Research Article |
Corresponding author: Tariq Saiff Ullah ( tariq.saiff@uokajk.edu.pk ) Academic editor: Lorenzo Peruzzi
© 2022 Tariq Saiff Ullah, Syeda Sadiqa Firdous, Wayne Thomas Shier, Abdul Nasir Khalid.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Ullah TS, Firdous SS, Shier WT, Khalid AN (2022) Ramaria barenthalensis a new record from western Himalayas, Azad Jammu and Kashmir, Pakistan. Italian Botanist 14: 133-143. https://doi.org/10.3897/italianbotanist.14.96790
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The genus Ramaria is globally distributed, inhabiting different substrates. In the present study, samples of R. barenthalensis were collected during a fungal survey from Neelum valley AJK, Pakistan. Identification was carried out through combined morpho-anatomical and molecular data from nrITS region, which confirmed the identification as R. barenthalensis. The taxon is a new record for fungi of AJK, Pakistan.
Coral fungi, ITS, Mushrooms of Kashmir, Neelum valley, nrDNA, Ramaria
Ramaria species are cosmopolitan in distribution and grow on living and dead hardwood, tree trunks, partially decomposed organic matter, and under conifers as mycorrhizal species (
Ramaria abietina (Pers.) Quél., R. aurea (Schaeff.) Quél., R. apiculata (Fr.) Donk, R. botrytis (Pers.) Bourdot, R. flava (Schaeff.) Quél., R. flavescentoides Hanif & Khalid, R. formosa (Pers.) Quél., R. pallida (Schaeff.) Ricken, R. soluta (P. Karst.) Corner and R. stricta (Pers.) Quél. have previously been reported from Pakistan (
The study area state of Azad Jammu and Kashmir lies between longitude 73–75°E and latitude 33–36°N and comprises an area of 13,297 km2 (
Fresh basidiomata were collected from selected sites in Neelum valley, AJK, Pakistan, during 2019–2020 through consecutive field surveys (Fig.
Slides were mounted in 5% KOH (w/v) and 1% Congo red to study anatomical features, examined by using a light microscope (MX4300H, Japan) at 100× magnification: size and shape of basidiospores, basidia, cystidia and other structures. For basidiospores and other structures at least 50 measurements were made and fungal specimen names with authorities were retrieved from Index Fungorum (http://www.indexfungorum.org).
DNA was extracted from dried samples using a modified CTAB method as proposed by
Consensus sequence was generated using the molecular tool BioEdit ver. 7.2.5 (
Basidiomata 5–12 cm in height, 4–9 cm wide with a well-developed branched mycelium. Branches vertically oriented, furcated, elongated to flattened, smooth, light brown when young and dark brown at maturity. Base variable, sometimes reduced, and well-developed, whitish to brown. Flesh whitish to light brown and soft. Odor is not distinctive and pleasant. Basidiospores [50/5/2] 6.5–9 × 3–4.6 µm, elliptical to ovoid, roughened. Basidia 45–59 × 7.5–9 µm, sterigmate. Cheilocystidia 46–62 × 7–8.5 µm, elongated. Clamp-connections are present. Trama hyphae are thick-walled with clamps. Hyphae in basal tomentum are smooth, with a tapering base (Fig.
ITS dataset (Fig.
Species | Strain | Location | Substrate | GenBank Accession Number |
---|---|---|---|---|
Ramaria sp. | SD125.4 | USA | Fir forest | DQ365646 |
Ramaria sp. | OSC 81622 | USA | Fir forest | EU652343 |
R. flavobrunnesens | AY102864 | USA | Fir forest | AY102864 |
R. barenthalensis | AMB17386 | Spain | mixed forest | MK493039 |
R. barenthalensis | AMB17381 | Spain | Abies and Piecea forest | MK493038 |
R. barenthalensis | T40 | Kashmir | Dead hardwood of conifers | ON209680 |
R. edwinii | ALV11173 | Spain | Abies and Piecea | MK493034 |
Ramaria sp. | RAM2 | Srinagar J&K | Abies forest | MH930937 |
Ramaria sp. | OSC 73311 | USA | Abies and Piecea mix wood forest | KP658154 |
Ramaria sp. | OSC 144044 | USA | Abies and Piecea mix wood forest | KF206335 |
Ramaria sp. | MHM312 | Mexico | NA | EU569259 |
Ramaria sp. | NVE 367 | Amazonia | NA | KF937356 |
Ramaria sp. | OSC 115803 | USA | NA | KP658139 |
Ramaria sp. | DB215-08/5 | USA | NA | KT968608 |
R. flava | AMB n. 17484 | Italy | Abies and Piecea | MK581224 |
R. flava | AMB 17393 | Italy | Abies and Piecea | MK493035 |
R. flava | AMB n. 17481 | Italy | Mix forest type | MK557953 |
R. flava | MA-Fungi 48072 | Spain | Abies and Piecea | AJ408367 |
R. flava | ZT Myc 55613 | Italy | NA | KY626146 |
R. pseudoflava | AMB 17390 | Italy | NA | MK493045 |
R. pseudoflava | AMB 17392 | Italy | NA | MK493046 |
R. pseudoflava | AMB 17391 | Italy | NA | MK493044 |
R. flava | MA-Fungi 48061 | Spain | Abies and Piecea | AJ408364 |
Ramaria sp. | OSC 61837 | USA | Mix forest | DQ365602 |
Ramaria sp. | OSC 134657 | USA | Mix forest | JX310403 |
R. magnipes | WTU-F-063057 | USA | Mix forest | MK169351 |
R. magnipes | WTU 063057 | Italy | Fir forest | MK493040 |
R. gracilis | OSC 134659 | USA | Abies and Piecea mix wood forest | JX310399 |
R. gracilis | OSC 112168 | USA | Abies and Piecea mix wood forest | KY354745 |
Hysterangium crassirahis | OSC 4860 | USA | Abies and Piecea mix wood forest | MN809540 |
Morphoanatomical comparisions of different Ramaria species with R. barenthalensis.
In mixed coniferous forest, associated with Abies pindrow and Betula jacquemontii, solitary or scattered and gregarious, common in the temperate zone of AJK, Pakistan.
Pakistan, Neelum valley (Azad Jammu and Kashmir) collected from a mixed coniferous forest on decayed hardwood, in late summers, 1524 m a.s.l, August 2020, Tariq Saiff Ullah, TS-40.
The newly generated ITS sequence of Pakistani Ramaria (T40) specimen, yielded a fragment of 702 base pairs. In initial BLAST search results, our sequence showed 99.33% similarity with R. barenthalensis MK493039, with 98% query cover, and 0.0 E value. The phylogram comprised 30 sequences and Hysterangium crassirh Zeller & C.W. Dodge was chosen as an out-group taxon. The final aligned dataset consist of 421 conserved sites, 256 were variable, 167 were informative and 86 were singletons. Our sequence Ramaria barenthalensis (T40) grouped with R. barenthalensis (MK493038 and MK493029) with strong bootstrap value (Fig.
Molecular Phylogenetic tree of Ramaria barenthalensis (ON209680) by maximum likehood method based on ITS sequences. The sequence generated in this study represents in bold.
In present study, a specimen of genus Ramaria was studied on the basis of morpho-anatomical and molecular approach. Our study is consistent with the original specimen description given by
The analyzed sample of R. barenthalensis (T-40) has similar morphological features, but with slight differences in basidiomata to other previously descibed species of Ramaria (
The first author is thankful to Javaid Qayum Swati and Muhammad Usman Mughal for their help during field and lab work, respectively.