The first report of Helvella crispa (Ascomycota, Pezizales), a rare fungal species in Sri Lanka

: Two striking, stipitate fruiting bodies of a fungus were found in Hantana area, bordering the Dunumadalawa Forest Reserve within the Kandy District (Central Province) of Sri Lanka. The apothecium was capitate, cap whitish and irregularly saddle-shaped; the stipe simple, shallowly ribbed, tapering and internally solid. The fruiting bodies were identified, using morphological characteristics, as Helvella crispa belonging to the family Helvellaceae, Order Pezizales and Phylum Ascomycota. This is the first record of H. crispa or the genus Helvella in Sri Lanka. The genus could also be rather rare in Sri Lanka (then Ceylon) as it has not been encountered over 230 year-long history of mycological work in the country.


INTRODUCTION
Sri Lanka is a small island of the Indian ocean, with a land area of 6,570,134 ha., located closer to the southern part of the Indian sub-continent, between 5 o 54' and 9° 52' North Latitude and 79 o 39' and 81 o 53' East Longitude. The island comprises of central mountains with an elevation of up to 2524 m, surrounded by the lowlands. The overall climate is considered tropical and the average temperature ranges between 28-32 ͦ C. Despite its small size, Sri Lanka is blessed with a variety of ecosystems and high biological diversity. Sri Lanka's native fungal flora is believed to be around 25,000 species of which only about less than 12% is known (Adikaram, 2004;Adikaram and Yakandawala, 2020). The earliest record of fungi in Ceylon was as far back as 1783 by Houttuyn. Mycological studies initiated by British scientists in the country from 1780s to late nineteen forties. The work continued even after 1940's, though not in the same intensity. The fact that none of the species belonging to the genus Helvella were reported to be encountered in the country during the long period of continuous mycological work in the country (Petch and Bisby, 1950;Karunarathna et al. 2012) shows that H. crispa might have been a rare species in Sri Lanka.
Helvella is a genus of apothecial ascomycetes, widespread in terrestrial biomes of the Northern and Southern Hemispheres (Kirk et al., 2008). The species in the genus are characterized by sub-sessile or stipitate fruiting body, cupulate to saddle-shaped and convex to campanulate apothecia, including species with folded and lobed caps seated on a simple ribbed or furrowed stipe (Skrede et al., 2017). The genus contains many of the macrofungi species of the order Pezizales and comprises a range of elaborate apothecia (Skrede et al., 2017).
In most of the past taxonomic circumscriptions, a broad species concept has been applied, allowing greater morphological variation among Helvella species that were accepted in their investigations (Afzelius, 1783;Fries, 1822;Dissing, 1966a, b;Weber, 1972Weber, , 1975Abbott and Currah, 1997). This has also resulted in listing a number of heterotypic synonyms for many Helvella species, mostly assessed based on character similarities of published descriptions rather than as a result of comparative studies of type specimens (Skrede et al., 2017). Skrede et al. (2017) have found the occurrence of a "crispa lineage", consisting of H. crispa, H. maculata and H. leucophaea, constituting a species assemblage of their own, in a combined morphological and molecular study, conducted using 183 Helvella specimens selected from 432 specimens, collected mostly from the Europe and a small sample from other regions of the world. Conversely, Zhao et al. (2015) concluded that the Chinese H. crispalike samples represented a species complex, containing at least six phylogenetic species. Helvella species have also been reported from Europe (Skrede et al., 2017) and North America (Kempton and Wells, 1970). Sixteen species of Helvella are recorded in India (Jamaluddin et al., 2004;Dorjey et al., 2013) of which eleven species have been reported from Jammu and Kashmir (Dorjey et al., 2013) with a temperate climate. Apothecial ascomycete (Pezizomycetes), H. crispa, is commonly known as white saddle, elfin saddle or false morel over the world.
There were no records of any of the members of the genus Helvella or even the Family Hevellaceae in Sri Lanka. The present study reports a species belonging to the genus Helvella Family Hevellaceae collected from Hantana, Kandy for the first time in Sri Lanka (Central Province).

Specimen collection
Two identical fruiting bodies of a macrofungus were found close to each other (4 cm apart) on 25 th July 2015, located in Hantana, at a site just opposite and in close proximity to the boundary of the Dunumandalawa Forest Reserve (N 07° 27.4848', E 080° 63.9493', 700 -1000 m elevation which was previously known as the Walker Estate, in the Kandy District (Central Province of Sri Lanka). The fruiting bodies, aboveground, were first examined, described and photographed at its natural habitat. Further observations and photographs of the fruiting bodies, with their underground portion, were made after uprooting.

Habitat
The collection site of the two stipitate fruiting bodies was located in Hantana area (Kandy District, Central Province, Sri Lanka), at an elevation of 700 -1000 m during the rainy season (South Western monsoon). They were growing on bare, sand-rich, soil with sparse ground cover closer to the vegetation of the lower montane forest. The conditions were slightly wet with moderate temperature, 25±2 o C, and 90% relative humidity.

Spore print
The fruiting body was longitudinally halved and examined for internal features and photographed, the observations were recorded. A spore print was obtained by keeping a longitudinally half-cut and exposed cap of the fruiting body, horizontally over a white paper for 6 h. The specimen was identified using morphological characteristics described by Fries (1822).

Specimen deposition
Specimen was deposited in a fungal collection at the Department of Botany, University of Peradeniya, Sri Lanka, the accession number is DB/UPDN/2015/03.

Identification
Using morphological characteristics of the ascocarps, the fungus was unambiguously identified as Helvella crispa (Scop.) Fr. belonging to the crispa lineage.
The stem or the stipe prominent, the aboveground portion 5.5 cm in height and white color, the outer surface shallowly ribbed over the entire length, continuing to the receptacle margin, the stipe internally solid, fleshy and white color. Stipe, at its broadest point at soil level, 4.2 cm in diameter, and tapering to about 2.2 cm at the cap level. Stipe extends to 7 cm underground (Fig. 1), brown color and broader to begin with at ground level and tapers down over the initial 1/3 rd to 2 cm and continues with the same diameter (Fig. 1).

Taxonomy
The current taxonomic rank of the fungus is Kingdom  on macroscopic characteristics of the apothecium, shape, color and outer surface characteristics, the hymenium, excipulum and stipe.
While most of the morphological features of the cap, stipe or the receptacle of the specimens used in the present study tallied entirely with the currently available records and descriptions, certain features, such as the "flower-like appearance" of the cap, was complex and even difficult to be accurately described. These features could not be compared with any of the existing records of H. crispa. This again reiterates the greater morphological diversity within the species. A recent and extensive study, that gathered morphology and phylogenetic evidence from 4 loci, hsp, tef, rpb2 and LSU of Helvella species, using large numbers of samples from Europe and smaller sample numbers from other regions of the world, had shown very little "within species divergence" (Skrede et al., 2017). Nevertheless, some species showed more intraspecific diversity than others. Among them, H. costifera and H. crispa had highest "within-species diversity" (Skrede et al., 2017).
Morphological features and the habitat-ecology of the two species (Skrede et al., 2017), H. maculata and H. leucophaea, that make a "cripa lineage", together with H. crispa, were distinct from the fungal specimens subjected to the present study. Helvella leucophaea resembles H. crispa in size and stature, the stipe is whitish but appears deeply ribbed. Helvella maculata is a temperate species that features an elaborately ribbed and pocketed stem, a brownish cap that is loosely lobed, and a fuzzy, whitish undersurface. The species appears under conifers and hardwoods from Alaska to California, usually in fall, but also over winter or in spring in coastal areas (Weber, 1972).
Neither any species belonging to the genus Helvella nor the genus itself has been recorded in Sri Lanka during its 230-year long history of intense Mycological investigations. The occurrence of the specimens described in the present paper in the country could be considered rare.

CONCLUSIONS
Fungal fruiting bodies collected from Hantana, Kandy, Sri Lanka were morphologically identified as Helvella crispa. This is the first record of H. crispa or the genus Helvella in Sri Lanka.