Isolation Of Endophytic Fungi Research Articles

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.

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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<0.001), followed by SAF51 (R2=0.9432; p<0.001) and SAF41 (R2=0.9351; p<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.

Potential Endophytic Fungi from the Leaves of Syzygium zeylanicum (L.) and Their Secondary Metabolite

Endophytic fungi coexist without harms in the host in some parts or all parts of its lifecycle. Endophytic fungi excrete specialized bioactive compounds beneficial for of its host; the compound itself can be different from what can be found on their host. Endophytic fungi are capable to excrete biologically significant secondary metabolites acting as antidiabetic, antioxidant, antimicrobial, and cytostatic agents. Endophytic fungi existence are common across different plants; including Syzygium zeylanicum that can be found in Indonesia. This study aims to isolate endophytic fungi found in leaves of Syzygium zeylanicum, L., to elucidate their characterized morphologically, and to examine its antimicrobial and antioxidant properties in addition to their chemical structures. Characterization of endophytic fungi was conducted by their macroscopic and microscopic features, followed by molecular characterization of highly bioactive metabolites. Antimicrobial activities were measured by disc diffusion method. Antioxidant properties were measured with DPPH. Secondary metabolites were chromatographically isolated and identified with spectroscopy techniques (NMR ID and 2D). Four endophytic fungi isolates were obtained: Penicillium citrinum (SZ1), Colletotrichum lindemuthianum (SZ2), Aspergillus nidulan (SZ3), Scopulariopsis asperula (SZ4). Penicillium citrinum (SZ1) showed antimicrobial activities against four different bacteria (71.3% against E. coli; 74.1% against S. aureus; 76.2% against S. typhi; and dan76.9% against B. subtilis). Antioxidant activity ini all ekstrak of endophytic fungi showed very activity (IC50 SZ3 extract = 3.85 µg/mL). Potential endophytic fungi SZ1 was molecularly identified as Penicillium citrinum. Extracts from SZ1 fungi contains bioactive 4-hydroxy-2-(4-hydroxyphenyl)-ɣ-butyrolactone-3-yl) methyl acetate. The newly obtained substance could be developed into antimicrobial and antioxidant agents in further studies.

Diversity of fungal endophytes isolated from the invasive plant Solanum rostratum

A culture-dependent method was used to isolate fungal endophytes from the leaves, stems, and roots of the invasive plant Solanum rostratum Dunal. growing in Xinjiang Province, China. All isolates were identified according to ITS (internal transcribed spacer) region of ribosomal DNA sequences and analyzed by Nucleotide BLAST according to NCBI GenBank and Mycobank database. Altogether 176 endophytic fungal isolates corresponding to 44 OTUs were identified, which were classified into 12 genera, with Penicillium (59.66%) and Aspergillus (23.29%) being the highly dominant genera. Ten endophytic isolates (OTU1, OTU15, OTU16, OTU21, OTU23, OTU25, OTU26, OTU30, OTU37 and OTU44) were identified as potential new species.

Isolation of endophytic fungi from Cotoneaster multiflorus and screening of drought-tolerant fungi and evaluation of their growth-promoting effects.

In the context of climate change and human factors, the drought problem is a particularly serious one, and environmental pollution caused by the abuse of chemical fertilizers and pesticides is increasingly serious. Endophytic fungi can be used as a protection option, which is ecologically friendly, to alleviate abiotic stresses on plants, promote plant growth, and promote the sustainable development of agriculture and forestry. Therefore, it is of great significance to screen and isolate endophytic fungi that are beneficial to crops from plants in special habitats. In this study, endophytic fungi were isolated from Cotoneaster multiflorus, and drought-tolerant endophytic fungi were screened by simulating drought stress with different concentrations of PEG-6000, and the growth-promoting effects of these drought-tolerant strains were evaluated. A total of 113 strains of endophytic fungi were isolated and purified from different tissues of C. multiflorus. After simulated drought stress, 25 endophytic fungi showed strong drought tolerance. After ITS sequence identification, they belonged to 7 genera and 12 species, including Aspergillus, Fusarium, Colletotrichum, Penicillium, Diaporthe, Geotrichum, and Metarhizium. According to the identification and drought stress results, 12 strains of endophytic fungi with better drought tolerance were selected to study their abilities of dissolving inorganic phosphorus and potassium feldspar powder and producing indole-3-acetic acid (IAA). It was found that the amount of dissolved phosphorus in 7 strains of endophytic fungi was significantly higher than that of CK, and the content of soluble phosphorus was 101.98-414.51 μg. ml-1; 6 endophytic fungi had significantly higher potassium solubilization than CK, and the content of water-soluble potassium ranged from 19.17 to 30.94 mg·l-1; 6 strains have the ability to produce IAA, and the yield of IAA ranged between 0.04 and 0.42 mg. ml-1. This study for the first time identified the existence of endophytic fungi with drought tolerance and growth-promoting function in C. multiflorus, which could provide new direction for plant drought tolerance and growth promotion fungi strain resources. It also provides a theoretical basis for the subsequent application of endophytic fungi of C. multiflorus in agricultural and forestry production to improve plant tolerance.

Isolation and identification of endophytic fungi involved in the promotion of sesquiterpene accumulation in Syringa pinnatifolia

Abstract Background The stem of Syringa pinnatifolia is used as traditional Mongolian medicine for the treatment of cardiopulmonary diseases, in which sesquiterpenes are regarded as the primary medicinal compounds. However, only S. pinnatifolia from natural stands is able to produce medicinal-grade stems when compared with that from cultivated stands. Despite the need to develop an effective technique to promote sesquiterpene accumulation in cultivated stands to ensure sustainable resource production, studies on the potential use of fungal endophytes as fungal elicitors were limited. Objective This study explored the potential of selected endophytic fungi isolated from the stems of wild and cultivated S. pinnatifolia and further examined their efficacy in promoting sesquiterpene production in S. pinnatifolia calli. Results Among the 11 fungal groups, Valsa sp, isolated from wild S. pinnatifolia, emerged as a promising fungal elicitor for promoting sesquiterpene accumulation in S. pinnatifolia. This conclusion is drawn from the production of a humulane-type sesquiterpene, known to be one of the most prevalent subtypes of sesquiterpenes in S. pinnatifolia. In addition, it was demonstrated that methyl jasmonate and salicylic acid displayed little effect on sesquiterpene accumulation in S. pinnatifolia. Conclusions The findings of this study lay a foundation for using fungal endophytes to promote sesquiterpene production in S. pinnatifolia. This contribution could subsequently advance the development of the local traditional medicine industry and aid in the protection of wild S. pinnatifolia.

Identification and Characterization of Endophytic Fungi from Garcinia atroviridis for Potential Antagonistic Against Phytopathogenic, Colletotrichum gloeosporioides

Biological control is referred to as the “use of natural or modified organisms, genes” to minimize the effects of undesirable pests, pathogenic microorganisms, and diseases on plant crops. This measure has become a suitable and safe alternative for chemical fungicides in plant disease management. Endophytic fungi have received much attention as biological control agents against many plant pathogens through antibiosis, parasitism, invading spores, mycelium, and cells of the pathogen, and secreting bioactive metabolites. While the therapeutic properties of Garcinia atroviridis have been studied, the existence of microbial endophytes and their properties is still less documented. In this research, G. atroviridis endophytic fungi were isolated and identified by fungal colony morphology observation combined with the PCR-amplified fungal internal transcribed spacer (ITS) sequence analyses. Fungal endophytes were assessed for their biocontrol potential against Colletotrichum gloeosporioides. In total, 111 endophytic fungal isolates harboring inside the leaf, branch, and fruit of G. atroviridis belonged to 5 different species with 3 different genera and two unidentified genera. All the endophytic fungal species isolated were evaluated using an in vitro dual culture assay against C. gloeosporioides, a common pathogen that causes anthracnose disease. The results of the present study clearly showed that seven species of isolated fungal endophytes were capable of inhibiting the mycelial colony growth of C. gloeosporioides with an inhibition percentage between 54.67% to 87.94%. Among these species, Nigrospora sphaerica recorded the highest PIRG with 87.94%. Our work indicates that endophytic fungi isolated from G. atroviridis have a biocontrol effect on C. gloeosporioides and are expected to be a potential source of bioactive metabolites.

One stone for two birds: Endophytic fungi promote maize seedlings growth and negatively impact the life history parameters of the fall armyworm, Spodoptera frugiperda.

The fall armyworm (FAW) Spodoptera frugiperda, is a voracious pest of cereals native to the Americas and which invaded Africa in 2016. Chemical control is the main management option, which however remains ineffective and unsustainable. Fungal endophytes are increasingly used as alternative for the management of insect pests of economic importance. This study assessed the potential of eight endophytic fungal isolates to colonize maize plant and their ability to promote seedlings growth through seed and foliar inoculations, as well as their suppressive effects on FAW. Fungal colonization rates of different plant parts by the endophytes varied as per the inoculation methods. Beauveria bassiana ICIPE 279 colonized more than 60% of all the seedling parts while B. bassiana G1LU3 only colonized stem (25%) and leaf (5%) tissues through foliar inoculation. Trichoderma atroviride F2S21, T. asperellum M2RT4, T. harzianum F2R41, Trichoderma sp. F2L41, Hypocrea lixii F3ST1 and Fusarium proliferatum F2S51 successfully colonized all the plant parts and therefore were selected and further evaluated through seed inoculation for their endophytic persistence, effect on plant growth, and pathogenicity to Spodoptera frugiperda immature and adult stages. Weekly assessment showed varied effect of the endophytes on maize plant growth parameters compared to the control. During the first week, percentage colonization of the plant parts ranges between 90%-100%, 65%-100%, and 60%-100%, in the roots, stems, and leaves, respectively for all the five tested isolates. However, the colonization pattern/rates significantly decreased over time for H. lixii F3ST1 in the stems and leaves, and for T. harzianum F2R41 in the leaves and for T. asperellum M2RT4 in the roots. In addition, T. harzianum F2R41 outperformed all the other isolates in boosting the plant height, whereas H. lixii F3ST1 and T. asperellum M2RT4 outperformed all the other isolates in increasing the wet and dry shoots weight. Furthermore, the number of egg masses laid on endophytically-colonized maize plants varied among the treatments. Trichoderma asperellum M2RT4 and H. lixii F3ST1 endophytically-colonized maize plants significantly reduced the number of egg masses and the defoliation/feeding rates of the pest compared to the control. Additionally, T. harzianum F2R41 had the highest negative impact on the pupation and adult emergence of S. frugiperda with a female-biased sex ratio. Our findings indicate that T. asperellum M2RT4, T. harzianum F2R41, and H. lixii F3ST1 hold a potential to be developed as endophytic-fungal-based biopesticides for sustainable management of S. frugiperda and as plant growth promoters.

Detection of bacterial pathogen on corn plants and test the potential of endophytic microbes to controlling stewart wilt disease in vitro

The use of synthetic pesticides to increase maize production sustainably contributes to increasing the development of pathogens and tends to be less effective in environmental conditions that support pathogens. In addition, the emergence of environmental pollution due to the use of chemicals causes the need for environmentally-friendly control technologies. Endophytic microbes are biological agents that can be used to plant disease control that is environmentally friendly and practical in their application. The study aimed to detect pathogenic bacteria that cause wilt and blight in maize and then examine the potential of several endophytic microbial isolates in controlling Stewart wilt pathogens in maize in vitro. The research was carried out from February to July 2021 at the Cereal Crop Research Institute Laboratory in Maros for morphological identification and microbial potential testing, while serological tests were carried out at the Makassar Plant Quarantine Laboratory, South Sulawesi. The study began with detecting pathogenic bacteria serologically using the Enzyme-Linked Immunosorbent Assay (ELISA) method. The results showed that blight symptoms on corn isolated from the experimental garden of IP2TP Bajeng Gowa Regency PS-4 and PS-5 were detected as bacteria Pantoea stewartii subsp. stewartii. The antagonistic test of endophytic bacterial isolates I.A1, II.D1, III.A2, IV.B2, and endophytic fungi isolates AC-1, AC-3, and DC-5 was able to suppress the growth of pathogenic bacteria isolates PS-4 and PS-5 The growth suppression of pathogenic bacteria was observed by forming inhibitory zones of antibiosis compounds. Pathogenic bacteria Pantoea stewartii subsp. stewartii can be controlled using endophytic microbes.

Antibacterial Activity of Ethanol Extract of Purple Kencana Leaves (Ruellia tuberosa L.) Using TLC-Bioautography and Agar Diffusion Methods

Tunjuk Langit Plant (Helminthostachys zeylanica (L.) Hook) contains a lot of secondary metabolite compounds, one of which is found in the root which contain the flavonoids ugonin and quercetin. The purpose of this study was to determine the antibacterial activity of compound 4 isolates IFATL-05 endophytic fungi from Tunjuk Langit (Helminthostachys zeylanica (L.) Hook) root against bacterial gastrointestinal tract infection. The results of purification of endophytic fungi with IFATL-05 code on macroscopic examination showed blackish-white fungus color, round with scalloped colony shapes, wavy edges and drop-like elevation angles. The process of separating the compounds using KLTP with n-hexane:ethyl acetate (1:1) as an eluent resulted in four bands of active compounds in UV lamps of 254 and 366 nm (TL-1, TL-2, TL-3 and TL-4). Test results for compound 4 isolates of IFATL-05 endophytic fungi from Tunjuk Langit root (Helminthostachys zeylanica (L.) Hook) using the Bioautographic TLC method using n-hexane:ethyl acetate (6:1) as eluent on Escherichia coli, Salmonella thypi, Shigella dysenteriae and Vibrio cholerae with Rf value = 0.90.

Isolation and characterization of endophytic fungi associated with Ephedra alata (macroscopic and microscopic characterization)

Ephedra alata, known in Algeria as Alanda, is a medicinal plant belonging to the Ephedracea family, it looks like a shrub without leaves. This species can grow in semi-arid and desert conditions thanks to its high tolerance to water shortage. It is used in traditional medicine for its therapeutic properties, in particular to treat the bronchi, the circular system, digestive system disorders, kidneys, oedema, fever, headaches, allergies, and to relieve asthma attacks as well as bacterial and fungal infections this is due to its production of secondary metabolites with a wide range of biological activities. On the other hand, This plant represents a host that allows endophytic fungi to feed, protect and spread; in return, this plant also benefits from certain advantages of endophytes. Despite its importance, no studies on its endophytic fungi has been conducted to date. This study aimed to isolate and identify the fungal endophyte community associated with the plant Ephedra alata growing in arid environments. the harvest of the Ephedra alata plant (aerial part) was made in the region of Bechar (southwest algeria). Isolation and purification of fungal strains were carried out on PDA medium supplemented with lactic acid. The identification of the isolated strains was made on the basis of morphological and cultural criteria. Ephedra alata presents a wide diversity of fungal species with a high load. A total of twelve fungal strains were isolated and the predominant genera were Aspergillus, then Penicillium, Mucor, and unidentified structures in small percentages. The most common phylum of fungal genera identified was Ascomycota, followed by Zygomycota and small percentages of unidentified structures (SNI).

Isolation and Characterization of Fungal Endophytes from Petiveria alliacea and Their Antimicrobial Activities in South Florida

Microorganisms associated with medicinal plants are of great interest as they are the producers of important bioactive compounds effective against common and drug-resistant pathogens. The characterization and biodiversity of fungal endophytes of the Petiveria alliacea plant and their antimicrobial production potential are of great interest as they are known for their antimicrobial and anticancer properties. In this study, we investigated the endophytic fungal microbiome associated with P. alliacea, and the endophytic fungal isolates were classified into 30 morphotypes based on their cultural and morphological characteristics. Ethyl acetate extract of fungal endophytes was obtained by liquid–liquid partitioning of culture broth followed by evaporation. The crude extract dissolved in dimethyl sulfoxide was screened for antimicrobial activity against three bacterial strains (Escherichia coli ATTC 25902, Staphylococcus aureus ATTC 14775, Bacillus subtilis NRRL 5109) and two fungal strains (Candida albicans ATTC 10231 and Aspergillus fumigatus NRRL 5109). Among the crude extracts from endophytes isolated from leaves, 65% of them showed antimicrobial activity against the bacteria tested. Similarly, 71 and 88% of the fungal crude extracts from endophytes isolated from root and stem, respectively, showed inhibitory activities against at least one of the bacterial strains tested. Crude extracts (at a concentration of 10 mg/mL) from ten of the fungal isolates have shown a zone of inhibition of more than 12 mm against both Gram-positive and negative bacteria tested. Sequenced data from isolates showing strong inhibitory activity revealed that Fusarium solani, F. proliferatum, and Fusarium oxysporium are the major endophytes responsible for bioactive potential. These results indicate that Petiveria alliacea harbors fungal endophytes capable of producing antimicrobial metabolites. Future studies need to focus on testing against drug-resistant bacteria (ESKAPE group) and other pathogenic bacteria and fungi.

Comparison of Fungal Genera Isolated from Cucumber Plants and Rhizosphere Soil by Using Various Cultural Media.

Plant endophytic fungi and rhizosphere soil fungi are often reported as biocontrol agents against plant pathogens or with plant growth promotion potential. Four treatments were performed in field and greenhouse experiments where cucumber plants were inoculated with Trichoderma harzianum and Fusarium oxysporum in 2022. The roots, stems and leaves of cucumber plants and their rhizosphere soil were collected twice individually from the field and greenhouse for isolation of cucumber endophytic and rhizosphere soil fungi. All fungal strains were identified through sequence similarity of the ITS1-5.8s-ITS2 rDNA region. The potato dextrose agar (PDA) media yielded the highest number of genera isolated from cucumber plants, rhizosphere soil and both compared to other media. There were no significant differences among the four media for the isolation of all cucumber endophytic fungi. However, in the roots, the number of endophytic fungi isolated by MRBA was significantly higher than that isolated on malt extract agar (MEA), while in the stems, the number of fungi isolated with PDA was significantly higher than that isolated with Martin's rose bengal agar medium (MRBA). PDA had significantly higher isolation efficiency for the rhizosphere soil fungi than MRBA. The 28 fungal genera had high isolation efficiency, and the endophytic Trichoderma strains were significantly more isolated by MEA than those of MRBA. It is suggested that PDA can be used as a basic medium, and different cultural media can be considered for specific fungal genera.