Plant Growth-promoting Fungi Research Articles

Plant growth-promoting fungi improve tobacco yield and chemical components by reassembling rhizosphere fungal microbiome and recruiting probiotic taxa

BackgroundTobacco production faces ongoing challenges due to soil degradation, leading to a persistent decline in yield. Plant growth-promoting fungi (PGPF) have been recognized as an environmentally friendly agricultural strategy. However, many commercial PGPF products exhibit instability due to insufficient environmental compatibility.ResultsIn this study, Penicillium sp. PQxj3 was isolated and assessed for its potential to enhance tobacco productivity under field conditions. The results demonstrated that Penicillium sp. PQxj3 treatment significantly promoted the tobacco growth and improved the crop yield. The height of tobacco in Penicillium sp. PQxj3 treatment group significantly increased by 50.19% and 24.05% compared with CK at exuberant and maturity period (P < 0.05). The average yield of tobacco significantly increased by 36.16% compared to CK (P < 0.05). Fungal microbiome analysis revealed that phylogenetically similar probiotic taxa were recruited by Penicillium sp. PQxj3 and reassembled tobacco rhizosphere fungal microbiome. The key chemical indicators of tobacco such as alkaloid, total sugar, and phosphorus were significantly enhanced in Penicillium sp. PQxj3 treatment. The recruited probiotic taxa (Penicillium brasilianum, Penicillium simplicissimum, Penicillium macrosclerotiorum and Penicillium senticosum) were significantly associated with alkaloid, total sugar etc. (P < 0.05), which were identified as the key drivers for improving the chemical components of tobacco. Transcriptome analysis indicated that Penicillium sp. PQxj3 promoted up-regulation of key functional genes involved in alkaloid, indoleacetic, and gibberellin biosynthesis pathways.ConclusionIn summary, this study assessed the biopromotion mechanism of PGPF Penicillium sp. PQxj3 linking chemical traits, rhizosphere fungal microbiome, and transcriptome profiling. The findings provide a fundamental basis and a sustainable solution for developing fungal fertilizers to enhance agricultural sustainability.

Sonic restoration: acoustic stimulation enhances plant growth-promoting fungi activity.

Ecosystem restoration interventions often utilize visible elements to restore an ecosystem (e.g. replanting native plant communities and reintroducing lost species). However, using acoustic stimulation to help restore ecosystems and promote plant growth has received little attention. Our study aimed to assess the effect of acoustic stimulation on the growth rate and sporulation of the plant growth-promoting fungus Trichoderma harzianum Rifai, 1969. We played a monotone acoustic stimulus (80 dB sound pressure level (SPL) at a peak frequency of 8 kHz and a bandwidth at -10 dB from the peak of 6819 Hz-parameters determined via review and pilot research) over 5 days to T. harzianum to assess whether acoustic stimulation affected the growth rate and sporulation of this fungus (control samples received only ambient sound stimulation less than 30 dB). We show that the acoustic stimulation treatments resulted in increased fungal biomass and enhanced T. harzianum conidia (spore) activity compared to controls. These results indicate that acoustic stimulation influences plant growth-promoting fungal growth and potentially facilitates their functioning (e.g. stimulating sporulation). The mechanism responsible for this phenomenon may be fungal mechanoreceptor stimulation and/or potentially a piezoelectric effect; however, further research is required to confirm this hypothesis. Our novel study highlights the potential of acoustic stimulation to alter important fungal attributes, which could, with further development, be harnessed to aid ecosystem restoration and sustainable agriculture.

Microbial Fertilizers: A Study on the Current Scenario of Brazilian Inoculants and Future Perspectives.

The increasing need for sustainable agricultural practices, combined with the demand for enhanced crop productivity, has led to a growing interest in utilizing microorganisms for biocontrol of diseases and pests, as well as for growth promotion. In Brazilian agriculture, the use of plant growth-promoting rhizobacteria (PGPR) and plant growth-promoting fungi (PGPF) has become increasingly prevalent, with a corresponding rise in the number of registered microbial inoculants each year. PGPR and PGPF occupy diverse niches within the rhizosphere, playing a crucial role in soil nutrient cycling and influencing a wide range of plant physiological processes. This review examines the primary mechanisms employed by these microbial agents to promote growth, as well as the strategy of co-inoculation to enhance product efficacy. Furthermore, we provide a comprehensive analysis of the microbial inoculants currently available in Brazil, detailing the microorganisms accessible for major crops, and discuss the market's prospects for the research and development of novel products in light of current challenges faced in the coming years.

Draft genome sequence of Penicillium citrinum B9, a plant growth-promoting symbiont from barley rhizosphere.

Penicillium citrinum strain B9 is a plant growth-promoting fungus isolated from Barley (Hordeum vulgare) rhizosphere. We report the first draft genome of P. citrinum B9 assembled using single-molecule real-time sequencing and Illumina reads. The assembled genome spans 31.3 Mb comprising nine contigs and 10,106 protein-encoding genes.

Purpureocillum lilacinum as an Agent of Nematode Control and Plant Growth-Promoting Fungi

Plants support numerous microorganisms within their tissues and the rhizosphere, and these microorganisms, known as the microbiota, can influence plant growth and health. Up to 40% of a plant’s photosynthetic metabolism may be invested in the rhizosphere. The microbiota are considered an extra genome that can be modulated to meet plant needs. Researchers have identified a set of genes from these microorganisms, known as the microbiome, which can be manipulated to enhance plant growth and health, improve nutrient absorption, reduce the need for chemical fertilizers, increase resistance to pathogens and pests, and increase stress tolerance. In particular, fungi exhibit large genetic and metabolic diversity and are often used to promote plant growth. For example, the fungus Purpureocillum lilacinum has been employed primarily as a biocontrol agent to manage nematodes, but some studies have suggested that it may also promote plant growth by increasing the efficiency of the plant in absorbing nutrients from the soil and providing phytohormones to plants. Therefore, the current review aims to summarize the existing literature on the use of this fungus in agriculture as nematodes control, and discuss its potential as a plant growth-promoter.

Trichoderma spp. isolates stimulate rice seedling growth of Sertani 13 variety

Trichoderma has become one of the most studied filamentous fungi to be used as a greener and more sustainable solution for improving the production and growth of numerous crops, due to its capability to form symbiotic associations with plants. This study aimed to evaluate the effect of Trichoderma isolates obtained from the rhizosphere of organic rice fields in Sukabumi, Indonesia, in enhancing rice germination and seedling growth. A laboratory experiment used a completely randomized design consisting of seed treatments of 21 Trichoderma isolates (T1-T21) and a control treatment without Trichoderma (C). The inoculation was employed to elucidate any potential effects of Trichoderma isolates. Results showed that five isolates, i.e., T5, T7, T9, T10, and T14 stimulated the highest seedling vigor index, root and shoot length, and fresh weight and dry weight. These findings exhibited the potential of these five isolates as plant growth-promoting fungi to improve rice seedling growth and contribute to our understanding of the role of symbiotic fungi in sustainable rice crop production. Keywords: plant growth promoting fungi; seed treatment; plant-microbe interactions, symbiotic fungi

Isolation and Molecular Profiling of Halotolerant Plant Growth Promoting Rhizosphere Fungi from Salt affected Agroforestry Plantation

Soil salinity is a major abiotic stress that adversely affects plant growth, and productivity. About 20% of irrigated lands are affected by salinity worldwide; In India, there are 6.74 million hectares of salt-affected lands. Salt-tolerant Plant Growth Promoting (PGP) microorganisms can enhance the growth of plants in such salt-stressed areas. Therefore, this study aimed to investigate the diversity of beneficial fungal communities, screen for their ability to support plant growth, and evaluate the production of various essential compounds in view of plant growth in salt-stressed lands. A total of 68 fungal colonies were isolated from 5 different agroforestry plantation sites in Karur, Tamil Nadu, South India at quarterly intervals. The isolates were screened for sodium chloride (NaCl) tolerance (0%, 5%, 10%, 15%, and 20% concentration). A total of 7 isolates showed considerable salt tolerance and were tested qualitatively in-vitro, for PGP traits such as phosphate, potassium, and zinc solubilization, nitrogen fixation, hydrogen cyanide production, siderophore production, and ACC deaminase production. Finally, 5 isolates with maximum values for PGP properties under 20% NaCl concentration were tested for the quantity of Indole-3-Acetic Acid (IAA) and Exo-polysaccharide (EPS) production. All 5 isolates were identified up to the species level using 18S rRNA gene sequencing. To the best of our knowledge, this is the first report on isolating saline-tolerant PGP Fungi (PGPF) from the rhizosphere region of Casuarina equisetifolia and Eucalyptus camaldulensis in Karur, Tamil Nadu, India. In the future, the bioformulation of PGPF and its application will boost the cultivation of tree saplings in this salt affected regions.

Trichoderma guizhouense NJAU4742 augments morphophysiological responses, nutrient availability and photosynthetic efficacy of ornamental Ilex verticillata.

Winterberry holly (Ilex verticillata [L.] A. Gray), a deciduous shrub producing glossy bright red berries, is a valuable ornamental and medicinal plant with good market prospects. However, the growth and development of I. verticillata are significantly affected by various stresses, and environmentally hazardous agrochemicals are often used to mitigate them. Trichoderma spp., ubiquitous soil-borne eco-friendly plant growth-promoting fungi, are potent biostimulants and biofertilizers and viable alternatives to agrochemicals for healthy and sustainable agriculture. In this study, the temporal efficacy of different dosages of the filamentous fungus Trichoderma guizhouense NJAU4742 in promoting morphophysiological responses of I. verticillata and the physicochemical properties and enzymatic activities of the substrate were investigated. Different concentrations of the strain T. guizhouense NJAU4742 spore suspension (C [0%], T1 [5%, v/m], T2 [10%, v/m] and T3 [15%, v/m]) were injected in the substrate contained in a pot in which 1-year-old I. verticillata was planted for temporal treatment (15, 45 and 75 days) under open-air conditions. The beneficial effects of T2 and/or T3 treatment for a long duration (75 days) were evident on the different root, aerial and photosynthetic traits; total contents of nitrogen (N), phosphorus (P) and potassium (K) in different tissues and the physicochemical properties of the substrate and its enzymatic activities (urease and invertase). Overall, the study revealed the potency of strain T. guizhouense NJAU4742 as a sustainable solution to improve the growth and development and ornamental value of I. verticillata.

The Effects of Salinity and Genotype on the Rhizospheric Mycobiomes in Date Palm Seedlings.

Salinity severely affects the health and productivity of plants, with root-associated microbes, including fungi, potentially playing a crucial role in mitigating this effect and promoting plant health. This study employed metagenomics to investigate differences in the structures of the epiphyte mycobiomes in the rhizospheres of seedlings of two distinct date palm cultivars with contrasting salinity tolerances, the susceptible cultivar, 'Zabad', and the tolerant cultivar, 'Umsila'. Next-generation sequencing (NGS) of the internal transcribed spacer (ITS) rRNA was utilized as a DNA barcoding tool. The sequencing of 12 mycobiome libraries yielded 905,198 raw sequences of 268,829 high-quality reads that coded for 135 unique and annotatable operational taxonomic units (OTUs). An OTU analysis revealed differences in the rhizofungal community structures between the treatments regardless of genotype, and non-metric dimensional scaling (N-MDS) analyses demonstrated distinct separations between the cultivars under saline stress. However, these differences were not detected under the control environmental conditions, i.e., no salinity. The rhizospheric fungal community included four phyla (Ascomycota, Basidiomycota, Chytridiomycota, and Mucoromycota), with differences in the abundances of Aspergillus, Clonostachys, and Fusarium genera in response to salinity, regardless of the genotype. Differential pairwise comparisons showed that Fusarium falciforme-solani and Aspergillus sydowii-versicolor increased in abundance under saline conditions, providing potential future in vitro isolation guidelines for plant growth-promoting fungi. This study highlights the intricate dynamics of the rhizosphere microbial communities in date palms and their responses to salt stress. Additionally, we found no support for the hypothesis that indigenous epiphytic fungal communities are significantly involved in salinity tolerance in date palms.

Fungi beyond limits: The agricultural promise of extremophiles.

Global climate changes threaten food security, necessitating urgent measures to enhance agricultural productivity and expand it into areas less for agronomy. This challenge is crucial in achieving Sustainable Development Goal 2 (Zero Hunger). Plant growth-promoting microorganisms (PGPM), bacteria and fungi, emerge as a promising solution to mitigate the impact of climate extremes on agriculture. The concept of the plant holobiont, encompassing the plant host and its symbiotic microbiota, underscores the intricate relationships with a diverse microbial community. PGPM, residing in the rhizosphere, phyllosphere, and endosphere, play vital roles in nutrient solubilization, nitrogen fixation, and biocontrol of pathogens. Novel ecological functions, including epigenetic modifications and suppression of virulence genes, extend our understanding of PGPM strategies. The diverse roles of PGPM as biofertilizers, biocontrollers, biomodulators, and more contribute to sustainable agriculture and environmental resilience. Despite fungi's remarkable plant growth-promoting functions, their potential is often overshadowed compared to bacteria. Arbuscular mycorrhizal fungi (AMF) form a mutualistic symbiosis with many terrestrial plants, enhancing plant nutrition, growth, and stress resistance. Other fungi, including filamentous, yeasts, and polymorphic, from endophytic, to saprophytic, offer unique attributes such as ubiquity, morphology, and endurance in harsh environments, positioning them as exceptional plant growth-promoting fungi (PGPF). Crops frequently face abiotic stresses like salinity, drought, high UV doses and extreme temperatures. Some extremotolerant fungi, including strains from genera like Trichoderma, Penicillium, Fusarium, and others, have been studied for their beneficial interactions with plants. Presented examples of their capabilities in alleviating salinity, drought, and other stresses underscore their potential applications in agriculture. In this context, extremotolerant and extremophilic fungi populating extreme natural environments are muchless investigated. They represent both new challenges and opportunities. As the global climate evolves, understanding and harnessing the intricate mechanisms of fungal-plant interactions, especially in extreme environments, is paramount for developing effective and safe plant probiotics and using fungi as biocontrollers against phytopathogens. Thorough assessments, comprehensive methodologies, and a cautious approach are crucial for leveraging the benefits of extremophilic fungi in the changing landscape of global agriculture, ensuring food security in the face of climate challenges.

Papiliotrema flavescens, a plant growth-promoting fungus, alters root system architecture and induces systemic resistance through its volatile organic compounds in Arabidopsis

The current trend in agricultural development is the establishment of sustainable agricultural systems. This involves utilizing and implementing eco-friendly biofertilizers and biocontrol agents as alternatives to conventional fertilizers and pesticides. A plant growth-promoting fungal strain, that could alter root system architecture and promote the growth of Arabidopsis seedlings in a non-contact manner by releasing volatile organic compounds (VOCs) was isolated in this study. 26S rDNA sequencing revealed that the strain was a yeast-like fungus, Papiliotrema flavescens. Analysis of plant growth-promoting traits revealed that the fungus could produce indole-3-acetic acid and ammonia and fix nitrogen. Transcriptome analysis in combination with inhibitor experiments revealed that P. flavescens VOCs triggered metabolic alterations, promoted auxin accumulation and distribution in the roots, and coordinated ethylene signaling, thus inhibiting primary root elongation and inducing lateral root formation in Arabidopsis. Additionally, transcriptome analysis and fungal infection experiments confirmed that pretreatment with P. flavescens stimulated the defense response of Arabidopsis to boost its resistance to the pathogenic fungus Botrytis cinerea. Solid-phase microextraction, which was followed by gas chromatography-mass spectrometry analysis, identified three VOCs (acetoin, naphthalene and indole) with significant plant growth-promoting attributes. Their roles were confirmed using further pharmacological experiments and upregulated expression of auxin- and ethylene-related genes. Our study serves as an essential reference for utilizing P. flavescens as a potential biological fertilizer and biocontrol agent.

Bioprospecting the roles of Trichoderma in alleviating plants’ drought tolerance: Principles, mechanisms of action, and prospects

Drought-induced stress represents a significant challenge to agricultural production, exerting adverse effects on both plant growth and overall productivity. Therefore, the exploration of innovative long-term approaches for addressing drought stress within agriculture constitutes a crucial objective, given its vital role in enhancing food security. This article explores the potential use of Trichoderma, a well-known genus of plant growth-promoting fungi, to enhance plant tolerance to drought stress. Trichoderma species have shown remarkable potential for enhancing plant growth, inducing systemic resistance, and ameliorating the adverse impacts of drought stress on plants through the modulation of morphological, physiological, biochemical, and molecular characteristics. In conclusion, the exploitation of Trichoderma's potential as a sustainable solution to enhance plant drought tolerance is a promising avenue for addressing the challenges posed by the changing climate. The manifold advantages of Trichoderma in promoting plant growth and alleviating the effects of drought stress underscore their pivotal role in fostering sustainable agricultural practices and enhancing food security.

Plant growth-promoting fungi and rhizobacteria control Fusarium damping-off in Mason pine seedlings by impacting rhizosphere microbes and altering plant physiological pathways

Plant growth-promoting fungi and rhizobacteria control Fusarium damping-off in Mason pine seedlings by impacting rhizosphere microbes and altering plant physiological pathways

Enhanced use of chemical fertilizers and mitigation of heavy metal toxicity using biochar and the soil fungus Bipolaris maydis AF7 in rice: Genomic and metabolomic perspectives

Chemical fertilizers are the primary source of crop nutrition; however, their increasing rate of application has created environmental hazards, such as heavy metal toxicity and eutrophication. The synchronized use of chemical fertilizers and eco-friendly biological tools, such as microorganisms and biochar, may provide an efficient foundation to promote sustainable agriculture. Therefore, the current study aimed to optimize the nutrient uptake using an inorganic fertilizer, sulfate of potash (SOP) from the plant growth-promoting fungus Bipolaris maydis AF7, and biochar under heavy metal toxicity conditions in rice. Bioassay analysis showed that AF7 has high resistance to heavy metals and a tendency to produce gibberellin, colonize the fertilizer, and increase the intake of free amino acids. In the plant experiment, the co-application of AF7 +Biochar+MNF+SOP significantly lowered the heavy metal toxicity, enhanced the nutrient uptake in the rice shoots, and improved the morphological attributes (total biomass). Moreover, the co-application augmented the glucose and sucrose levels, whereas it significantly lowered the endogenous phytohormone levels (salicylic acid and jasmonic acid) in the rice shoots. The increase in nutrient content aligns with the higher expression of the OsLSi6, PHT1, and OsHKT1 genes. The plant growth traits and heavy metal tolerance of AF7 were validated by whole-genome sequencing that showed the presence of the heavy metal tolerance and detoxification protein, siderophore iron transporter, Gibberellin cluster GA4 desaturase, and DES_1 genes, as well as others that regulate glucose, antioxidants, and amino acids. Because the AF7 +biochar+inorganic fertilizer works synergistically, nutrient availability to the crops could be improved, and heavy metal toxicity and environmental hazards could be minimized.

Isolation and Inoculation Effect of Trichoderma reesei on Growth and Yield of Barley

In today’s world, usage of chemical fertilizers has become necessity for betterment of crop yield; however they have negative impact on the environment, quality of soil and human health. Therefore, involvement of substitute for chemical fertilizers is an essential requirement of present time. Plant growth promoting fungi (PGPF) serves as a best alternative in place of chemicals to enhance plant growth, crop productivity and improve nutrients availability for plants. They also show an involvement in ceasing growth of plant pathogens, hence acting as a bio control agent. The aim of our study is to screen and examine plant growth promoting fungi effect on barley crop. A total seven fungal strains were isolated from wheat rhizosphere. The isolated fungal strains were screened for their In vitro plant growth promotional traits. Among total, one isolate 14F found positive for five different plant growth promoting traits. This isolate was further identified at molecular level by amplification and sequencing of ITS gene region and was identified as Trichoderma reesei. Trichoderma reesei was inoculated with barley seeds and its effects were analyzed. Significant increase was observed in terms of plant height (root length and shoot length), plant weight (Dry and wet weight) as compared to un-inoculated plants. This Trichoderma strain could become a fantastic bio-fertilizer for sustainable agriculture.

Synergistic Effect of Trichoderma reesei and Fusarium solani on growth and yield of Barley plant

Chemical fertilizers are known to improve plant yield but on the cost of environment, soil health as well as human health. Thus, alternatives of chemical fertilizers are the demand of time. Plant growth-promoting fungi (PGPF) are using nowadays as an alternative of chemical fertilizers to improve the growth, yield, and plant nutrient uptake. They are also involved in the improvement of rhizosphere fertility, without compromising environment and human health. A total of seven fungal isolates were obtained from rhizospheric soil of two crop plants viz., turnip and wheat. Fungal strains were examined for their In vitro plant growth promotional traits. A total of two strains found positive for 4 or more plant growth promoting traits. These potential fungal strains were characterized by amplification and sequencing of ITS region and were identified as Trichoderma reesei, and another was identified as Fusarium solani. Our current study is aimed to screen the effect of single inoculation as well as paired fungal inoculations and to provide evidence for the synergistic effects of paired isolates having high impact on growth and development of barley plant.

Plant growth-promoting fungi from desert soil

Plant growth-promoting fungi from desert soil

Rhamnolipids supplement in salinized soils improves cotton growth through ameliorating soil properties and modifying rhizosphere communities

Soil salinization is a global environmental concern due to its great restriction on land use efficiencies and crop production. Rhamnolipids demonstrate great potential in ameliorating soils and improving plant growth. However, the exact performance and underlying mechanisms still remain unclear. Herein, a field-scale study was conducted to investigate the effect of rhamnolipids addition on cotton growth in soils with different degrees of salinization (i.e., slightly, moderately, and highly salinized soils). The drip-irrigated rhamnolipids solution (300 mg/L) effectively reduced the salinity in rhizosphere soils, with desalination rates of 9.7 %, 4.5 %, and 2.5 % in slightly, moderately, and highly salinized soils, respectively, and mitigated the salt stress on cotton plants. The analysis of microbial community in highly salinized soils showed that the rhizosphere community was modified by the rhamnolipids, with enriched populations of plant growth-promoting fungi and decreased abundance of plant pathogens. Further investigation of functional genes related to carbon, nitrogen, sulfur, and phosphorus cycling suggested that the rhamnolipids efficiently drove the nutrient cycling and promoted the interconnection of functional microorganisms. The ameliorated soil environment and strengthened microecological functions led to the enhanced photosynthetic process and improved cotton growth and yield (up to 24.4 %). Our study is the first to demonstrate the feasibility of applying rhamnolipids to improve cotton growth in salinized areas and disclose the underlying mechanisms.

Biocontrol of Fusarium wilt disease in pepper plant by plant growth promoting Penicillium expansum and Trichoderma harzianum

Plant growth promoting fungi (PGPF) were employed in the present study to biocontrol Fusarium wilt disease in pepper plants. Two of the five fungal isolates were chosen based on biochemical characteristics such as their production of hydrocyanic acid, siderophores, and IAA, phosphate solubilization, and in vitro antifungal activities. The most potent fungal isolates were identified as Penicillium expansum (P. expansum) and Trichoderma harzianum (T. harzianum). Using GC-MS, it was found that PGPF extracts contain compounds with antifungal activity, antioxidants, and plant growth stimulators. The combined effect of T. harzianum and P. expansum increased the protection against fusarial wilt by (76.74%), followed by T. harzianum by (50%), then P. expansum by (17.64%). Significant improvement because of using the mixture (T. harzianum and P. expansum) showed an increase in shoot length (59.4%), root length (129%), and number of leaves (52.6%). Chlorophyll A and B levels in infected plants were consistently raised by 28.71% and 67.58%, respectively; as a result of application the mixture (T. harzianum and P. expansum). Also, there was an increase in soluble proteins and carbohydrates in infected plants treated with (T. harzianum) by 25.42% and 31.78% over untreated infected plants, respectively. It could be recommended that the use of targeted PGPF strains, especially a mixture of T. harzianum and P. expansum could be commercially used as therapeutic nutrients against Fusarium wilt of pepper plants.

Application of Rhizopus microsporus and Aspergillus oryzae to enhance the defense capacity of eggplant seedlings against Meloidogyne incognita

Several phytopathogens attack eggplant, causing crop damage. One of the most destructive plant diseases, Root-Knot Nematode (RKN), causes significant damage to eggplant seedlings. Finding safe and effective biological alternatives to prevent eggplant root nematode disease, which significantly limits plant productivity, is the innovative aspect of this study. Six isolates of plant growth-promoting fungus (PGPF) were tested in the current work for improving biochemical defense and physio-biochemical performance in eggplant seedlings under the Meloidogyne incognita challenge. PGPF isolates were tested in vitro for some biochemical traits such as Siderophores and HCN production. Besides, the antagonistic efficacy of PGPF filtrates against M. incognita was tested in vitro. The best isolates capable of producing HCN were F5 and F3 respectively. Also, F5 followed by F3 exhibited the maximum mortality proportions of 74.20% and 60.35% mortality in nematode juveniles after 96 hours respectively. Moreover, F5 has the highest level of antioxidant activity, with IC50 145 µg/mL followed by F3 with IC50 350 µg/mL. thus, we identified F5 and F3 completely as Rhizopus microsporus (OQ291571.1 and Aspergillus oryzae OQ291572.1. Implementing R. microsporus and A. oryzae collectively in vivo study was the most successful therapy, limiting nematode recordings as 95.23%, 86.98%, 80.35%, 80%, and 68.78% reduction in females, galls, developmental stage, egg masses, second juveniles, respectively, in diseased seedlings. It could be suggested that the use of ethyl acetate extracts (EAE) of A. oryzae and R. microsporus might be commercially applied as a stimulator of eggplant and or anti-nematodes against M. incognita.