Endophytic Species Research Articles

2,4-Di-tert-butylphenol (2,4-DTBP) purified from Streptomyces sp. KCA1 from Phyllanthus niruri: Isolation, characterization, antibacterial and anticancer properties

ObjectivesThe present study reports the bioactivity of 2,4-Di-tert-butylphenol (2,4-DTBP) purified from an endophytic Streptomyces species KCA1 isolated from the leaves of Phyllanthus niruri and its antibacterial and anticancer properties. MethodsThe extracellular metabolites were produced from the strain KCA1 by submerged fermentation using ethyl acetate. The crude extract was further evaluated for antibacterial activity against set of pathogens. Active metabolite from the extract was purified using chromatography techniques and detected its antibacterial activity by disc diffusion assay. The molecular structure of the active molecule was identified through various spectral study. Moreover, the bioactive metabolite 2,4-DTBP was analyzed antibacterial and anti-proliferative activity. ResultsStrain KCA1 was identified as Streptomyces sp. In the preliminary screening, the crude extract exhibited broad spectrum activity against various bacterial pathogens. Based on the spectral properties, the active metabolite was identified as 2,4-Di-tert-butylphenol. The MIC of active compound 2,4-DTBP inhibited E. coli ATCC 25922 and S. aureus ATCC 29213 at 50 μg/ml and 0.78 μg/ml, respectively. The IC50 value of 2,4-DTBP was found to be 11.0 µg/ml and 116.8 µg/ml, against breast cancer cell line (MCF7) and normal VERO cell line, respectively. ConclusionsThis study concluded that 2,4-DTBP, produced from the endophytic Streptomyces sp. KCA1, is the potential candidate to develop as promising antibacterial and anticancer agent.

Phytochemical Screening and Bioactivity Studies of Endophytes Cladosporium sp. Isolated from the Endangered Plant Vateria Indica Using In Silico and In Vitro Analysis.

Vateria indica is persistent tree used in Unani sources for the medication and classified as critically endangered. Thus, endophytes for alternative methods to explore these endangered Plants having rich source pharmaceuticals' active molecules for drug development and production. Endophytes comprises unexplored microbes as a potential source of rich pharmaceutically bioactive compounds attributable to their relationship with the host. In the current study, we have isolated endophyte fungi Cladosporium from the plant Vateria indica and performed phytochemical screening of its ethanolic extract to detect the phytochemicals using thin layer chromatography (TLC), gas chromatography-mass spectrometry (GC-MS), high-performance liquid chromatography (HPLC), UV-visible spectrophotometry (UV-VIS), and Fourier transform infrared spectroscopy (FTIR). GC-MS analysis revealed the presence of an anticancer compound hydroxymethyl colchicine, antioxidant compound benzoic acid, and antimicrobial 2-(4-chlorophenoxy)-5-nitro in endophyte fungal extract of plant Vateria indica. Moreover, in silico analysis of bioactive compounds identified by GC-MS analysis using the Autodock Vina and SwissADME confirmed excellent anticancer activity methanone, [4-amino-2-[(phenylmethyl) amino]-5-thiazolyl] (4-fluorophenyl)- and hydroxymethyl colchicine against 6VO4 (Bfl-1 protein) as per Lipinski rule. Furthermore, we also demonstrated the excellent antioxidant of endophytic extract compared to plant extract by DPPH and ABTS assay, as well as antimicrobial activity against both Gram (+ ve) and Gram (- ve) bacteria. Moreover, the endophytic extract also showed its antimitotic activity with a mitotic index of 65.32, greater than the plant extract of 32.56 at 10mg/ml. Thus endophytic fungi Cladosporium species isolated from plant Vateria indica might be used as a potential source for phytochemical anticancer hydroxymethyl colchicine, an antioxidant benzoic acid, and antimicrobial 2-(4-chlorophenoxy)-5-nitro.

Use of endophytic fungi in turfgrasses: Difficulties in delivery to the market

AbstractThe agronomic benefits of endophytic Epichloë fungi for the performance of ryegrass (Lolium spp.) and fescue (Festuca spp.) species were first reported in the 1980s. Various Epichloë endophyte species and strains have been shown to produce a series of different alkaloids with considerable impact on insect pests and grazing animals, some effects on nematodes and diseases, and improvements in plant stress tolerance. However, the impact of these alkaloids on plant performance and turf resilience varies across environments, and often relates to specific insect pressures and stressful summer conditions. Over 40 insect pests and diseases have been documented as responding to endophyte infection.As many Epichloë fungi are seed transmitted, endophyte transmission to the seed is controlled by the levels found in the parent plants and how well the endophyte is maintained during seed production. Endophyte viability in seed lots can drop during storage and transport, especially when stored under high temperatures and humidity. Endophyte growth into seed during seed production can be influenced by the weather, use of fungicides and growth regulators, and the timing of combining after swathing. In the United States, the endophyte infection levels in breeders’ seed of the cultivars used in the National Turfgrass Evaluation Program trials have been reported but currently, no standards exist for labeling cultivars delivered with high endophyte levels. Without suitable protocols and standards, challenges exist in delivering high endophyte infection levels to tall fescue (Festuca arundinacea Schreb.), ryegrass, and fine fescues.

Endophytic Fungal Species in Tropical Trees: A Review

Pathogenic fungi are common in forest ecosystems which cause diseases and sometimes death of plants, while some fungi live inside trees harmlessly without causing issues. Sometimes, plants benefit from the presence of those endophytic fungi, such as gaining resistance to environmental stresses, protection from harmful pathogens etc. Numerous studies have been conducted on such relationships between endophytic fungi and short-term agricultural crops. However, such studies are rare in the literature on tropical tree species which bear timber and non-timber values. This study illustrates the studies conducted on endophytic fungi in tropical trees and explores the potential use of such fungi for obtaining benefits.

Endophytic Fungus from Opuntia ficus-indica: A Source of Potential Bioactive Antimicrobial Compounds against Multidrug-Resistant Bacteria.

Endophytic Aspergillus species represent an inexhaustible source for many medicinally important secondary metabolites. The current study isolated the endophytic Aspergillus niger (OL519514) fungus from Opuntia ficus-indica fruit peels. The antibacterial activities were reported for both Aspergillus species and Opuntia ficus-indica fruit peel extract. Extraction of the endophytic fungal metabolites using ethyl acetate and fractionation was performed, yielding dihydroauroglaucin (C1), isotetrahydroauroglaucin (C2), and cristatumin B (C3). Resistant bacterial strains were used to investigate the efficiency of the total fungal ethyl acetate extract (FEA) and the isolated compounds. FEA showed promising wide spectrum activity. (C3) showed excellent activity against selected Gram-negative resistant bacteria; However, (C2) exhibited tremendous activity against the tested Gram-positive resistant strains; conversely, (C1) possessed the lowest antibacterial activity compared to the two other compounds. An in silico virtual molecular docking demonstrated that cristatumin B was the most active antimicrobial compound against the selected protein targets. In conclusion, the active metabolites newly isolated from the endophytic fungus Aspergillus niger (OL519514) and present in plants’ waste can be a promising antimicrobial agent against multidrug-resistant bacteria.

Bioactive Compounds from Terrestrial and Marine-Derived Fungi of the Genus Neosartorya †

Fungi comprise the second most species-rich organism group after that of insects. Recent estimates hypothesized that the currently reported fungal species range from 3.5 to 5.1 million types worldwide. Fungi can grow in a wide range of habitats, from the desert to the depths of the sea. Most develop in terrestrial environments, but several species live only in aquatic habitats, and some live in symbiotic relationships with plants, animals, or other fungi. Fungi have been proved to be a rich source of biologically active natural products, some of which are clinically important drugs such as the β-lactam antibiotics, penicillin and cephalosporin, the immunosuppressant, cyclosporine, and the cholesterol-lowering drugs, compactin and lovastatin. Given the estimates of fungal biodiversity, it is easy to perceive that only a small fraction of fungi worldwide have ever been investigated regarding the production of biologically valuable compounds. Traditionally, fungi are classified primarily based on the structures associated with sexual reproduction. Thus, the genus Neosartorya (Family Trichocomaceae) is the telemorphic (sexual state) of the Aspergillus section known as Fumigati, which produces both a sexual state with ascospores and an asexual state with conidiospores, while the Aspergillus species produces only conidiospores. However, according to the Melbourne Code of nomenclature, only the genus name Aspergillus is to be used for both sexual and asexual states. Consequently, the genus name Neosartorya was no longer to be used after 1 January 2013. Nevertheless, the genus name Neosartorya is still used for the fungi that had already been taxonomically classified before the new rule was in force. Another aspect is that despite the small number of species (23 species) in the genus Neosartorya, and although less than half of them have been investigated chemically, the chemical diversity of this genus is impressive. Many chemical classes of compounds, some of which have unique scaffolds, such as indole alkaloids, peptides, meroterpenes, and polyketides, have been reported from its terrestrial, marine-derived, and endophytic species. Though the biological and pharmacological activities of a small fraction of the isolated metabolites have been investigated due to the available assay systems, they exhibited relevant biological and pharmacological activities, such as anticancer, antibacterial, antiplasmodial, lipid-lowering, and enzyme-inhibitory activities.

​Diversity of Actinomycetes in Tomato Plants

Background: Tomato plant holds great biodiversity of actinomycetes. This diversity can be explored for new potential actinomycetes strains. Methods: Two different methods were followed for actinomycetes isolation (Serial dilution and direct streaking method). The direct streaking method was found to be the best method where it gave a higher number of actinomycetes compared to the plant extract method. All the actinomycetes were isolated by ISP2 medium supplemented with nystatin and cycloheximide (at 50 µg/ml). All the endophytic actinomycetes strains were isolated and purified based on the difference in appearances such as colony morphology, growth pattern, aerial hyphae growth, filamentous appearance and pigment production. Result: The direct streak method, yielded 192 isolates and only 48 isolates by plant extract method. Thus, a total of 240 strains were isolated. Plants from Madurai yielded a maximum of 130 isolates (54%) and a minimum from Tuticorin 20 isolates (8%). The highest viable count of 80.01±14.24 (CFU g-1) x 103 were recorded in the agro- fields of Dindugal followed by Madurai, Theni, Tirunelveli and Tuticorin. Out of 240 strains, 49.5% exhibited different aerial hyphae and 36.25% showed different filamentous appearances of growth and 5.8% produced pigments. The endophytic actinomycetes species isolated from tomato plants in different locations possess significant morphologically varied actinobacterial diversity. We can explore further on how these diverse actinomycetes are involved in plant growth promotion and protection against pathogens.

Ability Test of IAA (Indole-3-Acetic Acid) Hormone-Producing Endophytic Bacteria from Lamongan Mangrove

Most of the plant-associated bacteria can synthesize active biological components of phytohormones such as auxin. This study aimed to examine the potency of 61 endophytic bacteria isolates from the mangroves at Kutang Beach, Lamongan in producing IAA hormone and to identify types of isolates effecting the concentration of IAA, morphological characteristics of isolates, as well as endophytic bacterial species that have the most potential to produce IAA hormone. Screening of endophytic bacteria isolates was performed using the colorimetric method and the production of IAA was carried out using the spectrophotometric method. IAA production by endophytic bacteria was analyzed descriptively and statistically. One-Way ANOVA was employed to determine the effect of the isolate type on the concentration of IAA. The most potential isolates to produce IAA hormone are identified by 16s rRNA gene marker. The screening results showed that 12 isolates of endophytic bacteria have the potential to produce IAA hormones (2.0-9.3 ppm), coded with LMG 7, 15, 31, 32, 43, 53, 54, 55, 56, 57, 62, and 63. The results of the One-Way ANOVA test suggested that the type of isolates did not affect the concentration of IAA produced by endophytic bacteria. The twelve isolates had different morphological characters and those were Gram-positive bacilli with cell sizes ranging from 1.5 µm - 3 µm. The highest concentration of IAA was produced by LMG 15 (9.3 ppm). LMG 15 was identified as Bacillus cereus strain LMG 15, having 99.33% similarity to Bacillus cereus strain IAM 12605.

A New Species in Pseudophialophora From Wild Rice and Beneficial Potential.

Wild rice (Oryza granulata) is a natural resource pool containing abundant unknown endophytic fungi species. There are few reports on the endophytic fungi in wild rice. Here, one isolate recovered from wild rice roots was identified as a new species Pseudophialophora oryzae sp. nov based on the molecular phylogeny and morphological characteristics. Fluorescent protein-expressing P. oryzae was used to monitor the fungal colonization pattern. Hyphae invaded the epidermis to the inner cortex but not into the root stele. The inoculation of P. oryzae promoted the rice growth, with the growth parameters of chlorophyll content, shoot height, root length, fresh shoot weight, fresh root weight and dry weight increasing by 24.10, 35.32, 19.35, 90.00, 33.3, and 79.17%, respectively. P. oryzae induced up-regulation of nitrate transporter OsPTR9 and potassium transporter OsHAK16 by 7.28 ± 0.84 and 2.57 ± 0.80 folds, promoting nitrogen and potassium elements absorption. In addition, P. oryzae also conferred a systemic resistance against rice blast, showing a 72.65 and 75.63% control rate in sterile plates and potting conditions. This systemic resistance was mediated by the strongly up-regulated expression of resistance-related genes NAC, OsSAUR2, OsWRKY71, EL5, and PR1α. Since P. oryzae can promote rice growth, biomass and induce systemic disease resistance, it can be further developed as a new biogenic agent for agricultural production, providing a new approach for biocontrol of rice blast.

Fungal endophytic community associated with Hevea spp.: diversity, enzymatic activity, and biocontrol potential.

Plants of the genus Hevea present a great diversity of endophytic fungal species, which can provide bioactive compounds and enzymes for biotechnological use, and antagonist agents for plant disease biological control. The diversity of endophytic fungi associated with leaves of Hevea spp. clones in western Amazonia was explored using cultivation-based techniques, combined with the sequencing of the ITS rRNA-region. A total of 269 isolates were obtained, and phylogenetic analysis showed that they belong to 47 putative species, of which 24 species were unambiguous. The phylum Ascomycota was the most abundant (95.4%), with predominance of the genera Colletotrichum and Diaporthe, followed by the phylum Basidiomycota (4.6%), with abundance of the genera Trametes and Phanerochaete. Endophytic composition was influenced by the clones, with few species shared among them, and the greatest diversity was found in clone C44 (richness: 26, Shannon: 14,15, Simpson: 9.11). The potential for biocontrol and enzymatic production of endophytes has been investigated. In dual culture tests, 95% of the isolates showed inhibitory activity against C. gloeosporioides, and 84% against C. cassiicola. Efficient inhibition was obtained with isolates HEV158C and HEV255M (Cophinforma atrovirens and Polyporales sp. 2) for C. gloeosporioides, and HEV1A and HEV8B (Phanerochaete sp. 3 and Diaporthe sp. 4) for C. cassiicola. The endophytic isolates were positive for lipase (69.6%), amylase (67.6%), cellulase (33.3%), and protease (20.6%). The enzyme index ≥ 2 was found for amylase and lipase. The isolates obtained from rubber trees showed good antimicrobial and enzymatic potential, which can be tested in the future for use in the industry, and in the control of plant pathogens.

First report of Cercospora dianellicola as an endophyte of Paspalum notatum (Poaceae)

The aim of this work was to study the fungal diversity in Paspalum notatum, in order to determine the potential biological control of disease-causing pathogens and their effects on growth promotion. Two isolates of Cercospora dianellicola were identified from asymptomatic P. notatum leaves, as an endophytic species. The possible benefits for the host plant remain to be analyzed.

Study the stability of chickpea endophytic actinobacteria species on broth and agar culture media

The actinobacteria acts by colonising plant roots and increases the nitrogen fixation capacity of the rhizobial partner. In this study, endophytic actinobacterial strains CP21A2, CP56, CP84B, and CP200B isolated from chickpea were evaluated for the sporulation rate in solid and liquid media. These resultant spores were evaluated for their stability at different pH and temperature. Calcium carbonate in the liquid broth and MS medium in solid agar media can be used to increase the sporulation rate of the actinobacteria. Additionally, we found out that almost all spore-producing strains were stable at 70°C 4 minutes but temperatures greater than that were lethal to the spores obtained from both types of media. In addition, the tested spore strains were more sensitive and prone to lysis at alkaline pH rather than acidic. Furthermore, our study suggested that CP56 spores obtained from liquid media and CP84B from solid media can be the best performers in promoting the overall growth of plants and nodules. However, further detailed investigations need to be carried out in order to determine their influence on the growth and development of legume plants which can be useful to increase the yield in the agricultural industry.

Novel endophytic species of Coniochaeta and Preussia from the botanical tribe Triticeae in Iran

Novel endophytic species of Coniochaeta and Preussia from the botanical tribe Triticeae in Iran

Fungal Endophytes of Vitis vinifera-Plant Growth Promoters or Potentially Toxinogenic Agents?

Fungal endophytes occurring in grapevine (Vitis vinifera L.) are usually important sources of various compounds with biological activities with great potential for use in agriculture. Nevertheless, many species isolated from this plant belong to the genera Fusarium, Alternaria, or Aspergillus, all of which are well-known to produce mycotoxins. Our study is focused on the assessment of the toxinogenic potential of fungal endophytes isolated from vineyards in the Czech Republic. In total, 20 endophytic fungal species were cultivated in wine must, and 57 mycotoxins of different classes were analysed by liquid chromatography coupled with mass spectrometry. As a result, alternariol, tentoxin, meleagrin, roquefortine C, gliotoxin, and verruculogen were detected in the culture medium, of which verruculogen followed by gliotoxin were the most frequent (present in 90 and 40% of samples, respectively) and most concentrated (up to thousands ng/mL). The alternaria mycotoxins alternariol and tentoxin were detected not only in Alternaria sp. cultures, but traces of these mycotoxins were also quantified in the Diatripe and Epicoccum cultures. Meleagrin and roquefortine C were detected in Didymella sancta and Penicillium crustosum, gliotoxin was detected in Alternaria sp., Didymella sp., Aureobasidium pullulans, Cladosporium herbarum, Penicillium crustosum and Pleurophoma ossicola, and verruculogen was quantified in 99% of endophytic isolates investigated. The potential of endophytes to produce mycotoxins should be carefully checked, specifically in cases where they are intended for the purpose of V. vinifera growing.

Endophytic Colletotrichum Species from Aquatic Plants in Southwest China.

Colletotrichum species are plant pathogens, saprobes, and endophytes in many economically important hosts. Many studies have investigated the diversity and pathogenicity of Colletotrichum species in common ornamentals, fruits, and vegetables. However, Colletotrichum species occurring in aquatic plants are not well known. During the investigation of the diversity of endophytic fungi in aquatic plants in southwest China, 66 Colletotrichum isolates were obtained from aquatic plants there, and 26 of them were selected for sequencing and analyses of actin (ACT), chitin synthase (CHS-1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), the internal transcribed spacer (ITS) region, and β-tubulin (TUB2) genomic regions. Based on morphological characterization and multi-locus phylogenetic analyses, 13 Colletotrichum species were recognized, namely, C. baiyuense sp. nov., C. casaense sp. nov., C. demersi sp. nov., C. dianense sp. nov., C. fructicola, C. garzense sp. nov., C. jiangxiense, C. karstii, C. philoxeroidis sp. nov., C. spicati sp. nov., C. tengchongense sp. nov., C. vulgaris sp. nov., C. wuxuhaiense sp. nov. Two species complexes, the C. boninense species complex and C. gloeosporioides species complex, were found to be associated with aquatic plants. Pathogenicity tests revealed a broad diversity in pathogenicity and aggressiveness among the eight new Colletotrichum species.

Synergistic Association With Root Endophytic Fungi Improves Morpho-Physiological and Biochemical Responses of Chenopodium quinoa to Salt Stress

Symbiotic associations with microbes can contribute to mitigating abiotic environmental stress in plants. In this study, we investigated individual and interactive effects of two root endophytic fungal species on physiological and biochemical mechanisms of the crop Chenopodium quinoa in response to salinity. Fungal endophytes (FE) Talaromyces minioluteus and Penicillium murcianum, isolated from quinoa plants that occur naturally in the Atacama Desert, were used for endophyte inoculation. A greenhouse experiment was developed using four plant groups: (1) plants inoculated with T. minioluteus (E1+), (2) plants inoculated with P. murcianum (E2+), (3) plants inoculated with both fungal species (E1E2+), and (4) non-inoculated plants (E-). Plants from each group were then assigned to either salt (300 mM) or control (no salt) treatments. Differences in morphological traits, photosynthesis, stomatal conductance, transpiration, superoxide dismutase (SOD), ascorbate peroxidase (APX), peroxidase, (POD), phenylalanine ammonia-lyase (PAL), phenolic content, and lipid peroxidation between plant groups under each treatment were examined. We found that both endophyte species significantly improved morphological and physiological traits, including plant height, number of shoots, photosynthesis, stomatal conductance, and transpiration, in C. quinoa in response to salt, but optimal physiological responses were observed in E1E2+ plants. Under saline conditions, endophyte inoculation improved SOD, APX, and POD activity by over 50%, and phenolic content by approximately 30%, with optimal enzymatic responses again observed in E1E2+ plants. Lipid peroxidation was significantly lower in inoculated plants than in non-inoculated plants. Results demonstrate that both endophyte species enhanced the ability of C. quinoa to cope with salt stress by improving antioxidative enzyme and non-enzyme systems. In general, both FE species interacting in tandem yielded better morphological, physiological, and biochemical responses to salinity in quinoa than inoculation by a single species in isolation. Our study highlights the importance of stress-adapted FE as a biological agent for mitigating abiotic stress in crop plants.

Inoculação de sementes com espécies endofíticas de Induratia na produtividade do feijoeiro

ABSTRACT Common bean (Phaseolus vulgaris L.) is a leguminous species of great economic and nutritional importance worldwide, and thus the development of management strategies to promote plant growth and increase grain yield of legumes is of great interest to farmers. The growth-promoting effects of the symbiosis of endophytic microorganisms in plants have been exploited in several crops. This study investigated the effects of Induratia sp. endophytic fungi on the growth and grain yield of seed-inoculated common bean plants. The experiments were conducted in a greenhouse during two growing seasons using a randomized complete block design with three replicates. Growth and yield-related parameters such as plant height, root length, leaf index, number of days for flowering, number of pods per plant, number of grains per pod, mass of 100 grains, and grain yield were evaluated. The grain yield of plants inoculated with Induratia coffeana strains CML 4018 and CML 4020 and Induratia sp. strain CML 4015 increased by 52.5%, 48.9%, and 56.4% compared to the non-inoculated control, respectively. Seed inoculation of common bean plants with endophytic fungi species of the genus Induratia showed a beneficial interaction as indicated by the recorded increase in grain yield. The present pioneering study could provide the means for developing crop management strategies for enhancing common bean crop productivity.

Extracellular Enzymes of Endophytic Fungi Hosted Salt Marsh Plants in the South Eastern, Algeria

Endophyte microorganisms have great biotechnological interest, with features applicable to different areas and are potentially useful in agriculture. In the current study, the most dominant and representative endophytic fungal species of seven halophytic plants prevalent in the southeastern Algeria, Touggourt, were screened for their ability to produce four extracellular enzymes namely: cellulase, amylase, laccase, and lipase. Zygophyllum album came first by hosting diverse endophytic species among all the tested plants with eight species followed by Tamarix boveana and Limoniastrum guyonianum (46.66%) as well as Phragmites communis with 40% and Haloxylon articulatum and Aeluropus littoralis with 33.33%. Unlike J. effusus, it was the less diverse plant where only three species (20%) were identified. In term of enzymatic activities of the selected fungi, the two species of Chaetomium presented cellulase, amylase and lipase activity. Unlike, Trichoderma harzianum involved in lipase, Ulocladium sp. in cellulase, Bipolaris sp. and Botryostimphylium sp. in Amylase. Unlike, the two species of Chaetomium involved in cellulase, amylase and lipase. On the other hand, no enzyme activity was recorded in the colonies of Fusarium sp.

Multigene Phylogeny, Diversity and Antimicrobial Potential of Endophytic Sordariomycetes From Rosa roxburghii.

Rosa roxburghii Tratt. is widely applied in food, cosmetics, and traditional medicine, and has been demonstrated to possess diverse bioactivities. Plant endophytic fungi are important microbial resources with great potential for application in many fields. They not only establish mutualistic symbiosis with host plants but also produce a variety of bioactive compounds. Therefore, in the present study, endophytic fungi were isolated from R. roxburghii, the diversity and antimicrobial activities were evaluated. As a result, 242 strains of endophytic Sordariomycetes were successfully isolated. Multigene phylogenetic analyses showed that these isolates included eight orders, 19 families, 33 genera. The dominant genera were Diaporthe (31.4%), Fusarium (14.4%), Chaetomium (7.9%), Dactylonectria (7.0%), Graphium (4.5%), Colletotrichum (4.1%), and Clonostachys (4.1%). For different tissues of R. roxburghii, alpha diversity analysis revealed that the diversity of fungal communities decreased in the order of root, fruit, stem, flower, leaf, and seed, and Clonostachys and Dactylonectria exhibited obvious tissue specificity. Meanwhile, functional annotation of 33 genera indicated that some fungi have multitrophic lifestyles combining endophytic, pathogenic, and saprophytic behavior. Additionally, antimicrobial activities of endophytic Sordariomycetes against Lasiodiplodia theobromae, Botryosphaeria dothidea, Colletotrichum capsici, Pyricularia oryzae, Rhizoctonia solani, Fusarium oxysporum, Pseudomonas syringae, Pantoea agglomerans, Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa were screened. Dual culture test assays showed that there were 40 different endophytic species with strong inhibition of at least one or moderate inhibition of two or more against the 12 tested strains. The results from the filter paper diffusion method suggested that extracellular metabolites may be more advantageous than intracellular metabolites in the development of antimicrobial agents. Eleven isolates with good activities were screened. In particular, Hypomontagnella monticulosa HGUP194009 and Nigrospora sphaerica HGUP191020 have shown promise in both broad-spectrum and intensity. Finally, some fungi that commonly cause disease have been observed to have beneficial biological activities as endophytic fungi. In conclusion, this study showed the species composition, alpha diversity, and lifestyle diversity of endophytic Sordariomycetes from R. roxburghii and demonstrated these isolates are potential sources for exploring antimicrobial agents.

Production of Extracellular Enzymes, Antimicrobial and other Agriculturally Important Metabolites by Fungal Endophytes of Litsea glutinosa (Lour.) C.B.Rob. a Medicinal Plant

The present paper deals with the endophytic fungi of Litsea glutinosa, a medicinal plant highly valued in Ayurvedic system. A total of 21 fungal endophytes were extracted from different parts of the plant. The following eight species viz, Aspergillus oryzae, A. terreus, Curvularia sp, Fusarium oxysporum, Gliocladium solani, Penicillium citrinum, Trichoderma viride and Verticillium dahliae were selected for testing the production of secondary metabolites including hydrolytic enzymes. All the eight endophytic fungal species produced amylase, cellulase, protease and lipase. Except for Curvularia species, others did not elaborate laccase. All the test fungi produced antibacterial and antifungal metabolites; however, the crude extracts exhibited poor antimicrobial activity. The present endophytic fungi also produced IAA, ammonia and phosphate solubilizing substances. They were also capable to produce IAA with or without adding the tryptophan. An affect was also made to assess the influence of crude extracts on the root growth of maize seedlings. These studies revealed the enhancement effect of extracts on root growth. Based on these observations, discussions and conclusions were made.