Author(s): Madhara K. Wimalasena
Institute: Faculty of Graduate Studies, Sabaragamuwa University of Sri Lanka, Belihuloya, 70140, Sri Lanka (MPhil Registered University) Department of Bioprocess Technology, Faculty of Technology, Rajarata University, Mihintale, 50300, Sri Lanka (Place of laboratory work were conducted)
Principal Investigator: Prof. Nalin N. Wijayawardene, Prof. T.C. Bamunuarachchige, Prof. Dong-Qin Dai, Dr. Heethaka K. S. de Zoysa, Prof. Darbhe J. Bhat and Dr. Turki M. Dawoud
Published: August 29, 2025
Category: Research Projects
Abstract: Open Publication
📄 Download Research Document

Abstract

This project focuses on exploring the diversity of **freshwater fungi in Sri Lanka**, with special attention to freshwater hyphomycetes (Ingoldian fungi). It aims to discover new fungal species, record novel host and habitat associations, and document fungal biodiversity through taxonomic checklists and culture collections. The research combines morphological and molecular techniques, contributing not only to global fungal taxonomy and ecology but also to building local research capacity by mentoring young scientists.

Full Description

About research:

Research topic:

Taxonomy and Phylogeny of Plant Material Associated Freshwater Fungi in Anuradhapura and Ratnapura Districts and Screening Their Extracellular Enzymes

Conducted by:

Madhara K. Wimalasena, Prof. Nalin N. Wijayawardene, Prof. T.C. Bamunuarachchige, Prof. Dong-Qin Dai, Dr. Heethaka K. S. de Zoysa, Prof. Darbhe J. Bhat and Dr. Turki M. Dawoud

Collaborations:

  1. Center for Yunnan Plateau Biological Resources, Protection and Utilization & Yunnan International Joint Laboratory of Fungal Sustainable Utilization in South and Southeast Asia, College of Biology and Food Engineering, Qujing Normal University, Qujing, Yunnan, 655011, P.R. China.
  2. Department of Bioprocess Technology, Faculty of Technology, Rajarata University of Sri Lanka, Mihintale 50300, Sri Lanka.
  3. Faculty of Graduate Studies, Sabaragamuwa University of Sri Lanka, Belihuloya 70140, Sri Lanka.
  4. Department of Botany and Microbiology, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia.
  5. Formerly, Department of Botany, Goa University, Goa 403206, India.
Funded by:
  1. Science and Technology Human Resource Development Project, Ministry of Higher Education, Sri Lanka, funded by the Asian Development Bank (Grant number R3-RJ2)
  2. Tropical Microbiology Research Foundation, 96/N/10, Meemanagoda Road, Pannipitiya 10230, Sri Lanka.

Objectives:

Main objective
  1. To study the taxonomy and classification of plant material associated freshwater fungi in lotic and lentic freshwater ecosystems in the districts of Anuradhapura and Ratnapura
Specific objectives
  1. To collect and isolate the plant material associated with freshwater fungi from Anuradhapura and Ratnapura districts
  2. To classify and identify freshwater fungi using DNA-based phylogenetic studies
  3. To screen and identify the freshwater fungi which are producing lignocellulolytic, cellulolytic, and hemicellulolytic enzymes in freshwater ecosystems
  4. To maintain the fungal repository of living and dry cultures for the freshwater fungi at Rajarata University of Sri Lanka

Background research:

This study focused on the taxonomy and classification of plant materials associated with freshwater fungi in the Anuradhapura and Ratnapura districts of Sri Lanka, along with their extracellular enzymatic potentials. To identify freshwater fungi, both lentic and lotic freshwater habitats were selected as sampling locations in the Anuradhapura and Ratnapura districts, which represent the country’s dry and wet climatic zones, respectively. A total of 35 sampling sites were designated, with 22 in Anuradhapura and 13 in Ratnapura, ensuring thorough geographic coverage. Samples of submerged dead plant materials, healthy freshwater plant species, and foam formations were collected to isolate freshwater fungi. Various isolation techniques were employed, including single-spore isolation, direct isolation, and the surface sterilization method.

A total of 130 fungal strains were isolated and identified morphologically, with approximately 50 species confirmed through molecular identification using the ITS gene region. Multi-gene molecular identification was performed for 15 species, and multi-gene phylogenetic analyses were conducted using ITS, LSU, SSU, ACT, CHS-1, GAPDH, tub2, rpb2, and tef1-α gene regions. As a result of this study, a significant outcome is the discovery of two novel fungal taxa, ten new host records, and eleven new geographical records of saprobic and endophytic fungi in Sri Lanka.

The novel species Ectophoma salviniae and Neottiosporina mihintaleensis were isolated from Salvinia spp. The number of endophytic fungal species isolated from plant tissues was greater in the Ratnapura district. In contrast, saprobic fungi were more frequently isolated from submerged plant materials in the Anuradhapura district. Furthermore, the identified fungal species, characterized through both morphological and molecular analyses, were evaluated for extracellular enzymatic activities, including amylase, cellulase, and laccase, using qualitative assays. These findings help to fill the gap in knowledge of fungal species in Sri Lanka and reveal their enzymatic capabilities, offering promising avenues for future biotechnological applications.

Research outcomes:
  1. Introduced three novel species: Colletotrichum hydei, Ectophoma salviniae, and Neottiosporina mihintaleensis
  2. Introduced ten new host records for the following species: Colletotrichum chlorophyti, coccodes, C. gloeosporioides, C. queenslandicum, C. siamense, C. truncatum, C. vittalense, Daldinia eschscholtzii, Nemania primolutea, and Nigrospora lacticolonia
  3. Introduced eleven new geographical records for the following species: Aureobasidium melanogenum, Chaetomella raphigera, Colletotrichum chlorophyti, coccodes, C. vittalense, Coniochaeta velutina, Lasiodiplodia crassispora, L. pseudotheobromae, Nemania primolutea, Nigrospora lacticolonia, and Rhytidhysteron neorufulum
  4. Laccase enzymatic activity was exhibited by freshwater fungi, including Aureobasidium melanogenum, Chaetomella raphigera, Colletotrichum siamense, Colletotrichum, Colletotrichum truncatum, Ectophoma salviniae sp. nov., Lasiodiplodia pseudotheobromae, Microdiplodia sp., Montagnula donacina, Neottiosporina mihintaleensis sp. nov., Nemania primolutea, Nigrospora sphaerica, Paraconiothyrium archidendri, Phyllosticta capitalensis, Rhytidhysteron neorufulum, Talaromyces pinophilus, and Talaromyces stipitatus
  5. Cellulase enzymatic activity was exhibited by freshwater fungi, including Chaetomella raphigera, Colletotrichum siamense, Colletotrichum truncatum, Ectophoma salviniae nov., Nemania primolutea, Phyllosticta capitalensis, Talaromyces pinophilus, and Talaromyces stipitatus
  6. Amylase enzymatic activity was exhibited by freshwater fungi, including Colletotrichum siamense, Colletotrichum, Ectophoma salviniae sp. nov., Nemania primolutea, and Phyllosticta capitalensis
  7. The highest amylase, cellulase, and laccase production was exhibited by Nemania primolutea, Chaetomella raphigera, and Neottiosporina mihintaleensis nov., respectively
  8. Freshwater saprobic and endophytic fungal species identified through morphological and molecular analyses are listed below
1.       Aureobasidium melanogenum 1.       Ectophoma multirostrata 1.       Neottiosporina mihintaleensis
2.       Chaetomella raphigera 2.       Ectophoma salviniae 2.       Neurospora crassa
3.       Colletotrichum chlorophyti 3.       Hypoxylon lenormandii 3.       Nigrospora lacticolonia
4.       Colletotrichum coccodes 4.       Lasiodiplodia crassispora 4.       Paraconiothyrium sp.
5.       Colletotrichum gloeosporioides 5.       Lasiodiplodia pseudotheobromae 5.       Phyllosticta capitalensis
6.       Colletotrichum hydei 6.       Lentinus tigrinus 6.       Rhytidhysteron neorufulum
7.       Colletotrichum queenslandicum 7.       Microdiplodia sp. 7.       Rhytidhysteron spp.
8.       Colletotrichum scovillei 8.       Microdochium musae 8.       Talaromyces pinophilus
9.       Colletotrichum siamense 9.       Montagnula donacina 9.       Talaromyces stipitatus
10.    Colletotrichum vittalense 10.    Nemania primolutea 10.    Talaromyces verruculosus
11.    Colletotrichum truncatum 11.    Neopestalotiopsis clavispora 11.    Trichoderma harzianum
12.    Coniochaeta velutina 12.    Neopestalotiopsis saprophytica 12.    Trichoderma koningii
13.    Daldinia eschscholtzii 13.    Neottiosporina cylindrical 13.    Trichoderma sp.
14.    Alternaria spp. 14.    Corynespora spp. 14.    Curvularia spp.

Publications:

Published research articles in SCI-indexed journals:
  1. Wimalasena, M. K., Wijayawardene, N. N., Bamunuarachchige, T. C., Zhang, G.-Q., Jayalal, R. G. U., Bhat, D. J., Dawoud, T. M., de Zoysa, H. K. S., & Dai, D.-Q. (2025). Ectophoma salviniae nov., Neottiosporina mihintaleensis sp. nov., and four other endophytes associated with aquatic plants from Sri Lanka and their extracellular enzymatic potential. Frontiers in Cellular and Infection Microbiology, 14, 1475114. https://doi.org/10.3389/fcimb.2024.1475114 | Quartile score: Q1
  2. Wimalasena, M. K., Bamunuarachchige, T. C., Jayalal, U., Sci, C. J. M., Jayalal, R. U., Bhat, D. J., Dawoud, T. M., Rathnayaka, R. K., & Dai, D.-Q. (2024). Ongoing study reveals high aquatic fungal diversity: An untouched component in Sri Lankan biodiversity. Chiang Mai Journal of Science, 51(3), 1–31. https://doi.org/10.12982/CMJS.2024.039 | Quartile score: Q3
Ongoing research manuscripts:
  1. Madhara K. Wimalasena, Nalin N. Wijayawardene, Susan B. Dharmarathne, Nimesha Gunasekara, Manavi S. Ekanayake, R.G. Udeni Jayalal, Jayarama D. Bhat, Turki M. Dawoud, Gui-Qing Zhang, Damith C. Weerasinghe, Heethaka K. S. de Zoysa, Dong-Qin Dai, Haohan Wang, Thushara C. Bamunuarachchige. (2025). In vitro Evaluation of Synthetic Dye Decolorization by Fungi Isolated from Freshwater Environments in Sri Lanka. Frontiers in Cellular and Infection Microbiology. Manuscript ID:1650835 | Quartile score: Q1 | In review
  2. Madhara K. Wimalasena, Nalin N. Wijayawardene, Thushara C. Bamunuarachchige, Heethaka K. S. de Zoysa, Gui-Qing Zhang, Darbhe J. Bhat, Turki M. Dawoud, Sajeewa S. N. Maharachchikumbura, Dong-Qin Dai, Udeni Jayalal. (2025). Colletotrichum hydei nov. and New Geographical and Five New Records of Endophytic Fungi Associated with Freshwater Plants in Sri Lanka. New Zealand Journal of Botany | Quartile score: Q3 | In prep.
Published abstracts:
  1. Wimalasena, S.D.M.K. et al. (2025). Exploring the Untouched Diversity of Freshwater Hyphomycetous in Lentic Freshwater Habitats in Anuradhapura District, Sri Lanka. 4th International Symposium on Agriculture (ISA 2025). 6th of March 2025. Eastern University of Sri Lanka
  2. Wimalasena, S.D.M.K. et al. (2024). Revealing the Endophytic Fungal Diversity Associated with Freshwater Plants in Anuradhapura and Ratnapura Districts, Sri Lanka. Graduate Colloquium 2024 (ARS-24). 20th December 2024. Sabaragamuwa University of Sri Lanka
  3. Wimalasena, S.D.M.K. et al. (2024). Lasiodiplodia crassispora & pseudotheobromae isolated in Sri Lanka show mycoremediation abilities for decolorizing crystal violet. The 50th International Congress on Science, Technology & Technology-based Innovation (STT 50). 27th of November 2024. Chiang Mai University, Thailand
  4. Wimalasena, S.D.M.K. et al. (2024). Exploring laccase-producing fungi isolated from freshwater ecosystems in the Anuradhapura & Ratnapura Districts, Sri Lanka. Proceedings of the 44th Annual Sessions, Institute of Biology Sri Lanka (IOBSL). 28th of September 2024. Institute of Biology Sri Lanka
  5. Wimalasena, S.D.M.K. et al. (2023). Morphological & molecular identification of Rhytidhysteron neorufulum found in aquatic environments of Anuradhapura District, Sri Lanka. Young Scientists’ Conference on Multidisciplinary Research (YSCMR). 09th of November 2024. National Institute of Fundamental Studies, Sri Lanka
  6. Wimalasena, S.D.M.K. et al. (2023). Diversity of freshwater fungi in Anuradhapura District, Sri Lanka. 1st International Conference on Technological Research & Innovation (ICTRI – 2023). 28th of September 2023. Faculty of Technology, Eastern University, Sri Lanka
  7. Wimalasena, S.D.M.K. et al. (2022). Screening & identification of the cellulolytic, hemicellulolytic & lignocellulolytic enzymes of freshwater fungi in the Anuradhapura District, Sri Lanka. International Conference on Development & Utilization of Fungal Resources (ICDUFR 2022). 26th October 2022. Qujing Normal University, China
  8. Wimalasena, S.D.M.K. et al. (2022). Freshwater fungal diversity in selected Anuradhapura lakes of Sri Lanka. 3rd International Eurasian Mycology Congress (EMC’22). 09th of September 2022. Van YYU Mushroom Application and Research Center, Van, Türkiye