Isolation Of Endophytic Fungi Research Articles

Isolation and molecular identification of endophytic fungi associated with Campomanesia adamantium, a Brazilian Cerrado plant.

This work presents the isolation of endophytic fungi from the leaves of Campomanesia adamantium (Cambess.) O. Berg (Myrtaceae), a native species found in Brazil and popularly known as "guabiroba-do-campo", with abundant distribution in the Brazilian Cerrado. It has been popularly used for its anti-rheumatic, anti-inflammatory, antidiarrheal, blood cholesterol-reducing, urinary antiseptic, and depurative properties. Theese fungi are microorganisms that live inside higher plants, at least for a period of their life cycle, occupying the intercellular spaces of plant tissues such as leaves and stems. These fungi are harmless to the host plant, and their secondary metabolites promote protection, regulate growth, combat bacteria, viruses, and fungi, and promote resistance to abiotic stress, as well as insecticidal effects. Endophytic fungi associated with the leaves of C. adamantium were isolated using the culture medium isolation technique. After growth, the fungi were divided into groups based on morphotypes. Fungal genomic DNA was extracted, and a polymerase chain reaction (PCR) was conducted to amplify the ITS1-5.8S-ITS2 regions of rRNA, and the nucleotide sequences obtained were compared with those available in the GenBank database for molecular identification of the isolates. The phylogenetic tree was constructed using MEGA 11 software. The results showed representatives of the Ascomycota phylum, and it was possible to identify at the genus level 18 fungi of the genera Colletotrichum, Diaporthe, Epicoccum, and Neofusicoccum.

Fungal Endophytes - An Alternative Source of Bioactive Compounds from Plant Species for Prospective Applications – Review

Plant derived compounds have a strong antagonistic activity on pathogenic microorganisms using tremendous natural bioactive compounds accumulated in different parts of plants. Limonoids are important group of metabolically altered triterpene in Meliaceae. Out of 300 limonoids known today, Melia azedarach (melianone, meliarachin, meliacine), Melia dubia (nimbin, meliacarpin) and Azadirachta indica (azadirachtin, nimbolide) contributes about one-third of limonoids. Endophytic fungi are plant symbionts that act as an elicitor for the production of secondary metabolites which serves as an excellent source of drugs for various ailments. It has a great potential to synthesize several novel bioactive compounds which paved a way for commercial exploitation of secondary metabolites for pharmaceutical and agricultural sectors. Many dominant endophytic fungi isolated from these plants includes Colletotrichum sp, Trichoderma sp, Penicillium citrinum, Fusarium solani, Aspergillus sp, Nigrospora sp and Alternaria sp which acts on human pathogenic microorganisms such as Escherichia coli Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae and Candida albicans common causative agents for Urinary Tract Infections (UTI) predominantly occurring in women. This review aims to delineate the overview on sources of limonoids present in the leaves and the endophytic fungi isolated from these leaves of Meliaceae. The isolated endophytic fungi from the leaves exhibited antagonistic activity on UTI causing pathogens which often leads to infertility among young women. Thus, the sources of limonoids and isolation of endophytic fungi can be enhanced by reinforcing the production of commercial bioactive secondary metabolites that can be substantially utilized by pharmaceutical and agricultural industries to safeguard human beings.

Molecular Characterization and Antibacterial Potential of Endophytic Fungal Isolates from Selected Mangroves along the Coastline of Kenya.

The increasing emergence and re-emergence of resistant pathogenic microbes causes a health threat to the human population. Scientists have been striving to find novel bioactive compounds and drugs to overcome these obstacles. This study aimed to characterize mangrove endophytic fungi and evaluate their antibacterial activity. Heritiera littoralis, Rhizophora mucronata, Bruguiera gymnorrhiza, Avicennia marina, and Xylocarpus granatum species were collected from Tudor Creek, Mida Creek, and Gazi Bay. A total of 30 fungal isolates were subjected to molecular identification based on analysis of their ITS gene region. The isolates in the inferred phylogenetic trees were affiliated with the genus Aspergillus. Ethyl acetate and butanol crude extracts of 38.2% of the 76 isolated fungal endophytes and eight mycelia samples were screened for antibacterial activity against Staphylococcus aureus (ATCC 27853), Escherichia coli (ATCC 25922), and Pseudomonas aeruginosa (ATCC 25923) using the disc diffusion method. A. marina and R. mucronata harbored the most fungal endophytes that showed the highest antibacterial activity. Seven fungal broth extracts exhibited higher antibacterial activities against the tested microorganisms than the positive control. The minimum inhibitory concentration (MIC) activity for the isolates demonstrated that the ethyl acetate extract of a root endophytic fungal isolate (RC6) (3.31 ± 0.01) of A. marina is a strong inhibitor since it showed significantly lower MIC activity compared to the positive control (3.84 ± 0.00) against Pseudomonas aeruginosa (P < 0.05). Therefore, this study confirms that mangrove species harbor fungal isolates that have antibacterial activity and hence could serve as a novel source of antibiotics. It is recommended that the pure compounds from these extracts be isolated for further bioactivity tests and structural elucidation for consideration as lead molecules in drug discovery. In addition, the genes responsible for the enhanced bioactivity in these isolates can be characterized and bioengineered for pharmaceutical application.

Endophytic fungi from medicinal plant Garcinia cowa Roxb. ex Choisy and their antibacterial activity

Endophytic fungi grow inside plant tissues and produce biologically active substances without harming the host plant. Endophytic fungi isolated from the roots, barks, and leaves of Garcinia cowa Roxb. ex Choisy, an ethnomedicinal plant commonly used in West Sumatera, Indonesia, are the focus of this research, which aims to evaluate their antibacterial activity. Traditional medicine has made extensive use of this plant’s various parts. In total, 14 endophytic fungal isolates were obtained from plant fragments of G. cowa that had undergone surface sterilization. After four to six weeks of cultivation in rice medium, the fungus isolate was extracted with ethyl acetate (EtOAc). Each EtOAc extract of fungus was tested for antimicrobial activity using Kirby–Bauer diffusion against Staphylococcus aureus, Escherichia coli, Methicillin Resistant S. aureus, and Candida albicans. The measurement results of the inhibition zone indicate that the GCA3 isolate of endophytic fungi exhibited significant inhibitory diameters and demonstrated potential as an antibacterial agent. The GCA3 strain showed an inhibition diameter zone of 15.05 } 0.51 mm against S. aureus, 14.68 } 0.5 mm against E. coli, and 13.48 } 0.15 mm against MRSA. None of the fungi extracts can inhibit the growth of C. albicans. The fungus GCA3 has been determined to be identical to Penicillium citrinum through macroscopic, microscopic, and molecular identification methods. The findings indicate that endophytic fungi from the Garcinia plant can generate antibacterial compounds. Hence, additional investigation is required to separate these bioactive compounds.

Chemical characterization, pathway enrichments and bioactive potentials of catechin-producing endophytic fungi isolated from tea leaves.

Endophytes acquire flavonoid biosynthetic genes from the host medicinal plants. Despite tea (Camellia sinensis (L.) Kuntze) being the major source of bioactive catechins, catechin-producing endophytic fungi have never been reported from the tea plant. Here, we report the isolation and characterization of catechin-producing endophytic fungi isolated from tea leaves, their chemical characterization, and associated bioactivities. Among the nine isolated endophytes, two (CSPL6 and CSPL5b) produced catechin (381.48 and 166.40 μg per mg extract) and epigallocatechin-o-gallate (EGCG; 484.41 and 281.99 μg per mg extract) as quantified by high-performance liquid chromatography (HPLC). The isolates were identified as Pseudopestalotiopsis camelliae-sinensis and Didymella sinensis based on molecular and morphological characterization. Untargeted metabolomics using gas-chromatography mass spectroscopy (GCMS) revealed the presence of several bioactive phytochemicals mostly belonging to tyrosols, pyridoxines, fatty acids, aminopyrimidine, and benzenetriol classes. Metabolic pathways pertaining to the biosynthesis of unsaturated fatty acids (UFAs), butanoate metabolism, and linoleic acid metabolism were highly enriched in both catechin-producing isolates. The isolates were able to differentially scavenge intracellular O2 and N2 free-radicals, but CSPL5b demonstrated relatively superior bioactivities compared to CSPL6. Both isolates stimulated the growth of various probiotic strains, indicating prebiotic effects that are otherwise known to be associated with catechins. Collectively, the current study demonstrated that fungal endophytes CSPL6 and CSPL5b, isolated from tea leaves, could be used as alternative sources of catechins, and hold promising potential in evidence-based therapeutics.

Isolation, identification, and antibacterial activity of endophytic fungus from Acanthus ilicifolius

Abstract Antimicrobial-resistant bacteria pose serious public health risks, necessitating bioprospecting for novel antimicrobial drugs. The endophytic fungi of the mangrove ecosystem are hotspots for discovering new bioactive chemical compounds. In this context, an investigation was designed to determine the isolation of the major endophytic fungi inhabiting the leaves of Acanthus ilicifolius, a mangrove plant with a long history of traditional use in the Chinese and Indian medical systems. Based on the morphological characterizations and molecular analysis of internal transcribed spacer rDNA sequence data, the study identified three unique endophytic fungal species, namely, Periconia macrospinosa, Coprinopsis cinerea, and Alternaria sp. Of these, P. macrospinosa was identified as the most dominant one, with the highest relative frequency (35.22%). The antibacterial activity of P. macrospinosa isolate (CMFRI/fPM-01) was evaluated by the well and diffusion method against six human pathogens, viz., Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus. The results demonstrated a high and wide spectrum of antimicrobial action of the isolate against all the tested human pathogens, with no significant difference (P &gt; 0.05) in the activity between the pathogens. The antibacterial activity was further confirmed by determining the fungal culture supernatant's minimum inhibitory concentration and minimum bactericidal concentration. Although the studied fungi are known from other sources, this is the first report of P. macrospinosa and C. cinerea as endophytes in A. ilicifolius leaves. The outcomes also showed that the P. macrospinosa isolate could be used to discover effective antibacterial drugs against various human diseases.

Antibacterial and antioxidant activity of endophytic fungi extracts isolated from the petiole of sungkai plant (Peronema canescens)

Abstract. Oktiansyah R, Elfita, Widjajanti H, Salni, Setiawan A. 2023. Antibacterial and antioxidant activity of endophytic fungi extracts isolated from the petiole of sungkai plant (Peronema canescens). Biodiversitas 24: 6516-6526. Indonesia has a diverse number of medicinal plants that are very helpful in preventing infectious diseases. Sungkai (Peronema canescens) is a medicinal plant of the family Verbenaceae that is often used and found in Indonesia. Its leaves possessed antibacterial and antioxidant properties that boost immunity and lessen the signs of infectious disorders. This study explored the types of endophytic fungi found in sungkai petioles and examined their relationship to previously reported sungkai leaf endophytic fungal isolates and spectrum of compounds contained them. Endophytic fungal were isolated from fresh leaf petioles of host plants, and their morphological and molecular characteristics were determined. Endophytic fungal extracts were tested for antibacterial and antioxidant properties. Utilizing the paper disk diffusion technique, the antibacterial characteristics were ascertained, while the DPPH method was used to determine antioxidant activity. Molecular identification was carried out on the fungal isolate with the most potential, and chemical compounds were isolated using column chromatography. The structures of the compounds were determined using spectroscopy, including 1D and 2D NMR. Eight endophytic fungal isolates were obtained from sungkai leaf stalks (RA1-RA8) of various species of the genus Trichoderma. Differences in the diversity of endophytic fungi found in leaf stalks and endophytic fungi from leaves were identified. RA1 showed the strongest antibacterial and antioxidant activity and was molecularly identified as T. harzianum. Spectroscopic analysis showed that the pure compound contained within was 9-hydroxy-7-methylenebenzo[c]oxepin-3(7H)-one (1), which had never previously been found in sungkai plants and endophytic fungi. The antibacterial activity of Compound 1 is in the strong category (MIC = 64 µg/mL) but is not active as an antioxidant. In terms of producing medicinal ingredients from endophytic fungi, T. harzianum in extract form has more potential to be developed.

Antagonistic Endophytic Fungi from Papaya Fruit Against Anthracnose Causing Pathogens (Colletotrichum gloeosporioides) on Papaya Fruit

Anthracnose disease caused by Colletotrichum gloeosporioides considered a major disease on papaya fruit. One way to control plant diseases is to use antagonistic fungi as biocontrol agents. Several antagonistic fungi can be found in plant tissues (endophytic fungi). This study aims to get endophytic fungi from papaya fruit antagonistic to the fungus C. gloeosporioides. The research used a Completely Randomized Design (CRD) consisting of four treatments and five replications on in-vitro and in-vivo tests. The results showed that three isolates of endophytic fungi found from papaya fruit were Fusarium sp., Aureobasidium sp., and Acremonium sp., which had an inhibition of 63.5%, 67.86%, and 7.52%, respectively. Fusarium sp. and Aureobasidium sp. are potentially considered antagonist fungi in controlling the fungus C. gloeosporioides in in-vitro testing based on the inhibition results were more than 60%. Aureobasidium sp. is considered potential antagonist fungi according to the colonization or growth of C. gloeosporioides mycelium inhibition that emerges on papaya up to 97%.

Isolasi, Identifikasi, dan Uji Aktivitas Antibakteri Fungi Endofit Daun Alpukat (Persea Americana Mill) Terhadap Pseudomonas aeruginosa dan Staphylococcus aureus

Avocado (Persea americana Mill) is a plant that contains antibacterial substances, especially in its leaves. Flavonoids, saponins and alkaloids are active compounds in avocado leaves which can prevent some bacterial growth. This study aims to isolate, identify and determine the antibacterial activity of avocado leaf endophytic fungal isolates growing P. aeruginosa and S. aureus. This type of research, namely experimental research, isolates endophytic fungi using avocado leaves inoculated on PDA media which has been added with 0.005% chloramphenicol in a petri dish and incubated at a temperature of 25oC for 5-7 days. Antibacterial activity was tested using a paper disk with the agar diffusion method. This research showed that young avocado leaves produced 3 isolates, namely isolate 1 was suspected to be Trichoderma sp., isolate 2 was suspected to be Rhizoctonia sp., and isolate 3 was suspected to be Fusarium sp. From the results obtained, it was concluded that the endophytic fungi of avocado leaves from isolate 2, and isolate 3 provided an inhibition zone against P. aeruginosa and isolate 1, and isolate 3 provided an inhibition zone against S. aureus.

Control of Root-knot Nematodes (Meloidogyne spp.) on Tomato (Lycopersicum esculentum Mill.) using Antagonist Fungi

Nematodes Meloidogyne spp. is the most important phytonematode in the world, especially in agriculture in tropical areas. The level of root damage to tomato plants caused by the nematode Meloidogyne spp. can reach 68.3%. In general, control of plant parasitic nematodes is still carried out using pesticides in the form of insecticides which can also be used as nematicides. Continuous use of chemicals to control nematodes can cause environmental pollution, resurgence and nematode resistance to chemicals. One alternative control is biological. One of the biological agents that can be used to control Meloidogyne nematodes is the endophytic fungus Beauveria bassiana and Trichoderma asperellum. The research aims to obtain the best isolates of endophytic fungi for controlling rootknot nematodes (Meloidogyne spp.). The isolates tested were 2 Beauveria bassiana isolates and 2 Trichoderma asperellum isolates. The results showed that the fungus isolate Beauveria bassiana isolate TD312 had a higher ability to control Meloidogyne spp on tomatoes.&#x0D;

Optimization of Temperature and pH for the Protease Production By Endophytic Fungi From Mangrove Sonneratia alba

The research about optimization of temperature and pH for the protease production from mangrove Sonneratia alba endophytic fungi in Mandeh Area, Pesisir Selatan District was conducted from March to June 2023 at Biotechnology Laboratory, Andalas University, Padang. This research aims to determine the optimum temperature and pH of endophytic fungal isolates of mangrove plants Sonneratia alba from the Mandeh area, Pesisir Selatan District in producing protease and to determine the enzyme production after optimization. The research method used in this study was experimental and descriptive.Protease activity was determined using the Takami method. The results of the study showed that the optimum temperature for isolates EUA-124 and EUA-126 in producing protease is 34°C the optimum pH for isolate EUA-124 in producing protease is 7 and the optimum pH for isolate EUA-126 in producing protease is 6. Protease production by isolates EUA-124 increased by 8.621% and protease production by isolate EUA-126 increased by 3.334%.

Isolation and Molecular identification of Endophytic fungi from Michelia champaca Linn. and screening for its Extracellular Enzyme Production

Fungal endophytes are the group of microorganisms that occupy parts and colonize living, internal tissues of plants without causing any immediate, negative effect or noticeable injury to the host. In this study, fungal endophytes were targeted for isolation from young and healthy leaves of Michelia champaca Linn. from Guwahati region, Assam. The colony morphology was screened for observation of spores/ conidia or fruiting bodies under light compound microscope after staining. The endophyte was screened for the presence of various enzymatic activities like Amylolytic, Lipolytic, Cellulolytic, Proteolytic and catalase activity. Results showed that the isolated endophyte was a potential source of amylolytic, cellulolytic and Lipolytic activities whereas it showed negative result for catalase and proteolytic activities. It was further characterized using 1, 5.8S ribosomal RNA gene ITS sequencing and phylogenetic analysis. The molecular characterization identified it as Cladosporium pseudocladosporioides. Hence, this study showed that fungal isolate from M. champaca could be a potential source for enzymes with biological activities.

Harnessing the Potentialities of Arsenic Resistant Fungal Endophytes for Improved Growth Promoting Traits under in vitro Condition

Endophytic fungi enhance the plant’s ability to tolerate stressful conditions including heavy metal stress via secretion of numerous secondary metabolites. However, the role of heavy metal resistant fungal endophytes in growth promotion of plants in extreme environments is need to be understood. Therefore, eight endophytic fungal isolates having arsenic tolerance potential up to 2000 ppm explored from the arsenic stressed plants were subjected to various in vitro growth promoting traits viz., phytohormones production, phosphate solubilization, siderophore production, ammonia production, HCN production and ACC deaminase activity under normal and arsenic stress (100 ppm) condition. Indole acetic acid produced by fungal endophytes ranged between 120-610 μg/ mL, which was reduced to 50-340 μg/ mL when they were grown on arsenic induced medium. In the siderophore production test, fungal isolate S3P1S1 produced significantly higher siderophore (96 ± 0.002 μmol) compared to other isolates and reference culture. In addition, HCN production was observed in only one isolate. Therefore, present study clearly identified specific traits in the fungal endophytes, which make them good candidates as PGPR and might contribute to plant adaption to arsenic contaminated soils. These fungal endophytes, possessing metal tolerance as well growth promoting properties under in vitro conditions could have vital implications for the agricultural sector if used as biofertilizer.

Diversity of endophytic fungi isolated from tree spinach (Cnidoscolus aconitifolius) (Euphorbiaceae) and their bioactivities

Abstract. Praptiwi, Palupi KD, Ilyas M, Marlina L, Fathoni A. 2023. Diversity of endophytic fungi isolated from tree spinach Cnidoscolus aconitifolius (Euphorbiaceae) and their bioactivities. Biodiversitas 24: 6229-6239. Endophytic fungi associated with medicinal plants have been known as a reservoir for therapeutic compounds. Cnidoscolus aconitifolius I.M. Johns (Euphorbiaceae), which possesses several local names, including papaya jepang in Indonesia, has been historically used as traditional medicine. However, research on the bioactivity of endophytic fungi associated with it has yet to be conducted. The purposes of the study were to isolate endophytic fungi from C. aconitifolius and determine their bioactivity, i.e., antibacterial activity against Escherichia coli and S. aureus and free radical scavenging activity representing antioxidant activity. Identification of endophytic fungi was carried out based on their morphological characteristics. Total phenolic and flavonoid contents were determined by the spectrophotometric method. The qualitative antibacterial and antioxidant bioactivity screening was performed using the TLC-bioautography method. The quantitative analysis of antibacterial and antioxidant activity was conducted by microdilution to obtain IC50 and MIC values. In total, 14 endophytic fungal isolates were successfully isolated. Antioxidant analysis showed that one isolate had strong antioxidant activity, and one had moderate growth inhibition activity against E. coli. The total phenolic content strongly correlates with the AAI value as an indicator of antioxidant activity. The findings of this study reveal that Phomopsis PjBg-1, an endophytic fungus from C. aconitifolius, is identified as Phomopsis sp. LH243 possesses strong antioxidant activity and moderate antibacterial activity against E. coli. Endophytic fungi from C. aconitifolius have the prospect of being an alternative to antioxidants and antibacterials. The isolation of compounds responsible for bioactivities needs to be further studied.

Evaluation of Antibacterial Properties from Endophytic Fungi of Chrysanthemum indicum (L.) Flowers against Escherichia coli and Staphylococcus aureus

Uncovering the therapeutic potential of secondary metabolites produced by plants, animals, and microbes constitutes the foundation for the development of novel medications. The objective of this investigation is to discern the classes of secondary metabolites and assess the antibacterial properties of endophytic fungal extracts obtained from Chrysanthemum indicum L. flowers. Through the isolation process, five isolates designated as JEC1, JEC2, JEC3, JEC4, and JEC5 were identified. The cultivation of endophytic fungal isolates spanned a three-week period before undergoing extraction with ethyl acetate. The phytochemical tests revealed the presence of alkaloids, flavonoids, steroids, terpenoids, saponins, and tannins in the ethyl acetate extract. Antibacterial activity was determined using the agar well diffusion method, with ciprofloxacin serving as a positive control. Notably, all ethyl acetate extracts from endophytic fungi exhibited antibacterial activity. The most substantial inhibitory diameter against Staphylococcus aureus was recorded as 19.1±0.8 mm for the JEC3 endophytic fungi, while Escherichia coli exhibited an inhibitory diameter of 16±1.1 mm for the JEC2 endophytic fungi.

The antioxidant activity of endophytic fungal extract of nutmeg plant (Myristica fragrans Houtt.) from Cimande West Java, Indonesia

The bioproduction of bioactive metabolites from nutmeg endophytic fungi from Cimande, Wesh Java Indonesia was successful. In this research, bioactive antioxidant metabolites were extracted from nutmeg endophytic fungi. Endophytic fungi from nutmeg plant parts were grown on CMMA media. Each fungus was isolated by growing it on PDA media. A total of 21 endophytic fungi isolates were grown on PDB media before being extracted with ethyl acetate as a solvent. The morphology of 21 endophytic fungal isolates was identified based on morphological characteristics. There are 11 Phomopsis sp., 1 Fusarium oxysporum, 5 Colletotrichum sp., 1 Schizophyllum sp., 1 Dematiaceae sp., 1 Coelomycetes sp., and 1 Xylaria sp. The bioautographic TLC method and DPPH-free radical scavenging activity were used to determine antioxidant activity. The antioxidant activity analysis showed that 14 of the 21 endophytic fungus extracts were active. One nutmeg pericarp endophytic fungus showed strong antioxidant activity, with an IC50 of 30.0 g mL-1 and AAI of 1.04. Furthermore, the IC50 values for two fungal extracts from the leaf and pistil nutmeg were less than 100 g.mL-1. The three fungi with IC50 less than 100 g.mL-1 were identified as Phomopsis sp.

DIVERSITY OF ENDOPHYTIC FUNGI FROM SHALLOTS AS A FUSARIUM OXYSPORUM BIOLOGICAL CONTROL AGENT

Fusarium oxysporum is the primary cause of Fusarium wilt, one of the major diseases that harm shallots. 50% of yield can be lost due to Fusarium wilt. It is anticipated that endophytic fungus will inhibit the growth of Fusarium wilt disease in shallots. The endophytic fungi from shallot plants that have the potential to act as biological control agents for the fungal disease F. oxysporum are suggested to be chosen in this study. Using Malt Extract Agar (MEA) medium, endophytic fungi were isolated from healthy shallot plant components (roots, tubers, stems, and leaves). Using dual culture and volatile techniques, the antagonistic potential of endophytic fungi against F. oxysporum was examined. Using shallot seeds, pathogenicity studies for endophytic fungus were conducted in vitro. Twelve endophytic fungal isolates were able to impede the growth of F. oxysporum using the dual culture method; the percentage of inhibition reached 72.52% at seven days post-inoculation. The endophytic fungus was also able to stop F. oxysporum mycelial growth using the volatile compound test, with a percentage inhibition ranging from 4.08 - 27.37%. The findings of the pathogenicity test demonstrated that the four isolates posed a risk to the shallot seeds. The most successful isolate in preventing the growth of F. oxysporum was the endophytic fungus AB4D1, which is a member of the Trichoderma asperellum species.

English

Salinity and drought are among the major abiotic stresses, leading to decreased crop productivity. Fungi are notorious for their detrimental effects on crop growth and development. In light of these circumstances, the primary objective of this project is to assemble a collection of endophytic fungi to develop new organic bio-stimulants that can effectively combat salinity and drought. To achieve this goal, diverse soil types, including cultivated, desert, and salt marshes, were sampled (five samples each) from different habitats and locations in the southern part of Sinai, the Saint Katherine Protectorate (SKP) is part of the upper Sinai massif, spanning specific coordinates (33°550 to 34°300 E and 28°300 to 28°350 N). Additionally, 80 samples of the predominant medicinal plant species from ten localities in the SKP were collected. The results of survey showed that the highest abundance of endophytic fungi was observed in Artemisia judaica L. and Alkanna orientalis (L.) Boiss., with both species exhibiting 61.1 % endophytic fungi. Five isolates of fungi labeled as SAF41, SAF46, SAF51, SAF57, and SAF63, were assessed for their tolerance to salinity stress by exposing them to different NaCl concentrations. Isolate SAF63 had the most substantial negative and significant correlation with salinity stress (R2=0.9565; p&lt;0.001), followed by SAF51 (R2=0.9432; p&lt;0.001) and SAF41 (R2=0.9351; p&lt;0.001). The results indicate that different fungi isolates exhibited distinct responses to varying salinity levels and drought stress, as measured by their growth in different NaCl concentrations. Isolate SAF63 demonstrated the most significant negative correlation with salinity stress, followed by SAF51 and SAF41. These findings suggest that specific endophytic fungi isolates exhibit enhanced tolerance to both salinity and drought stress, underscoring their promising potential in mitigating metabolic and physiological damage in date palm plants, ultimately leading to increased growth and yields

Characterization of Endophytic Fungi in Robusta Coffee (Coffea canephora L.) Beans Through 18S rRNA Gene Sequencing and Evaluation of Antioxidant Activity and Chlorogenic Acid Content

Utilizing endophytic fungi to discover metabolites by characterizing the fungi produced from robusta coffee beans can be highly beneficial without causing harm to the plants. Additionally, this approach aids in identifying raw materials for medical drug development, specifically compounds like chlorogenic acid, which hold promise as potential treatments for diabetes mellitus. This study aimed at characterizing the endophytic fungi present in Robusta coffee (Coffea canephora L.) beans through 18S rRNA gene sequencing and evaluation of antioxidant activity and chlorogenic acid content found in these endophytic fungi. Green, unripe Robusta coffee beans were harvested from a coffee plantation in Tana Toraja and isolated using Potato Dextrose Agar medium. The isolation process yielded three isolates, designated RF1, RF2, and RF3. Subsequently, the isolates were subjected to testing for endophytic fungi activity, and the isolate that exhibited activity was further characterized using 18S rRNA gene sequencing. Following that, the isolates were fermented using a Potato Dextrose Broth medium, resulting in two phases: filtrate and biomass. The filtrate was then extracted using ethyl acetate as the solvent. The extracted compound was subsequently evaluated for antioxidant activity using ABTS and DPPH methods, and the concentration of chlorogenic acid was measured. From the three isolates of endophytic fungi obtained from green Robusta coffee beans with the codes RF1, RF2, and RF3, all of them exhibited antibacterial activity against Escherichia coli and Staphylococcus aureus. The 18S rRNA gene sequencing analysis revealed that isolate RF1 was identified as Fusarium sp. For the evaluation of antioxidant activity using the ABTS method, an IC50 value of 53.49 µg/mL was obtained, while the DPPH method resulted in an IC50 value of 54.03. Furthermore, the measurement of chlorogenic acid content in the extract of endophytic fungi from Robusta coffee beans indicated a concentration of 11.58%. The characterization result of the endophytic fungi isolates from green Robusta coffee beans revealed the presence of Fusarium sp. It exhibited high antioxidant activity and a chlorogenic acid content of 11.58%.

Isolation and Characterization of Novel Huperzine-Producing Endophytic Fungi from Lycopodiaceae Species.

Huperzine A (HupA) is an important drug for treating Alzheimer's disease (AD) and is primarily extracted from the Huperzia serrata (Lycopodiaceae). Failures in the chemical synthesis of Hup and in vitro culture have put H. serrata in danger of extinction, and there is a need for an extensive investigation of Hup from alternative perspectives. The aim of this study is to identify endophytic fungi that produce high Hup or simultaneously produce many types of Hup and have high genetic stability derived from other Lycopodiaceae species as a source of materials for natural Hup production. In this work, Hup-producing endophytic fungi were isolated from three species: Lycopodium clavatum, Phlegmariurus squarrosus, and P. phlegmaria. Of these, L. clavatum and P. squarrosus were confirmed as novel sources of Hup-producing fungi. Based on morphological characteristics and nuclear ribosomal DNA ITS sequences, four endophytic fungi Colletotrichum siamense THG1-17, Epicoccum sorghinum THG01-18, Phoma sp. TKH3-2, and Phyllosticta sp. THG2-27 were firstly isolated from these Lycopodiaceae plants, which were capable of simultaneously producing both HupA and HupB, as evidenced by high-performance liquid chromatography analysis. The four strains showed stability in Hup yield over 50 generations of culture with an in vitro storage period of 3 months. These isolated fungi will provide a new source of materials for further research to develop drugs containing HupA as well as HupB for AD treatment in the future.