Antioxidant activity and myco-chemical profiling of Campylospora parvula Kuzuha isolated from the roots of Rubus ellipticus

Antioxidant activity and myco-chemical profiling of Campylospora parvula Kuzuha isolated from the roots of Rubus ellipticus | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Antioxidant activity and myco-chemical profiling of Campylospora parvula Kuzuha isolated from the roots of Rubus ellipticus Prabha Pant, Parikshit Kumar, Suresh Chandra Sati This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4683858/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 3 You are reading this latest preprint version Abstract Endophytic fungi are important sources of various bioactive natural compounds. Present work elucidates the antioxidant activity and chemical characterization of Campylospora parvula Kuzuha isolated from the roots of ethno medicinal plant Rubus ellipticus . Antioxidant activities were tested using DPPH, MCA and FRAP assays, total phenolic and flavonoid content by Folin-Ciocalteu and aluminum chloride method, respectively. Chemical profile of ethyl acetate extract was illustrated by using Gas chromatography and mass spectrometry (GC-MS) analysis. The ethyl acetate extract of C. parvula presents IC50, 47.75±3.6 µg/ml for DPPH, 53.36±2.6 µg/ml for MCA and 360.54±8.6 AAE/g dry extract for FRAP assay. GCMS analysis revealed the presence of Octanal, 2-(phenylmethylene)- (16.24%), 1-(4-Iopropylphenyl)-2-methylpropyl acetate (12.91%), Benzoic acid, 2-hydroxy-, phenylmethyl ester (11.85%) as major compounds. Findings of the present study are promising and keenly hopeful for further research on individual compound to assess their potential, especially in the pharmacology sector. Aquatic hyphomycetes endophyte Campylospora parvula antioxidant myco-chemicals Figures Figure 1 Figure 2 Figure 3 Figure 4 INTRODUCTION Plants as well as microbes have always been considered a treasure house of several natural products with a wide range of entities like bio-control agents, catalyst, drugs, enzymes, and immune-suppressants. They are prominent source for the exploration of novel drugs (Bull and Stach 2007). Plants and endophytic fungi have an advantageous symbiotic alliance in which the host plants provide essential nutrients to endophytic fungi and endophytic fungi support host plants by improving resistance and tolerance to various biotic and abiotic stresses (Khare et al. 2018 ). Endophytic fungi produce wide range of secondary metabolites which are the rich source of potent bioactive metabolites and are also considered as potential antioxidant resource (Huang et al. 2007). Aquatic hyphomycetes (Ingoldian fungi, amphibious fungi or freshwater hyphomycetes) are a phylogenetically heterogeneous group of micro fungi flourish in well oxygenated water. These fungi are morphologically, physiologically, ecologically, phylogenetically and chemically diverse group of organisms found in freshwater ecosystem as saprobes as well as endophytes. The endophytic nature of these fungi reveals that aquatic hyphomycetous fungi play an important role in plant health by producing various bioactive compounds. For the last few years, these fungi have been studied for their various biological activities, such as antimicrobial, nutrient solubilizer, plant growth promoter etc, but very less studies have demonstrated their antioxidant activity. Rubus ellipticus Sm., commonly known as the yellow Himalayan raspberry is an evergreen shrub belonging to the Rosaceae family. There are around 600–800 species of Rubus in which, R. ellipticus is the most abundant and wild-growing fruit species on the Indian subcontinent (Badhani et al. 2015 ). All of the parts of this plant have been used in traditional medicine to treat various diseases as well as this plant has been found to be rich in various secondary metabolites, including flavonoids, polyphenols, terpenoids, tannins, and anthocyanins (Lamichhane et al. 2023 ). The present study narrates the isolation of a fungal endophyte, Campylospora parvula , from the healthy roots of R. ellipticus , and to evaluate the antioxidant activity along with chemical profiling by GCMS technique. To the best of our knowledge there was no previous report on the antioxidant activity and chemical characterization of C. parvula Kuzuha. MATERIAL AND METHODS 1. Isolation of root endophytic aquatic Hyphomycetes Roots of riparian plants were collected from the streams and their catchment areas of Nainital district of Kumaun Himalayas, Uttarakhand, India. Fresh and healthy roots from the living plants were sampled, kept in pre- sterilized bags, and brought to the laboratory for further processing. Root samples were thoroughly washed under running tap water (2–4 hours), then with sterile water (5–7 min), and finally with 2% sodium hypochlorite solution (1–2 min) to remove adhering free living microbes of the root surfaces. These root samples were then cut into small pieces (1–2 cm long) and placed into Petri dishes containing 20 mL of sterile water. The root segments in Petri dishes were incubated at 20 ± 2°C for 3–7 days and examined regularly. Observations were made with compound microscope to confirm mycelial growth and conidial liberation into the water. Floating conidia were picked up by a sterile needle and studied further by making semi-permanent slides. Pure cultures were initiated by transferring conidia onto 2% malt extract agar (MEA). Germinated conidia were allowed to grow into colonies and maintained on MEA slants at 4°C (Sati and Belwal 2005). The isolates were identified with the help of relevant monographs and literature. Pure cultures of the isolates have been submitted in Kumaun University Mycological Society (KUMS) collection of the Department of Botany, Kumaun University, Nainital, India. 2. Preparation of fungal extracts For the preparation of crude fungal extracts the selected REAH were inoculated into 250 ml of conical flask containing 100 ml of malt extract broth. These flasks were incubated at 28°C for up to 15 days in a BOD incubator. After 15 days, the cultures were filtered with the help of Whatman paper no. 1 and centrifuged at 5000 rpm for the separation of fungal mycelia. The mycelia and fungal broth were extracted by 100 ml of methanol and ethyl acetate, respectively. These two extracts containing secondary metabolites were combined, filtered and evaporated to dryness. 3. Antioxidant activity Three different assays of fungal extracts was estimated using 1,1-diphenyl-2-picryl-hydrazyl (DPPH) assay, ferric ion reducing antioxidant power (FRAP) and ferrous ion scavenging (Metal Chelating) assay. 3.1: DPPH Assay The antioxidant activity of ethyl acetate extracts of C. parvula and T. setigerum were assessed on the basis of their scavenging effect of the DPPH (2, 2-diphenyl-1-picryl-hydrazyl) free radical. DPPH radical scavenging ability was evaluated based on the method given by Xie et al. (2010) with minor modifications. A fresh solution of DPPH (0.1mM) was prepared in methanol and 2 mL of this solution was mixed with 2 mL of the extract at different concentrations (10–100 µg/ mL). The mixture of 2mL DPPH in 2mL methanol was used as control. The reaction mixture was incubated in dark for 30 minutes at room temperature and analyzed spectrophotometrically at 517 nm against blank. Ascorbic acid was used as positive control. The free radical scavenging potential (%) of the fungal extract was calculated with formula given by Blois (1958). % Inhibition \(=\frac{\text{A}\text{b}\text{s}\text{o}\text{r}\text{b}\text{a}\text{n}\text{c}\text{e} \text{o}\text{f} \text{C}\text{o}\text{n}\text{t}\text{r}\text{o}\text{l} - \text{A}\text{b}\text{s}\text{o}\text{r}\text{b}\text{a}\text{n}\text{c}\text{e} \text{o}\text{f} \text{t}\text{e}\text{s}\text{t} \text{S}\text{a}\text{m}\text{p}\text{l}\text{e}}{\text{A}\text{b}\text{s}\text{o}\text{r}\text{b}\text{a}\text{n}\text{c}\text{e} \text{o}\text{f} \text{C}\text{o}\text{n}\text{t}\text{r}\text{o}\text{l}}\times 100\) IC 50 value of test sample and standard was calculated from the graph of inhibition percentage plotted against concentration. 3.2: FRAP Assay Ferric reducing antioxidant power (FRAP) assay was performed by the using method of Cai et al. (2004), with minor modification. The reductive potential of C. parvula and T. setigerum were determined based on the chemical reduction of Fe 3+ to Fe 2+ . 10µl of sample extract was taken and make it up to 100µl with double distilled water and was mixed with 1.5mL of freshly prepared and pre-warmed (37ºC) FRAP reagent and kept at 37 0 C for 10 minutes. Absorbance was taken at 593 nm. Standard was prepared by using ascorbic acid. For control absorbance of FRAP reagent (300 mM acetate buffer, pH = 3.6, 10 mM tripyridyl-s-triazine (TPTZ) in 40 mM HCl and 20 mM FeCl 3 .6H 2 O in the ratio of 10:1:1) was taken without adding sample extract. Results were expressed in mg ascorbic acid equivalent per g dry weight of the sample extract. 3.3: MCA Assay The Fe 2+ chelation by Ferrozinein fungal extracts was investigated by the method as described by Dinis et al. (1994). EDTA was used as the reference compound. Six different concentrations (10–100 µg/ml) of fungal extracts and EDTA were prepared. Different each dilution was added with 1 ml solution of 2mM ferrous sulphate solution. This supplying of Fe2 + in sample dilution forms the interaction between them and Fe2 + ions are chelated by the sample. The reaction was initiated by addition of 1ml of ferrozine (0.25 mM) and the mixture was stirred vigorously. After the incubation for 20 minutes at room temperature, absorbance of the Fe 2+ -ferrozine complex in reaction mixture was measured at 562 nm. 4. GCMS Analysis Detection and quantification of the presence of secondary metabolites in plant extracts was carried out by gas chromatography coupled with mass spectrometry (Shimadzu QP2010 Plus) equipped with a Rtx- 5 MS capillary column (0.25 mm film thickness, 0.25 mm internal diameter, and 30 m in length). The oven temperature was set at 100°C for 2 min, then increased to 260°C with a rate of 5°C per minute, and finally to 280°C with a rate of 15°C per minute. One µl of each sample was injected to the column in split mode (split ratio 10) with helium as the carrier gas with a flow rate of 1.21 ml per minute. The presence of distinctive peak fragmentation patterns for various metabolites was detected by an MS detector in full scan mode. Identification of metabolites was confirmed by comparing the spectral data of peaks with the corresponding standard mass spectra from the library database [National Institute of Standards and Technology library (NIST) and Wiley 8]. RESULTS Morphological features of isolated REAH Root endophytic fungus with branched septate mycelium. Conidiophore 12–15 µm long and 1.5-2 µm wide unbranched. Conidia hyaline, tetraradiate arise from conidiophores singly. Conidial main axis consisting of smaller part, 3–4 celled, 8–10 µm long and 5–6 µm wide allantoid and larger part 10-12.5 × 5.5-7 µm longitudinally, 2–4 septate and transversely two septate. Appendages are 7-9.5 × 2–3 µm tapering to 0.5 to 0.01 µm at their tips (Fig. 1 A). The isolated fungus was identified as Campylospora parvula Kuzuha isolated from the roots of Rubus elipticus (KUMS 007). Antioxidant Activity: The antioxidant activity of ethyl acetate extract of C. parvula was performed by DPPH, FRAP and MCA assay. The free radical scavenging activity of C. parvula was 40.75µg/mL and 53.36µg/mL in DPPH and MCA assay respectively. The reducing activity of tested isolate was 360.54 in terms of mg AAE gm − 1 dry extract in FRAP assay (Table 1 ). Table 1 Antioxidant activity of C. parvula Extracts IC 50 value (µg/mL) of radical scavenging *FRAP activity (mg AAE/g of dry extract) DPPH scavenging MCA CP 47.75 ± 3.6 53.36 ± 2.6 360.54 ± 8.6 Ascorbic acid 19.84 ± 0.7 12.56 ± 1.5 *FRAP- Ferric reducing antioxidant power, mg Ascorbic acid equivalent (AAE)/g of dry extract GCMS Analysis Outcome of the GC-MS results affirm the presence of different bioactive compounds. A total of 18 compounds were detected in ethyl acetate extract of C. parvula (Table 2 ). Octanal, 2-(phenylmethylene)- (16.24%) was reported as a major compound followed by 1-(4-Iopropylphenyl)-2-methylpropyl acetate (12.91%), Benzoic acid, 2-hydroxy-, phenylmethyl ester (11.85%), Benzoic acid, 2-hydroxy-, methyl ester (2.16%). Table 2 Myco-chemical constituents of ethyl acetate extract of C. parvula Name of Compound Molecular Formula Nature of of Compound Retention Time % of Total Molecular Weight Pentadecafluorooctanoic acid, octyl ester C 16 H 17 F 15 O 2 Ester 5.004 0.59 526 Benzoic acid, 2-hydroxy-, methyl ester C 8 H 8 O 3 Ester 5.182 2.16 152 1,3-Hexadiene, 3-ethyl-2-methyl-, (Z)- C 9 H 16 Alkene 7.414 0.28 124 1-hexadecene C 16 H 32 Alkene 7.697 0.91 224 Phenol, 3,5-bis(1,1-dimethylethyl)- C 14 H 22 O Phenol 9.274 0.63 206 2(3H)-furanone, 5-heptyldihydro- C 11 H 20 O 2 Ketone 10.066 1.80 184 7a-Isopropenyl-4,5-dimethyloctahydroindene-4-carboxylic acid C 15 H 24 O 2 Carboxylic acid 10.737 0.82 236 3-Cyclohexene-1-carboxaldehyde, 4-(4-hydroxy-4-methylpentyl)- C 13 H 22 O 2 Ester 11.112 1.79 210 1-(4-isopropylphenyl)-2-methylpropyl acetate C 15 H 22 O 2 Ester 11.175 12.91 234 1H-Indene, 2,3,3a,4,7,7a-hexahydro-2,2,4,4,7,7-hexamethyl C 15 H 26 Alkene 11.551 1.33 206 Octanal, 2-(phenylmethylene)- C 15 H 20 O Aldehyde 12.077 16.24 216 1-Nonadecene C 19 H 38 Alkene 12.484 0.93 266 8-Ethyl-4,6,6,8-tetramethyl-3,4,6,7-tetrahydro-1H-cyclopenta(G)-2-benzopyran C 18 H 26 O Ketone 12.714 1.03 258 N2,N2,N4,N4,N5,8-Hexamethylquinoline-2,4,5-triamine C 15 H 22 N 4 Amine 13.259 1.01 258 Benzoic acid, 2-hydroxy-, phenylmethyl ester C 14 H 12 O 3 Ester 13.478 11.85 228 1-docosanol C 22 H 46 O Alcohol 14.532 0.73 326 n-Pentadecanol C 15 H3 2 O Alcohol 16.398 0.63 228 1-Nonadecene C 19 H 38 Alkene 18.282 0.28 266 DISCUSSION Fungi are dynamic producers of secondary metabolites and have been extensively investigated in the search of new secondary metabolites for years (Bills and Gloer 2016 ). They represent an essential source of natural substances with advantageous bioactive properties harboring immense potential for biotechnological applications (Hyde et al. 2019 ; Suryanarayanan et al., 2009 ). Analyzing fungal secondary metabolites (FSMs) from fungal extracts is a preliminary process to know the chemical composition of fungi. Presence of FSMs in fungal crude extract indicates that they could be used as precursors in the development and progression of synthetic medicines. The present study supports the previous findings for aquatic hyphomycetous fungi having biologically active FSMs (El-Elimat et al. 2021 ; Pant et al. 2023). Major secondary metabolites observed in this study are well documented in earlier reports. Based on morphological structure the root endophytic fungus (KUMS 007) was identified as Campylospora parvula Kuzuha. Previously C. parvula was reported for its antibacterial, phosphate solubilization and plant growth promoting activity (Sati and Pant 2020 ; Pant and Sati 2021 ; Sati et al. 2023 ). However, information available on the antioxidant activity and chemical composition of the tested species is scanty. In the present study C. parvula recovered from healthy roots of Rubus elipticus growing in riparian area of Nainital, Kumaun Himalaya, India was investigated for its antioxidant potentiality and myco-chemical composition. Antioxidants have become the subject of interest in recent years. Fungal endophytes represent an abundant and reliable source of novel antioxidant compounds. A number of antioxidants are known to provide protection against several diseases. Epidemiological studies have demonstrated that higher intake of antioxidants results in reduced risk of heart disease and many other diseases. This is the reason for the strong interest in natural antioxidants and their role in human health and nutrition (Bahera et al. 2006). The antioxidant activity of ethyl acetate extract of C. parvula was evaluated by DPPH radical scavenging, ferric reducing antioxidant power assay (FRAP) and metal chelating assay (MCA). The results summarized in Table 1 clearly suggest that C. parvula is a potential source of natural antioxidants. To investigate the chemical composition of ethyl acetate extract of C. parvula GCMS technique was applied. GCMS analysis of the extract showed the various myco chemicals which have antimicrobial and antioxidant properties. Ester and alkene (28%) were detected as highest chemical group in the ethyl acetate extract of tested fungi followed by ketone and alcohol (12%), phenol, carboxylic acid, aldehyde and amine (5%) (Fig. 4 ). Octanal, 2-(phenylmethylene)- (16.24%) was reported as a major compound followed by, 1-(4-Iopropylphenyl)-2-methylpropyl acetate (12.91%), Benzoic acid, 2-hydroxy-, phenylmethyl ester (11.85%), benzoic acid, 2-hydroxy-, methyl ester (2.16%). To the best of our knowledge this is the first study on the antioxidant activity and GCMS analysis of C. parvula . Present work will serve as a platform for future research on endophytic aquatic hyphomyceteous fungi for the production of bioactive compounds having various biological activities. CONCLUSION Endophytes are evidently an affluent and unswerving source of bioactive and chemically unique substances with significant therapeutic and agricultural potential but these unseen repositories of microorganisms hidden within the host plants are inadequately studied. This is the first report on the antioxidant activity and myco-chemical profiling of endophytic fungi C. parvula Kuzuha isolated from the riparian area of Nainital, Kumaun Himalaya. Present study revealed the presence of biologically active compounds which provide an alternative source of developing drug like metabolites. Used fungus is also a good source of antioxidants which can be used to find to find some new antioxidants. This study will also provide a foreword to more comprehensive work on bioactive compounds produced by these endophytic hyphomyceteous fungi. Declarations Author Contribution PP performed the experimental work, data collection, analysed the data and wrote the manuscript, and both the co-authors PK and SCS helped in conceptualization and corrected the manuscript. DISCLOSURE STATEMENT The authors declare that they have no conflict of interest. References Badhani A, Rawat S, Bhatt ID, Rawal RS (2015) Variation in Chemical Constituents and Antioxidant Activity in Yellow Himalayan ( Rubus ellipticus Smith) and Hill Raspberry ( Rubus niveus Thunb). J Food Biochem 39:663–672 Behera BC, Verma N, Sonone A, Makhija U (2006) Determination of antioxidative potential of lichen Usnea ghattensis in vitro. LWT Food Sci Technol 39(1):80–85. 10.1016/j. lwt.2004.11.007 Bills GF, Gloer JB (2016) Biologically Active Secondary Metabolites from the Fungi. Microbiol Spectrum 2016, 4 (6) FUNK-0009- 2016. 10.1128/microbiolspec.FUNK-0009-2016 El-Elimat T, Raja HA, Figueroa M, Al Sharie AH, Bunch RL, Oberlies NH (2021) Freshwater fungi as a source of chemical diversity: a review. J Nat Prod 84(3):898–916 Hyde KD, Xu J, Rapior S, Jeewon R, Lumyong S, Niego AGT, Abeywickrama PD, Aluthmuhandiram JVS, Brahamanage RS, Brooks S et al (2019) The amazing potential of fungi: 50 ways we can exploit fungi industrially. Fungal Divers 97(1). 10.1007/s13225-019-00430-9 Khare E, Mishra J, Arora NK (2018) Multifaceted interactions between endophytes and plant: Developments and prospects. Frontiers in Microbiology, 9 , 2732. https://doi.org/10.3389/fmicb . 2018.02732 Lamichhane A, Lamichhane G, Devkota HP (2023) Yellow Himalayan Raspberry ( Rubus ellipticus Sm.): Ethnomedicinal, Nutraceutical, and Pharmacological Aspects. Molecules 28:6071 Pant P, Sati SC (2021) Antibacterial activity of root endophytic aquatic hyphomycetes against plant pathogenic bacteria. Vegetos 34:785–789 Sati SC, Pant P, Vassilev N (2023) Significance of root endophytic aquatic hyphomycetes in phosphate solubilization. Sydowia 75:99–105 Sati SC, Pant P (2020) Two root endophytic aquatic hyphomycetes Campylospora parvula and Tetracladium setigerum as plant growth promoters. Asian J Agricultural Res 14:28–33 Suryanarayanan TS, Thirunavukkarasu N, Govindarajulu MB, Sasse F, Jansen R, Murali TS (2009) Fungal endophytes and bioprospecting. Fungal Biology Reviews 23(1–2):9–19. 10.1016/j.fbr.2009.07.001 Additional Declarations No competing interests reported. Supplementary Files GraphicalAbstract.png Graphical Abstract Cite Share Download PDF Status: Under Review Version 1 posted Editor assigned by journal 05 Jul, 2024 Submission checks completed at journal 05 Jul, 2024 First submitted to journal 04 Jul, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4683858","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":323263490,"identity":"8506c2f3-c443-4133-908e-aad72a63e61d","order_by":0,"name":"Prabha Pant","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA90lEQVRIiWNgGAWjYBCDBAMG5sMPPgBZbOyEVTM2QLSwpRnOAGlhJl4Lj4E0D4hPSIs5+9njjwv32OWZs58xMLb5tU2ej5mB8cPHHNxaLHvyEptnPEsutuxJK3ic23fbsI2ZgVly5jbcWgwO5Bg28xxgTtxwIHmDcW7PbUagFjZmXnxazr8BaalP3HD+gYG0Zc9te8JaboBtOZy44UaKgTTDj9uJBLVYznhjOHvGgePFBjeepRn2NtxObmNmbMbrF3P+HIPPBQeq8wzOJx9+8OPPbdv57c0HP3zE5zAG5IhgbAOTDbjVY2hh+INX8SgYBaNgFIxQAACcjlcuUDqcEgAAAABJRU5ErkJggg==","orcid":"","institution":"Kumaun University","correspondingAuthor":true,"prefix":"","firstName":"Prabha","middleName":"","lastName":"Pant","suffix":""},{"id":323263491,"identity":"23097731-e54b-4f9e-9a5e-af3113c363cd","order_by":1,"name":"Parikshit Kumar","email":"","orcid":"","institution":"Harsh Vidya Mandir (P.G.) College","correspondingAuthor":false,"prefix":"","firstName":"Parikshit","middleName":"","lastName":"Kumar","suffix":""},{"id":323263492,"identity":"f918422c-f698-4ce1-beee-47490e074ca2","order_by":2,"name":"Suresh Chandra Sati","email":"","orcid":"","institution":"Kumaun University","correspondingAuthor":false,"prefix":"","firstName":"Suresh","middleName":"Chandra","lastName":"Sati","suffix":""}],"badges":[],"createdAt":"2024-07-04 05:18:54","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4683858/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4683858/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":61359887,"identity":"2c2ba539-4b82-445f-8ff3-0667cae65c8d","added_by":"auto","created_at":"2024-07-29 21:48:54","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":158589,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eA:\u003c/strong\u003e Conidia of \u003cem\u003eC. parvula\u003c/em\u003e; \u003cstrong\u003eB:\u003c/strong\u003e Pure culture of \u003cem\u003eC. parvula\u003c/em\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4683858/v1/2ad34b68697d8b602be07420.png"},{"id":61359340,"identity":"b618631d-422f-4151-a6b8-6a379313d880","added_by":"auto","created_at":"2024-07-29 21:32:54","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":47824,"visible":true,"origin":"","legend":"\u003cp\u003eGCMS Chromatogram of the ethyl acetate extract of \u003cem\u003eC. parvula\u003c/em\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-4683858/v1/e8a9e71da8aac209880948f7.png"},{"id":61359342,"identity":"eb31fa05-c875-4d8c-a18c-46f48441c10a","added_by":"auto","created_at":"2024-07-29 21:32:54","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":358991,"visible":true,"origin":"","legend":"\u003cp\u003eChemical structure of major compounds detected in the extract of \u003cem\u003eC. parvula\u003c/em\u003e- Comp. \u003cstrong\u003e1:-\u003c/strong\u003e Benzoic acid, 2-hydroxy-, methyl ester; Comp. \u003cstrong\u003e2:-\u003c/strong\u003e 2(3H)-furanone, 5-heptyldihydro-; Comp. \u003cstrong\u003e3:-\u003c/strong\u003e 1-(4-isopropylphenyl)-2-methylpropyl acetate; Comp. \u003cstrong\u003e4:-\u003c/strong\u003e Octanal, 2-(phenylmethylene)-; Comp. \u003cstrong\u003e5:-\u003c/strong\u003e Benzoic acid, 2-hydroxy-, phenylmethyl ester\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-4683858/v1/a509c24a95548d9dedb285e1.png"},{"id":61359341,"identity":"dea23463-86b3-470f-82e3-766efcaa168b","added_by":"auto","created_at":"2024-07-29 21:32:54","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":62966,"visible":true,"origin":"","legend":"\u003cp\u003ePresence of different chemical groups in the ethyl acetate extract of \u003cem\u003eC. parvula\u003c/em\u003e\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-4683858/v1/f99775a93bb51ef303985a95.png"},{"id":61360493,"identity":"10c40874-30d8-438f-978a-8cbabc7dc83c","added_by":"auto","created_at":"2024-07-29 21:56:55","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1260146,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4683858/v1/1e82e6e9-8311-419e-b498-9ae2ae0ca3f1.pdf"},{"id":61359676,"identity":"0ec7f780-696e-4af0-a0a7-e67fa08895c7","added_by":"auto","created_at":"2024-07-29 21:40:54","extension":"png","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":153944,"visible":true,"origin":"","legend":"\u003cp\u003eGraphical Abstract\u003c/p\u003e","description":"","filename":"GraphicalAbstract.png","url":"https://assets-eu.researchsquare.com/files/rs-4683858/v1/07c84c5856a0df00a91239c1.png"}],"financialInterests":"No competing interests reported.","formattedTitle":"Antioxidant activity and myco-chemical profiling of Campylospora parvula Kuzuha isolated from the roots of Rubus ellipticus","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003ePlants as well as microbes have always been considered a treasure house of several natural products with a wide range of entities like bio-control agents, catalyst, drugs, enzymes, and immune-suppressants. They are prominent source for the exploration of novel drugs (Bull and Stach 2007). Plants and endophytic fungi have an advantageous symbiotic alliance in which the host plants provide essential nutrients to endophytic fungi and endophytic fungi support host plants by improving resistance and tolerance to various biotic and abiotic stresses (Khare et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Endophytic fungi produce wide range of secondary metabolites which are the rich source of potent bioactive metabolites and are also considered as potential antioxidant resource (Huang et al. 2007).\u003c/p\u003e \u003cp\u003eAquatic hyphomycetes (Ingoldian fungi, amphibious fungi or freshwater hyphomycetes) are a phylogenetically heterogeneous group of micro fungi flourish in well oxygenated water. These fungi are morphologically, physiologically, ecologically, phylogenetically and chemically diverse group of organisms found in freshwater ecosystem as saprobes as well as endophytes. The endophytic nature of these fungi reveals that aquatic hyphomycetous fungi play an important role in plant health by producing various bioactive compounds. For the last few years, these fungi have been studied for their various biological activities, such as antimicrobial, nutrient solubilizer, plant growth promoter etc, but very less studies have demonstrated their antioxidant activity.\u003c/p\u003e \u003cp\u003e \u003cem\u003eRubus ellipticus\u003c/em\u003e Sm., commonly known as the yellow Himalayan raspberry is an evergreen shrub belonging to the Rosaceae family. There are around 600\u0026ndash;800 species of Rubus in which, \u003cem\u003eR. ellipticus\u003c/em\u003e is the most abundant and wild-growing fruit species on the Indian subcontinent (Badhani et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). All of the parts of this plant have been used in traditional medicine to treat various diseases as well as this plant has been found to be rich in various secondary metabolites, including flavonoids, polyphenols, terpenoids, tannins, and anthocyanins (Lamichhane et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe present study narrates the isolation of a fungal endophyte, \u003cem\u003eCampylospora parvula\u003c/em\u003e, from the healthy roots of \u003cem\u003eR. ellipticus\u003c/em\u003e, and to evaluate the antioxidant activity along with chemical profiling by GCMS technique. To the best of our knowledge there was no previous report on the antioxidant activity and chemical characterization of \u003cem\u003eC. parvula\u003c/em\u003e Kuzuha.\u003c/p\u003e"},{"header":"MATERIAL AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e1. Isolation of root endophytic aquatic Hyphomycetes\u003c/h2\u003e \u003cp\u003eRoots of riparian plants were collected from the streams and their catchment areas of Nainital district of Kumaun Himalayas, Uttarakhand, India. Fresh and healthy roots from the living plants were sampled, kept in pre- sterilized bags, and brought to the laboratory for further processing. Root samples were thoroughly washed under running tap water (2\u0026ndash;4 hours), then with sterile water (5\u0026ndash;7 min), and finally with 2% sodium hypochlorite solution (1\u0026ndash;2 min) to remove adhering free living microbes of the root surfaces. These root samples were then cut into small pieces (1\u0026ndash;2 cm long) and placed into Petri dishes containing 20 mL of sterile water. The root segments in Petri dishes were incubated at 20\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C for 3\u0026ndash;7 days and examined regularly. Observations were made with compound microscope to confirm mycelial growth and conidial liberation into the water. Floating conidia were picked up by a sterile needle and studied further by making semi-permanent slides. Pure cultures were initiated by transferring conidia onto 2% malt extract agar (MEA). Germinated conidia were allowed to grow into colonies and maintained on MEA slants at 4\u0026deg;C (Sati and Belwal 2005).\u003c/p\u003e \u003cp\u003eThe isolates were identified with the help of relevant monographs and literature. Pure cultures of the isolates have been submitted in Kumaun University Mycological Society (KUMS) collection of the Department of Botany, Kumaun University, Nainital, India.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2. Preparation of fungal extracts\u003c/h2\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eFor the preparation of crude fungal extracts the selected REAH were inoculated into 250 ml of conical flask containing 100 ml of malt extract broth. These flasks were incubated at 28\u0026deg;C for up to 15 days in a BOD incubator. After 15 days, the cultures were filtered with the help of Whatman paper no. 1 and centrifuged at 5000 rpm for the separation of fungal mycelia. The mycelia and fungal broth were extracted by 100 ml of methanol and ethyl acetate, respectively. These two extracts containing secondary metabolites were combined, filtered and evaporated to dryness.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e3. Antioxidant activity\u003c/h2\u003e \u003cp\u003eThree different assays of fungal extracts was estimated using 1,1-diphenyl-2-picryl-hydrazyl (DPPH) assay, ferric ion reducing antioxidant power (FRAP) and ferrous ion scavenging (Metal Chelating) assay.\u003c/p\u003e \u003cp\u003e3.1: DPPH Assay\u003c/p\u003e \u003cp\u003eThe antioxidant activity of ethyl acetate extracts of \u003cem\u003eC. parvula\u003c/em\u003e and \u003cem\u003eT. setigerum\u003c/em\u003e were assessed on the basis of their scavenging effect of the DPPH (2, 2-diphenyl-1-picryl-hydrazyl) free radical. DPPH radical scavenging ability was evaluated based on the method given by Xie et al. (2010) with minor modifications. A fresh solution of DPPH (0.1mM) was prepared in methanol and 2 mL of this solution was mixed with 2 mL of the extract at different concentrations (10\u0026ndash;100 \u0026micro;g/ mL). The mixture of 2mL DPPH in 2mL methanol was used as control. The reaction mixture was incubated in dark for 30 minutes at room temperature and analyzed spectrophotometrically at 517 nm against blank. Ascorbic acid was used as positive control. The free radical scavenging potential (%) of the fungal extract was calculated with formula given by Blois (1958).\u003c/p\u003e \u003cp\u003e% Inhibition \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(=\\frac{\\text{A}\\text{b}\\text{s}\\text{o}\\text{r}\\text{b}\\text{a}\\text{n}\\text{c}\\text{e} \\text{o}\\text{f} \\text{C}\\text{o}\\text{n}\\text{t}\\text{r}\\text{o}\\text{l} - \\text{A}\\text{b}\\text{s}\\text{o}\\text{r}\\text{b}\\text{a}\\text{n}\\text{c}\\text{e} \\text{o}\\text{f} \\text{t}\\text{e}\\text{s}\\text{t} \\text{S}\\text{a}\\text{m}\\text{p}\\text{l}\\text{e}}{\\text{A}\\text{b}\\text{s}\\text{o}\\text{r}\\text{b}\\text{a}\\text{n}\\text{c}\\text{e} \\text{o}\\text{f} \\text{C}\\text{o}\\text{n}\\text{t}\\text{r}\\text{o}\\text{l}}\\times 100\\)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e value of test sample and standard was calculated from the graph of inhibition percentage plotted against concentration.\u003c/p\u003e \u003cp\u003e3.2: FRAP Assay\u003c/p\u003e \u003cp\u003eFerric reducing antioxidant power (FRAP) assay was performed by the using method of Cai et al. (2004), with minor modification. The reductive potential of \u003cem\u003eC. parvula\u003c/em\u003e and \u003cem\u003eT. setigerum\u003c/em\u003e were determined based on the chemical reduction of Fe\u003csup\u003e3+\u003c/sup\u003e to Fe\u003csup\u003e2+\u003c/sup\u003e. 10\u0026micro;l of sample extract was taken and make it up to 100\u0026micro;l with double distilled water and was mixed with 1.5mL of freshly prepared and pre-warmed (37\u0026ordm;C) FRAP reagent and kept at 37\u003csup\u003e0\u003c/sup\u003eC for 10 minutes. Absorbance was taken at 593 nm. Standard was prepared by using ascorbic acid. For control absorbance of FRAP reagent (300 mM acetate buffer, pH\u0026thinsp;=\u0026thinsp;3.6, 10 mM tripyridyl-s-triazine (TPTZ) in 40 mM HCl and 20 mM FeCl\u003csub\u003e3\u003c/sub\u003e.6H\u003csub\u003e2\u003c/sub\u003eO in the ratio of 10:1:1) was taken without adding sample extract. Results were expressed in mg ascorbic acid equivalent per g dry weight of the sample extract.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e3.3: MCA Assay\u003c/h2\u003e \u003cp\u003eThe Fe\u003csup\u003e2+\u003c/sup\u003e chelation by Ferrozinein fungal extracts was investigated by the method as described by Dinis et al. (1994). EDTA was used as the reference compound. Six different concentrations (10\u0026ndash;100 \u0026micro;g/ml) of fungal extracts and EDTA were prepared. Different each dilution was added with 1 ml solution of 2mM ferrous sulphate solution. This supplying of Fe2\u0026thinsp;+\u0026thinsp;in sample dilution forms the interaction between them and Fe2\u0026thinsp;+\u0026thinsp;ions are chelated by the sample. The reaction was initiated by addition of 1ml of ferrozine (0.25 mM) and the mixture was stirred vigorously. After the incubation for 20 minutes at room temperature, absorbance of the Fe\u003csup\u003e2+\u003c/sup\u003e-ferrozine complex in reaction mixture was measured at 562 nm.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e4. GCMS Analysis\u003c/h2\u003e \u003cp\u003eDetection and quantification of the presence of secondary metabolites in plant extracts was carried out by gas chromatography coupled with mass spectrometry (Shimadzu QP2010 Plus) equipped with a Rtx- 5 MS capillary column (0.25 mm film thickness, 0.25 mm internal diameter, and 30 m in length). The oven temperature was set at 100\u0026deg;C for 2 min, then increased to 260\u0026deg;C with a rate of 5\u0026deg;C per minute, and finally to 280\u0026deg;C with a rate of 15\u0026deg;C per minute. One \u0026micro;l of each sample was injected to the column in split mode (split ratio 10) with helium as the carrier gas with a flow rate of 1.21 ml per minute. The presence of distinctive peak fragmentation patterns for various metabolites was detected by an MS detector in full scan mode. Identification of metabolites was confirmed by comparing the spectral data of peaks with the corresponding standard mass spectra from the library database [National Institute of Standards and Technology library (NIST) and Wiley 8].\u003c/p\u003e \u003c/div\u003e"},{"header":"RESULTS","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eMorphological features of isolated REAH\u003c/h2\u003e \u003cp\u003eRoot endophytic fungus with branched septate mycelium. Conidiophore 12\u0026ndash;15 \u0026micro;m long and 1.5-2 \u0026micro;m wide unbranched. Conidia hyaline, tetraradiate arise from conidiophores singly. Conidial main axis consisting of smaller part, 3\u0026ndash;4 celled, 8\u0026ndash;10 \u0026micro;m long and 5\u0026ndash;6 \u0026micro;m wide allantoid and larger part 10-12.5 \u0026times; 5.5-7 \u0026micro;m longitudinally, 2\u0026ndash;4 septate and transversely two septate. Appendages are 7-9.5 \u0026times; 2\u0026ndash;3 \u0026micro;m tapering to 0.5 to 0.01 \u0026micro;m at their tips (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA).\u003c/p\u003e \u003cp\u003eThe isolated fungus was identified as \u003cem\u003eCampylospora parvula\u003c/em\u003e Kuzuha isolated from the roots of \u003cem\u003eRubus elipticus\u003c/em\u003e (KUMS 007).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eAntioxidant Activity:\u003c/h2\u003e \u003cp\u003eThe antioxidant activity of ethyl acetate extract of \u003cem\u003eC. parvula\u003c/em\u003e was performed by DPPH, FRAP and MCA assay. The free radical scavenging activity of \u003cem\u003eC. parvula\u003c/em\u003e was 40.75\u0026micro;g/mL and 53.36\u0026micro;g/mL in DPPH and MCA assay respectively. The reducing activity of tested isolate was 360.54 in terms of mg AAE gm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e dry extract in FRAP assay (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAntioxidant activity of \u003cem\u003eC. parvula\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eExtracts\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e value (\u0026micro;g/mL) of radical scavenging\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e*FRAP activity\u003c/p\u003e \u003cp\u003e(mg AAE/g of dry extract)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eDPPH scavenging\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMCA\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCP\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e47.75\u0026thinsp;\u0026plusmn;\u0026thinsp;3.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e53.36\u0026thinsp;\u0026plusmn;\u0026thinsp;2.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e360.54\u0026thinsp;\u0026plusmn;\u0026thinsp;8.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAscorbic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e19.84\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e12.56\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003e*FRAP- Ferric reducing antioxidant power, mg Ascorbic acid equivalent (AAE)/g of dry extract\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eGCMS Analysis\u003c/h2\u003e \u003cp\u003eOutcome of the GC-MS results affirm the presence of different bioactive compounds. A total of 18 compounds were detected in ethyl acetate extract of \u003cem\u003eC. parvula\u003c/em\u003e (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Octanal, 2-(phenylmethylene)- (16.24%) was reported as a major compound followed by 1-(4-Iopropylphenyl)-2-methylpropyl acetate (12.91%), Benzoic acid, 2-hydroxy-, phenylmethyl ester (11.85%), Benzoic acid, 2-hydroxy-, methyl ester (2.16%).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eMyco-chemical constituents of ethyl acetate extract of \u003cem\u003eC. parvula\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eName of Compound\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMolecular Formula\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNature of of Compound\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eRetention\u003c/p\u003e \u003cp\u003eTime\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e% of Total\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMolecular Weight\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePentadecafluorooctanoic acid, octyl ester\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003csub\u003e16\u003c/sub\u003eH\u003csub\u003e17\u003c/sub\u003eF\u003csub\u003e15\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eEster\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.004\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e526\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBenzoic acid, 2-hydroxy-, methyl ester\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003csub\u003e8\u003c/sub\u003eH\u003csub\u003e8\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eEster\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.182\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e152\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1,3-Hexadiene, 3-ethyl-2-methyl-, (Z)-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003csub\u003e9\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAlkene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e7.414\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e124\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1-hexadecene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003csub\u003e16\u003c/sub\u003eH\u003csub\u003e32\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAlkene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e7.697\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e224\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePhenol, 3,5-bis(1,1-dimethylethyl)-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003csub\u003e14\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eO\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePhenol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e9.274\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e206\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2(3H)-furanone, 5-heptyldihydro-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003csub\u003e11\u003c/sub\u003eH\u003csub\u003e20\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eKetone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10.066\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1.80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e184\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7a-Isopropenyl-4,5-dimethyloctahydroindene-4-carboxylic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCarboxylic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10.737\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e236\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3-Cyclohexene-1-carboxaldehyde, 4-(4-hydroxy-4-methylpentyl)-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003csub\u003e13\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eEster\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e11.112\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e210\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1-(4-isopropylphenyl)-2-methylpropyl acetate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eEster\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e11.175\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e12.91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e234\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1H-Indene, 2,3,3a,4,7,7a-hexahydro-2,2,4,4,7,7-hexamethyl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e26\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAlkene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e11.551\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e206\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOctanal, 2-(phenylmethylene)-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e20\u003c/sub\u003eO\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAldehyde\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e12.077\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e16.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e216\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1-Nonadecene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003csub\u003e19\u003c/sub\u003eH\u003csub\u003e38\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAlkene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e12.484\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e266\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8-Ethyl-4,6,6,8-tetramethyl-3,4,6,7-tetrahydro-1H-cyclopenta(G)-2-benzopyran\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e26\u003c/sub\u003eO\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eKetone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e12.714\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e258\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN2,N2,N4,N4,N5,8-Hexamethylquinoline-2,4,5-triamine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eN\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAmine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e13.259\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e258\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBenzoic acid, 2-hydroxy-, phenylmethyl ester\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003csub\u003e14\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eEster\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e13.478\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e11.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e228\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1-docosanol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003csub\u003e22\u003c/sub\u003eH\u003csub\u003e46\u003c/sub\u003eO\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAlcohol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e14.532\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e326\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003en-Pentadecanol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH3\u003csub\u003e2\u003c/sub\u003eO\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAlcohol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e16.398\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e228\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1-Nonadecene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC\u003csub\u003e19\u003c/sub\u003eH\u003csub\u003e38\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAlkene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e18.282\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e266\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e\u003c/h2\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eFungi are dynamic producers of secondary metabolites and have been extensively investigated in the search of new secondary metabolites for years (Bills and Gloer \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). They represent an essential source of natural substances with advantageous bioactive properties harboring immense potential for biotechnological applications (Hyde et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Suryanarayanan et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Analyzing fungal secondary metabolites (FSMs) from fungal extracts is a preliminary process to know the chemical composition of fungi. Presence of FSMs in fungal crude extract indicates that they could be used as precursors in the development and progression of synthetic medicines. The present study supports the previous findings for aquatic hyphomycetous fungi having biologically active FSMs (El-Elimat et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Pant et al. 2023). Major secondary metabolites observed in this study are well documented in earlier reports.\u003c/p\u003e \u003cp\u003eBased on morphological structure the root endophytic fungus (KUMS 007) was identified as \u003cem\u003eCampylospora parvula\u003c/em\u003e Kuzuha. Previously \u003cem\u003eC. parvula\u003c/em\u003e was reported for its antibacterial, phosphate solubilization and plant growth promoting activity (Sati and Pant \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Pant and Sati \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Sati et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). However, information available on the antioxidant activity and chemical composition of the tested species is scanty. In the present study \u003cem\u003eC. parvula\u003c/em\u003e recovered from healthy roots of \u003cem\u003eRubus elipticus\u003c/em\u003e growing in riparian area of Nainital, Kumaun Himalaya, India was investigated for its antioxidant potentiality and myco-chemical composition.\u003c/p\u003e \u003cp\u003eAntioxidants have become the subject of interest in recent years. Fungal endophytes represent an abundant and reliable source of novel antioxidant compounds. A number of antioxidants are known to provide protection against several diseases. Epidemiological studies have demonstrated that higher intake of antioxidants results in reduced risk of heart disease and many other diseases. This is the reason for the strong interest in natural antioxidants and their role in human health and nutrition (Bahera et al. 2006). The antioxidant activity of ethyl acetate extract of \u003cem\u003eC. parvula\u003c/em\u003e was evaluated by DPPH radical scavenging, ferric reducing antioxidant power assay (FRAP) and metal chelating assay (MCA). The results summarized in Table \u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e clearly suggest that \u003cem\u003eC. parvula\u003c/em\u003e is a potential source of natural antioxidants.\u003c/p\u003e \u003cp\u003eTo investigate the chemical composition of ethyl acetate extract of \u003cem\u003eC. parvula\u003c/em\u003e GCMS technique was applied. GCMS analysis of the extract showed the various myco chemicals which have antimicrobial and antioxidant properties. Ester and alkene (28%) were detected as highest chemical group in the ethyl acetate extract of tested fungi followed by ketone and alcohol (12%), phenol, carboxylic acid, aldehyde and amine (5%) (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eOctanal, 2-(phenylmethylene)- (16.24%) was reported as a major compound followed by, 1-(4-Iopropylphenyl)-2-methylpropyl acetate (12.91%), Benzoic acid, 2-hydroxy-, phenylmethyl ester (11.85%), benzoic acid, 2-hydroxy-, methyl ester (2.16%). To the best of our knowledge this is the first study on the antioxidant activity and GCMS analysis of \u003cem\u003eC. parvula\u003c/em\u003e. Present work will serve as a platform for future research on endophytic aquatic hyphomyceteous fungi for the production of bioactive compounds having various biological activities.\u003c/p\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eEndophytes are evidently an affluent and unswerving source of bioactive and chemically unique substances with significant therapeutic and agricultural potential but these unseen repositories of microorganisms hidden within the host plants are inadequately studied. This is the first report on the antioxidant activity and myco-chemical profiling of endophytic fungi \u003cem\u003eC. parvula\u003c/em\u003e Kuzuha isolated from the riparian area of Nainital, Kumaun Himalaya. Present study revealed the presence of biologically active compounds which provide an alternative source of developing drug like metabolites. Used fungus is also a good source of antioxidants which can be used to find to find some new antioxidants. This study will also provide a foreword to more comprehensive work on bioactive compounds produced by these endophytic hyphomyceteous fungi.\u003c/p\u003e "},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003ePP performed the experimental work, data collection, analysed the data and wrote the manuscript, and both the co-authors PK and SCS helped in conceptualization and corrected the manuscript.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eDISCLOSURE STATEMENT\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no conflict of interest.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eBadhani A, Rawat S, Bhatt ID, Rawal RS (2015) Variation in Chemical Constituents and Antioxidant Activity in Yellow Himalayan (\u003cem\u003eRubus ellipticus\u003c/em\u003e Smith) and Hill Raspberry (\u003cem\u003eRubus niveus\u003c/em\u003e Thunb). J Food Biochem 39:663\u0026ndash;672\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBehera BC, Verma N, Sonone A, Makhija U (2006) Determination of antioxidative potential of lichen \u003cem\u003eUsnea ghattensis\u003c/em\u003e in vitro. LWT Food Sci Technol 39(1):80\u0026ndash;85. 10.1016/j. lwt.2004.11.007\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBills GF, Gloer JB (2016) Biologically Active Secondary Metabolites from the Fungi. Microbiol Spectrum 2016, 4 (6) FUNK-0009- 2016. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1128/microbiolspec.FUNK-0009-2016\u003c/span\u003e\u003cspan address=\"10.1128/microbiolspec.FUNK-0009-2016\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEl-Elimat T, Raja HA, Figueroa M, Al Sharie AH, Bunch RL, Oberlies NH (2021) Freshwater fungi as a source of chemical diversity: a review. J Nat Prod 84(3):898\u0026ndash;916\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHyde KD, Xu J, Rapior S, Jeewon R, Lumyong S, Niego AGT, Abeywickrama PD, Aluthmuhandiram JVS, Brahamanage RS, Brooks S et al (2019) The amazing potential of fungi: 50 ways we can exploit fungi industrially. Fungal Divers 97(1). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s13225-019-00430-9\u003c/span\u003e\u003cspan address=\"10.1007/s13225-019-00430-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKhare E, Mishra J, Arora NK (2018) Multifaceted interactions between endophytes and plant: Developments and prospects. Frontiers in Microbiology, \u003cem\u003e9\u003c/em\u003e, 2732. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fmicb\u003c/span\u003e\u003cspan address=\"10.3389/fmicb\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. 2018.02732\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLamichhane A, Lamichhane G, Devkota HP (2023) Yellow Himalayan Raspberry (\u003cem\u003eRubus ellipticus\u003c/em\u003e Sm.): Ethnomedicinal, Nutraceutical, and Pharmacological Aspects. Molecules 28:6071\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePant P, Sati SC (2021) Antibacterial activity of root endophytic aquatic hyphomycetes against plant pathogenic bacteria. Vegetos 34:785\u0026ndash;789\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSati SC, Pant P, Vassilev N (2023) Significance of root endophytic aquatic hyphomycetes in phosphate solubilization. Sydowia 75:99\u0026ndash;105\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSati SC, Pant P (2020) Two root endophytic aquatic hyphomycetes \u003cem\u003eCampylospora parvula\u003c/em\u003e and \u003cem\u003eTetracladium setigerum\u003c/em\u003e as plant growth promoters. Asian J Agricultural Res 14:28\u0026ndash;33\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSuryanarayanan TS, Thirunavukkarasu N, Govindarajulu MB, Sasse F, Jansen R, Murali TS (2009) Fungal endophytes and bioprospecting. Fungal Biology Reviews 23(1\u0026ndash;2):9\u0026ndash;19. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.fbr.2009.07.001\u003c/span\u003e\u003cspan address=\"10.1016/j.fbr.2009.07.001\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"archives-of-microbiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"aomi","sideBox":"Learn more about [Archives of Microbiology](https://www.springer.com/journal/203)","snPcode":"203","submissionUrl":"https://submission.nature.com/new-submission/203/3","title":"Archives of Microbiology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Aquatic hyphomycetes, endophyte, Campylospora parvula, antioxidant, myco-chemicals","lastPublishedDoi":"10.21203/rs.3.rs-4683858/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4683858/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eEndophytic fungi are important sources of various bioactive natural compounds. Present work elucidates the antioxidant activity and chemical characterization of \u003cem\u003eCampylospora parvula\u003c/em\u003e Kuzuha isolated from the roots of ethno medicinal plant \u003cem\u003eRubus ellipticus\u003c/em\u003e. \u0026nbsp;Antioxidant activities were tested using DPPH, MCA and FRAP assays, total phenolic and flavonoid content by Folin-Ciocalteu and aluminum chloride method, respectively. Chemical profile of ethyl acetate extract was illustrated by using Gas chromatography and mass spectrometry (GC-MS) analysis. \u0026nbsp;The ethyl acetate extract of\u003cem\u003e C. parvula\u003c/em\u003e presents IC50, 47.75±3.6 µg/ml for DPPH, \u0026nbsp;53.36±2.6 µg/ml for MCA and 360.54±8.6 AAE/g dry extract for FRAP assay. GCMS analysis revealed the presence of Octanal, 2-(phenylmethylene)- (16.24%), 1-(4-Iopropylphenyl)-2-methylpropyl acetate (12.91%), Benzoic acid, 2-hydroxy-, phenylmethyl ester (11.85%) as major compounds. Findings of the present study are promising and keenly hopeful for further research on individual compound to assess their potential, especially in the pharmacology sector.\u003c/p\u003e","manuscriptTitle":"Antioxidant activity and myco-chemical profiling of Campylospora parvula Kuzuha isolated from the roots of Rubus ellipticus","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-07-29 21:32:49","doi":"10.21203/rs.3.rs-4683858/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"editorAssigned","content":"","date":"2024-07-05T16:26:12+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-07-05T07:25:23+00:00","index":"","fulltext":""},{"type":"submitted","content":"Archives of Microbiology","date":"2024-07-04T05:17:41+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"archives-of-microbiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"aomi","sideBox":"Learn more about [Archives of Microbiology](https://www.springer.com/journal/203)","snPcode":"203","submissionUrl":"https://submission.nature.com/new-submission/203/3","title":"Archives of Microbiology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"cc641745-751f-4656-ad7d-54ba795ead45","owner":[],"postedDate":"July 29th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2024-07-29T21:32:49+00:00","versionOfRecord":[],"versionCreatedAt":"2024-07-29 21:32:49","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4683858","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4683858","identity":"rs-4683858","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}