Journal of Threatened Taxa | www.threatenedtaxa.org | 26 March 2026 | 18(3): 28594–28599

 

ISSN 0974-7907 (Online) | ISSN 0974-7893 (Print) 

https://doi.org/10.11609/jott.9846.18.3.28594-28599

#9846 | Received 11 April 2025 | Final received 09 February 2026| Finally accepted 28 February 2026

 

 

Range extension of the lichenized ascomycete, Cladonia fruticulosa Kremp., 1882 (Lecanoromycetes: Lecanorales: Cladoniaceae), from Similipal Biosphere Reserve of Odisha

 

Shubham Pradhan 1  , Satyabrata Dash 2 , Bijayananda Sahoo 3   & Biswajit Rath 4          

 

1–4 Department of Biotechnology, Maharaja Sriram Chandra Bhanja Deo University, Sriram Chandra Vihar, Takatpur, Baripada,

Odisha 757003, India.

1 shubham9668518446@gmail.com, 2 satya9132@gmail.com, 3 bsahoo41293@gmail.com,

4 brath_2000@yahoo.com (corresponding author)

 

 

Abstract: The lichen diversity of the Similipal Biosphere Reserve in Odisha remains underexplored, particularly for fruticose lichens genus Cladonia P. Browne. The present study aims to document a specimen collected from the high-altitude region of the reserve. Morphological observations, chemical analysis, and molecular phylogenetic methods were employed for species identification. Detailed descriptions, illustrations, habitat, and ecology are provided. Previously, no specific fruticose lichen species was reported from Similipal, and the present identification evidence confirms the first record of this species from Odisha.

 

Keywords: Diversity, documentation, fruticose, habitat, high altitude, identification, lichen, morphology, phylogeny, species.

 

Editor: Vishal Chauhan, University of Jammu, Jammu, India.       Date of publication: 26 March 2026 (online & print)

 

Citation: Pradhan, S., S. Dash, B. Sahoo & B. Rath (2026). Range extension of the lichenized ascomycete, Cladonia fruticulosa Kremp., 1882 (Lecanoromycetes: Lecanorales: Cladoniaceae), from Similipal Biosphere Reserve of Odisha. Journal of Threatened Taxa 18(3): 28594–28599. https://doi.org/10.11609/jott.9846.18.3.28594-28599

  

Copyright: © Pradhan et al. 2026. Creative Commons Attribution 4.0 International License. JoTT allows unrestricted use, reproduction, and distribution of this article in any medium by providing adequate credit to the author(s) and the source of publication.

 

Funding: The research work is financially supported by the Science & Technology Department, Govt. of Odisha under

                 Biju Pattnaik Research Fellowship (BPRF) [Grant no. ST-BT-MIS-0026-2023-45/ST dt.04.01.2024].

 

Competing interests: The authors declare no competing interests.

 

 

The Similipal Biosphere Reserve (SBR) situated in the northern part of Odisha in eastern India represents a significant centre of biodiversity, characterized by diverse flora and fauna (Sahu et al. 2023). The reserve encompasses heterogeneous vegetation types, high relative humidity, and largely undisturbed forest ecosystems, including stream corridors, woodland patches, valleys, and mountainous terrains. The altitude gradient and microclimatic conditions within the landscape create favourable conditions for lichen colonization and succession (Rinas 2023). Lichens, the symbiotic associations between a mycobiont (fungus) and a photobiont (green alga and/or cyanobacterium), are widely recognized as reliable bioindicators of air quality and climatic conditions (Nash 2008). The moist tropical climate of Similipal, together with its complex topography and stable forest canopy, supports a rich assemblage of epiphytic, saxicolous, and terricolous lichen taxa (Singh & Sinha 2010; Singh & Kumar 2012). Despite the ecological importance of lichens, systematic studies on lichen diversity in Similipal Biosphere Reserve remain limited, especially for fruticose species of the genus Cladonia. Many areas of the reserve, particularly the core forest zones, remain poorly explored for their lichen flora. Considering the diverse habitats and favourable climate, the region may harbour previously unreported lichen taxa. Therefore, the present study aimed to survey lichen diversity in the core zone of the reserve and the fruticose lichen Cladonia fruticulosa was recorded from terricolous and muscicolous substrates representing a new addition to the lichen biota of Odisha.

 

Materials And Methods

Study area and lichen collection:

The Similipal Biosphere Reserve (SBR), located in Mayurbhanj District, Odisha, India (21.116°′–22.200° N  85.966°–86.700° E), represents a heterogeneous landscape comprising dry and moist deciduous forests, grasslands, riverine systems, valleys, and undulating mountainous terrain. The elevation range is between a minimum of 40 m and a maximum of 1,178 m. The reserve harbours rich and diverse flora and fauna, including a significant assemblage of lichenized fungi. The area experiences a moist tropical climate with annual rainfall ranging 1,200–2,000 mm. The mean maximum temperature reaches 38–44 °C during summer (May–Jun), while minimum temperatures decline to about 08–12 °C in winter (Dec–Jan). Such weather conditions, together with topographic heterogeneity and stable forest cover, provide favourable microhabitats that support lichen colonisation and succession, resulting in varied distribution patterns across substrates.

       Lichen specimens were collected from different substrates using random sampling technique from transition, buffer, and core zones of SBR. The transition zone included Lulung, Sitakund, Kachudahan, Palasibeda, Digdiga, Namtidar, Tunki, Taldiha, Devkund, and Upparbarakamuda. The buffer zone comprised Bhajam, Chandanchaturi, Champajhar, Khadkei, Nigirdha, Tulasibani, Manada, and Brundeiposi. The core zone encompasses Dholabani, Kailani, Jamuani, Andharajuli, Bakua, Ghar Similipal, Jodapal, Dhudruchampa, Kukurbhuka, Hatibandha, Utras, Bariooan, Ektali, Upparbarakamuda, Jenabil, Bhanjabasa, and Meghasani.

 

Morphological and chemical characterization

Lichen specimens were critically examined and differentiated based on morphological, anatomical, and chemical characters following Awasthi (2007). Macromorphological characters were analysed under a stereomicroscope (Stemi 305, Carl Zeiss), while anatomical features were analysed using a dissecting microscope (Dewinter, DGI 1000). Routine reagents, i.e., K reagent (Potassium hydroxide, KOH 10%), C reagent (Potassium hypochlorite, KClO, 5.25%) and paraphenylenediamine (PD) were used for spot tests. The voucher specimens are deposited in the National Botanical Research Institute, Lucknow (LWG).

 

Molecular characterization

Molecular characterization was carried out through rRNA sequencing, including 18s, 28s, 5.8s, 1.5s, and ITS sequences. Genomic DNA was extracted from 10 mg of lichen thallus following the precipitation method of Park et al. (2014). The thallus was placed in micro vial along with three to four sterile 2.5 mm glass beads, under liquid nitrogen and disrupted the powder by using a mini-beadbeater-24 for 30 s. This freeze-thaw disruption cycle was repeated until the sample was ground into a fine powder. Subsequently, 300 µL of KCl extraction buffer was added, and the tube was vigorously inverted by hand approximately 20 times to ensure thorough mixing. An equal volume (300 µL) of chloroform was then added, and the sample was gently inverted about 20 times before centrifugation at 12,000 rpm for one minute at room temperature. The upper aqueous phase was transferred to a 1.5 ml microcentrifuge tube, and DNA was precipitated by adding 180 µL of chilled isopropanol (60%), followed by gentle inversion. The mixture was centrifuged again at 12,000 rpm for one minute, and the DNA pellet was washed with 300 µL of chilled 70% ethanol. After drying the pellet at 50–65 °C then resuspended in 100 µL of 1× TE buffer and incubated again at 50–65 °C for five minutes to ensure complete dissolution. The purified DNA was stored at 4 °C for short-term use or at -20 °C for long-term preservation.

 

PCR amplification and sequencing

PCR amplification was performed following the protocol of White et al. (1990) using universal fungal primers targeting the small subunit (SSU), large subunit (LSU), and internal transcribed spacer (ITS) regions. For SSU amplification, primers SR1R (5′-TACCTGGTTGATCCTGCCAGT-3′) and SR7 (5′-GTTCAACTACGAGCTTTTTAA-3′) were used. LSU was amplified using primers LS1 (5′-GTACCCGCTGAACTTAAGC-3′) and LS5 (5′-TCCTGAGGGAAACTTCG-3′). The ITS region was amplified using ITS5 (5′-GGAAGTAAAAGTCGTAACAAGG-3′) and ITS4 (5′-TCCTCCGCTTATTGATATGC-3′) (White et al. 1990), along with the fungal-specific primer nu-SSU-1583-59 (5′-CAACGAGGAATTCCTAGT-3′) (DePriest 1993). PCR products were electrophoresed on 1.5% agarose gel at 120 V for 25 min to confirm amplicon size and integrity. Amplified products were purified using Exonuclease I and recombinant Shrimp Alkaline Phosphatase, and sequenced bidirectionally on an ABI 3730 Capillary Electrophoresis Genetic Analyzer. Forward and reverse sequences were edited and assembled into a consensus sequence (481 bp) using MEGA 11 and Sequencher v5 software. To align the sequences with other sequences obtained from NCBI GenBank, Clustal W was employed (Tamura et al. 2021)  using MEGA 11.

 

Results

Cladonia fruticulosa Kremp., Verh. Zool.-Bot. Ges. Wien 30: 331 (1880)      

Thallus fruticose; basal squamules persistent, 2–4 mm long, 1–2 mm in diam., faintly to deeply lacinate, rarely granular-sorediate on borders or below; podetia growing from basal squamules, 0.5–2 cm rarely 4 cm tall, 0.5–1.0 mm diam., simple, rarely branched, scyphose, often partly squamulose, corticate or upper part ecorticate or almost completely ecorticate; ecorticate area partly granular-sorediate; scyphi 0.5–4 mm in diam., deformed or well defined, often with marginal scyphi; apothecia 1–2 mm in in diameter, convex, pedicellate or marginal on scyphi, pale brown to dark brown; pycnidia on scyphi, 0.3–0.5 × 0.2–0.3 mm, flask-shaped, dark brown (Image 1).

Spot tests: K–, UV–, C–, I– (All tests negative).

Specimen examined: India, Odisha, Mayurbhanj District; elevation 1,133 m; substrate-moss, rock and red soil; date of collection22.ii.2024; collected by Shubham Pradhan, Satyabrata Dash, Bijayananda Sahoo and Biswajit Rath; Accession No-LIC-69571; Herbarium acronym-LWG.

 

Phylogenetic analysis

The evolutionary history was inferred by using the maximum likelihood method. The bootstrap consensus tree inferred from 1,000 replicates is taken to represent the evolutionary history of the taxa analysed. The percentage of replicate trees in which the associated taxa clustered together in the bootstrap test 1,000 replicates) are shown below the branches. Initial tree(s) for the heuristic search were obtained automatically by applying Neighbor-Join and BioNJ algorithms to a matrix of pairwise distances estimated using the Tamura-Nei model. This analysis involved 26 nucleotide sequences. There was a total of 535 positions in the final dataset. Evolutionary analyses were conducted in MEGA11 (Tamura et al. 2021) (Figure 1).

 

Discussion

The family Cladoniaceae has been comprehensively investigated in the Indian subcontinent by several authors, notably Upreti (1987) and Awasthi (1988). According to the updated account of Singh & Sinha (2010), the family in India comprises four genera: Cladia (one species), Cladonia (58 species), Gymnoderma (one species), and Pilophorus (one species). A significant contribution to the taxonomy of Indian Cladonia was made by Ahti et al. (2002), who provided the first detailed account of several species from India, including the description of Cladonia singhii Ahti & Dixit from the eastern Himalayan region. In addition, seven species of Cladonia corniculata, C. kanewskii, C. laii, C. luteoalba, C. mauritiana, C. mongolica, and C. rei were newly incorporated into the recent checklist of lichens of the Indian subcontinent. Furthermore, the occurrence of Cladonia cartilaginea in India was confirmed by Ahti et al. (2002), thereby refining the known distributional range of the genus in the region. A detailed account of the altitudinal distribution and edaphic preferences of several Cladonia species in India was provided by Řídká et al. (2014). Cladonia furcata was recorded on soil substratum at 3,250 m from Tungnath (Rudraprayag District), Uttarakhand, whereas Cladonia rangiferina occurred on terricolous substrata at 2,553 m between Bogdiyar and Naher Devi in Pithoragarh, Uttarakhand. Cladonia praetermissa was collected on soil at 1,665 m in Anuppur District, Madhya Pradesh. Cladonia scabriuscula exhibited a broader ecological amplitude, growing on soil substratum at 1,014 m in North Cachar (Assam) and at 1,410 m in Satara (Maharashtra). Cladonia verticillata was reported as terricolous at 1,890 m in Champawat, Uttarakhand, while Cladonia fruticulosa was documented on soil at 2,607 m in the Nilgiri Hills of Tamil Nadu. These records emphasize the predominance of terricolous (soil-inhabiting) habitats among Indian Cladonia species and demonstrate their adaptability across a wide altitudinal gradient from subtropical to alpine ecosystems.

Additional regional records further substantiate the expanding distributional range of Cladonia species in India. Reports from Sikkim by Sinha & Ram (2011) contributed to the floristic documentation of the genus in northeastern India. Cladonia coniocraea and C. fruticulosa were reported from the Mandi District of Himachal Pradesh by Thakur et al. (2020). Subsequently, Gogoi et al. (2022) documented C. fruticulosa, C. scabriuscula, C. subradiata, and C. cervicornis from Assam, indicating a broader ecological amplitude of the genus across the Indo-Burma biodiversity region.

The present detection of C. fruticulosa at 1,133 m altitude on red soil substratum in northern Odisha represents the first confirmed record from this phytogeographical sector. Previous diversity assessments within the Similipal Biosphere Reserve and adjoining habitats by Pradhan et al. (2025, 2026) did not include this species in their respective checklists. Therefore, the present report significantly augments the known distribution of C. fruticulosa in India and contributes to a more comprehensive understanding of Cladonia diversity in eastern habitats.

 

Conclusions

The present study records Cladonia fruticulosa Kremp. for the first time from the Similipal Biosphere Reserve, extending its known distribution in eastern India. Its occurrence at higher elevations indicates a preference for well-drained, open forest microhabitats with specific edaphic and microclimatic conditions. This record refines the regional species inventory of Cladonia and contributes to the taxonomic study of fruticose lichens in tropical forest ecosystems.

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