Isolation Of Endophytic Fungi Research Articles

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.

Isolation and Phytochemical Testing of Endophytic Fungi from Imperata cylindrica (L.) Beauv plants.

High drug prices and increasing antibiotic resistance have prompted exploration of natural resources as a source of new antibiotics. The Imperata cylindrica (L.) Beauv. plant is one of the plants used in traditional medicine. Secondary metabolites as bioactive compounds in plants can be obtained through endophytic fungi without having to extract them from plants. Secondary metabolites produced by endophytic fungi can function as raw materials for medicines. This research aims to isolate endophytic fungi and test phytochemicals from Imperata plants to determine the type of secondary metabolites produced. The methods used in this research include isolation and purification of endophytic fungi, identification of endophytic fungi, production of secondary metabolites of endophytic fungi, and screening of phytochemicals resulting from endophytic fungi fermentation. The isolation results showed that there were 7 isolates of endophytic fungi, consisting of 1 isolate from the flower, 3 isolates from the stem, and 3 isolates from the roots. Identification of endophytic fungus using macroscopic and microscopic observations including the genera Fusarium sp., Mucor sp., Penicillium sp., Trichoderma sp., Aspergillus sp., Phytophthora sp. The results of the phytochemical test for endophytic fungi showed that the secondary metabolites produced were flavonoid and saponin compounds. Based on this research, it can be concluded that the endophytic fungi of the Imperata cylindrica (L.) Beauv. plant produces secondary metabolites which have the potential to be developed as medicinal raw material compounds.

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.

Antagonistic potential of endophytic fungi against Meloidogyne enterolobii, M. incognita and Nacobbus aberrans sensu lato

Root-knot nematodes limit horticultural crop production in Mexico where management is mainly based on nematicides that represent a risk to health and the environment. Thus, biological alternatives, such as the use of endophytic fungi, are currently being developed. The objective of this research was to isolate, identify and evaluate the antagonistic potential of endophytic fungi against the root-knot nematodes Meloidogyne enterolobii, M. incognita and the false root-knot nematode Nacobbus aberrans sensu lato (s.l.) occurring in Mexico. Endophytic fungi from chili pepper and tomato plants with a low root galling index were isolated from three localities in Sinaloa state (Villa Juárez, Valle de Culiacan and La Cruz de Elota) and one in Mexico City (Xochimilco). Fourteen isolates from leaves and roots were evaluated to assess: i) the ability to promote growth in chili pepper and tomato plants, ii) the degree of parasitism on nematode eggs and second-stage juveniles (J2), and iii) the nematostatic and nematicidal effect of their filtrates on the J2. In the trials, the three most effective endophytic fungal isolates were Chaetomium globosum (S1TH2), Echria macrotheca (S3RT2) and Stagonospora trichophoricola (XRC3). All three fungal isolates increased growth on chili pepper plants after application, but only two (S3RT2 and XRC3) did so on tomato plants. S1TH2 parasitized M. enterolobii eggs; S3RT2 and XRC3 eggs of N. aberrans s.l.; S3RT2 also parasitized M. incognita eggs and J2 of N. aberrans s.l. The S1TH2 filtrate was nematostatic for M. incognita and N. aberrans s.l., the XRC3 filtrate was nematostatic for N. aberrans s.l., and the S3RT2 filtrate was nematostatic and nematicidal for the J2 of N. aberrans s.l. The results obtained indicate that these isolates could be involved in natural regulation and biological control of root-knot nematodes.

Phenolic composition and biological activity of endophytic fungi isolates inhabited <i>Acacia nilotica</i>

This study aimed to screen the phenolic profile, antimicrobial and antiradical activities of endophytic fungi residing in the tissues of <i>Acacia nilotica</i> grown in Sudan. Isolates of endophytic fungi were isolated from fruit, leaf and twig. Methanolic extracts were prepared by maceration. Antimicrobial activity was determined by the disc diffusion method. Antiradical activity was evaluated by the 1, 1-diphenyl-2-picryl hydrazyl (DPPH) assay. Ten endophytic fungi were recovered from the twig (6), leaf (2) and fruit (2) of <i>A. nilotica</i>. Extracts revealed better antifungal activity, with isolates T3, L1 and F1 exerting the highest inhibition zones (20.6, 20 and 18.3 mm, respectively) against <i>Aspergillus niger</i>. Isolates T4 and T5 revealed the highest antiradical effect with IC<sub>50</sub> values of 302 and 478 µg/mL respectively. Chemical analysis revealed that gallic acid and naringenin were the dominant compounds. Their pattern divided the endophytic fungi into two categories; gallic acid- rich endophytes including isolates L1 and L2 (isolated from the leaf) as well as isolates T4 and T2 (isolated from the twig). The other category was naringenin- rich endophytes comprised isolates F1 and F2, which were isolated from the fruit, and isolates T1, T3, T5 and T6 isolated from the twig. Moreover, extracts accumulated gallic acid were devoid of naringenin and the opposite was true. This behavior was not detected in the host plant where these two compounds coexist and identified in all extracts.

Potential of endophytic fungi as a pathogenic biocontrol agent and growth promoters in corn seedlings

BackgroundUse of endophytic fungi, as pathogen control of Bipolaris maydis and Curvularia sp., is an alternative method of control without the use of synthetic pesticides that are more environmentally friendly. This study aimed to determine the potential of endophytic fungi in controlling the growth of pathogens B. maydis and Curvularia sp. in vitro and in spurring the growth of corn plants. It was consisted of three types of testing three endophytic fungal species (Aspergillus_1, Fusarium_2, and Trichoderma_11), namely (1) testing the antagonistic activity of endophytic fungal against pathogens by double culture method, (2) physiological characterization of endophytic fungal as phosphate solvents and chitinase producers, and (3) testing of corn seed vigor with the blotter test method.ResultsThe results of testing endophytic fungal isolates against B. maydis pathogens showed that the three isolates were able to suppress the development of B. maydis, whereas the Trichoderma_11 isolate showed higher suppression results than others. The isolate that showed the best ability to dissolve phosphates is Fusarium_2 with a dissolving index of 1.9 and their effectiveness up to 91.5%. Meanwhile, Trichoderma_11 was able to produce the highest chitinase activity index of 1.9 with an effectiveness of 90.6%. The best corn root lengths and plant height were shown on Fusarium_2 treatment. Similar outcomes were observed when Curvularia sp. was tested. The whole isolates were able to suppress the growth of the pathogen by 16.43–40.44% on the 4th day after incubation. Trichoderma sp. isolate was 72.50% more effective at suppressing than the other two isolates. On day 11, the isolate of Aspergillus sp. was suppressed by 62.50%, while Fusarium sp. showed the lowest suppression of 59.17%.ConclusionsTrichoderma_11 isolate was potentially the best biocontrol agent against maydis leaf blight and Curvularia leaf spot in vitro. Meanwhile, the Fusarium_2 isolate had promoted the growth of the corn seedlings.

Isolation and molecular identification of endophytic fungi associated with Ziziphora tenuior L. and their biological potential

Endophytic fungi are microorganisms found in the inter- or intracellular compartments of plant tissue and can improve plant performance through a variety of mechanisms. They are considered a potential source of biological compounds such as terpenoids, phenols, alkaloids, and fatty acids, which are extensively used in various industries. The present study was conducted to investigate the molecular identification, fatty acid profile analysis, and antibacterial properties of endophytic fungi isolated from the aerial parts of Ziziphora tenuior (L.). Phylogenetic analysis was performed based on the Internal Transcribed Spacer (ITS) region and seven fungal isolates were classified into six genera as follows: Acremonium, Aspergillus, Cladosporium, Neocamarosporium, Alternaria, and Stemphylium. The analysis of fatty acids in the n-hexane extracts of Z. tenuior and mycelia of endophytic fungi with GC device showed that the amount of fatty acids is between 32.7-55.5%. A. infectoria (55.5%) and A. ochraceus (54.3%) had the highest amount of fatty acids among studied isolated. γ–Linolenic acid (C18:3), linoleic acid (C18:2), palmitic acid (C16:0), stearic acid (C18:0), and oleic acid (C18:1) were the main fatty acids. Also, the n-hexane extract of A. ochraceus showed antibacterial activity against Bacillus cereus, B. subtilis, Escherichia coli, and Pseudomonas aeruginosa. The results reveal that endophytic fungi are a source of natural bioactive metabolites, which could be applied in different industries. This is the first report of endophytic fungi diversity in Z. tenuior, which provides knowledge on their future use on biotechnological processes.

Antiphytopathogenic applications of Endophytic Fungus, Talaromyces purpureogenus J8, isolated from Woodfordia fruticosa

The apprehension surrounding the utilisation of pesticides and other chemical agents in agriculture has prompted a search for alternative, non-chemical methodologies for the management of diseases. Biological control agents are highly specialised and pose minimal risk to non-target species. The study involves the isolation of novel endophytic fungi Talaromyces purpureogenus J8 from Woodfordia fruticosa, a medicinal plant with multifarious applications. The culture was identified using ITS sequencing and phylogenetic analysis. The sequence was subsequently deposited into the NCBI database, resulting in the generation of an accession number (OP355300). The culture and the metabolites were evaluated for the potential of biocontrol against different plant pathogens using direct confrontation on solid media and the culture exhibited significant inhibition against Exserohilum turcicum, 84.3% direct confrontation and 78.36% more than the positive control in the case of metabolite. Phytochemical assays such as auxin estimates, total phenolics and flavonoids were carried out. J8 displayed antioxidant activity with an IC50 value of 65.36 μg/mL in the DPPH radical scavenging assay. Cultural conditions were optimised to enhance the production. GC-MS analysis was performed leading to the identification of many bioactive compounds responsible for antibiotic, antioxidant and anticancer properties.

Enhancement of Anti-MRSA Potential Produced by an Endophytic Fungus Ceratobasidium Ramicola IBRLCM127 via Submerged Fermentation System

Introduction: Exploring endophytic fungi isolated from medicinal herbs could be a turning point in the research of secondary metabolites biosynthesis, as these endophytic fungi are capable of synthesizing the similar compounds as their host plant. The advantages of manipulating endophytic fungi for bioactive compound production are the reduction of dependency rate on slow-growing and rare plants, cost-effective, continuous process, environmentally friendly and high yield in a short period. Thus, the current study envisages investigating the influence of culture conditions against the anti-MRSA potential production of the endophytic fungal isolate, Ceratobasidium ramicola IBRLCM127 isolated from the local medicinal plant Curcuma mangga Valeton & Zijp. Methods: The endophytic fungal isolate was used to produce fungal metabolites through submerged fermentation. The physical parameter improvement was investigated using the ‘one-factor-at-a-time’ technique. The fungal fermentative broth was subjected to an anti-MRSA assay using Lorian method, whereas the growth of a fungus was determined based on the cell growth weight. Results: The highest anti-MRSA potential of 42.50±0.1 U/ml and 5.49±0.1 g/L of mycelial growth was observed after improving the basal medium containing yeast extract sucrose broth incorporated with water extract from the host plant, 6 days old of inoculum age, 2 agar plugs of mycelia, incubation temperature of 25 0C and 12 days of cultivation 12 days of cultivation shaken at 120 rpm in the absence of light. Conclusion: The improved culture conditions shorten the incubation period and yield a significant enhancement of anti-MRSA potential and fungal growth with 13.27% and 10.91%, respectively.

Production, bioprocessing and antiproliferative activity of camptothecin from Aspergillus terreus, endophyte of Cinnamomum camphora: restoring their biosynthesis by indigenous microbiome of C. camphora

Fungal producing potency of camptothecin (CPT) raise the hope for their usage to be a platform for industrial production of CPT, nevertheless, attenuation of their productivity of CPT with the subculturing and preservation is the challenge. So, screening for novel endophytic fungal isolates with a reliable CPT-biosynthetic stability was the objective. Among the isolated endophytic fungi from the tested medicinal plants, Aspergillus terreus OQ642314.1, endophyte of Cinnamomum camphora, exhibits the highest yield of CPT (89.4 μg/l). From the NMR, FT-IR and LC–MS/MS analyses, the extracted CPT from A. terreus gave the same structure and molecular mass fragmentation pattern of authentic CPT (349 m/z). The putative CPT had a significant activity against MCF7 (0.27 µM) and HEPG-2 (0.8 µM), with a strong affinity to inhibits the human Topoisomerase 1 activity (IC50 0.362 μg/ml) as revealed from the Gel-based DNA relaxation assay. The purified CPT displayed a strong antimicrobial activity for various bacterial (E. coli and B. cereus) and fungal (A. flavus and A. parasiticus) isolates, ensuring the unique tertiary, and stereo-structure of A. terreus for penetrating the microbial cell walls and targeting the topoisomerase I. The higher dual activity of the purified CPT as antimicrobial and antitumor, emphasize their therapeutic efficiency, especially with growth of the opportunistic microorganisms due to the suppression of human immune system with the CPT uses in vivo. The putative CPT had an obvious activity against the tumor cell (MCF7) metastasis, and migration as revealed from the wound healing assay. The overall yield of A. terreus CPT was maximized with the Blackett-Burman design by twofolds increment (164.8 μg/l). The CPT yield by A. terreus was successively diminished with the multiple fungal subculturing, otherwise, the CPT productivity of A. terreus was restored, and increased over the zero culture upon coculturing with C. camphora microbiome (1.5% w/v), ensuring the restoring of CPT biosynthetic potency of A. terreus by the plant microbiome-derived chemical signals “microbial communication”. This is the first report exploring the feasibility of A. terreus “endophyte of C. camphora” to be a preliminary platform for commercial production of CPT with a reliable sustainability upon uses of indigenous C. camphora microbiome.

Isolation and Identification of Endophytic Fungi of Beech Trees from some Forests in the Suburbs of Sari

Isolation and Identification of Endophytic Fungi of Beech Trees from some Forests in the Suburbs of Sari

Antibacterial Potential of Endophytic Fungi Isolated from Mangrove Rhizophora apiculata Blume Species at Tanjung Api-Api, South Sumatra, Indonesia

Endophytics fungi are found in Rhizophora apiculata mangrove. They are known to produce bioactive compounds that act as an antibacterial, which can be a solution for sustainable use without destroying existing resources. This study aims to determine the types of endophytic fungi on the species of R. apiculata as antibacterial. The method used was the isolation and identification of endophytic fungi from the species of R. apiculata and the antibacterial activity test using the Kirby Bauer method. The results obtained three types of fungi from five pure isolates, namely Aspergillus sp., Cladosporium sp., and Penicillium sp. Three genera of endophytic fungi were found to have antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa bacteria. The rate of inhibition against S. aureus and P. aeruginosa from Cladosporium sp. extract respectively were 17.45 ± 1.15 mm and 2.85 ± 0.08 mm, Aspergillus sp. was 14.61 ± 1.07 mm and 2.6 ± 0.20 mm, and Penicillium sp. was 14.11 ± 0.08 mm and 2.7 ± 0.20 mm. These three isolates showed inhibitory activity against both bacteria. However, the level of inhibition was stronger against S. aureus (gram positive) than P. aeruginosa (gram negative), especially by marine Cladosporium extract found in this study.

Diversity of Endophytic Fungi from Lime Plants (Citrus aurantifolia, Swingle)

Lime (Citrus aurantifolia, Swingle) is a plant with medicinal properties. This plant is needed on a large scale to produce biomass. Medicinal plants with decreasing numbers and populations can be developed through scientific technology by isolating endophytic fungi. Endophytic fungi are microorganisms that are found in plant tissues without causing damage to their host plants. This study uses the exploratory method, namely searching and finding. The purpose of this study was to identify endophytic fungi in lime plants. The study's results obtained ten isolates of endophytic fungi from different plant organs (leaves, stems, bark, roots, fruits). Endophytic fungi were identified through macroscopic and microscopic characteristics and then compared with an identification book of endophytic fungi. Eight genera were found, including Beltrania and Nigrospora on leaves, Fusarium and Chaetomium on fruit, Pythium on stems and bark, Paecilomyces on bark, Mucor and Mortierella on roots.