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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<p>Amandeep, K., Atri, N.S. & Munruchi, K. (2015) Diversity of species of the genus <em>Conocybe </em>(Bolbitiaceae, Agaricales) collected on dung from Punjab, India. <em>Mycosphere </em>6 (1): 19–42. https://doi 10.5943/mycosphere/6/1/4</p>
<p>Arnolds, E.E.F. (2005) <em>Conocybe</em> Fayod. <em>Flora Agaricina Neerlandica</em> 6: 120–179.</p>
<p>Atri, N.S., Amandeep, K. & Munruchi, K. (2009) Three new records of <em>coprophilous</em> mushrooms of family Bolbitiaceae from India. <em>Mushroom Research</em> 18 (2): 51–56.</p>
<p>Cagli, G., Ozturk, A. & Kocak, M.Z.G. (2019) Two new basidiomycete records for the Mycobiota of Turkey. <em>Anatolian Journal of Botany</em> 3 (2): 40–43. https://doi:10.30616/ajb.487507</p>
<p>Dentinger, B.T., Didukh, M.Y. & Moncalvo, J.M. (2011) Comparing COI and ITS as DNA barcode markers for mushrooms and allies (Agaricomycotina). <em>Public library of Science</em> 6 (9): e25081. https://doi.org/10.1371/journal.pone.0025081</p>
<p>Enderle, M. & Hübner, H.J. (1999) Studies in <em>Conocvbe</em> and <em>Pholiotina</em> VIII. <em>Zeitschrift Fur Mykologie</em> 65: 3–22.</p>
<p>Enderle, M. (1994) <em>Conocybe-Pholiotina</em> studies V. <em>Zeitschrift Fur Mykologie</em> 60 (1): 35–48.</p>
<p>Gardes, M. & Bruns, T.D. (1993) ITS primers with enhanced specificity for basidiomycetes—application to the identification of mycorrhizae and rusts. <em>Molecular ecology</em> 2 (2): 113–118. https://doi.org/10.1111/j.1365-294X.1993.tb00005.x</p>
<p>Gui, Y., Zhu, G.S., Callac, P., Hyde, K.D., Parra, L.A., Chen, J. & Liu, Z.Y. (2015) <em>Agaricus </em>section <em>Arvenses</em>: three new species in highland subtropical Southwest China. <em>Fungal biology</em> 119 (2–3): 79–94. https://doi.org/10.1016/j.funbio.2014.10.005</p>
<p>Hallen, H.E., Watling, R. & Adams, G.C. (2003) Taxonomy and toxicity of <em>Conocybe lactea</em> and related species. <em>Mycological research</em> 107 (8): 969–979. https://doi.org/10.1017/S0953756203008190</p>
<p>Hausknecht, A. & Contu, M. (2007) Interesting species of <em>Conocybe</em> (Agaricales, Bolbitiaceae) from Gallura (NE Sardinia, Italy). <em>Osterreichische Zeitschrift fur Pilzkunde</em> 16: 157–166.</p>
<p>Hausknecht, A. (2005) Type studies in north American species of Bolbitiaceae belonging to the genera <em>Conocybe</em> and <em>Pholiotina</em>. <em>Austrian Journal of Mycology</em> 13: 153–235.</p>
<p>Hausknecht, A., Kalamees, K., Knudsen, H. & Mukhin, V. (2009) The genera <em>Conocybe</em> and <em>Pholiotina</em> (Agaricomycotina, Bolbitiaceae) in temperate Asia. <em>Folia Cryptogamica Estonica</em> 45: 23–47.</p>
<p>Izhar, A., Bashir, H. & Khalid, A.N. (2019) A new species of <em>Conocybe</em> (Bolbitaceae) from Punjab, Pakistan. <em>Phytotaxa </em>402 (5): 251–258. https://doi.org/10.11646/phytotaxa.402.5.4</p>
<p>Jukes, T.H. & Cantor, C.R. (1969) Evolution of protein molecules. <em>In</em>: Munro, H.N. (Ed.) <em>Mammalian protein metabolism, </em>3<em>.</em> Academic Press, New York, pp. 21–132.</p>
<p>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. <em>Journal of Forestry Research</em> 31 (4): 1147–1158. https://doi.org/10.1007/s11676-019-00893-1</p>
<p>Khalid, A.N. (2022) A checklist of macrofungi of Pakistan published from 1998–2020. <em>Pakistan Jouranl of Botany </em>54 (5): 1947–1962.</p>
<p>Khan, M. & Hussain, F. (2013) Conservation status of plant species in Tehsil Takht-e-Nasrati, District Karak, Khyber Pakhtun Khawa, Pakistan. <em>International Journal of Biodiversity and Conservation</em> 5 (1): 20–26.</p>
<p>Khan, M., Hussain, F. & Musharaf, S. (2013) Ethnobotanical profile of Tehsil Takht-e-Nasratti, District Karak, Pakistan. <em>Journal of Medicinal Plants Research</em> 7 (22): 1636–1651.</p>
<p>Khan, M., Musharaf, S. & Shinwari, Z.K. (2011) Ethnobotanical importance of halophytes of Noshpho salt mine, District Karak, Pakistan. <em>Research In Pharmaceutical Biotechnology</em> 3 (4): 46–52.</p>
<p>Khattak, N.S., Nouroz, F., Rahman, I.U. & Noreen, S. (2015) Ethno veterinary uses of medicinal plants of District Karak, Pakistan. <em>Journal of ethnopharmacology</em> 171: 273–279. https://doi.org/10.1016/j.jep.2015.05.048</p>
<p>Kumar, M., Harsh, N.S.K. & Prasad, R. (2014) <em>Conocybe apala</em> Fr.: Fr. Arnolds, <em>Helvella solitaria</em> P. Karst. and <em>Hygrocybe miniata</em> Fr. P. Kumm.; three new mushrooms from India. Current <em>Research in Environmental and Applied Mycology</em> 4 (1): 100–105.</p>
<p>Liu, J. & Bau, T. (2018) New species and new records in the genus <em>Conocybe</em> (Bolbitaceae) from China. <em>Phytotaxa </em>357 (4): 261–274. https://doi.org/10.11646/phytotaxa.357.4.2</p>
<p>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. <em>Mycologia</em> 98 (6): 982–995. https://doi.org/10.1080/15572536.2006.11832627</p>
<p>Melo, R.F.R., Chikowski, R.D.S., Miller, A.N. & Maia, L.C. (2016) <em>Coprophilous</em> Agaricales (Agaricomycetes, Basidiomycota) from Brazil. <em>Phytotaxa</em> 266 (1): 1–14. https://doi.org/10.11646/phytotaxa.266.1.1</p>
<p>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. <em>Molecular phylogenetics and evolution</em> 23 (3): 357–400. https://doi.org/10.1016/S1055-7903(02)00027-1</p>
<p>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. <em>Systematic Biology</em> 60 (3): 303–317. https://doi.org/10.1093/sysbio/syr005</p>
<p>Nemcsics, A. (1994) Spacing in the Munsell Color System Relative to the Coloroid Color Systems. <em>Color Research & Application</em> 19 (2): 122–125. https://doi.org/10.1111/j.1520-6378.1994.tb00071.x</p>
<p>Pomilio, A.B., Battista, M.E. & Vitale, A.A. (2006) Naturally-occurring cyclopeptides: structures and bioactivity. <em>Current Organic Chemistry</em> 10 (16): 2075–2121. https://doi.org/10.2174/138527206778742669</p>
<p>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. <em>Journal of ethnopharmacology</em> 165: 251–259. https://doi.org/10.1016/j.jep.2015.02.035</p>
<p>Sengul, Demirak., M.S., Turkekul, I. & Isik, H. (2021) <em>Conocybe romagnesii</em> and <em>Gerronema subclavatum</em> (Basidiomycota: Agaricales) in the Central Black Sea Region of Turkey<em>. Nordic Journal of Botany</em> 39 (12): 1–9. https://doi.org/10.1111/njb.03439</p>
<p>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. <em>Molecular biology and evolution</em> 28 (10): 2731–2739. https://doi.org/10.1093/molbev/msr121</p>
<p>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. <em>Public library of Science</em> 8 (2): e56143. https://doi.org/10.1371/journal.pone.0056143</p>
<p>Ullah, T., Saba, M. & Syed, M.F. (2022) <em>Coprinopsis afrocinerea</em> (Agaricales, Psathyrellaceae): First record from Asia (Pakistan). <em>Microscopy Research and Technique</em> 85 (10): 1–8. https://doi.org/10.1002/jemt.24192</p>
<p>Watling, R. (1971) Observations on the Bolbitiaceae—IV. Developmental studies on <em>Conocybe</em> with particular reference to the annulate species. <em>Persoonia-Molecular Phylogeny and Evolution of Fungi </em>6 (2): 281–289.</p>
<p>White, T.J., Bruns, T.D., Lee, S.B., Taylor, J.W., Innis, N. & Gelfand, D. (1990) <em>PCR-protocols and applications-a laboratory manual</em>. Academic Press, New York, pp. 315–322.</p>
<p>Amandeep, K., Atri, N.S. & Munruchi, K. (2015) Diversity of species of the genus <em>Conocybe </em>(Bolbitiaceae, Agaricales) collected on dung from Punjab, India. <em>Mycosphere </em>6 (1): 19–42. https://doi 10.5943/mycosphere/6/1/4</p>
<p>Arnolds, E.E.F. (2005) <em>Conocybe</em> Fayod. <em>Flora Agaricina Neerlandica</em> 6: 120–179.</p>
<p>Atri, N.S., Amandeep, K. & Munruchi, K. (2009) Three new records of <em>coprophilous</em> mushrooms of family Bolbitiaceae from India. <em>Mushroom Research</em> 18 (2): 51–56.</p>
<p>Cagli, G., Ozturk, A. & Kocak, M.Z.G. (2019) Two new basidiomycete records for the Mycobiota of Turkey. <em>Anatolian Journal of Botany</em> 3 (2): 40–43. https://doi:10.30616/ajb.487507</p>
<p>Dentinger, B.T., Didukh, M.Y. & Moncalvo, J.M. (2011) Comparing COI and ITS as DNA barcode markers for mushrooms and allies (Agaricomycotina). <em>Public library of Science</em> 6 (9): e25081. https://doi.org/10.1371/journal.pone.0025081</p>
<p>Enderle, M. & Hübner, H.J. (1999) Studies in <em>Conocvbe</em> and <em>Pholiotina</em> VIII. <em>Zeitschrift Fur Mykologie</em> 65: 3–22.</p>
<p>Enderle, M. (1994) <em>Conocybe-Pholiotina</em> studies V. <em>Zeitschrift Fur Mykologie</em> 60 (1): 35–48.</p>
<p>Gardes, M. & Bruns, T.D. (1993) ITS primers with enhanced specificity for basidiomycetes—application to the identification of mycorrhizae and rusts. <em>Molecular ecology</em> 2 (2): 113–118. https://doi.org/10.1111/j.1365-294X.1993.tb00005.x</p>
<p>Gui, Y., Zhu, G.S., Callac, P., Hyde, K.D., Parra, L.A., Chen, J. & Liu, Z.Y. (2015) <em>Agaricus </em>section <em>Arvenses</em>: three new species in highland subtropical Southwest China. <em>Fungal biology</em> 119 (2–3): 79–94. https://doi.org/10.1016/j.funbio.2014.10.005</p>
<p>Hallen, H.E., Watling, R. & Adams, G.C. (2003) Taxonomy and toxicity of <em>Conocybe lactea</em> and related species. <em>Mycological research</em> 107 (8): 969–979. https://doi.org/10.1017/S0953756203008190</p>
<p>Hausknecht, A. & Contu, M. (2007) Interesting species of <em>Conocybe</em> (Agaricales, Bolbitiaceae) from Gallura (NE Sardinia, Italy). <em>Osterreichische Zeitschrift fur Pilzkunde</em> 16: 157–166.</p>
<p>Hausknecht, A. (2005) Type studies in north American species of Bolbitiaceae belonging to the genera <em>Conocybe</em> and <em>Pholiotina</em>. <em>Austrian Journal of Mycology</em> 13: 153–235.</p>
<p>Hausknecht, A., Kalamees, K., Knudsen, H. & Mukhin, V. (2009) The genera <em>Conocybe</em> and <em>Pholiotina</em> (Agaricomycotina, Bolbitiaceae) in temperate Asia. <em>Folia Cryptogamica Estonica</em> 45: 23–47.</p>
<p>Izhar, A., Bashir, H. & Khalid, A.N. (2019) A new species of <em>Conocybe</em> (Bolbitaceae) from Punjab, Pakistan. <em>Phytotaxa </em>402 (5): 251–258. https://doi.org/10.11646/phytotaxa.402.5.4</p>
<p>Jukes, T.H. & Cantor, C.R. (1969) Evolution of protein molecules. <em>In</em>: Munro, H.N. (Ed.) <em>Mammalian protein metabolism, </em>3<em>.</em> Academic Press, New York, pp. 21–132.</p>
<p>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. <em>Journal of Forestry Research</em> 31 (4): 1147–1158. https://doi.org/10.1007/s11676-019-00893-1</p>
<p>Khalid, A.N. (2022) A checklist of macrofungi of Pakistan published from 1998–2020. <em>Pakistan Jouranl of Botany </em>54 (5): 1947–1962.</p>
<p>Khan, M. & Hussain, F. (2013) Conservation status of plant species in Tehsil Takht-e-Nasrati, District Karak, Khyber Pakhtun Khawa, Pakistan. <em>International Journal of Biodiversity and Conservation</em> 5 (1): 20–26.</p>
<p>Khan, M., Hussain, F. & Musharaf, S. (2013) Ethnobotanical profile of Tehsil Takht-e-Nasratti, District Karak, Pakistan. <em>Journal of Medicinal Plants Research</em> 7 (22): 1636–1651.</p>
<p>Khan, M., Musharaf, S. & Shinwari, Z.K. (2011) Ethnobotanical importance of halophytes of Noshpho salt mine, District Karak, Pakistan. <em>Research In Pharmaceutical Biotechnology</em> 3 (4): 46–52.</p>
<p>Khattak, N.S., Nouroz, F., Rahman, I.U. & Noreen, S. (2015) Ethno veterinary uses of medicinal plants of District Karak, Pakistan. <em>Journal of ethnopharmacology</em> 171: 273–279. https://doi.org/10.1016/j.jep.2015.05.048</p>
<p>Kumar, M., Harsh, N.S.K. & Prasad, R. (2014) <em>Conocybe apala</em> Fr.: Fr. Arnolds, <em>Helvella solitaria</em> P. Karst. and <em>Hygrocybe miniata</em> Fr. P. Kumm.; three new mushrooms from India. Current <em>Research in Environmental and Applied Mycology</em> 4 (1): 100–105.</p>
<p>Liu, J. & Bau, T. (2018) New species and new records in the genus <em>Conocybe</em> (Bolbitaceae) from China. <em>Phytotaxa </em>357 (4): 261–274. https://doi.org/10.11646/phytotaxa.357.4.2</p>
<p>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. <em>Mycologia</em> 98 (6): 982–995. https://doi.org/10.1080/15572536.2006.11832627</p>
<p>Melo, R.F.R., Chikowski, R.D.S., Miller, A.N. & Maia, L.C. (2016) <em>Coprophilous</em> Agaricales (Agaricomycetes, Basidiomycota) from Brazil. <em>Phytotaxa</em> 266 (1): 1–14. https://doi.org/10.11646/phytotaxa.266.1.1</p>
<p>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. <em>Molecular phylogenetics and evolution</em> 23 (3): 357–400. https://doi.org/10.1016/S1055-7903(02)00027-1</p>
<p>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. <em>Systematic Biology</em> 60 (3): 303–317. https://doi.org/10.1093/sysbio/syr005</p>
<p>Nemcsics, A. (1994) Spacing in the Munsell Color System Relative to the Coloroid Color Systems. <em>Color Research & Application</em> 19 (2): 122–125. https://doi.org/10.1111/j.1520-6378.1994.tb00071.x</p>
<p>Pomilio, A.B., Battista, M.E. & Vitale, A.A. (2006) Naturally-occurring cyclopeptides: structures and bioactivity. <em>Current Organic Chemistry</em> 10 (16): 2075–2121. https://doi.org/10.2174/138527206778742669</p>
<p>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. <em>Journal of ethnopharmacology</em> 165: 251–259. https://doi.org/10.1016/j.jep.2015.02.035</p>
<p>Sengul, Demirak., M.S., Turkekul, I. & Isik, H. (2021) <em>Conocybe romagnesii</em> and <em>Gerronema subclavatum</em> (Basidiomycota: Agaricales) in the Central Black Sea Region of Turkey<em>. Nordic Journal of Botany</em> 39 (12): 1–9. https://doi.org/10.1111/njb.03439</p>
<p>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. <em>Molecular biology and evolution</em> 28 (10): 2731–2739. https://doi.org/10.1093/molbev/msr121</p>
<p>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. <em>Public library of Science</em> 8 (2): e56143. https://doi.org/10.1371/journal.pone.0056143</p>
<p>Ullah, T., Saba, M. & Syed, M.F. (2022) <em>Coprinopsis afrocinerea</em> (Agaricales, Psathyrellaceae): First record from Asia (Pakistan). <em>Microscopy Research and Technique</em> 85 (10): 1–8. https://doi.org/10.1002/jemt.24192</p>
<p>Watling, R. (1971) Observations on the Bolbitiaceae—IV. Developmental studies on <em>Conocybe</em> with particular reference to the annulate species. <em>Persoonia-Molecular Phylogeny and Evolution of Fungi </em>6 (2): 281–289.</p>
<p>White, T.J., Bruns, T.D., Lee, S.B., Taylor, J.W., Innis, N. & Gelfand, D. (1990) <em>PCR-protocols and applications-a laboratory manual</em>. Academic Press, New York, pp. 315–322.</p>