Molecular Identification Of Endophytic Fungi Research Articles

Investigation the biological activities and the metabolite profiles of endophytic fungi isolated from Gundelia tournefortii L.

Endophytic fungi are microorganisms that are considered as a potential source of natural compounds, and can be applied in various industries. The aims of this research were molecular identification of endophytic fungi isolated from the Gundelia tournefortii stems, and investigation their biological activities as well as phenolic and fatty acid profile. Surface sterilized stems of G. tournefortii were placed on potato dextrose agar (PDA) to isolate the fungal endophytes. Genomic DNA was extracted by CTAB method, and PCR amplification was performed by ITS 1 and ITS 4 as primers. The enzyme production of endophytic fungi was determined based on the formation of a clear zone that appeared around the colonies of fungus. The anti-oxidant activity was evaluated by measuring the amount of free radicals DPPH. Also, the total phenol and flavonoid contents were measured obtained by Folin-Ciocalteu and aluminum chloride colorimetric methods, respectively. Moreover, the separation and identification of phenolic acids and fatty acids were done by HPLC and GC, respectively. Phylogenetic analysis was done based on the Internal Transcribed Spacer (ITS) region, and five isolates were identified as following: Aspergillus niger, Penicillium glabrum, Alternaria alternata, A. tenuissima, and Mucor circinelloides. Evaluation of the enzymatic properties showed that P. gabrum (31 ± 1.9 mm), and A. niger (23 ± 1.7) had more ability for producing pectinase and cellulase. The anti-oxidant activity of isolates showed that A. alternata extract (IC50 = 471 ± 29 µg/mL) had the highest anti-oxidant properties, followed by A. tenuissima extract (IC50 = 512 ± 19 µg/mL). Also, the extract of A. alternata had the greatest amount of total phenols and flavonoids contents (8.2 ± 0.4 mg GAL/g and 2.3 ± 0.3 mg QE/g, respectively). The quantification analysis of phenolic acid showed that rosmarinic acid, para-coumaric acid, and meta-coumaric acid (42.02 ± 1.31, 7.53 ± 0.19, 5.41 ± 0.21 mg/g, respectively) were the main phenolic acids in the studied fungi. The analysis of fatty acids confirmed that, in all fungi, the main fatty acids were stearic acid (27.9–35.2%), oleic acid (11.3–17.3%), palmitic acid (16.9–23.2%), linoleic acid (5.8–11.6%), and caprylic acid (6.3–10.9%). Our finding showed that endophytic fungi are a source of bioactive compounds, which could be used in various industries. This is the first report of endophytic fungi associated with G. tournefortii, which provides knowledge on their future use on biotechnological processes.

Antibacterial activities and molecular identification of endophytic fungi isolated from mangrove Avicennia marina

Abstract. Mulyani Y, Wulandari AP, Sinaga SE, Safriansyah W, Azhari A, Purbaya S, Abdullah FF, Farabi K, Shiono Y, Supratman U. 2023. Antibacterial activities and molecular identification of endophytic fungi isolated from mangrove Avicennia marina. Biodiversitas 24: 6923-6933. This research explored the antibacterial potential of endophytic fungi from the Avicennia marina's (Forssk.) Vierh leaves, stem bark, and root, located in Blanakan, Subang District, West Java. The screening process for antibacterial activity potential was conducted using the Kirby Bauer disk diffusion method. Subsequently, the molecular screening results of the most promising fungi species were identified using Internal Transcribed Spacer (ITS) markers. The results revealed the isolation of 30 fungal isolates. Among these, seven endophytic fungi exhibited significant antibacterial activity against Staphylococcus aureus ATCC 29213 and Vibrio harveyi ATCC 5339, with inhibition zones ranging from 7.88±1.52 to 23.60±0.77 mm. Through molecular identification, 7 endophytic fungi were identified, including Penicillium chrysogenum, Cladosporium anthropophilum, Trichosporon asahii, Cladosporium sphaerospermum, Fusarium verticillioides, Meyerozyma carpophila, and Penicillium steckii. Notably, C. anthropophilum exhibited the highest inhibition zones against S. aureus and V. harveyi measuring 23.60±0.77 and 21.80±0.26 mm, and showed (Minimum Inhibitory Concentration) MIC values of 15.625±0.98 and 31.25±0.39 ?g/mL, respectively. In this study, endophytic fungi isolated from different parts of A. marina exhibited promising antibacterial activity, with Cladosporium anthropophilum from the stem bark showing the highest potency against S. aureus and V. harveyi, suggesting their potential as a source of antibacterial agents. To the best of our knowledge, this is the inaugural study uncovering the isolation and antibacterial potential of endophytic fungi from the genera Cladosporium, Trichosporon, Fusarium, and Meyerozyma. These fungi were extracted from the bark, leaves, and roots of A. marina, situated within the unique mangrove ecosystem of Blanakan Sub-district, Subang District, West Java, Indonesia.

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.

Molecular Identification of Endophytic Fungi from Javanese Ginseng (Talinum paniculatum Gaertn.) and their Potential against Phytopathogens

Endophytes from medicinal plants have been demonstrated to produce bioactive chemicals with antibacterial capabilities. Previous research found many endophytes in Javanese ginseng roots. Identifying and comprehending the benefits of endophyte microorganisms is critical for boosting plant growth and defense. The aim of this study was to identify endophytic microbes from Javanese ginseng with antifungal activity. In this present study, two isolates the GJ-9 and FN-1, were analyzed using NL1/NL4 and ITS1/4 primers. The amplification and sequencing of the ITS and the D1-D2 LSU rRNA gene were performed. The D1-D2 LSU rRNA sequence analysis results showed that GJ-9 has similarities with Candida Spencermartinsiae with 99.82% percentage identity, while FN-1 has been identified as Aspergillus flavus according to the ITS sequence analysis result with 99.83% percentage identity. Antagonistic assays showed both isolates displayed considerable antagonistic ability against Rhizopus stolonifera, Pythium sp., Botrytis sp., and Fusarium sp, with overall inhibition rate were over 50%. The Aspergillus flavus showed the better ability to inhibit the growth of Phytium sp. (74.90 %) and Rhizopus stolonifera (60 %), while the Candida Spencermartinsiae is better for controlling Botrytis sp. (55.68%) and Fusarium sp. (60.39 %). The result suggests the endophyte from Javanese ginseng can be used as an agent for controlling various plant diseases; furthermore, it could lead to a reduction of agrochemical inputs.

Screening and molecular identification of endophytic fungi promoting accumulation of flavonoids in callus of Scutellaria baicalensis

To screen and identify the endophytic fungal strains that could promote the accumulation of flavonoids in the callus of Scutellaria baicalensis. Seventeen endophytic fungal strains from S. baicalensis were used to prepare mycelium elicitors and fermentation broth elicitors. Their effects on flavonoid accumulation in S. baicalensis callus were then determined. The results showed that the fermentation broth elicitors of two strains(CL79, CL105) promoted the accumulation of flavonoids. The fermentation broth elicitor of CL79 significantly promoted accumulation of baicalin, wogonoside, baicalein, and wogonin, with the maximum levels increased by 37.8%, 40.4%, 44.7%, and 42.2%(vs. blank), respectively. Similarly, the fermentation broth elicitor of CL105 significantly promoted the accumulation of baicalin, wogonoside, baicalein, and wogonin, with the maximum levels increased by 78.1%, 140.9%, 275.6%, and 208.5%(vs. blank), respectively. CL79 was identified as Alternaria alternata, and CL105 as Fusarium solani. The fermentation broth elicitors of A. alternata CL79 and F. solani CL105 were able to promote the flavonoid accumulation in the callus of S. baicalensis, which enriched the resources of endophytic fungi and provided candidate strains for the development of microbial fertili-zers for improving the quality of S. baicalensis.

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.

Fermentation, bioactivity and molecular identification of endophytic fungi isolated from mangrove Ceriops tagal

Abstract. Putra IPYA, Utami KS, Hardini J, Wirasuta IMAG, Ujam NT, Ariantari NP. 2023. Fermentation, bioactivity and molecular identification of endophytic fungi isolated from mangrove Ceriops tagal. Biodiversitas 24: 3091-3098. Mangrove-associated-endophytic fungi are recognized as a considerable source of bioactive compounds in drug discovery. This study evaluated the antimicrobial activity and toxicity of endophytic fungi isolated from Ceriops tagal. Four endophytic fungal isolates, documented as SU-1-4, were obtained. In addition, isolate SU-3 was further identified as Hypoxylon mangrovei according to the comparison of the internal transcribed spacer region. Each fungal isolate was fermented on a solid rice medium, then extracted with ethyl acetate at the end of fermentation. After that, the crude ethyl acetate extracts were partitioned between aqueous methanol and n-hexane, and the methanolic phases were subjected to phytochemical screening and antimicrobial and toxicity assays. Preliminary phytochemical identification suggested the presence of alkaloids, triterpenoids, and steroids in all fungal methanolic extracts. Among them, H. mangrovei SU-3 extract showed the most pronounced antimicrobial effects with minimum inhibitory concentration (MIC) values of 125 and 250 µg.mL-1 towards Staphylococcus aureus ATCC 6538 and Staphylococcus epidermidis ATCC 12228. Meanwhile, the toxicity assay employing the brine shrimp lethality test revealed that all fungal methanolic extracts were toxic, with LC50 values ranging from 173.09 to 227.76 µg.mL-1. Isolation of antibacterial compounds and evaluation of their cytotoxicity on human cancer cells will be promising for further studies.

Morphological and Molecular Identification of Endophytic Fungi from Roots of Tomato and Evaluation of Their Antioxidant and Cytotoxic Activities

Morphological and Molecular Identification of Endophytic Fungi from Roots of Tomato and Evaluation of Their Antioxidant and Cytotoxic Activities

Molecular identification of endophytic fungi in lawn grass (Axonopus compressus) and their pathogenic ability

Lawn grass (Axonopus compressus) is a widely distributed grass species from the family Poaceae that is ubiquitous in Malaysia. We isolated endophytic fungi from the leaves of A. compressus and molecularly identified them as Fusarium parceramosum, Colletotrichum siamense, C. gigasporum, C. endophyticum, Curvularia lunata, Stagonospora bicolor, Calonectria gracilis, and Albifimbria verrucari. These fungal endophytes are considered host generalists, as they have been isolated from other plants and have also been reported to be latent plant pathogens. We tested the pathogenicity of selected endophytic fungal isolates on A. compressus leaves, chili (Capsicum annum), and tomato (Solanum lycopersicum), and found that they were pathogenic to wounded A. compressus leaves with low to moderate virulence, and several were pathogenic to wounded and unwounded chili and tomato fruits. This indicated that the endophytes could infect both vegetable fruits with low to very high virulence. Pathogenicity tests demonstrated that endophytic fungi from the leaves of A. compressus can become pathogenic and infect the host and other plant species. The findings also indicated that leaves of A. compressus may harbor pathogens with latent ability that can become active due to changes in environmental conditions, thereby disrupting the balance between host-endophyte antagonism.

Molecular Identification of Fungal Endophytes of Medicinal Plant Citrullus colocynthis (L.) Schrad as a Medicinal Plant: Role of Tissue Type and Sampling Location on the Diversity.

Endophytic fungi are an important group of organisms in association with plants which are able to colonize all plant internal tissues and improve their fitness. The present research aims to isolate and identify endophytic fungi of Citrullus colocynthis plant and then investigate the effects of sampling location and tissue type on the fungal endophyte diversity of this plant. To do so, a sampling program was done in 11 geographically isolated C. colocynthis growing areas of Hormozgan province, Iran. For molecular identification of endophytic fungi of C. colocynthis, the internal transcribed spacer region (ITS1-5.8S-ITS4), as a universal DNA barcode marker for fungi, was amplified using primer sets. Totally, 12 taxa (Alternaria solani, Cladosporium halotolerans, Setosphaeria rostrata, Aspergillus niger, A. allahabadii, A. terreus, A. occultus, A. cristatus, Penicillium chrysogenum, Talaromyces purpureogenus, Fusarium sp., and Pseudozyma flocculosa) were isolated. Our findings also showed that the diversity of fungal endophytes isolated from C. colocynthis was affected by the tissue type and sampling site. Accordingly, the leaves and seeds were found to have the highest and lowest rates of endophyte colonization and richness in all sampling seasons, respectively. Simpson's diversity index of 0.8165 in root tissue indicated the high diversity of endophytes in this organ. In addition, Shannon's diversity index in the root (1.846) was higher than that in the other organs. The highest Shannon's and Simpson's indices were observed in Khoon Sorkh and Minab regions. Generally, at least two factors (region and type of tissue) played the most important roles in determining the composition of fungal endophytes in C. colocynthis.

Molecular identification of endophytic fungi associated with Coleus forskohlii (Willd.) Briq.

Bioactivity assessment of endophytic fungi associated with Centella asiatica and Murraya koengii Archana Nath, Jyoti Pathak and SR Joshi

Isolation, antibacterial activity, and molecular identification of endophytic fungi from Pogostemon cablin

Pogostemon cablin is the medicinal plants that produces patchouli essential oils as secondary metabolites which has multiple functions including antibacterial ability. The secondary metabolites in plants mostly associated with their endophytic fungi. In this study we isolated endophytic fungi from Pogostemon cablin’s leaves and examined antibacterial activity of the endophytic fungi against Escherichia coli and Staphylococcus aureus, as well as find out the identity of most potential isolate based on Internal Transcribe Spacer (ITS) region. The isolation of endophytic fungi was performed using surface sterilization method on Malt Extract Agar (MEA) medium. The antibacterial activity was tested using paper disc on Muller Hinton Agar (MHA) medium and molecular identification was amplified using ITS 4 and ITS 5 primers. The isolation process resulted in 5 isolates of endophytic fungi. The antibacterial assay indicated one potential isolate with the highest antibacterial activity when tested against E. coli and S. Aureus, exhibited 20.9 mm and 19 mm clear zone respectively. Molecular identification from ITS region database depicted that the potential isolate has high homology with Nigrospora sp. by 99% similarity. This result suggested that the antibacterial ability of essential oils from the Pogostemon cablin’s leaves might has high correlation with the occurrence of endophytic fungi.

Molecular identification of endophytic fungi associated with orchids from Mount Cameroon region

The orchidaceae (orchid family) is the second largest family of flowering plants after the Asteraceae. Orchids are important in herbal medicine, the food industry, perfumery industry and as ornamentals. They rely on mycorrhizal fungi to provide the carbon sources for seed germination and seedling establishment. The Mount Cameroon Region has a rich diversity of orchids which is under threats from land use patterns. This study was aimed at (i) identifying fungi associated with orchid mycorrhiza; and (ii) identifying non-mycorrhizal endophytic fungi. Nine species of orchids were selected for this study, three each from the different life forms. Selection criteria were based on vulnerability, scarcity and abundance. Mycorrhizal and non-myccorrhizal endophytic fungi were isolated from dissected single pelotons and from non-peloton root tissues respectively. Identification of fungi was based on morphological and sequencebased molecular methods. A total of 18 fungi species belonging to 12 genera were identified with Penicillium being the most abundant. The inferred phylogenetic tree grouped all endophytes into 9 major clusters belonging to 2 phyla. Clustering was independent of whether endophytes were mycorrhizal or non-mycorrhizal. The results of this study could contribute to orchid conservation and for the discovery of bioactive compounds.

Molecular identification of endophytic fungi from torch ginger (Etlingera elatior) antagonist to phytopathogenic fungi

Abstract. Lutfia A, Munir E, Yurnaliza. 2020. Molecular identification of endophytic fungi from torch ginger (Etlingera elatior) antagonist to phytopathogenic fungi. Biodiversitas 21: 2681-2689. Medicinal plants are valuable sources in exploration of endophytic microorganisms. The utilization of culturable endophytes, particularly endophytic fungi in pharmacological aspects has been reported in many literature. Zingiberaceae, being known as one of medicinal plant families, is commonly used in tropical countries, e.g Indonesia to treat health disorders and ailments. However, little information has been reported upon their associative endophytic fungi that may be used as biocontrol agents against phytopathogenic fungi. Efforts to elaborate on the symbiotic relationship between endophytic fungi and host to explain the antifungal ability from endophytes are presented in this study. Six fungal morphotypes were recovered from the rhizome fragment of torch ginger (Etlingera elatior (Jack) RM Smith) sampled from Sibayak forest, North Sumatra, Indonesia. All isolates displayed potential antifungal activities against Fusarium oxysporum, Ganoderma boninense, and Rigidoporus lignosus as representative phytopathogens in a dual culture plate assay. All fungal morphotypes were identified based on their 18S rDNA-ITS regions. Molecular identification revealed that most isolates were related to Trichoderma spp. while one isolate was identified as Pestaliotopsis sp. The endophytic assemblage of mostly Trichoderma strains in our study may reveal a mutual relationship of these soil fungi as root colonizer displaying biocontrol properties against phytopathogenic fungi to host.

Molecular Identification of Endophytic Fungi Isolated from Medicinal Plant

Endophytes are microorganisms that colonize the tissues of live host plants usually showing no apparent symptoms. These endophytes, both bacteria and fungus, can produce a variety of secondary metabolites having a wide range of essential properties. Thus, they can be used for various medicinal, environmental and agronomic purposes. Argemone mexicana, a plant of the family Papaveraceae is widespread across India, and blooms in the month of March. The study was carried out to isolate endophytic fungus living in the shoot and root areas of A. mexicana from Northeast India, and to identify and characterize these endophytes. A total of 20 types of fungal species were isolated from both the root and shoot parts of the plant. Endophytes were identified by 18SrDNA sequencing using ITS1 and ITS4 primers. Genomic DNA was isolated using Cetyl trimethyl ammonium bromide method of the endophytes. The endophytic fungi were identified as belonging to Aspergillus and Penicillium sps by using sequence analysis of the internal transcribed spacer region and Basic local alignment search tool. Aspergillus oryzaem A. niger, and A. flavus showed 100% relative abundance in root and shoot. A. versicolor, A. sydowii, Penicillium chrysogenum were found in root only and A. striatus, A. tubingensis, Emericella qinqixianii and E. striata were found in the shoot of A. mexicana.

Molecular Identification of Endophytic Fungi from Local Rice and Growth Test on Several Types of Culture Media

Local rice is rice that has been cultivated for generations by the community and commonly cultivated without using chemical inputs. Endophytic fungi are fungi that live in the plant tissue and does not cause disease symptoms in the host plants. This study aimed to molecular identifying isolates of MDTA and MDTB endophytic fungi which have been isolated from the local Pulu Mandoti rice plant tissue and growth test on the four types of culture media those were synthetic PDA, natural PDA, MPA, and MEA. The fungi DNA isolation using DNesay Kit. DNA sequencing analysis using the mega BLAST program showed that the MDTB fungus has similarities to Podoscypha bolleana strain 32034 no accession JQ675334 and Podoscypha bolleana strain 32032 no accession JQ675332, whereas the MDTA fungus has similarities to Coprinopsis cinerea A2S3-5 isolate and Coprinopsis cinerea strain CNRMA / F 07-32. The best culture media and sporulation of endophytic fungi is MPA media. This research is the first study to molecular identifying with endophytic fungi from local rice and viability test on the four types of culture media. The results of this study contribute to the diversity of local rice endophytic fungi in Sulawesi.

Molecular identification and phytochemical screening of endophytic fungi isolated from Lithospermum officinale L. roots: A new source of shikonin

Endophytic fungi are microorganisms located in the inter- or intracellular compartments of plant tissues but with no harmful effects. They are considered a potential source of biological compounds. The present study was conducted to investigate the molecular identification of endophytic fungi isolated from the roots of Lithospermum officinale and their potential production of shikonin. Phylogenetic analysis was performed based on the Internal Transcribed Spacer (ITS) region and the isolates were classified into five genera as follows: Alternaria, Chaetosphaeronema, Fusarium, Mucor, and Trichoderma. The study on the methanol extracts of endophytic fungi indicated that total polyphenol content had a positive relationship with antioxidant activities and the highest antioxidant activity belonged to the methanol extracts of Fusarium tricinctum and Alternaria altenata. Then, to investigate the ability of the fungal isolates to produce shikonin, a naphthoquinone compound with high biological activity, the extracts were subjected to HPLC. The results obtained from HPLC-mass spectrometry showed that shikonin could be produced only by F. tricinctum. Thus, F. tricinctum isolated from the roots of L. officinale can be presented as a new source of shikonin.

Molecular Identification of Endophytic Fungi from Banana Leaves (Musa spp.).

Endophytic fungi are part of microbial community found in various types of plant tissues including the leave, and display a range of symbiotic interactions with the plant host. In this study, endophytic fungi isolated from banana leaves were identified using ITS (Internal Transcribed Spacer region) sequences of which 10 genera comprising 17 species were molecularly identified. Endophytic fungal species identified were Nigrospora oryzae, Nigrospora sphaerica, Colletotrichum gloeosporioides, Colletotrichum siamense, Fusarium equiseti, Fusarium chlamydosporum, Phoma sorghina, Pestalotiopsis oxyanthi, Pestalotiopsis theae, Pestalotiopsis eugeniae, Penicillium steckii, Penicillium purpurogenum, Bipolaris papendorfii, Bipolaris sp., Lasidiodiplodia theobromae, Cochliobolus intermedius dan Aspergillus niger. The present study showed that several endophytic fungal genera/species are common plant pathogen and there is a possibility that these endophytes can become pathogenic. Some of the fungal endophyte might be mutualist or saprophyte.

Molecular Identification of Endophytic Fungi from Bark of Raru (Cotylelobium melanoxylon) that Produce the Antibacterial Compounds

<p class="IsiAbstrakIndo"><span lang="EN-GB">The Dipterocarpaceae plant, raru (</span><em><span lang="EN-GB">Cotylelobium melanoxylon</span></em><span lang="EN-GB">) is widespread in Southeast Asia. The bark of raru has been used by local communities in North Sumatera as antidiarrheal drugs due to its antibacterial compounds. The antibacterial activity of the </span><em><span lang="EN-GB">raru’s</span></em><span lang="EN-GB"> bark is induced by endophytic fungi that live in the region of the bark. This study aimed to identify the endophytic fungi-producer of antibacterial compounds in the bark of raru (</span><em><span lang="EN-GB">C. melanoxylon</span></em><span lang="EN-GB">) by means of molecular analysis. In general, endophytic fungi have the ability to inhibit the growth of pathogenic bacteria. Thirty-eight isolates of endophytic fungi were isolated from the bark of raru. Selection of isolates for antibacterial activity against </span><em><span lang="EN-GB">Escherichia coli</span></em><span lang="EN-GB"> ATCC 35218 and </span><em><span lang="EN-GB">Staphylococcus aureus</span></em><span lang="EN-GB"> ATCC 25923 used the dual culture assay. Selection using the dual culture assay yielded 6 endophytic fungal isolates that have the ability to inhibit the growth of test bacteria. EF10A sample was the most powerful isolate inhibiting the growth of both bacteria test. Those six bacteria molecularly identified used a sequence generated from ITS rDNA region. Based on rDNA ITS region sequences, isolate, the producers of the antibacterial compound were identified as </span><em><span lang="EN-GB">Talaromyces cellulolyticus, Penicillium purpurogenum, Aspergillus</span></em><span lang="EN-GB"> sp., </span><em><span lang="EN-GB">Trichoderma harzianum</span></em><span lang="EN-GB">, and </span><em><span lang="EN-GB">Aspergillus orizae</span></em><span lang="EN-GB">. The results of this study can be used by researchers to explore more potential endophytic fungi in raru plants (</span><em><span lang="EN-GB">C. melanoxylon</span></em><span lang="EN-GB">) as a source of medicine. The data obtained need to be supported by further research to isolate the bioactive compounds that can inhibit the growth of microbial pathogens.</span></p>

SCREENING AND IDENTIFICATION OF HEAVY METAL-TOLERANT ENDOPHYTIC FUNGI LASIODIPLODIA THEOBROMAE FROM BOSWELLIA OVALIFOLIOLATA AN ENDEMIC PLANT OF TIRUMALA HILLS

ABSTRACTAim: The aim of this study was to evaluate the heavy metal resistance potentiality of endophytic fungi isolated from the leaves of Boswellia ovalifoliolata,an endemic medicinal plant of Tirumala Hills.Methods: Initially, isolation of fungal endophytes was carried out. Isolated fungi were screened for the heavy metal resistance against Co, Cd, Cuand Zn using growth and evaluated their maximum tolerant capacity. Molecular identification of endophytic fungi was carried out by 18S rRNA geneamplification and Sanger’s nucleotide sequencing. Phylogenetic tree was constructed using NCBI Clustal W.Results: Ten different endophytic fungi were isolated from the leaves of B. ovalifoliolata. Among the isolated endophytic fungi, five showed resistanceto Co, Cd, Cu, and Zn. The most resistant fungus was identified as Lasiodiplodia theobromae based on 18S rRNA gene sequencing.Conclusions: L. theobromae was isolated from B. ovalifoliolata and identified as one of the useful fungi involved in mycoremediation against heavymetal toxicity.Keywords: Heavy metals, Endophytic fungi, Endemic plant, Bioremediation.