Journal of Threatened Taxa |
www.threatenedtaxa.org | 26 October 2022 | 14(10): 22021–22025
ISSN 0974-7907 (Online) | ISSN 0974-7893
(Print)
https://doi.org/10.11609/jott.7881.14.10.22021-22025
#7881 | Received 14 February 2022 | Final
received 03 August 2022 | Finally accepted 30 September 2022
First report of Gymnopilus
ochraceus Høil. 1998 (Agaricomycetes:
Agaricales: Hymenogastraceae)
from India and determination of bioactive components
1 dhirajanj@gmail.com,
2 sushant.bornak94@gmail.com (corresponding author)
Abstract: A rare Gymnopilus
species G. ochraceus Høil.
collected from live Ficus platyphylla Del. tree is described and illustrated for
the first time from India. Morphological and microscopic characters with
molecular and biochemical analysis has been discussed. To indicate that, 25
species of Gymnopilus have previously been
recorded from India and the species Gymnopilus
ochraceus is hereby reported for the first time,
thus making a total of 26 species from India.
Keywords: Biochemical analysis, Ficus
platyphylla, fungi, microscopic characters,
mushroom, taxonomy.
Editor: Richard Mibey,
University of Nairobi, Kenya Date
of publication: 26 October 2022 (online & print)
Citation: Patil, A.R. & S.I. Bornak (2022). First report of Gymnopilus ochraceus Høil. 1998
(Agaricomycetes: Agaricales: Hymenogastraceae)
from India and determination of bioactive components. Journal of Threatened Taxa 14(10): 22021–22025. https://doi.org/10.11609/jott.7881.14.10.22021-22025
Copyright: © Patil & Bornak 2022. 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: None.
Competing interests: The authors declare no competing
interests.
The genus Gymnopilus (family Hymenogastraceae) is represented by 200 species in the
world (Kirk et al. 2008). Members of the genus are characterized by
yellow to brown, ferruginous or purple fruiting bodies, saprotrophic nature, cortinoid to membranous veil and a rusty brown spore print
(Kaur et al. 2015). The genus was confirmed by roughened basidiospores that
range from verrucose to rugulose; capitate to
sub-capitate, ventricose cheilocystidia and clamp
connections present on almost all kinds of hyphae (Khan et al. 2017). Shape and
size of the spores and cystidia are considered important characters for distinction
among the species (Rees et al. 2004). Based on the pigments and the
non-mycorrhizal habit, Kühner (1984) classified Gymnopilus together with Galerina
Earle in the Strophariaceae family. Later, Singer
(1986) placed it in family Cortinariaceae due to the
ornamentation and lack of germinal pore of the basidiospores. Currently,
according to the results of phylogenetic analysis, Gymnopilus
forms an independent clade called “Gymnopilae”
which is not related to any of the two families mentioned above by Matheny et
al. (2006) and is currently placed in the family Hymenogastraceae
(Kirk et al. 2008). 25 species of Gymnopilus
have been reported from India (Kulkarni 1990; Thomas et al. 2003; Ministry of
Environment and Forests 2011; Farook et al. 2013; Kumar et al. 2014; Kaur et
al. 2015). Two species of Gymnopilus have been
described from Maharashtra viz. G. karnalensis
S.M.Kulk. and G. chrysopellus
(Berk. & Crutis) Murril
(Senthilarasu 2014; Patil
2019).
During the present study, mature fruiting body of G. ochraceus were collected on the living tree of Ficus platyphylla
Del. from Kate Bhogaon, Panhala,
Kolhapur district (M.S.), India. Morphological and microscopic analysis have
been described in this paper. Analysis of ITS rDNA sequence was done to
evaluate phylogenetic relationships and Gas Chromatography–Mass Spectrometry
(GC-MS) was done for determination of bioactive components.
Material and Methods
a)
Morphological and microscopic analysis
Morphological and ecological characters were noted in the field.
Microscopic observations of fresh fruiting body was done with the help of 1.5% phloxine B staining and Lawrence & Mayo
N-300M research microscope.
b)
Identification of fungal strains
The identification of isolates was carried out at the sequencing facility
of National Centre for Microbial Resource (NCMR), National Centre for Cell
Science, Pune. At the facility, genomic DNA was isolated by the standard
phenol/chloroform extraction method (Sambrook et al. 1989), followed by PCR
amplification of the ITS regions using universal primers ITS1 [5’-TCC GTA GGT
GAA CCT GCG G-3’] and ITS4 [5’-TCC TCC GCT TAT TGA TAT GC -3’]. The amplified
ITS PCR product was purified by PEG-NaCl
precipitation and directly sequenced on an ABI® 3730XL automated DNA sequencer
(Applied Biosystems, Inc., Foster City, CA) as per manufacturer’s instructions.
Essentially, sequencing was carried out from both ends so that each position
was read at least twice. Assembly was carried out using Lasergene
package followed by NCBI BLAST against sequences from type material for
tentative identification (Boratyn et al. 2013).
c)
GC-MS analysis
10% methanolic extract of dried fruiting bodies was sonicated for 60
minutes at 35°C followed by centrifugation at 10,000 rpm for five minutes.
Supernatant was used for GC-MS analysis using Shimadzu, Japan TQ 8050 plus HS
20. Helium was used as the carrier gas at a flow rate of 1ml/min and an
injection volume of 1.0 µL. Injector temperature was 250˚C; ion source
temperature 250˚C. The oven temperature was 60˚C isothermal for 2.0 min, with
an increase of 10˚C/min to 250˚C, then 5˚C/min to 275˚C, ending with 10 min.
isothermal at 275˚C. Detector voltage was 0.7ev.
Results and Discussion
Habitat:
Growing on live F. platyphylla stem in a cluster (Image 1 A–G)
Pileus 2–8 cm wide, convex to plane, scaly, with appressed squamules at the centre, surface
pale brown, honey brown, pale ochraceous to pale yellow brown, squamules pale to honey brown; lamellae adnexed
to adnate, dark yellow brown; stipe 2–9 x 0.5–1.2 cm, cylindrical to somewhat
clavate, fibrillose, concolourous with pileus, with
an membranous pale yellow brown ring, context yellowish to pale buff, with
bitter taste; Basidiospores (5.5) 5.6–7.1 (7.5) x (4) 4.2–5.3 (5.5) µm,
ellipsoid to ovoid, verrucose to punctate, pale to yellow brown, dextrinoid;
Basidia 16–26 x 5–6 µm with four sterigmata; Cheilocystidia 15–18 x 4.5–7 µm, ventricose-rostrate with a clavate or rounded apex with
3–3.5 µm broad content; Pleurocystidia not seen; hyphae of pileipellis
6–18 µm broad, smooth, yellow brown, hyaline, clamp connections present (Figure
1 A–D).
GC-MS
analysis (Table 1 & Figure 2)
In GC-MS analysis three components are detected viz. 4,4’-Bipyridine,
9,12-Octadecadienoic acid, methyl ester and 9,12-Octadecadienoic acid (Z,Z).
Application of 4,4’- 4,4’-bipyridine is a prototypical bridging ligand and an
ideal connector between the transition metal atoms (Biradha
et al. 2006). Heufler et. al. (1987) stated that
4,4’- bipyridne derivate orellanine
causes acute renal failure in man. 9,12-Octadecadienoic acid (Z,Z)- methyl
ester is used as fuel and fuel additive. It has potential cancer preventive,
anti-inflammatory and anti-arthritic activities (Hagr
et al. 2018).
Conclusion
The above described species G. ochraceus is
reported for the first time from India.
It clearly indicates that G. ochraceus
is extremely rare species. Morphological and microscopic study along with ITS
rDNA analysis confirms the species authentication. Many species of Gymnopilus are bitter or foul to taste, some are
hallucinogenic and edibility of majority is unknown. There are many species and
little consensus on identifying them. Microscopically they can be confused with
Pholiota species and with the deadly Galerina marginata
complex. Microscopically, spores of Gymnopilus species
are finely roughened (warty) and lack an apical pore.
Table 1. GC-MS analysis of Gymnopilus ochraceus.
|
Peak# |
R.Time |
I.Time |
F.Time |
Molecular formula |
Area% |
Name |
|
1 |
16.395 |
16.070 |
16.875 |
C10H8N2 |
95.47 |
4,4’-Bipyridine |
|
2 |
24.655 |
24.580 |
24.715 |
C19H34O2 |
3.41 |
9,12-Octadecadienoic acid, methyl ester |
|
3 |
25.590 |
25.530 |
25.605 |
C19H34O2 |
1.12 |
9,12-Octadecadienoic acid (Z,Z)- |
|
|
|
|
|
|
100 |
|
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