Abstract
Endophytes are microbes that live inside the tissues of plants without causing any disease. Many of these belonging to fungi have been exploited earlier for their biological activities. This study focused on the exploration and characterization of culturable endophytic fungi inhabiting the medicinal plant Dillenia indica l. during four different seasons (summer, monsoon, autumn, and winter) from 2018 to 2019. A total of 2360 segments from different parts (leaves, fruits, and stem) were screened to isolate endophytic fungi. During the study, 25 species of fungi belonging to 20 genera were isolated from the selected plant. The identification of these fungi was validated at morphological, microscopic, and molecular levels. Results indicate the plant has the highest affinity for Daldinia eschscholtzii, followed by Colletotrichum gloeosporioides and Cladosporium cladosporioides. Further, the percent frequency was highest in leaves, followed by stem and fruits. The results were further supported by a similar trend of colonization rate for different plant parts. The monsoon season had the highest number of isolates (312), followed by summer (208), winter (164), and autumn (114). Species diversity was highest during the monsoon season and lowest during the winter. These fungi also produce amylase, lipase, protease, asparaginase, cellulase, and ligninolytic enzymes. This study focused only on culturable fungal endophytes, yet the scope can be extended for other non-culturable microbes and their interaction by using high-throughput genomics and novel next-generation sequencing (NGS) tools. The results indicate that Dillenia indica l. harbors novel endophytic fungi having industrial applications.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
Data are included in this article.
Code availability
Not applicable.
References
Archibald FS (1992) A new assay for lignin-type peroxidases employing the dye azure B. Appl Environ Microbiol 58(9):3110–3116. https://doi.org/10.1128/aem.58.9.3110-3116.1992
Arora P, Wani ZA, Ahmad T, Sultan P, Gupta S, Riyaz-Ul-Hassan S (2019) Community structure, spatial distribution, diversity and functional characterization of culturable endophytic fungi associated with Glycyrrhiza glabra L. Fungal Biol 123(5):373–383. https://doi.org/10.1016/j.funbio.2019.02.003
Barman D, Dkhar MS (2020) Seasonal variation influence endophytic actinobacterial communities of medicinal plants from tropical deciduous forest of meghalaya and characterization of their plant growth-promoting potentials. Curr Microbiol 77(8):1689–1698. https://doi.org/10.1007/s00284-020-01988-3
Blackwell M (2011) The fungi: 1, 2, 3… 5.1 million species? Am J Bot 98(3):426–438. https://doi.org/10.3732/ajb.1000298
Brakhage AA, Schroeckh V (2011) Fungal secondary metabolites–strategies to activate silent gene clusters. Fungal Genet Biol 48(1):15–22. https://doi.org/10.1016/j.fgb.2010.04.004
Dhouib A, Hamza M, Zouari H, Mechichi T, Hmidi R, Labat M, Martinez MJ, Sayadi S (2005) Screening for ligninolytic enzyme production by diverse fungi from Tunisia. World J Microbiol Biotechnol 21(8):1415–1423. https://doi.org/10.1007/s11274-005-5774-z
Elango D, Manikandan V, Jayanthi P, Velmurugan P, Balamuralikrishnan B, Ravi AV, Shivakumar MS (2020) Selection and characterization of extracellular enzyme production by an endophytic fungi Aspergillus sojae and its bio-efficacy analysis against cotton leaf worm Spodoptera Litura. Curr Plant Biol 23:100153. https://doi.org/10.1016/j.cpb.2020.100153
Fang W, Yang L, Zhu X, Zeng L, Li X (2013) Seasonal and habitat dependent variations in culturable endophytes of Camellia sinensis. J Plant Pathol Microbiol 4(3):1000169. https://doi.org/10.4172/2157-7471.1000169
Ferreira MC, Vieira MDLA, Zani CL, de Almeida Alves TM, Junior PAS, Murta SM, Vital MJS (2015) Molecular phylogeny, diversity, symbiosis and discover of bioactive compounds of endophytic fungi associated with the medicinal Amazonian plant Carapa guianensis Aublet (Meliaceae). Biochem Syst Ecol 59:36–44. https://doi.org/10.1016/j.bse.2014.12.017
Ferreira MC, de Assis JCS, Rosa LH (2020) Diversity of endophytic fungi associated with Carapichea ipecacuanha from a native fragment of the Atlantic Rain Forest. S Afr J Bot 134:225–229. https://doi.org/10.1016/j.sajb.2019.12.031
Fuchs B, Krischke M, Mueller MJ, Krauss J (2017) Plant age and seasonal timing determine endophyte growth and alkaloid biosynthesis. Fungal Ecol 29:52–58. https://doi.org/10.1016/j.funeco.2017.06.003
Gopane B, Tchatchouang CK, Regnier T, Ateba CN, Manganyi MC (2021) Community diversity and stress tolerance of culturable endophytic fungi from black seed (Nigella sativa L.). South African J Botany 137:272–277
Goveas SW, Madtha R, Nivas SK, Souza D, L, (2011) Isolation of endophytic fungi from Coscinium fenestratum- a red listed endangered medicinal plant. EurAsian J Biosci 5:48–53. https://doi.org/10.5053/ejobios.2011.5.0.6
Hallmann J, Berg G, Schulz B (2006) Isolation procedures for endophytic microorganisms. In Microbial root endophytes, pp 299–319. Springer, Berlin. https://doi.org/10.1007/3-540-33526-9_17
Hoffman MT, Arnold AE (2008) Geographic locality and host identity shape fungal endophyte communities in cupressaceous trees. Mycol Res 112(3):331–344. https://doi.org/10.1016/j.mycres.2007.10.014
Hyde KD, Bussaban B, Paulus B, Crous PW, Lee S, Mckenzie EHC, Wipornpan P, Lumyong S (2007) Diversity of saprobic microfungi. Biodivers Conserv 16(1):7–35. https://doi.org/10.1007/s10531-006-9119-5
Jagannath S, Konappa N, Lokesh A, Dasegowda T, Udayashankar AC, Chowdappa S, Jogaiah S (2020) Bioactive compounds guided diversity of endophytic fungi from Baliospermum montanum and their potential extracellular enzymes. Anal Biochem. https://doi.org/10.1016/j.ab.2020.114024
Jin Z, Li D, Liu T, Yu F, Zhang Z, Su C, Wang Y, Guo Q, Liu Z (2017) Cultural endophytic fungi associated with Dendrobium officinale: identification, diversity estimation and their antimicrobial potential. Curr Sci 112(8):1690–1697
Joppa LN, Roberts DL, Pimm SL (2010) How many species of flowering plants are there? Proc R Soc B Biol Sci 278(1705):554–559. https://doi.org/10.1098/rspb.2010.1004
Juybari HZ, Tajick Ghanbary MA, Rahimian H, Karimi K, Arzanlou M (2019) Seasonal, tissue and age influences on frequency and biodiversity of endophytic fungi of Citrus sinensis in Iran. For Pathol 49(6):e12559. https://doi.org/10.1111/efp.12559
Kumar V, Prasher IB (2021) Phytochemical analysis and antimicrobial potential of Nigrospora sphaerica (Berk. & Broome) Petch, a fungal endophyte isolated from Dillenia indica L. Adv Tradit Med. https://doi.org/10.1007/s13596-021-00619-x
Kumar V, Prasher IB (2022) Antimicrobial potential of endophytic fungi isolated from Dillenia indica l. and identification of bioactive molecules produced by Fomitopsis meliae (Undrew.) Murril. Nat Prod Res. https://doi.org/10.1080/14786419.2022.2043855
Kusari P, Kusari S, Spiteller M, Kayser O (2013) Endophytic fungi harbored in Cannabis sativa L: diversity and potential as biocontrol agents against host plant-specific phytopathogens. Fungal Divers 60(1):137–151. https://doi.org/10.1007/s13225-012-0216-3
Li WC, Zhou J, Guo SY, Guo LD (2007) Endophytic fungi associated with lichens in Baihua mountain of Beijing, China. Fungal Divers 25:69–80
Liu AR, Chen SC, Jin WJ, Zhao PY, Jeewon R, Xu T (2012) Host specificity of endophytic Pestalotiopsis populations in mangrove plant species of South China. Afr J Microbiol Res 6(33):6262–6269. https://doi.org/10.5897/AJMR12.766
Malinowski DP, Belesky DP (2006) Ecological importance of Neotyphodium spp. grass endophytes in agroecosystems. Grassl Sci 52(1):1–14. https://doi.org/10.1111/j.1744-697X.2006.00041.x
Martinez-Arias C, Sobrino-Plata J, Ormeno-Moncalvillo S, Gil L, Rodriguez-Calcerrada J, Martin JA (2020) Endophyte inoculation enhances Ulmus minor resistance to Dutch elm disease. Fungal Ecol 50:101024. https://doi.org/10.1016/j.funeco.2020.101024
Mishra A, Gond SK, Kumar A, Sharma VK, Verma SK, Kharwar RN, Sieber TN (2012) Season and tissue type affect fungal endophyte communities of the Indian medicinal plant Tinospora cordifolia more strongly than geographic location. Microb Ecol 64(2):388–398. https://doi.org/10.1007/s00248-012-0029-7
Naik BS, Shashikala J, Krishnamurthy YL (2009) Study on the diversity of endophytic communities from rice (Oryza sativa L.) and their antagonistic activities in vitro. Microbiol Res 164(3):290–296. https://doi.org/10.1016/j.micres.2006.12.003
Nishad JH, Singh A, Gautam VS, Kumari P, Kumar J, Yadav M, Kharwar RN (2021) Bioactive potential evaluation and purification of compounds from an endophytic fungus Diaporthe longicolla, a resident of Saraca asoca (Roxb.) Willd. Arch Microbiol 203(7):4179–4188. https://doi.org/10.1007/s00203-021-02390-8
Oita S, Ibáñez A, Lutzoni F, Miadlikowska J, Geml J, Lewis LA, Hom EHY, Carbone I, U’Ren JM, Arnold AE (2021) Climate and seasonality drive the richness and composition of tropical fungal endophytes at a landscape scale. Commun Biol 4(1):1–11. https://doi.org/10.1038/s42003-021-01826-7
Patil MP, Patil RH, Maheshwari VL (2012) A novel and sensitive agar plug assay for screening of asparaginase-producing endophytic fungi from Aegle marmelos. Acta Biol Szeged 56(2):175–177
Rather RA, Srinivasan V, Anwar M (2018) Seasonal deviation effects foliar endophyte assemblage and diversity in Asparagus racemosus and Hemidesmus indicus. BMC Ecol 18(1):1–11. https://doi.org/10.1186/s12898-018-0211-y
Rodriguez RJ, White JF Jr, Arnold AE, Redman RS (2009) Fungal endophytes: diversity and functional roles. New Phytol 182(2):314–330. https://doi.org/10.1111/j.1469-8137.2009.02773.x
Saikkonen K, Mikola J, Helander M (2015) Endophytic phyllosphere fungi and nutrient cycling in terrestrial ecosystems. Curr Sci 109(1):121–126
Singh DK, Sharma VK, Kumar J, Mishra A, Verma SK, Sieber TN, Kharwar RN (2017) Diversity of endophytic mycobiota of tropical tree Tectona grandis Linn. f.: spatiotemporal and tissue type effects. Sci Rep 7(1):1–14. https://doi.org/10.1038/s41598-017-03933-0
Singh AK, Saha S (2019) Chemistry, therapeutic attributes, and biological activities of Dillenia indica Linn. In: Sobti R, Arora N, Kothari R (eds) Environmental biotechnology: for sustainable future. Springer, Singapore, pp 237–260. https://doi.org/10.1007/978-981-10-7284-0_9
Sopalun K, Iamtham S (2020) Isolation and screening of extracellular enzymatic activity of endophytic fungi isolated from Thai orchids. S Afr J Bot 134:273–279. https://doi.org/10.1016/j.sajb.2020.02.005
Sunayana N, Nalini MS, Sampath KKK, Prakash HS (2014) Diversity studies on the endophytic fungi of Vitex negundo L. Mycosphere 5(4):578–590. https://doi.org/10.5943/mycosphere/5/4/10
Suryanarayanan TS, Venkatesan G, Murali TS (2003) Endophytic fungal communities in leaves of tropical forest trees: diversity and distribution patterns. Curr Sci 85(4):489–493
Tan RX, Zou WX (2001) Endophytes: a rich source of functional metabolites. Nat Prod Rep 18(4):448–459. https://doi.org/10.1039/B100918O
Taufiq MMJ, Darah I (2018) Fungal endophytes isolated from the leaves of a medicinal plant, Ocimum sanctum Linn and evaluation of their antimicrobial activities. Afr J Microbiol Res 12(26):616–622. https://doi.org/10.5897/AJMR2018.8812
Tejesvi MV, Mahesh B, Nalini MS, Prakash HS, Kini KR, Subbiah V, Shetty HS (2005) Endophytic fungal assemblages from inner bark and twig of Terminalia arjuna W. & A. (Combretaceae). World J Microbiol Biotechnol 21(8):1535–1540. https://doi.org/10.1007/s11274-005-7579-5
Unterseher M, Reiher A, Finstermeier K, Otto P, Morawetz W (2007) Species richness and distribution patterns of leaf-inhabiting endophytic fungi in a temperate forest canopy. Mycol Prog 6(3):201–212. https://doi.org/10.1007/s11557-007-0541-1
Uzma F, Konappa NM, Chowdappa S (2016) Diversity and extracellular enzyme activities of fungal endophytes isolated from medicinal plants of Western Ghats. Karnataka Egyp J Basic Appl Sci 3(4):335–342. https://doi.org/10.1016/j.ejbas.2016.08.007
Vazquez de Aldana BR, Bills G, Zabalgogeazcoa I (2013) Are endophytes an important link between airborne spores and allergen exposure? Fungal Divers 60(1):33–42. https://doi.org/10.1007/s13225-013-0223-z
Wang Y, Gao BL, Li XX, Zhang ZB, Yan RM, Yang HL, Zhu D (2015) Phylogenetic diversity of culturable endophytic fungi in Dongxiang wild rice (Oryza rufipogon Griff), detection of polyketide synthase gene and their antagonistic activity analysis. Fungal Biol 119(11):1032–1045. https://doi.org/10.1016/j.funbio.2015.07.009
Win PM, Matsumura E, Fukuda K (2018) Diversity of tea endophytic fungi: cultivar-and tissue preferences. Appl Ecol Environ Res 16(1):677–695. https://doi.org/10.15666/aeer/1601_677695
Wu Z, Su Q, Cui Y, He H, Wang J, Zhang Y, Zhao Y, Abul H, Yang Y, Long Y (2020) Temporal and spatial pattern of endophytic fungi diversity of Camellia sinensis (cv. Shu Cha Zao). BMC Microbiol 20(1):1–11
Xu LL, Ling XF, Zhao SJ, Wang RF, Wang ZT (2020) Distribution and diversity of endophytic fungi in Gentiana rigescens and cytotoxic activities. Chin Herb Med 12(3):297–302. https://doi.org/10.1016/j.chmed.2020.03.007
Yuan ZL, Zhang CL, Lin FC, Kubicek CP (2010) Identity, diversity, and molecular phylogeny of the endophytic mycobiota in the roots of rare wild rice (Oryza granulate) from a nature reserve in Yunnan. China Appl Environ Microbiol 76(5):1642–1652. https://doi.org/10.1128/AEM.01911-09
Acknowledgements
The authors acknowledge the Department of Botany, Panjab University Chandigarh, India, for providing infrastructure and instrumentation. Vijay Kumar is also thankful for the Senior Research Fellowship (File No. 09/135(0854)/2019-EMR-I) by the Council of Scientific and Industrial Research (CSIR), India, during research work.
Funding
The research work was not funded by any agency.
Author information
Authors and Affiliations
Contributions
VK carried out experimental work and prepared the manuscript. IBP supervised the work and refined the manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no conflict of interest.
Ethics approval and consent to participate
Not applicable.
Consent for publication
The work is original; there is no plagiarism, and it has not been published anywhere.
Additional information
Communicated by Erko Stackebrandt.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Kumar, V., Prasher, I.B. Seasonal variation and tissues specificity of endophytic fungi of Dillenia indica l. and their extracellular enzymatic activity. Arch Microbiol 204, 341 (2022). https://doi.org/10.1007/s00203-022-02933-7
Received:
Revised:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00203-022-02933-7