Abstract
This research presents a new species in the genus Conocybe collected from the tropical semi-arid District Karak, Khyber Pakhtunkhwa, Pakistan. Morphological features, and DNA barcoding marker (ITS-region) were used for the systematic placement of Conocybe karakensis. The present species is characterized by having a medium-sized fruiting body, semi-ovate, orange-colored pileus, free, spaced, and regular lamellae with a central, light brown stipe, thick-walled, oblong basidiospores and clavate shaped, tetra-sterigamtic basidia. Molecularly, initial Blast analysis and the phylogenetic analyses showed that Pakistani collection is closely related to Conocybe velutipes. In the phylogenetic tree, ITS sequences of Pakistani collection clustered in a separate clade sister to Conocybe velutipes. This study provides full description, illustrations, and phylogenetic analyses to distinguish this novel taxon from its closest relatives.
References
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Atri, N.S., Amandeep, K. & Munruchi, K. (2009) Three new records of coprophilous mushrooms of family Bolbitiaceae from India. Mushroom Research 18 (2): 51–56.
Cagli, G., Ozturk, A. & Kocak, M.Z.G. (2019) Two new basidiomycete records for the Mycobiota of Turkey. Anatolian Journal of Botany 3 (2): 40–43. https://doi:10.30616/ajb.487507
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Enderle, M. (1994) Conocybe-Pholiotina studies V. Zeitschrift Fur Mykologie 60 (1): 35–48.
Gardes, M. & Bruns, T.D. (1993) ITS primers with enhanced specificity for basidiomycetes—application to the identification of mycorrhizae and rusts. Molecular ecology 2 (2): 113–118. https://doi.org/10.1111/j.1365-294X.1993.tb00005.x
Gui, Y., Zhu, G.S., Callac, P., Hyde, K.D., Parra, L.A., Chen, J. & Liu, Z.Y. (2015) Agaricus section Arvenses: three new species in highland subtropical Southwest China. Fungal biology 119 (2–3): 79–94. https://doi.org/10.1016/j.funbio.2014.10.005
Hallen, H.E., Watling, R. & Adams, G.C. (2003) Taxonomy and toxicity of Conocybe lactea and related species. Mycological research 107 (8): 969–979. https://doi.org/10.1017/S0953756203008190
Hausknecht, A. & Contu, M. (2007) Interesting species of Conocybe (Agaricales, Bolbitiaceae) from Gallura (NE Sardinia, Italy). Osterreichische Zeitschrift fur Pilzkunde 16: 157–166.
Hausknecht, A. (2005) Type studies in north American species of Bolbitiaceae belonging to the genera Conocybe and Pholiotina. Austrian Journal of Mycology 13: 153–235.
Hausknecht, A., Kalamees, K., Knudsen, H. & Mukhin, V. (2009) The genera Conocybe and Pholiotina (Agaricomycotina, Bolbitiaceae) in temperate Asia. Folia Cryptogamica Estonica 45: 23–47.
Izhar, A., Bashir, H. & Khalid, A.N. (2019) A new species of Conocybe (Bolbitaceae) from Punjab, Pakistan. Phytotaxa 402 (5): 251–258. https://doi.org/10.11646/phytotaxa.402.5.4
Jukes, T.H. & Cantor, C.R. (1969) Evolution of protein molecules. In: Munro, H.N. (Ed.) Mammalian protein metabolism, 3. Academic Press, New York, pp. 21–132.
Kamran, S., Khan, S.M., Ahmad, Z., Ur Rahman, A., Iqbal, M., Manan, F., Ul Haq, Z. & Ullah, S. (2020) The role of graveyards in species conservation and beta diversity: a vegetation appraisal of sacred habitats from Bannu, Pakistan. Journal of Forestry Research 31 (4): 1147–1158. https://doi.org/10.1007/s11676-019-00893-1
Khalid, A.N. (2022) A checklist of macrofungi of Pakistan published from 1998–2020. Pakistan Jouranl of Botany 54 (5): 1947–1962.
Khan, M. & Hussain, F. (2013) Conservation status of plant species in Tehsil Takht-e-Nasrati, District Karak, Khyber Pakhtun Khawa, Pakistan. International Journal of Biodiversity and Conservation 5 (1): 20–26.
Khan, M., Hussain, F. & Musharaf, S. (2013) Ethnobotanical profile of Tehsil Takht-e-Nasratti, District Karak, Pakistan. Journal of Medicinal Plants Research 7 (22): 1636–1651.
Khan, M., Musharaf, S. & Shinwari, Z.K. (2011) Ethnobotanical importance of halophytes of Noshpho salt mine, District Karak, Pakistan. Research In Pharmaceutical Biotechnology 3 (4): 46–52.
Khattak, N.S., Nouroz, F., Rahman, I.U. & Noreen, S. (2015) Ethno veterinary uses of medicinal plants of District Karak, Pakistan. Journal of ethnopharmacology 171: 273–279. https://doi.org/10.1016/j.jep.2015.05.048
Kumar, M., Harsh, N.S.K. & Prasad, R. (2014) Conocybe apala Fr.: Fr. Arnolds, Helvella solitaria P. Karst. and Hygrocybe miniata Fr. P. Kumm.; three new mushrooms from India. Current Research in Environmental and Applied Mycology 4 (1): 100–105.
Liu, J. & Bau, T. (2018) New species and new records in the genus Conocybe (Bolbitaceae) from China. Phytotaxa 357 (4): 261–274. https://doi.org/10.11646/phytotaxa.357.4.2
Matheny, P.B., Curtis, J.M., Hofstetter, V., Aime, M.C., Moncalvo, J.M., Ge, Z.W. & Bougher, N.L. (2006) Major clades of Agaricales: a multilocus phylogenetic overview. Mycologia 98 (6): 982–995. https://doi.org/10.1080/15572536.2006.11832627
Melo, R.F.R., Chikowski, R.D.S., Miller, A.N. & Maia, L.C. (2016) Coprophilous Agaricales (Agaricomycetes, Basidiomycota) from Brazil. Phytotaxa 266 (1): 1–14. https://doi.org/10.11646/phytotaxa.266.1.1
Moncalvo, J.M., Vilgalys, R., Redhead, S.A., Johnson, J.E., James, T.Y., Aime, M.C. & Miller Jr, O.K. (2002) One hundred and seventeen clades of euagarics. Molecular phylogenetics and evolution 23 (3): 357–400. https://doi.org/10.1016/S1055-7903(02)00027-1
Nagy, L.G., Walther, G., Hazi, J.U.D.I.T., Vagvolgyi, C. & Papp, T. (2011) Understanding the evolutionary processes of fungal fruiting bodies: correlated evolution and divergence times in the Psathyrellaceae. Systematic Biology 60 (3): 303–317. https://doi.org/10.1093/sysbio/syr005
Nemcsics, A. (1994) Spacing in the Munsell Color System Relative to the Coloroid Color Systems. Color Research & Application 19 (2): 122–125. https://doi.org/10.1111/j.1520-6378.1994.tb00071.x
Pomilio, A.B., Battista, M.E. & Vitale, A.A. (2006) Naturally-occurring cyclopeptides: structures and bioactivity. Current Organic Chemistry 10 (16): 2075–2121. https://doi.org/10.2174/138527206778742669
Rehman, K., Khan, M.A., Ullah, Z. & Chaudhary, H.J. (2015) An ethno botanical perspective of traditional medicinal plants from the Khattak tribe of Chonthra Karak, Pakistan. Journal of ethnopharmacology 165: 251–259. https://doi.org/10.1016/j.jep.2015.02.035
Sengul, Demirak., M.S., Turkekul, I. & Isik, H. (2021) Conocybe romagnesii and Gerronema subclavatum (Basidiomycota: Agaricales) in the Central Black Sea Region of Turkey. Nordic Journal of Botany 39 (12): 1–9. https://doi.org/10.1111/njb.03439
Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. & Kumar, S. (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular biology and evolution 28 (10): 2731–2739. https://doi.org/10.1093/molbev/msr121
Toth, A., Hausknecht, A., Krisai-Greilhuber, I., Papp, T., Vágvölgyi, C. & Nagy, L.G. (2013) Iteratively refined guide trees help improving alignment and phylogenetic inference in the mushroom family Bolbitiaceae. Public library of Science 8 (2): e56143. https://doi.org/10.1371/journal.pone.0056143
Ullah, T., Saba, M. & Syed, M.F. (2022) Coprinopsis afrocinerea (Agaricales, Psathyrellaceae): First record from Asia (Pakistan). Microscopy Research and Technique 85 (10): 1–8. https://doi.org/10.1002/jemt.24192
Watling, R. (1971) Observations on the Bolbitiaceae—IV. Developmental studies on Conocybe with particular reference to the annulate species. Persoonia-Molecular Phylogeny and Evolution of Fungi 6 (2): 281–289.
White, T.J., Bruns, T.D., Lee, S.B., Taylor, J.W., Innis, N. & Gelfand, D. (1990) PCR-protocols and applications-a laboratory manual. Academic Press, New York, pp. 315–322.
Amandeep, K., Atri, N.S. & Munruchi, K. (2015) Diversity of species of the genus Conocybe (Bolbitiaceae, Agaricales) collected on dung from Punjab, India. Mycosphere 6 (1): 19–42. https://doi 10.5943/mycosphere/6/1/4
Arnolds, E.E.F. (2005) Conocybe Fayod. Flora Agaricina Neerlandica 6: 120–179.
Atri, N.S., Amandeep, K. & Munruchi, K. (2009) Three new records of coprophilous mushrooms of family Bolbitiaceae from India. Mushroom Research 18 (2): 51–56.
Cagli, G., Ozturk, A. & Kocak, M.Z.G. (2019) Two new basidiomycete records for the Mycobiota of Turkey. Anatolian Journal of Botany 3 (2): 40–43. https://doi:10.30616/ajb.487507
Dentinger, B.T., Didukh, M.Y. & Moncalvo, J.M. (2011) Comparing COI and ITS as DNA barcode markers for mushrooms and allies (Agaricomycotina). Public library of Science 6 (9): e25081. https://doi.org/10.1371/journal.pone.0025081
Enderle, M. & Hübner, H.J. (1999) Studies in Conocvbe and Pholiotina VIII. Zeitschrift Fur Mykologie 65: 3–22.
Enderle, M. (1994) Conocybe-Pholiotina studies V. Zeitschrift Fur Mykologie 60 (1): 35–48.
Gardes, M. & Bruns, T.D. (1993) ITS primers with enhanced specificity for basidiomycetes—application to the identification of mycorrhizae and rusts. Molecular ecology 2 (2): 113–118. https://doi.org/10.1111/j.1365-294X.1993.tb00005.x
Gui, Y., Zhu, G.S., Callac, P., Hyde, K.D., Parra, L.A., Chen, J. & Liu, Z.Y. (2015) Agaricus section Arvenses: three new species in highland subtropical Southwest China. Fungal biology 119 (2–3): 79–94. https://doi.org/10.1016/j.funbio.2014.10.005
Hallen, H.E., Watling, R. & Adams, G.C. (2003) Taxonomy and toxicity of Conocybe lactea and related species. Mycological research 107 (8): 969–979. https://doi.org/10.1017/S0953756203008190
Hausknecht, A. & Contu, M. (2007) Interesting species of Conocybe (Agaricales, Bolbitiaceae) from Gallura (NE Sardinia, Italy). Osterreichische Zeitschrift fur Pilzkunde 16: 157–166.
Hausknecht, A. (2005) Type studies in north American species of Bolbitiaceae belonging to the genera Conocybe and Pholiotina. Austrian Journal of Mycology 13: 153–235.
Hausknecht, A., Kalamees, K., Knudsen, H. & Mukhin, V. (2009) The genera Conocybe and Pholiotina (Agaricomycotina, Bolbitiaceae) in temperate Asia. Folia Cryptogamica Estonica 45: 23–47.
Izhar, A., Bashir, H. & Khalid, A.N. (2019) A new species of Conocybe (Bolbitaceae) from Punjab, Pakistan. Phytotaxa 402 (5): 251–258. https://doi.org/10.11646/phytotaxa.402.5.4
Jukes, T.H. & Cantor, C.R. (1969) Evolution of protein molecules. In: Munro, H.N. (Ed.) Mammalian protein metabolism, 3. Academic Press, New York, pp. 21–132.
Kamran, S., Khan, S.M., Ahmad, Z., Ur Rahman, A., Iqbal, M., Manan, F., Ul Haq, Z. & Ullah, S. (2020) The role of graveyards in species conservation and beta diversity: a vegetation appraisal of sacred habitats from Bannu, Pakistan. Journal of Forestry Research 31 (4): 1147–1158. https://doi.org/10.1007/s11676-019-00893-1
Khalid, A.N. (2022) A checklist of macrofungi of Pakistan published from 1998–2020. Pakistan Jouranl of Botany 54 (5): 1947–1962.
Khan, M. & Hussain, F. (2013) Conservation status of plant species in Tehsil Takht-e-Nasrati, District Karak, Khyber Pakhtun Khawa, Pakistan. International Journal of Biodiversity and Conservation 5 (1): 20–26.
Khan, M., Hussain, F. & Musharaf, S. (2013) Ethnobotanical profile of Tehsil Takht-e-Nasratti, District Karak, Pakistan. Journal of Medicinal Plants Research 7 (22): 1636–1651.
Khan, M., Musharaf, S. & Shinwari, Z.K. (2011) Ethnobotanical importance of halophytes of Noshpho salt mine, District Karak, Pakistan. Research In Pharmaceutical Biotechnology 3 (4): 46–52.
Khattak, N.S., Nouroz, F., Rahman, I.U. & Noreen, S. (2015) Ethno veterinary uses of medicinal plants of District Karak, Pakistan. Journal of ethnopharmacology 171: 273–279. https://doi.org/10.1016/j.jep.2015.05.048
Kumar, M., Harsh, N.S.K. & Prasad, R. (2014) Conocybe apala Fr.: Fr. Arnolds, Helvella solitaria P. Karst. and Hygrocybe miniata Fr. P. Kumm.; three new mushrooms from India. Current Research in Environmental and Applied Mycology 4 (1): 100–105.
Liu, J. & Bau, T. (2018) New species and new records in the genus Conocybe (Bolbitaceae) from China. Phytotaxa 357 (4): 261–274. https://doi.org/10.11646/phytotaxa.357.4.2
Matheny, P.B., Curtis, J.M., Hofstetter, V., Aime, M.C., Moncalvo, J.M., Ge, Z.W. & Bougher, N.L. (2006) Major clades of Agaricales: a multilocus phylogenetic overview. Mycologia 98 (6): 982–995. https://doi.org/10.1080/15572536.2006.11832627
Melo, R.F.R., Chikowski, R.D.S., Miller, A.N. & Maia, L.C. (2016) Coprophilous Agaricales (Agaricomycetes, Basidiomycota) from Brazil. Phytotaxa 266 (1): 1–14. https://doi.org/10.11646/phytotaxa.266.1.1
Moncalvo, J.M., Vilgalys, R., Redhead, S.A., Johnson, J.E., James, T.Y., Aime, M.C. & Miller Jr, O.K. (2002) One hundred and seventeen clades of euagarics. Molecular phylogenetics and evolution 23 (3): 357–400. https://doi.org/10.1016/S1055-7903(02)00027-1
Nagy, L.G., Walther, G., Hazi, J.U.D.I.T., Vagvolgyi, C. & Papp, T. (2011) Understanding the evolutionary processes of fungal fruiting bodies: correlated evolution and divergence times in the Psathyrellaceae. Systematic Biology 60 (3): 303–317. https://doi.org/10.1093/sysbio/syr005
Nemcsics, A. (1994) Spacing in the Munsell Color System Relative to the Coloroid Color Systems. Color Research & Application 19 (2): 122–125. https://doi.org/10.1111/j.1520-6378.1994.tb00071.x
Pomilio, A.B., Battista, M.E. & Vitale, A.A. (2006) Naturally-occurring cyclopeptides: structures and bioactivity. Current Organic Chemistry 10 (16): 2075–2121. https://doi.org/10.2174/138527206778742669
Rehman, K., Khan, M.A., Ullah, Z. & Chaudhary, H.J. (2015) An ethno botanical perspective of traditional medicinal plants from the Khattak tribe of Chonthra Karak, Pakistan. Journal of ethnopharmacology 165: 251–259. https://doi.org/10.1016/j.jep.2015.02.035
Sengul, Demirak., M.S., Turkekul, I. & Isik, H. (2021) Conocybe romagnesii and Gerronema subclavatum (Basidiomycota: Agaricales) in the Central Black Sea Region of Turkey. Nordic Journal of Botany 39 (12): 1–9. https://doi.org/10.1111/njb.03439
Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. & Kumar, S. (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular biology and evolution 28 (10): 2731–2739. https://doi.org/10.1093/molbev/msr121
Toth, A., Hausknecht, A., Krisai-Greilhuber, I., Papp, T., Vágvölgyi, C. & Nagy, L.G. (2013) Iteratively refined guide trees help improving alignment and phylogenetic inference in the mushroom family Bolbitiaceae. Public library of Science 8 (2): e56143. https://doi.org/10.1371/journal.pone.0056143
Ullah, T., Saba, M. & Syed, M.F. (2022) Coprinopsis afrocinerea (Agaricales, Psathyrellaceae): First record from Asia (Pakistan). Microscopy Research and Technique 85 (10): 1–8. https://doi.org/10.1002/jemt.24192
Watling, R. (1971) Observations on the Bolbitiaceae—IV. Developmental studies on Conocybe with particular reference to the annulate species. Persoonia-Molecular Phylogeny and Evolution of Fungi 6 (2): 281–289.
White, T.J., Bruns, T.D., Lee, S.B., Taylor, J.W., Innis, N. & Gelfand, D. (1990) PCR-protocols and applications-a laboratory manual. Academic Press, New York, pp. 315–322.