Root-associated endophytic fungi modulate endogenous auxin and cytokinin levels to improve plant biomass and root morphology of trifoliate orange

Root-associated endophytic fungi like Serendipita indica and arbuscular mycorrhizal(AM) fungi can improve plant growth and root construction, but the potential mechanism is unclear. In this study, Funneliformis mosseae(an AM fungus) and S. indica, singly or in combination were inoculated into trifoliate orange(Poncirus trifoliata) seedlings, to assess changes in biomass and root morphological traits, coupled with auxins and cytokinins concentrations in leaves and roots and the expression of auxin synthesis and transporter protein genes. After 20 weeks of inoculation with these fungi, shoot and root biomass, root total length, taproot length, average diameter, surface area, volume, and the number of lateral roots in 1st-, 2nd-, and 3rd-order were improved, and S. indica showed a relatively greater effect than F. mosseae and dual inoculation. Endophytic fungal inoculation also significantly increased the concentration of indoleacetic acid, indole butyric acid, trans-zeatin,dihydrozeatin, and isopentenyl adenine in leaves and roots, whilst F. mosseae and S. indica exhibited relatively greater effects on leaves and roots, respectively. Correlation analysis revealed that both biomass and root morphological traits(except root projected area) were significantly positively associated with endogenous auxins and cytokinins. In addition, the inoculated plants recorded comparatively higher expression levels of indoleacetic acid synthesis genes(PtTAA1, PtTAR2, PtYUC3, PtYUC4, PtYUC6, and PtYUC8) and indoleacetic acid transporter protein genes(PtAUX1, PtLAX1, PtLAX2, PtLAX3, PtPIN1, PtPIN3, PtPIN4, PtABCB1, and PtABCB19) than the non-inoculated plants, among which F. mosseae and S. indica showed better effects in leaves and roots, respectively. These results suggest that root-associated endophytic fungi improved plant growth and root architecture, which were associated with changes in endogenous auxins and cytokinins.

Isolation and Screening of Black Spot-Resistant Endophytic Fungi Strains from Rosa chinensis

[Objective] This study aimed to isolate and screen black-spot-resistant en- dophytic fungi strains. [Method] Various species of healthy Rosa chinensis were col- lected from Xi＇an, Xianyang, Baoji and Weinan City in Shaanxi Province. Endophytic fungi was isolated from stems and leaves, and purified by 2-3 times of inoculation to screen endophytic fungi antagonistic to Marssonina rosae with modified punching method. [Result] Samples collected from Xianyang exhibited the highest colonization rate and isolation rate; endophytic fungi strains isolated from stems presented the highest colonization rate and isolation rate compared with leaves. A total of 67 en- dophytic fungi strains were isolated from Rosa chinensis, including 3 black spot-re- sistant strains which were all derived from Baoji. [Conclusion] This study laid the foundation for further screening candidate strains of biocontrol fungi and environ- ment-friendly fungal biological control.

Separation and Purification of Antioxidant Endophytic Fungi from Stevia Leaves

[Objective] The aim was to separate and purify antioxidant endophytic fun- gi from Stevia leaves. [Method] Stevia leaves were used as materials for endophytic fungi screening and purification. Then, liquid culture was conducted to test fermented supernatant abilities of antioxidant, DPPH free radical clearing and chemilumines- cence inhibition. [Result] Forty two endophytic fungi were separated from F01 to F42. F41 was proved to be the strongest in antioxidant aspect; F37 was the strongest in DPPH clearing; F02, F07, F20, F25, F37, and F416 stood out in a comprehensive comparison of the above aspects. Flow injection chemiluminescenct method was used to test the above six fermentation liquors, and the results showed that antioxidant abilities of the six were quite strong for all of chemiluminescence rates were over 70%. Through preliminary morphological identification, bacterial colony was found white or yellowy and only F02 was of dissepiment. [Conclusion] Fungi with high-antioxidation activities were separated, which paved the way for future research.

Identification of Taxus cuspidata Sieb.et Zucc endophytic fungi─ new recorded-genus-species of China and the metabolite

A total of 94 isolates of endophytic fungi were isolated from the bark of 200-yr.-old Taxus cuspidata Sieb. et Zucc. in the primeval forest of the Changbai Mountain Natural Reserve, and 19 species of endophytic fungi were identified. Among the taxa 2 new species and 1 new varied species as well as the 6 known species of China had been described in a previous article published on Journal of Forestry Research, Vol. 14 (4): 290-294 (2003). This paper made a detail description on the morpho-logical characteristics of the 9 new recorded-genus-species and 1 new record species with illustrations, including Staphy-lotrichum coccosporum Meyer et Nicot, Botryodema lateritium Papendoorf & Upadhyay, Oedocephalum glomerulosum (Rul-liard) Sacc., Trichosporonoides oedocephalis Haskins & Spencer, Beniowskia sphaeroidea (Kal Chbrenner et Caeke) Mason, Thermomyces verrucosus Pugh, Blakeman & Morgan-jones, Gilmaniella humicola Brron , Arthrinium sphaerosperma (Per.:Fr.) Ellis, Ambrosiella brunnea (Verrall) Batra, and Cytosporina notha Diedz (1 new record species). And the habitats, hosts, distribu-tions and fruiting periods were recorded for the new recorded-genus-species. The determination result of thin layer chroma-tography (TLC) showed that the liquid fermentation metabolites of the strains could not react positively with taxoids developer - the vanillic aldehyde and dilute bismuth potassium iodide.

Effect of Endophytic Fungi AL12 on the Metabolites Distribution in Organs of Atraetylodes lancea

[Objective] To examine the effect of endophytic fungi ALl2 （Gilmaniella sp.） on metabolites distribution in organs of Atractylodes lancea. [Method] Endophytic fungi ALl2 was inoculated on Atraetylodes lancea plantlets in tissue culture, and the distribution of cellulose, hemicellulose, lignin, soluble sugar in leaves and roots of the inoculated group were detected. The weight of leaves and roots were compared. Gas Chromatography was used to analyze the volatile oil components. [Result] Compared with the control group, the average fresh weight and dry weight of leaves and roots of A. lancea which had been symbiosed with ALl2 increased significantly. The content of lignin and soluble sugar increased in the leaves of the inoculated group, and the content of cellulose, hemicellulose, lignin, soluble sugar and volatile oil also increased in roots. [Conclusion] The results indicate that symbiosed with ALl2 is benefit for the development of A. lancea roots and can promote the transfer and accumulation of the medicinal components to the roots.

Identification and Bioactivities of Secondary Metabolites Derived from Endophytic Fungi Isolated from Ethnomedicinal Plants of Tujia in Hubei Province:A Review

Tujia is a national minority,inhabiting in the mountainous Wuling area in China.Since 1978,Tujia medicine has been studied,summarized and developed,leading to numerous achievements by Chinese researchers,such as the publishing of approximately 30 monographs of Tujia medicine.These publications are focused on summarizing and improving the theory of Tujia medicine and developing clinical therapies from this system of medicine.The shortage of natural medicinal plants used in Tujia medicine has created the need to discover new resources to replace them and protect endangered natural plant species.Endophytic fungi are one of the conservation options,are considered a source of new bioactive natural products,and are a renewable and inexhaustible source of new drugs and agrochemicals.This review summarizes 260 compounds from endophytic fungi that have been previously isolated from the medicinal plants of Tujia.These compounds include steroids,terpenoids,meroterpenoids,polyketides,alkaloids,peptides,aliphatic compounds,aromatic compounds,and heterocyclic compounds.

A pilot study of antioxidant potential of endophytic fungi from some Sudanese medicinal plants

Objective: To evaluate the total phenolic content and total antioxidant capacity of ethyl acetate extracts of 21 endophytic fungi isolated from five Sudanese medicinal plants: Calotropis procera, Catharanthus roseus, Euphorbia prostrate, Vernonia amygdalina and Trigonella foenumgraecum. Methods: Crude extracts of endophytic fungi and their host plants were tested by classical Folin-Ciocalteu colorimetric method to determine the total phenolic content, also total antioxidant capacity was estimated using 1,1-diphenyl-2-picrylhydrazyl free radical scavenging in vitro method. Results: Among the endophytes, endophytic fungus Aspergillus sp. from Trigonella foenum-graecum seeds demonstrated the highest both total phenolic content in term of gallic acid equivalent(GAE) [(89.9±7.1 mg GAE/g)] and antioxidant activity for 1,1-diphenyl-2-picrylhydrazyl radical scavenging assay [IC50:(18.0±0.1 μg/m L)]. A high positive linear correlation(R2=0.999 1) was found between total antioxidant capacity and total phenolic content of endophytic fungi isolated from Vernonia amygdalina. Conclusions: The present study revealed that some endophytic fungi from the five Sudanese medicinal plants could be a potential source of novel natural antioxidant compounds.

Levels of rhizome endophytic fungi fluctuate in Paris polyphylla var.yunnanensis as plants age

Paris polyphylla var. yunnanensis is an important medicinal plant with abundant saponins that are widely used in the pharmaceuticals industry. It is unclear why the levels of active ingredients increase as these plants age. We speculated that the concentrations of those components in the rhizomes are mediated by fungal endophytes. To test this hypothesis, we took both culture-dependent and-independent(metagenomics) approaches to analyze the communities of endophytic fungi that inhabit those rhizomes in plants of different age classes(four, six, and eight years old). In all, 147 isolates representing 18 fungal taxa were obtained from 270 segments(90 per age class). Based on morphological and genetic characteristics, Fusarium oxysporum(46.55% frequency of occurrence) was the predominant endophyte,followed by Leptodontidium sp.(8.66%) and Trichoderma viride(6.81%). Colonization of endophytic fungi was maximized in the eight-year-old rhizomes(33.33%) when compared with four-year-old(21.21%) and six-year-old(15.15%) rhizomes. Certain fungal species were present only at particular ages. For example,Alternaria sp., Cylindrocarpon sp., Chaetomium sp., Paraphaeosphaeria sporulosa, Pyrenochaeta sp., Penicillium swiecickii, T. viride, and Truncatella angustata were found only in the oldest plants. Analysis of(metagenomics) community DNA extracted from different-aged samples revealed that, at the class level,the majority of fungi had the highest sequence similarity to members of Sordariomycetes, followed by Eurotiomycetes and Saccharomycetes. These results were mostly in accord with those we obtained using culture methods. Fungal diversity and richness also changed over time. Our investigation is the first to show that the diversity of fungi in rhizomes of P. polyphylla var. yunnanensis is altered as plants age, and our findings provide a foundation for future examinations of useful compounds.

Purification of four strains of endophytic fungi from Astragalus and their optimized liquid fermentations

This study was designed to isolate endophytic fungi from A. mongholicus （growing in northeast China） to determine whether they can produce bioactive metabolites. Four strains of endophytic fungi （strains 16, 17, 23 and 75） were successfully isolated from A. mongholicus using the surface disinfection method. According to ITS-rDNA sequences analysis, strains 16 and 75 were identified as Fusarium oxysporum, and strains 17 and 23 were identified as Bionectria ochroleuca. We applied the Box-Behnken design （BBD） to optimize the liquid fermentation conditions and obtain the maximum cell dry weight （CDW） yield. Opti-mal parameters were obtained under the following experimental condi-tions：temperature of 28＆#176;C, potato dextrose agar （PDA） liquid medium of 80 mL and rotation speed of 150 rpm. The four isolated endophytic fungi did not produce astragalosides I-IV, flavonoids or polysaccharides. Iso-lation of additional species of endophytic fungi from A. mongholicus and determination of their capacity to produce biologically active substances are subjects in need of further research.

Three Novel Endophytic Fungi as Potential Candidates Producing Bioactive Metabolites Isolated from Metasequoia glyptostroboides

[Objectives]This study was conducted to isolate and investigate the endophytic fungi with antimicrobial activity from the healthy leaves and bark of Metasequoia glyptostroboides Hu&W.C.Cheng regarded as an important medicinal plant in China.[Methods]Endophytic fungi with visible antimicrobial activity were isolated from the healthy leaves and bark of M.glyptostroboides,and primarily screened using Escherichia coli and Staphylococcus aureus as test microorganisms.[Results]Three isolates with relatively higher antimicrobial activity,labeled as MG-01,MG-02,MG-09,were selected for study.Molecular and morphological analysis indicated that the three isolated endophytes belong to Penicillium sp.The ethyl acetate(EtOAc)extracts of the three endophytes were obtained and subjected to the disk diffusion assay,using E.coil,S.aureus,Pseudomonas aeruginosa,Enterococcus faecalis,Monilia albicanas,and Bacillus subtilis as test microorganisms.The minimum inhibitory activity(MIC)of the endophyte with relatively highest antimicrobial activity was determined using two-fold broth micro-dilution method.All the endophytes showed a broad spectrum antimicrobial activity.Among the three endophytes,the strain MG-09 exhibited relatively highest antimicrobial activity with the zones of inhibition(ZOI)ranging from 14.15 to 24.07 mm and MIC ranging from 416.67 to 65.11μg/ml.[Conclusions]The endophytic fungi colonized in M.glyptostroboides are capable of producing effective antimicrobial compounds which may be a potential source for drug discovery and biocide production.

Diversity of endophytic fungi associated with the medicinally important aromatic plant Gaultheria fragrantissima

A total of thirty three(33)endophytic fungi including sterile mycelia were isolated from the leaf,stem and roots of medicinally important aromatic plant Gaultheria fragrantissima.Majority of the fungi belonged to the phylum Ascomycota.The total percentage colonization frequency(%CF)of each endophytic fungal species was calculated of which Juxtiphoma eupyrena showed highest colonization frequency(20.14%)in the leaf whereas Globisporangium irregulare was found to have highest colonization frequency in the stem(14.58%)and in the roots,Trichoderma viride was observed to have the highest colonization frequency of 21.43%.Diversity of endophytic fungi in the roots was found to be higher as compared to the other plant tissues.

Anticancer activity test of ethyl acetate extract of endophytic fungi isolated from soursop leaf(Annona muricata L.)

Objective: To analyze anticancer activity of an ethyl acetate extract of endophytic fungi isolated from soursop leaf(Annona muricata L.). Methods: Anticancer activity of fungal extracts was determined by observing its toxicity against MCF-7(Michigan Cancer Foundation-7) cells in vitro by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay method. At an extract concentration of 100 μg/m L, 4 isolates out of 12 showed high activity against the cancer cell growth. The four isolates were then selected for further IC50 determination, by measuring the inhibition of cancer cell proliferation at extract concentration of 25 μg/m L, 50 μg/m L, 100 μg/m L, 200 μg/m L and 400 μg/m L. Results: Results showed that isolate Sir-G5 had the highest anticancer activity with an IC50 of 19.20 μg/m L. The best isolates were screened again using a normal cell(Chang cells) to determine its toxicity against normal cells. Results indicated that the extracts do not affect the proliferation of normal cells. Molecular identification showed that the fungal isolate Sir-G5 has a close relationship with Phomopsis sp. Conclusions: The endophytic fungi isolated from soursop leaf has the potential to be used as a source of anticancer agents.

Community Structure of Endophytic Fungi in Ginseng and Its Correlation Analysis with Active Components

[Objectives]The paper was to explore the correlation between active components of ginseng and community structure of endophytic fungi in ginseng under different cultivation methods.[Methods]Using ginsengs under two main planting patterns in Huanren County,Liaoning Province as the samples,the community structure of endophytic fungi was studied by molecular means,and the chemical components were identified and the content of chemical markers was measured via LC-MS.Moreover,the correlation between medicinal components of ginseng and diversity of endophytic fungal flora was analyzed.[Results]The diversity and active components of endophytic fungi of ginseng were different.The diversity analysis showed that the medicinal components of ginseng were positively correlated with some fungal groups in ginseng.[Conclusions]Artificial intervention in the planting process would affect the endophytic fungi in medicinal materials,and further affect the chemical components of ginseng medicinal materials.

Polyaromatic Hydrocarbon Degradation of Moss Endophytic Fungi Isolated from Macromitrium sp. in Sri Lanka

Polyaromatic hydrocarbons （PAHs）, which are the major by-products of fossil fuel burning, are released to the environment with the immense growth of urbanization and industrialization. These pollutants are subsequently deposited on many substrates including plant surfaces. Due to their toxicity, mutagenicity, carcinogenicity and recalcitrant nature, they can result in many hazardous effects on human health. Application of endophytes in bioremediation has shown much promise in removing these PAHs from contaminated substrates. In the present study, an attempt was made to isolate and identify endophytic fungi from the moss plant Macromitrium sp. （frequently available） in Sapugaskanda （highly polluted） and HettimuUa （less polluted） areas in Sri Lanka. Subsequently, their potential in degrading PAHs （naphthalene and phenanthrene） was investigated. Endophytes from the moss were isolated following the surface sterilization method, and their physiological roles in degrading naphthalene and phenanthrene were carried out using plate assays, spectrophotometric and high-performance liquid chromatography （HPLC） analysis. Most of the endophytie fungi isolated from Macromitrium sp. were able to grow in Bacto Bushnell-Haas （BBH） medium incorporated with naphthalene and phenanthrene, separately, displaying colony diameters more than 30 mm. As per the results obtained from spectrophotometric and HPLC analysis, Penicillium oxalicum, Nigrospora oryzae, Aspergillus oryzae, A. aculeatus, Penicillium sp.1, Penicillium sp.5, Eupenicillium sp.2 and Mortierella sp.1 degraded both naphthalene and phenanthrene more than 85%. The fmdings of the present investigation provide some insight into how these endophytic fungi could be used for bioremediation of PAHs in environmental sites where contamination prevails, and also open avenues for future research in the relevant field.

Endophytic Fungi Associated with a Holoparasitic Plant, Balanophora japonica (Balanophoraceae)

Holoparasitism is a special life cycle of flowering plants. All carbon resources are provided by photosynthetic host plants. A recent study revealed the presence of endophytic fungi in holoparasitic plants, but their ecological and evolutionary roles are still unknown. In this study, we examined endophytic fungi isolated from the holoparasitic plant Balanophora japonica (Balanophoraceae), collected from Kochi, Shikoku in western Japan. We isolated 23 fungal strains on inflorescences and tubers from three B. japonica plants at two locations and on one sample of the host plant (Symplocos lancifolia, Symplocaceae). Predominant isolates were Trichoderma-Hypocrea, Penicillium and Phialemonium. The first group was also predominant in the host plant. Fungal composition revealed in this study differed from the composition on B. harlandii or other root holoparasites with endophytic fungal (Rafflesia cantleyi) data. Those differences might be caused by various factors, including growth habits, location, phylogenetic position or host-parasite relationship.

Endophytic Fungi Isolated from Mangrove （Rhyzopora mucronata） and Its Antibacterial Activity on Staphylococcus aureus and Escherichia coli

Disc diffusion method was used to investigate the antibacterial activity of methanol extract of five endophytic fungi isolated from mangrove （Rhizopora mucronata） on Staphylococcus aureus ATCC 9144 and Escherichia coli ATCC 8739, Fungi were isolated from leaf, stem and root of R. mucronata. Potato dextrose agar and potato dextrose broth were used for fungi purification and fermentation. Fungal metabolites were extracted with methanol （1 ： 1 v/v） for 24 h before vacuum evaporated to yield the crude extracts. Antimicrobial activity test was done according to the Kirby-Bauer test. The inhibition zone around each disc was considered to measure antibacterial activity of endophytic fungi. Only three endophytic fungi showed inhibition activity on S. aureus ATCC 9144 and two endophytic fungi showed inhibition on E. coil ATCC 8739. The results suggested that the extracts have a high potential for enteric diseases treatment caused by foodbome pathogen.

Isolation, Identification and Pharmacological Activities of Endophytic Fungi from <i>Aponogeton undulatus</i>Roxb.

The objective of this study is to isolate, identify and investigate the pharmacological activities of the endophytic fungi from an aquatic plant Aponogeton undulatus Roxb. (A. undulatus). Endophytic fungi were isolated and identified based on morphological characters. The molecular identification of the fungal isolates was performed using by analyzing the DNA sequence based on mega BLAST program. Spectrums of antibacterial and antifungal activities were studied by Agar diffusion methods. Extracts of the endophytic fungal strains isolated from the plant A. undulatus were screened for probable cytotoxic activities using brine shrimp lethality bioassay. Six endophytic fungi, namely AULE-1, AULE-2, AULE-3, AURE-1, AURE-3 and AURE-4 were isolated and purified from the leaves and roots of the plant such as strains AULE-1 as Trichooderma sp., AULE-2 as Carvularia sp. AULE-3 as Penicillium sp. strains AURE-1 and AURE-4 as Fusarium sp. and AURE-3 as Mucor sp. Strain AURE-4 was further identified as Fusarium solani. At 300 μg/disc concentration, six ethyl acetate extracts of endophytic fungi of the plant A. undulates showed moderate to significant activities against most of the test bacteria and pathogenic fungi. The strain AURE-4 exhibited strong cytotoxic activities (10.18 μg/mL) compared to the standard antitumor agent vincristine sulphate and this positive result suggests that fungal extracts may contain antitumor or pesticidal compounds. This is the first study to report the isolation, identification, antimicrobial and cytotoxic properties of endophytic fungi of Aponogeton undulatus Roxb. in Bangladesh.

Potential use of cucumber (Cucumis sativus L.) endophytic fungi as seed treatment agents against root-knot nematode Meloidogyne incognita

Seed treatment with endophytic fungi has been regarded as an effective method for plant parasitic nematode control.Endophytic fungi from cucumber seedlings were isolated and screened for their potential to be used as seed treatment agents against Meloidogyne incognita.Among the 294 isolates screened,23 significantly reduced galls formed by M.incognita in greenhouse test.The 10 most effective isolates were Fusarium(5),Trichoderma(1),Chaetomium(1),Acremonium(1),Paecilomyces(1),and Phyllosticta(1) .Their control efficacies were repeatedly tested and their colonizations as well as in vitro activity against M.incognita were studied.They reduced the number of galls by 24.0%-58.4%in the first screening and 15.6%-44.3%in the repeated test,respectively.Phyllosticta Ph511 and Chaetomium Ch1001 had high colonizations on both the roots and the aboveground parts of cucumber seedlings.Fusarium isolates had colonization preference on the roots,their root colonizations ranging from 20.1%to 47.3%of the total root area.Trichoderma Tr882,Paecilomyces Pa972,and Acremonium Ac985 had low colonizations on both the roots and the aboveground parts.Acremonium Ac985,Chaetomium Ch1001,Paecilomyces Pa972,and Phyllosticta Ph511 produced compounds affecting motility of the second stage juveniles of M.incognita.Based on these results,Chae-tomium Ch1001 was considered to have the highest potential as a seed treatment agent for M.incognita biocontrol.

Antitumor and antimicrobial activities of endophytic fungi from medicinal parts of Aquilaria sinensis

The purpose of this study was to isolate and characterize endophytic fungi from the stem tissue which can produce fragrant ingredients in Aquilaria sinensis(also called agarwood) to determine their antitumor and antimicrobial activities.Twenty-eight fungal endophytes were isolated from agarwood by strict sterile sample preparation and were classified into 14 genera and 4 taxonomic classes(Sordariomycetes,Dothideomycetes,Saccharomycetes,and Zy-gomycetes) based on molecular identification.Of the 28 isolates,13(46.4%) showed antimicrobial activity against at least one of the test strains by the agar well diffusion method,and 23 isolates(82.1%) displayed antitumor activity against at least one of five cancer cell lines by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay.The diameters of inhibition zones of YNAS07,YNAS14,HNAS04,HNAS05,HNAS08,and HNAS11 were equal to or higher than 14.0 mm against Staphylococcus aureus,Escherichia coli,Bacillus subtilis,B.subtilis,Aspergillus fumigatus,and B.subtilis,respectively.The inhibition rates of YNAS06,YNAS08,and HNAS06 were not less than 60% to 293-T,293-T,and SKVO3 cells,respectively.These results suggest that the endophytic fungi associated with agarwood will provide us with not only useful micro-ecological information,but also potential antimicrobial and anti-tumor agents.

Endophytic fungi harbored in Cannabis sativa L.:diversity and potential as biocontrol agents against host plant-specific phytopathogens

The objective of the present work was isolation,phylogenetic characterization,and assessment of biocontrol potential of endophytic fungi harbored in various tissues(leaves,twigs,and apical and lateral buds)of the medicinal plant,Cannabis sativa L.A total of 30 different fungal endophytes were isolated from all the plant tissues which were authenticated by molecular identification based on rDNA ITS sequence analysis(ITS1,5.8S and ITS2 regions).The Menhinick’s index revealed that the buds were immensely rich in fungal species,and Camargo’s index showed the highest tissue-specific fungal dominance for the twigs.The most dominant species was Penicillium copticola that could be isolated from the twigs,leaves,and apical and lateral buds.A detailed calculation of Fisher’s log series index,Shannon diversity index,Simpson’s index,Simpson’s diversity index,and Margalef’s richness revealed moderate overall biodiversity of C.sativa endophytes distributed among its tissues.The fungal endophytes were challenged by two host phytopathogens,Botrytis cinerea and Trichothecium roseum,devising a dual culture antagonistic assay on five different media.We observed 11 distinct types of pathogen inhibition encompassing a variable degree of antagonism(%)on changing the media.This revealed the potential chemodiversity of the isolated fungal endophytes not only as promising resources of biocontrol agents against the known and emerging phytopathogens of Cannabis plants,but also as sustainable resources of biologically active and defensive secondary metabolites.