of ginger extract improved the seed germination rate, plant growth, and yield of Red beans, while also suppressing the growth of root-rotting pathogenic fungi. This was attributed to the presence of various bioactive compounds in ginger, including gingerol and shogaol, which have been shown to have antifungal and plant growth-promoting properties. The use of organic amendments, such as ginger, can have several benefits for plant growth and soil health. Organic amendments can improve soil fertility, increase microbial activity, and suppress the growth of soil-borne pathogens. This can lead to improved plant growth, yield, and quality, while also reducing the need for synthetic fertilizers and pesticides. Overall, the use of organic amendments like ginger can be an effective strategy for improving the growth and health of Red beans, while also promoting sustainable agriculture practices. An efficient method for promoting \u003cem\u003ePhaseolus vulgaris\u003c/em\u003e development and inhibiting root rotting fungal infections, including \u003cem\u003eM. phaseolina, F. oxysporum\u003c/em\u003e, and \u003cem\u003eR. solani\u003c/em\u003e is the application of powdered ginger. This purpose demonstrates that ginger (\u003cem\u003eZingiber officinale)\u003c/em\u003e promises a sustainable, natural, and environmentally friendly substitute for chemical fungicides in the treatment of legume diseases. In order to maximize ginger powder\u0026rsquo;s effectiveness, future studies should concentrate on adjusting its concentration and administration method under various agronomic circumstances. To evaluate the effect of recurrent ginger treatment on soil microbiota in group health, long-term research is also required. Additionally, combining ginger powder with other biological control medications may enhance its ability to inhibit illness.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval\u003c/strong\u003e Not applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to Participate\u003c/strong\u003e We confirm that the manuscript has been read and approved by all named authors and that there are no other people who satisfied the criteria for authorship but are not listed. We further confirm that all the authors listed in the manuscript have been approved by all of us.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors contribution\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eShazia Alam, Asma Hanif, Sumara Shaheen and Zainul Abideen: Conceptualization, Investigation, Formal analysis, Methodology. Shazia Alam, Min Zhu, Writing \u0026ndash; original draft. Supervision, Conceptualization and review \u0026amp; editing\u0026nbsp;Mohammad Mehdizadeh, Min Zhu and Zainul Abideen\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding \u003c/strong\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability statement\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eAkbar, R., Manzoor, S., Azad, R., Makai, G., Rahim, J., Sheikh, U.A.A., Ali, A., Aziz, T., Ahmad, H.I., Ahmed, M. and Du, D., 2024. Botanical Pesticides: Role of \u003cem\u003eRicinus communis\u003c/em\u003e in Managing \u003cem\u003eBactrocera zonata\u003c/em\u003e (Tephritidae: Diptera). Insects, 15(12), p.959.\u003c/li\u003e\n \u003cli\u003eAli, E. F., Al-Yasi, H. M., Issa, A. A., Hessini, K., \u0026amp; Hassan, F. A. (2022). Ginger extract and fulvic acid foliar applications as novel practical approaches to improve the growth and productivity of Damask Rose. \u003cem\u003ePlants\u003c/em\u003e, \u003cem\u003e11\u003c/em\u003e(3), 412.\u003c/li\u003e\n \u003cli\u003eAnderson, J. A., \u0026amp; Papavizas, G. C. (1965). A method for isolating \u003cem\u003eRhizoctonia solani\u003c/em\u003e from soil and plant tissue. \u003cem\u003ePhytopathology\u003c/em\u003e, \u003cem\u003e55\u003c/em\u003e, 1299\u0026ndash;1301.\u003c/li\u003e\n \u003cli\u003eBarratt, B. I. P., Moran, V. C., Bigler, F., \u0026amp; Van Lenteren, J. C. (2018). The status of biological control and recommendations for improving uptake for the future. \u003cem\u003eBioControl\u003c/em\u003e, \u003cem\u003e63\u003c/em\u003e, 155-167.\u003c/li\u003e\n \u003cli\u003eBeristain-Bauza, S. D. C., Hern\u0026aacute;ndez-Carranza, P., Cid-P\u0026eacute;rez, T. S., \u0026Aacute;vila-Sosa, R., RuizL\u0026oacute;pez, I. I., \u0026amp; Ochoa-Velasco, C. E. (2019). Antimicrobial activity of ginger (\u003cem\u003eZingiber Officinale\u003c/em\u003e) and its application in food products. \u003cem\u003eFood Reviews International\u003c/em\u003e, \u003cem\u003e35\u003c/em\u003e(5), 407-426.\u003c/li\u003e\n \u003cli\u003eB\u0026oacute;dalo, A., Borrego, R., Garrido, C., Bolivar-Anillo, H. J., Cantoral, J. M., Vela-Delgado, M. D., ... \u0026amp; Carb\u0026uacute;, M. (2023). In Vitro Studies of Endophytic Bacteria Isolated from Ginger (\u003cem\u003eZingiber officinale\u003c/em\u003e) as Potential Plant-Growth-Promoting and Biocontrol Agents against \u003cem\u003eBotrytis cinerea\u003c/em\u003e and \u003cem\u003eColletotrichum acutatum\u003c/em\u003e. \u003cem\u003ePlants\u003c/em\u003e, \u003cem\u003e12\u003c/em\u003e(23), 4032.Booth, C. (1971). \u003cem\u003eThe Genus Fusarium\u003c/em\u003e. Commonwealth Mycological Institute, Kew, UK.\u003c/li\u003e\n \u003cli\u003eCarolina, A., Herliyana, E. N., and Sulastri, H. (2019). Antifungal activity of castor (Ricinus communis) leaves methanolic extract on \u003cem\u003eAspergillus niger\u003c/em\u003e. \u003cem\u003eInternational Food Research Journal\u003c/em\u003e, \u003cem\u003e26\u003c/em\u003e(2):595-598.\u003c/li\u003e\n \u003cli\u003eCarolina, F. R., Lopez, S., \u0026amp; Morales, M. (2019). Biological activities of \u003cem\u003eRicinus communis\u003c/em\u003e plant parts: A review. \u003cem\u003eJournal of Medicinal Plants Research\u003c/em\u003e, 13(11), 255\u0026ndash;264.\u003c/li\u003e\n \u003cli\u003eChen, Y. C., Hsieh, S. L., \u0026amp; Hu, C. Y. (2020). Effects of red-bean tempeh with various strains of Rhizopus on GABA content and cortisol level in zebrafish. \u003cem\u003eMicroorganisms\u003c/em\u003e, \u003cem\u003e8\u003c/em\u003e(9), 1330.\u003c/li\u003e\n \u003cli\u003eChiejina, N. V., and Onaebi, C. N. (2016). Phytochemical constituents and antifungal properties of \u003cem\u003eChromolaenaodorata\u0026nbsp;\u003c/em\u003eand \u003cem\u003eMoringa oleifera Lam\u003c/em\u003e on fungal rot of cucumber (\u003cem\u003eCucumis sativus\u003c/em\u003e L.) fruit. \u003cem\u003eAsian Journal of Plant Sciences\u003c/em\u003e, \u003cem\u003e2\u003c/em\u003e(1): 10-15\u003c/li\u003e\n \u003cli\u003ede Borba, M. C., Garc\u0026eacute;s-Fiallos, F. R., \u0026amp; Stadnik, M. J. (2017). Reactions of black bean seedlings and adult plants to infection by \u003cem\u003eFusarium oxysporum\u003c/em\u003e f. sp. phaseoli. \u003cem\u003eCrop protection\u003c/em\u003e, \u003cem\u003e96\u003c/em\u003e, 221-227.\u003c/li\u003e\n \u003cli\u003eDebebe, S., 2022. Post-harvest losses of crops and its determinants in Ethiopia: tobit model analysis. Agriculture \u0026amp; food security, 11(1), pp.1-8.\u003c/li\u003e\n \u003cli\u003eDu, Q. J., Xiao, H. J., Li, J. Q., Zhang, J. X., Zhou, L. Y., \u0026amp; Wang, J. Q. (2021). Effects of different fertilization rates on growth, yield, quality and partial factor productivity of tomato under non-pressure gravity irrigation. \u003cem\u003ePLoS One\u003c/em\u003e, \u003cem\u003e16\u003c/em\u003e(3), e0247578.\u003c/li\u003e\n \u003cli\u003eEspinel-Ingroff, A., Arthington-Skaggs, B., Iqbal, N., Ellis, D., Pfaller, M. A., Messer, S., Rinaldi, M., Fothergill, A., Gibbs, D. L., \u0026amp; Wang, A. (2007). Multicenter evaluation of a new disk agar diffusion method for susceptibility testing of filamentous fungi with voriconazole, posaconazole, itraconazole, amphotericin B, and caspofungin. \u003cem\u003eJournal of clinical microbiology\u003c/em\u003e, \u003cem\u003e45\u003c/em\u003e(6), 1811\u0026ndash;1820.\u003c/li\u003e\n \u003cli\u003eFern\u0026aacute;ndez-Ruiz V., Morales P., Ruiz-Rodr\u0026iacute;guez B.M., TorijaIsasa E. (2017). Nutrients and bioactive compounds in wild fruits through different continents. \u003cem\u003eWild Plants Mushrooms Nuts Funct. Food Prop. Appl,\u0026nbsp;\u003c/em\u003e\u003cem\u003e17\u003c/em\u003e:263\u0026ndash;314.\u003c/li\u003e\n \u003cli\u003eGao, Y.; Lu, Y.; Zhang, N.; Udenigwe, C.C.; Zhang, Y.; Fu, Y. Preparation, pungency and bioactivity of gingerols from ginger (\u003cem\u003eZingiber officinale\u003c/em\u003e Roscoe): A review. \u003cem\u003eCrit. Rev. Food Sci. Nutr.\u003c/em\u003e 2022, \u003cem\u003e22\u003c/em\u003e, 1\u0026ndash;26.\u003c/li\u003e\n \u003cli\u003eGhosh T., Biswas M. K., Guin C., Roy P. (2018). A review on characterization, therapeutic approaches and pathogenesis of \u003cem\u003eMacrophomina phaseolina\u003c/em\u003e. \u003cem\u003ePlant Cell Biotechnol. Mol. Biol.\u003c/em\u003e \u003cem\u003e19\u003c/em\u003e, 72\u0026ndash;84\u003c/li\u003e\n \u003cli\u003eIbrahim, E., Ahmad, A. A., Abdo, E. S., Bakr, M. A., Khalil, M. A., Abdallah, Y., ... \u0026amp; Galal, A. A. (2024). Suppression of root rot fungal diseases in common beans (\u003cem\u003ePhaseolus vulgaris\u003c/em\u003e L.) through the application of biologically synthesized silver nanoparticles. \u003cem\u003eNanomaterials\u003c/em\u003e, \u003cem\u003e14\u003c/em\u003e(8), 710.\u003c/li\u003e\n \u003cli\u003eJabeen, S., Tariq, M., Abid, R., Hanif, A., Shehzad, K., Rehman, M.W.U., Dawar, S., Dias, D.A., Alarjani, K.M. and Abideen, Z., 2024. Application of powdered Medicago sativa L. enhances eco-physiological output and protect against root rot fungi disease in okra and cowpea. Scientia Horticulturae, 337, p.113458.\u003c/li\u003e\n \u003cli\u003eJamiołkowska, A. (2020). Natural compounds as elicitors of plant resistance against diseases and new biocontrol strategies. \u003cem\u003eAgronomy\u003c/em\u003e, \u003cem\u003e10\u003c/em\u003e(2), 173.\u003c/li\u003e\n \u003cli\u003eKlich, M. A. (2002). \u003cem\u003eIdentification of Common Aspergillus Species\u003c/em\u003e. Centraalbureau voor Schimmelcultures (CBS), Utrecht, The Netherlands.\u003c/li\u003e\n \u003cli\u003eLahouar, A., Crespo-Sempere, A., Mar\u0026iacute;n, S., Sa\u0026iuml;d, S., \u0026amp; Sanchis, V. (2015). Toxigenic molds in Tunisian and Egyptian sorghum for human consumption. \u003cem\u003eJournal of Stored Products Research\u003c/em\u003e, \u003cem\u003e63\u003c/em\u003e, 57-62.\u003c/li\u003e\n \u003cli\u003eLeslie, J. F., \u0026amp; Summerell, B. A. (2006). \u003cem\u003eThe Fusarium Laboratory Manual\u003c/em\u003e. Blackwell Publishing.\u003c/li\u003e\n \u003cli\u003eLi, Y., Xue, X., Guo, W., Wang, L., Duan, M., Chen, H., \u0026amp; Chen, F. (2019). Soil moisture and nitrate-nitrogen dynamics and economic yield in the greenhouse cultivation of tomato and cucumber under negative pressure irrigation in the North China Plain. \u003cem\u003eScientific Reports\u003c/em\u003e, \u003cem\u003e9\u003c/em\u003e(1), 4439.\u003c/li\u003e\n \u003cli\u003eMarquez, N., Giachero, M. L., Declerck, S., \u0026amp; Ducasse, D. A. (2021). \u003cem\u003eMacrophomina phaseolina\u003c/em\u003e: General characteristics of pathogenicity and methods of control. \u003cem\u003eFrontiers in Plant Science\u003c/em\u003e, \u003cem\u003e12\u003c/em\u003e, 634397.\u003c/li\u003e\n \u003cli\u003eMayo, S., Gutierrez, S., Malmierca, M. G., Lorenzana, A., Campelo, M. P., Hermosa, R., \u0026amp; Casquero, P. A. (2015). Influence of \u003cem\u003eRhizoctonia solani\u003c/em\u003e and \u003cem\u003eTrichoderma\u003c/em\u003e spp. in growth of bean (\u003cem\u003ePhaseolus vulgaris\u003c/em\u003e L.) and in the induction of plant defense-related genes. \u003cem\u003eFrontiers in plant science\u003c/em\u003e, \u003cem\u003e6\u003c/em\u003e, 685.\u003c/li\u003e\n \u003cli\u003eMej\u0026iacute;a-Garibay, B., Palou, E., \u0026amp; Pez-Malo, A. L. (2015). Composition, diffusion, and antifungal activity of black mustard (\u003cem\u003eBrassica nigra\u003c/em\u003e) essential oil when applied by direct addition or vapor phase contact. \u003cem\u003eJournal of food protection\u003c/em\u003e, \u003cem\u003e78\u003c/em\u003e(4), 843-848.\u003c/li\u003e\n \u003cli\u003eMichiels, J. A., Kevers, C., Pincemail, J., Defraigne, J. O., and Dommes, J. (2012). Extraction conditions can greatly influence antioxidant capacity assays in plant food matrices. \u003cem\u003eFood Chemistry, 130\u003c/em\u003e(4), 986-993.\u003c/li\u003e\n \u003cli\u003eNash, S.M., and Snyder, W.C. (1962). Quantitative estimations by plate counts of propagules of the bean root rot Fusarium in field soils.\u003cem\u003e\u0026nbsp;Phytopathology, 52\u003c/em\u003e: 567- 572.\u003c/li\u003e\n \u003cli\u003ePeng, H., Hu, H., Xi, K., Zhu, X., Zhou, J., Yin, J., ... \u0026amp; Zhu, Y. (2022). Silicon nanoparticles enhance ginger rhizomes tolerance to postharvest deterioration and resistance to \u003cem\u003eFusarium solani\u003c/em\u003e. \u003cem\u003eFrontiers in Plant Science\u003c/em\u003e, \u003cem\u003e13\u003c/em\u003e, 816143.\u003c/li\u003e\n \u003cli\u003ePitt, J. I., \u0026amp; Hocking, A. D. (2009). \u003cem\u003eFungi and Food Spoilage\u003c/em\u003e (3rd ed.). Springer.\u003c/li\u003e\n \u003cli\u003eSchafer, W., \u0026amp; Kiewnick, S. (2010). Morphological characteristics and molecular identification of \u003cem\u003eMacrophomina phaseolina\u003c/em\u003e. \u003cem\u003ePlant Pathology Journal\u003c/em\u003e, \u003cem\u003e26\u003c/em\u003e(3), 223\u0026ndash;229. https://doi.org/10.5423/PPJ.2010.26.3.223\u003c/li\u003e\n \u003cli\u003eSheikh, A.H., and Ghaffar, A. (1975). Population study of sclerotia of \u003cem\u003eMacrophomina phaseolina\u0026nbsp;\u003c/em\u003ein cotton field. \u003cem\u003ePakistan Journal of Botany, 7\u003c/em\u003e: 13-17.\u003c/li\u003e\n \u003cli\u003eShi, H., Li, J., Zhao, Y., Mao, J., Wang, H., \u0026amp; Zhu, J. (2023). Effect of \u003cem\u003eAspergillus flavus\u003c/em\u003e contamination on the fungal community succession, mycotoxin production and storage quality of maize kernels at various temperatures. \u003cem\u003eFood Research International\u003c/em\u003e, \u003cem\u003e174\u003c/em\u003e, 113662.\u003c/li\u003e\n \u003cli\u003eShi, Y., Liu, W., Wang, C., \u0026amp; Li, H. (2023). Traditional uses, phytochemistry, and pharmacological properties of \u003cem\u003eLantana camara\u003c/em\u003e: A review. \u003cem\u003eJournal of Ethnopharmacology\u003c/em\u003e, 301, 115789.\u003c/li\u003e\n \u003cli\u003eSilva, F., and Domingues, F. C. (2017). Antimicrobial activity of coriander oil and its effectiveness as food preservative. Critical reviews in \u003cem\u003efood science and nutrition, 57\u003c/em\u003e (1), 35-47.\u003c/li\u003e\n \u003cli\u003eSingh, B.K., Delgado-Baquerizo, M., Egidi, E., Guirado, E., Leach, J.E., Liu, H. and Trivedi, P., 2023. Climate change impacts on plant pathogens, food security and paths forward. Nature Reviews Microbiology, 21(10), pp.640-656.\u003c/li\u003e\n \u003cli\u003eSun, X.P., Qiu, H.J., Qiu, C.S., Wu, Z.M., Song, X., Tang, L., Liu, D., Cao, Y. and Wang, F., 2025. Research progress on biological prevention and control in tobacco green production. Cogent Food \u0026amp; Agriculture, 11(1), p.2448595.\u003c/li\u003e\n \u003cli\u003eTai, B., Chang, J., Liu, Y., \u0026amp; Xing, F. (2020). Recent progress of the effect of environmental factors on \u003cem\u003eAspergillus flavus\u0026nbsp;\u003c/em\u003egrowth and aflatoxins production on foods. \u003cem\u003eFood Quality and Safety\u003c/em\u003e, \u003cem\u003e4\u003c/em\u003e(1), 21-28.\u003c/li\u003e\n \u003cli\u003eWani, S., Nisa, Q., Fayaz, T., Nabi, N., Nabi, A., Lateef, I., ... \u0026amp; Padder, B. A. (2023). An Overview of Major Bean Diseases and Current Scenario of Common Bean Resistance. \u003cem\u003eDiseases in Legume Crops: Next Generation Breeding Approaches for Resistant Legume Crops\u003c/em\u003e, 99-123.\u003c/li\u003e\n \u003cli\u003eWhite, T. J., Bruns, T., Lee, S., \u0026amp; Taylor, J. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In \u003cem\u003ePCR Protocols: A Guide to Methods and Applications\u003c/em\u003e (pp. 315\u0026ndash;322). Academic Press.\u003c/li\u003e\n \u003cli\u003eWilhelm, S. (1955). Longevity of the \u003cem\u003eVerticillium\u003c/em\u003e wilt fungus in the laboratory and field. \u003cem\u003ePhytopathology, 45\u003c/em\u003e, 180\u0026ndash;181.\u003c/li\u003e\n \u003cli\u003eXi, K. Y., Xiong, S. J., Li, G., Guo, C. Q., Zhou, J., Ma, J. W., ... \u0026amp; Zhu, Y. X. (2022). Antifungal Activity of Ginger Rhizome Extract against \u003cem\u003eFusarium solani\u003c/em\u003e. \u003cem\u003eHorticulturae\u003c/em\u003e, \u003cem\u003e8\u003c/em\u003e(11), 983.\u003c/li\u003e\n \u003cli\u003eYadav, D., Gaurav, H., Yadav, R., Waris, R., Afzal, K., \u0026amp; Shukla, A. C. (2023). A comprehensive review on soft rot disease management in ginger (\u003cem\u003eZingiber officinale\u003c/em\u003e) for enhancing its pharmaceutical and industrial values. \u003cem\u003eHeliyon\u003c/em\u003e.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Root-rotting, Phytopathogens, Zone of Inhibition, Seed treatment, Soil drenching, Soil amendment","lastPublishedDoi":"10.21203/rs.3.rs-7054212/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7054212/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eChemical fungicides impart pollution, toxicity to non-target organisms and health risks. Utilization of plant derivatives hold potential as a valuable source of bioactive for both disease control and the promotion of plant growth. This study investigated the biological activity of \u003cem\u003eZingiber officinale\u003c/em\u003e (ginger) derived plant extract on \u003cem\u003eRicinus communis\u003c/em\u003e L. (castor), \u003cem\u003eBrassica\u003c/em\u003e spp., and \u003cem\u003eLantana camara\u003c/em\u003e L., using different solvents for extraction against major root-roting fungi, \u003cem\u003eMacrophomina phaseolina\u003c/em\u003e, \u003cem\u003eFusarium oxysporum\u003c/em\u003e and \u003cem\u003eAspergillus flavus\u003c/em\u003e, using disc diffusion and agar well diffusion methods. At 100% concentration, both fresh and dried ginger extracts displayed total suppression (100%) of \u003cem\u003eM. phaseolina\u003c/em\u003e, \u003cem\u003eF. oxysporum\u003c/em\u003e, and \u003cem\u003eA. flavus\u003c/em\u003e, with no apparent fungal growth. However, there was a little zone of inhibition (\u0026lt;\u0026thinsp;10%) for both dry, and fresh ginger, against \u003cem\u003eR. solani\u003c/em\u003e. In comparison to the disc approach, the agar well method consistently yielded a great zone of inhibition. On the other hand, fungal overgrowth was noted, and no antifungal activity was demonstrated by ethanol-based extracts of \u003cem\u003eLantana camara, Brassica nigra\u003c/em\u003e, and \u003cem\u003eRicinus communis\u003c/em\u003e. When compared to untreated controls, in \u003cem\u003evivo\u003c/em\u003e tests showed that seeds treated with 100% fresh and dried ginger extracts dramatically increased root rot pathogen colonization by over 80% (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01). A substantial decrease in pathogen colonization was also seen after 100% extracts were soaked into the soil (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05). When ginger-treated seeds were combined with soil amendments, growth indices, including biomass, shoot length, and root length significantly enhanced. For instance, dry ginger\u0026thinsp;+\u0026thinsp;fudan resulted in 38% increase in shoot length, whereas dry ginger\u0026thinsp;+\u0026thinsp;DAP resulted in a 41% rise in root weight (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05). When soil amendment with 0.5 fresh ginger powder, the greatest decrease in fungal colonization (up to 95%) was observed. These results demonstrate the potential of ginger extracts, especially at 100% concentration, as efficient, environmentally friendly treatments for \u003cem\u003ePhaseolus vulgaris\u003c/em\u003e L. root rot illnesses and enhancing plant development. Further research is recommended to optimize the application methods and explore the underlying mechanisms of ginger extract in promoting plant growth and disease suppression.\u003c/p\u003e","manuscriptTitle":"Enhancing growth and suppressing root rot in Phaseolus vulgaris using ginger powder derived natural antifungal agent for optimum disease control and crop health","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-08-04 16:56:33","doi":"10.21203/rs.3.rs-7054212/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"f5081760-f9e6-4e72-8933-87e9493df2cf","owner":[],"postedDate":"August 4th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-10-15T06:08:48+00:00","versionOfRecord":[],"versionCreatedAt":"2025-08-04 16:56:33","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7054212","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7054212","identity":"rs-7054212","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}