Bacterial Endophytes Research Articles

Enriching the endophytic bacterial microbiota of Ginkgo roots.

Bacterial endophytes of Ginkgo roots take part in the secondary metabolic processes of the fossil tree and contribute to plant growth, nutrient uptake, and systemic resistance. However, the diversity of bacterial endophytes in Ginkgo roots is highly underestimated due to the lack of successful isolates and enrichment collections. The resulting culture collection contains 455 unique bacterial isolates representing 8 classes, 20 orders, 42 families, and 67 genera from five phyla: Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria, and Deinococcus-Thermus, using simply modified media (a mixed medium without any additional carbon sources [MM)] and two other mixed media with separately added starch [GM] and supplemented glucose [MSM]). A series of plant growth-promoting endophytes had multiple representatives within the culture collection. Moreover, we investigated the impact of refilling carbon sources on enrichment outcomes. Approximately 77% of the natural community of root-associated endophytes were predicted to have successfully cultivated the possibility based on a comparison of the 16S rRNA gene sequences between the enrichment collections and the Ginkgo root endophyte community. The rare or recalcitrant taxa in the root endosphere were mainly associated with Actinobacteria, Alphaproteobacteria, Blastocatellia, and Ktedonobacteria. By contrast, more operational taxonomic units (OTUs) (0.6% in the root endosphere) became significantly enriched in MM than in GM and MSM. We further found that the bacterial taxa of the root endosphere had strong metabolisms with the representative of aerobic chemoheterotrophy, while the functions of the enrichment collections were represented by the sulfur metabolism. In addition, the co-occurrence network analysis suggested that the substrate supplement could significantly impact bacterial interactions within the enrichment collections. Our results support the fact that it is better to use the enrichment to assess the cultivable potential and the interspecies interaction as well as to increase the detection/isolation of certain bacterial taxa. Taken together, this study will deepen our knowledge of the indoor endophytic culture and provide important insights into the substrate-driven enrichment.

Impacts of Salt Stress on the Rhizosphere and Endophytic Bacterial Role in Plant Salt Alleviation

Salinity stress is among the key challenges for sustainable food production. It is continuously increasing against the backdrop of constant climate change and anthropogenic practices leading to a huge drop in soil, water, and cultivated crop quality and productivity. Halotolerant plants represent hot spots for endophytic bacteria which may have mechanisms to overcome salt stress. This research initiative aims to highlight the possible exploitation of bacterial endophytes as a microbial biotechnology tool in the productive success of plants exposed to saline stress. We started by solely studying the mechanisms of stress tolerance by plants and halotolerant bacteria. After that, we focused on the beneficial mechanisms of endophytic bacteria in salt stress mitigation. On one side, potent bacterium works by promoting plant performances by facilitating the plant’s nutrient uptake (P, K, Zn, N, and Fe) and by promoting the production of growth hormones (IAA and CKs). On the other side, they balance stress phytohormones (ABA, JA, GA, and ACC) produced by plants in case of soil salt augmentation. The selected potent endophytic bacteria could be exploited and applied to ameliorate the production and salt tolerance of food crops. Lastly, we elucidated deeper advanced technologies including (i) genomics unveiling the plant’s culture-dependent and culture-independent microbiomes, (ii) metabolomics focusing on genes’ metabolic pathways to discover novel secondary metabolites, (iii) transcriptomics studying gene expression, and (iv) proteomics delimiting proteins expressed in stress alleviation. These technologies have been used to understand the plant–bacterial mechanisms of interaction to combat salt stress.

In vivo and In vitro evaluation of the antifungal activity of the PGPR Bacillus amyloliquefaciens RaSh1 (MZ945930) against Alternaria alternata with growth promotion influences on Capsicum annuum L. plants

Alternaria alternata that threatens pepper production and causes major economic harm is responsible for the leaf spot/blight disease. Chemical fungicides have been widely employed; unfortunately, fungicidal resistance is a current concern. Therefore, finding new environmentally friendly biocontrol agents is a future challenge. One of these friendly solutions is the use of bacterial endophytes that have been identified as a source of bioactive compounds. The current study investigates the in vivo and in vitro fungicidal potential of Bacillus amyloliquefaciens RaSh1 (MZ945930) against pathogenic A. alternata. In vitro, the results revealed that RaSh1 exhibited strong antagonistic activity against A. alternata. In addition to this, we inoculated pepper (Capsicum annuum L.) plants with B. amyloliquefaciens RaSh1 and infected them with A. alternata. As a result of A. alternata infection, which generated the highest leaf spot disease incidence (DI), the plant's growth indices and physio-biochemical characteristics significantly decreased, according to our findings. Our results also showed the abnormal and deformed cell structure using light and electron microscopy of A. alternata-infected leaves compared with other treatments. However, DI was greatly reduced with B. amyloliquefaciens RaSh1 application (40%) compared to pepper plants infected with A. alternata (80%), and this led to the largest increases in all identified physio-biochemical parameters, including the activity of the defense-related enzymes. Moreover, inoculation of pepper plants with B. amyloliquefaciens RaSh1 decreased electrolyte leakage by 19.53% and MDA content by 38.60% as compared to A. alternata infected ones. Our results show that the endophyte B. amyloliquefaciens RaSh1 has excellent potential as a biocontrol agent and positively affects pepper plant growth.

Influence of bacterial endophytes on plant growth and fruit yield of tomato (Solanum lycopersicum L.)

Influence of bacterial endophytes on plant growth and fruit yield of tomato (Solanum lycopersicum L.)

Finger Millet (Eleusine coracana) Plant-Endophyte Dynamics: Plant Growth, Nutrient Uptake, and Zinc Biofortification.

Endophytic fungi and bacteria were isolated from finger millet and their effects on finger millet growth parameters and zinc and NPK contents in grains were studied. Out of 70 fungal and 112 bacterial endophytes, the two best fungal and bacterial isolates were selected on the basis of zinc solubilization and plant-growth-promoting attributes. The fungal isolates identified were Aspergillus terreus and Lecanicillium sp., and the bacterial isolates were Pseudomonas bijieensis and Priestia megaterium. The endophytic zinc, NPK mobilization, and plant-growth-promoting efficacy were determined in a pot experiment with zinc carbonate as the zinc source. Endophytic-primed plants showed enhanced shoot and root lengths compared to the unprimed control. Endophytes increased the zinc content in grains by between 12.12% and 18.80% compared to control plants. Endophytes also augmented the NPK concentrations in seeds compared to control plants and exhibited stability in a diverse range of pHs, temperatures, and NaCl concentrations, and exhibited growth on various carbohydrate and nitrogen sources. This is the first study reporting the interaction of Aspergillus terreus, Lecanicillium sp., Pseudomonas bijieensis, and Priestia megaterium with finger millet for grain Zn biofortification and NPK concentration enhancement. This study indicated that zinc-dissolving endophytes possess the potential for enhancing the zinc and NPK content in grains in addition to the plant-growth-promoting attributes.

Therapeutic compounds from medicinal plant endophytes: molecular and metabolic adaptations

During the last few decades, endophytes have attracted increased attention due to their ability to produce a plethora of bioactive secondary metabolites. These compounds not only help the endophytes to outcompete other plant-associated microbes or pathogens through quorum sensing, but also enable them to surmount the plant immune system. However, only a very few studies have described the interlink between various biochemical and molecular factors of host-microbe interactions involved in the production of these pharmacological metabolites. The peculiar mechanisms by which endophytes modulate plant physiology and metabolism through elicitors, as well as how they use transitional compounds of primary and secondary metabolism as nutrients and precursors for the synthesis of new compounds or enhancing existing metabolites, are still less understood. This study thus attempts to address the aspects of synthesis of such metabolites used in therapeutics by the endophytes in the light of their ecological significance, adaptation, and intercommunity interactions. Our study explores how endophytes adapt to the specific host environment, especially in medicinal plants that produce metabolites with pharmacological potential and simultaneously modulate host gene expression for the biosynthesis of these metabolites. We also discuss the differential interactions of fungal and bacterial endophytes with their hosts.

Endophytic Bacterium Lysobacter firmicutimachus Strain 5-7 Is a Promising Biocontrol Agent Against Rice Seedling Disease Caused by Pythium arrhenomanes in Nursery Trays.

Rice root rot disease caused by Pythium spp. is a highly destructive disease in rice nurseries. Biocontrol with endophytic bacteria was developed in this study to control rice seedling diseases. An in planta screening assay revealed that two bacterial endophytes, strains 5-7 and 6-4, displayed strong protection of rice seedlings from attack by Pythium arrhenomanes. Phylogenetic analysis indicated that strain 5-7 is Lysobacter firmicutimachus, while strain 6-4 belongs to the Kitasatospora genus. To quickly evaluate the disease severity of the root system damaged by Pythium spp. in nursery trays, a root surface area measurement assay was developed. By using this measurement, the control efficacy in nursery trays was evaluated, and L. firmicutimachus 5-7 showed promising biocontrol activity against Pythium disease. In a field trial, the two endophytes exhibited significant disease control efficacy on rice brown spot disease caused by Bipolaris oryzae naturally occurring in a commercial nursery field. The two endophytes exhibited multiple enzymatic activities and broad-spectrum antagonistic activities against multiple rice pathogens. The two endophytes colonized the root surface and inside of the root. L. firmicutimachus 5-7 primarily colonized the intercellular space and aerenchyma. Antibiosis is the major mechanism used by strain 5-7 to cause Bipolaris hyphal swelling and inhibit Pythium zoospore germination and sporangium formation, while a hyperparasitism-like phenomenon was found in the interaction of strain 6-4 with Pythium and Bipolaris hyphae. In conclusion, we report the promising biocontrol agent L. firmicutimachus 5-7 and the potential biocontrol agent Kitasatospora sp. 6-4 for disease control of rice seedlings in commercial nursery trays and their possible mechanisms of action.

Quantitative trait loci associated with apple endophytes during pathogen infection.

The plant phyllosphere is colonized by microbial communities that can influence the fitness and growth of their host, including the host's resilience to plant pathogens.There are multiple factors involved in shaping the assemblages of bacterial and fungal endophytes within the phyllosphere, including host genetics and environment. In this work, the role of host genetics in plant-microbiome assembly was studied in a full-sibling family of apple (Malus x domestica) trees infected with the fungal pathogen Neonectria ditissima. A Quantitative Trait Loci (QTL) analysis showed that there are multiple loci which influence the abundance of individual endophytic taxa, with the majority of QTL having a moderate to large effect (20-40%) on endophyte abundance. QTL regions on LG 1, 3, 4, 5, 10, 12, 13, 14 and 15 were shown to affect multiple taxa. Only a small proportion of the variation in overall taxonomic composition was affected by host genotype, with significant QTL hits for principal components explaining <8% and <7.4% of the total variance in bacterial and fungal composition, respectively. Four of the identified QTL colocalised with previously identified regions associated with tolerance to Neonectria ditissima. These results suggest that there is a genetic basis shaping apple endophyte composition and that microbe-host associations in apple could be tailored through breeding.

Drivers of bacterial and fungal root endophyte communities: understanding the relative influence of host plant, environment, and space

Bacterial and fungal root endophytes can impact the fitness of their host plants, but the relative importance of drivers for root endophyte communities is not well known. Host plant species, the composition and density of the surrounding plants, space, and abiotic drivers could significantly affect bacterial and fungal root endophyte communities. We investigated their influence in endophyte communities of alpine plants across a harsh high mountain landscape using high-throughput sequencing. There was less compositional overlap between fungal than bacterial root endophyte communities, with four 'cosmopolitan' bacterial OTUs found in every root sampled, but no fungal OTUs found across all samples. We found that host plant species, which included nine species from three families, explained the greatest variation in root endophyte composition for both bacterial and fungal communities. We detected similar levels of variation explained by plant neighborhood, space, and abiotic drivers on both communities, but the plant neighborhood explained less variation in fungal endophytes than expected. Overall, these findings suggest a more cosmopolitan distribution of bacterial OTUs compared to fungal OTUs, a structuring role of the plant host species for both communities, and largely similar effects of the plant neighborhood, abiotic drivers, and space on both communities.

Fungal and bacterial endophytes antagonistic to Phytophthora species causing root rot in Cinnamomum cassia

Endophytes were investigated for their potential to protect Cinnamomum cassia (cinnamon) from Phytophthora root rot. From healthy cinnamon trees in diseased plantations, 134 bacterial and fungal endophytes were isolated and screened in vitro for their ability to antagonise the root rot pathogens Phytophthora cinnamomi, P. multibullata and P. × vanyenensis. Seventeen endophytes exhibiting high levels of antagonism in vitro (more than 45%) were then tested in a glasshouse study for their ability to reduce the impact of Phytophthora infection in cinnamon seedlings. Trials using cinnamon seeds or seedlings inoculated with an endophyte and then infested with Phytophthora identified three endophytes expressing high levels of disease suppression (based on root damage reduction) and plant growth promotion. These were Penicillium citrinum, Xylaria curta and Clonostachys rosea. These endophytes can potentially be used in the biological control of root rot in cinnamon, but this must be explored further under field conditions.

Purification and characterization of lipase enzyme from endophytic Bacillus pumilus WSS5 for application in detergent industry

Lipolytic enzymes from endophytic microbes have been the focus of intense and growing research in view of the applicability of such enzymes in various industrial applications and endophytic microorganisms may emerge as a potential source of lipases with distinct characteristics. The current work involved purification and characterization of the lipase from an endophytic-bacteria isolated from the stem of Withania somnifera L. Dunal. Based on the analysis of morphology and 16S rDNA sequence, the endophytic bacteria was identified as Bacillus pumilus and designated as B. pumilus WSS5. Purified lipase from the B. pumilus WSS5 was found to be of 27KDa as revealed by SDS-PAGE with optimum activity at 37 °C and pH 8. The stability of the enzyme in presence of various metal ions, surfactants, inhibitors, and organic solvents suggested that this enzyme could be exploited in leather and detergent industries. In addition, the stain removal potential of the crude and purified lipase was determined on cotton fabric pieces stained with vegetable oil. Lipase added to water along with detergent significantly enhanced the stain removal efficiency of a commercially available detergent. The current findings suggest the potential of B. pumilusWSS5 lipase owing to promising properties such as the alkali-thermostability, organic solvent-tolerance, and broad substrate specificity that makes it a strong candidate for application in detergent formulations, biotransformation, industries, and medicine. To our knowledge, this is the first report on the purification and characterization of lipase from bacterial endophyte isolated from the stem of Withania somnifera L. Dunal.

Cadmium stimulated cooperation between bacterial endophytes and plant intrinsic detoxification mechanism in Lonicera japonica thunb

Due to the intimate association between plant physiology and metabolism, the internal colonizing microbe (endophytes) community must be adjusted to support plant productivity in response to cell damage in plants under stress. However, how endophytes coordinate their activities with plant intrinsic mechanisms such as antioxidative systems and detoxification pathways during Cd accumulation remains unknown. In this hydroponic pot study, we investigated how exposure of Lonicera japonica. thunb. to different levels of Cd (0.5, 2.5, 5, 10, and 20 mg kg−1) affected plant growth, metabolic pathways, and endophyte community structure and function. Although Cd accumulation increased at 5 mg kg−1 Cd, the biomass and height of L. japonica increased in association with elevated endophyte-involved plant detoxification activities. Endophytes, such as Sphingomonas, Klenkia, and Modestobacter, expressed major antioxidative regulators (superoxide dismutase and ascorbate acid) to detoxify Cd in L. japonica. Furthermore, L. japonica and its endophytes synergistically regulated the toxic effects of Cd accumulation via multiple plant metabolic defensive pathways to increase resistance to metal-induced stress.

Genomic, Molecular, and Phenotypic Characterization of Arthrobacter sp. OVS8, an Endophytic Bacterium Isolated from and Contributing to the Bioactive Compound Content of the Essential Oil of the Medicinal Plant Origanum vulgare L.

Medicinal plants play an important role in the discovery of new bioactive compounds with antimicrobial activity, thanks to their pharmacological properties. However, members of their microbiota can also synthesize bioactive molecules. Among these, strains belonging to the genera Arthrobacter are commonly found associated with the plant's microenvironments, showing plant growth-promoting (PGP) activity and bioremediation properties. However, their role as antimicrobial secondary metabolite producers has not been fully explored. The aim of this work was to characterize the Arthrobacter sp. OVS8 endophytic strain, isolated from the medicinal plant Origanum vulgare L., from molecular and phenotypic viewpoints to evaluate its adaptation and influence on the plant internal microenvironments and its potential as a producer of antibacterial volatile molecules (VOCs). Results obtained from the phenotypic and genomic characterization highlight its ability to produce volatile antimicrobials effective against multidrug-resistant (MDR) human pathogens and its putative PGP role as a producer of siderophores and degrader of organic and inorganic pollutants. The outcomes presented in this work identify Arthrobacter sp. OVS8 as an excellent starting point toward the exploitation of bacterial endophytes as antibiotics sources.

Isolation and Identification of Salt-Tolerant Mangrove Endophyte from Red Sea Coast Jeddah, Saudi Arabia

The aim of the study was to isolate and identify the most salt-tolerant endophytic bacteria from the mangrove plants (Avicennia marina) from the Red Sea Jeddah coastal region. In total, 21 endophytic bacteria were isolated from various parts of Avicennia marina cultured under various salt concentrations in laboratory conditions. Out of 21 endophytes, one isolate was selected based on its salt tolerance capacity, the isolate was identified as Bacillus halotolerans using 16S rRNA sequencing. The nucleotide sequence has been deposited under accession number MT858957. Bacillus halotolerans ymk 021 grows under high salinity and temperature. In response to high salinity stress (550 mM) mung bean (Vigna radiata L.) treated with B. halotolerans ymk021 showed significant results in morphological characteristics like fresh root and shoot weight, dry root and shoot weight, and root and shoot length. The ability of this salt-tolerant bacterial endophyte to produce antioxidants in plant may also resolve the major issue of oxidative damage during salinity stress. It may produce antibiotics, essential industrial enzymes too.

Antimicrobial Activity of Bioactive Compounds Isolated from Plant Endophytes

Abstract: To combat the escalating antimicrobial resistance, we need to develop new medications constantly. Recently, scientists more focused on endophytes to search for novel antimicrobial natural products. Endophytes are explored as 'bio-factories' of natural bioactive therapeutic compounds. These are microbes exhibited in plant living tissues without causing any harmful effect.Many therapeutic compounds isolated from endophytes are produced from biosynthetic pathways and belong to various structural groups like terpenoids, steroids, quinines, and phenols. Therapeutic compounds produced by endophytes exhibit different pharmacological properties. Fungal genera more commonly reside to plant tissue as compared to bacterial genera. The current review emphasizes comprehensive data on the antibacterial and antifungal activity and bio-therapeutic compounds produced by fungal and bacterial endophytes of some medicinal plants.

Endophytic Bacteria Isolated from Tea Leaves (Camellia sinensis var. assamica) Enhanced Plant-Growth-Promoting Activity

Tea (Camellia sinensis var. assamica) is a traditional and economically important non-alcoholic beverage-producing plant grown in large plantations in the northern region of Thailand and has a diverse community of endophytic bacteria. In this study, a total of 70 bacterial isolates were isolated from healthy asymptomatic samples of tea leaves from five different tea gardens in Chiang Mai, Thailand. Based on 16S rDNA sequence analysis, these bacterial isolates were taxonomically grouped into 11 different genera, namely Bacillus, Curtobacterium, Enterobacter Microbacterium, Moraxella, Neobacillus, Priestia, Pseudarthrobacter, Pseudomonas, Sporosarcina, and Staphylococcus. All these isolates were evaluated for their potential to produce indole-3-acetic acid (IAA), siderophores, and cellulolytic enzymes while having phosphate-solubilizing and tannin tolerance capacity. Most isolated bacterial endophytes belonged to the Bacillus genus and exhibited multiple plant-growth-promoting abilities. All bacterial endophytes could produce varied concentrations of the indole-related compounds, and the strain Curtobacterium citreum P-5.19 had the highest production of IAA at 367.59 µg/mL, followed by Pseudarthrobacter enclensis P-3.12 at 266.97 µg/mL. Seventy-eight percent (78%) of the total isolates solubilized inorganic phosphate, while 77%, 65%, and 52% were positive for extracellular proteases, cellulases, and pectinases, respectively. Remarkably, 80% of the isolates were capable of growth on nutrient agar supplemented with 1% (w/v) tannic acid. C. citreum P-5.19 and P. enclensis P-3.12 were selected for evaluation of plant growth promotion, and it was found that both bacterial endophytes enhanced seed germination rate and improved seedling growth parameters such as fresh and/or dry weight, root length, and shoot lengths of sunflower and tomato seeds. The selected bacterial endophytes isolated from tea leaves in this study could be used in bioformulation for plant growth promotion and advancing sustainable agricultural practices contributing to the decreased use of chemical inputs. This is the first report of an endophytic bacterium, Pseudarthrobacter enclensis, being isolated from C. sinensis.

Antibacterial and antibiofilm activity of extracts from sponge-associated bacterial endophytes

Sponges forms association with many bacteria that serve as sources of new bioactive compounds. The compounds are produced in response to environmental and nutritional conditions of the environment that enable them to protect their host from colonization. In this study, three sponge bacterial endophytes were isolated, identified, and subjected to solvent extraction processes. The identified bacteria are Bacillus amyloquifaciens, Bacillus paramycoides, and Enterobacter sp. The bacteria were cultured in two different fermentation media with varying nutritional composition for the extraction process. The extracts were evaluated for antibacterial and antibiofilm activity against microfouling bacteria and the chemical composition of each extract was analyzed via gas chromatography-mass spectrometry (GC-MS). The extract from the endophytes shows varying antibacterial and antibiofilm activity against the tested strains. Several compounds were detected from the extracts including some with known antibacterial/antibiofilm activity. The results showed variations in activity and secondary metabolite production between the extracts obtained under different nutritional composition of the media. In conclusion, this study indicated the role of nutrient composition in the activity and secondary metabolites production by bacteria associated with sponge Also, this study confirmed the role of sponge bacterial endophytes as producers of bioactive compounds with potential application as antifouling (AF) agents.

Endophytic microbiota of floating aquatic plants: recent developments and environmental prospects.

Plant-associated microorganisms play a critical role in plant survival and functional attributes. There are many studies on the taxonomical and functional aspects of microorganisms associated with terrestrial plants. However, the microbiome of aquatic plants is not much explored. This work details the studies on microbiomes and diversity in microbial communities inhabiting the three common free-floating aquatic plants of tropical regions viz. duckweed, water hyacinth and water lettuce, widely implicated for their bioremediation potential. Studies conducted till date reveal the prevalence and dominance of different Bacillus sp. Other genera, including Rhodanobacter, Pseudomonas, Rhizobium, Achromobacter, Serratia, Actinobacteria, Proteobacter, Klebsiella and Acidobacteria, have also been prominently reported. This lesser explored niche offers great bioprospecting opportunities to obtain taxonomically diverse and functionally distinct microorganisms. Bacterial endophytes from these aquatic plants have been primarily studied for their ability to produce indole acetic acid and degrade phenol. Limited studies reveal some fungal endophytes to have promising herbicidal effect. Not much is known on other functional attributes and hence microbial studies on these plants holds much promise for obtaining novel isolates or isolates with novel functions that would impact both aquatic and terrestrial ecosystems. This study proposes the need for exploring the role of endophytes as biocontrol agents and their potential to provide a pragmatic and robust solution to the aquatic weed menace in freshwater bodies. Bioprospecting of this lesser studied ecological niche hence is a promising field of research that has both environmental and economic potential.

Sustainable Applications of Endophytic Bacteria and Their Physiological/Biochemical Roles on Medicinal and Herbal Plants: Review.

Bacterial endophytes reside within the tissues of living plant species without causing any harm or disease to their hosts. These endophytes can be isolated, identified, characterized, and used as biofertilizers. Moreover, bacterial endophytes increase the plants' resistance against diseases, pests, and parasites, and are a promising source of pharmaceutically important bioactives. For instance, the production of antibiotics, auxins, biosurfactants, cytokinin's, ethylene, enzymes, gibberellins, nitric oxide organic acids, osmolytes, and siderophores is accredited to the existence of various bacterial strains. Thus, this manuscript intends to review the sustainable applications of endophytic bacteria to promote the growth, development, and chemical integrity of medicinal and herbal plants, as well as their role in plant physiology. The study of the importance of bacterial endophytes in the suppression of diseases in medicinal and herbal plants is crucial and a promising area of future investigation.

Exploration of Endophytes from Azadirachta indica for Antifungal Activity

Endophytes are the plant associated microbes that live within the living tissues of their host plants without causing any harm to them. Bioprospecting of endophytes from understudied ecosystems is a promising source for extracting novel antifungal metabolites useful for pharmaceutical/cosmetic purposes. The aim of present study was to explore endophytes for antifungal activity against Aspergillus species and Candida species. To achieve this, endophytes were isolated from Azadirachta indica leaves. Cross streak and agar overlay methods were followed for screening for antifungal activity. Secondary antifungal activity was carried out for positive antifungal isolates using agar well diffusion method. In present study, twenty four fungal endophytes and fourteen bacterial endophytes were isolated from leaves of Azadirachta indica by optimizing suitable surface sterilization protocol. Out of these six bacterial isolates were found to produce antifungal compound against Candida albicans and two isolates were found to be inhibitory towards Aspergillus species. In secondary antifungal assay, one bacterial isolate showed anticandida activity. The results of this study suggest that these endophytes producing antifungal compounds may have medicinal value. Further studies related to characterization and identification of these compounds may lead to the development of novel products beneficial to human kind.