Bull 89 BULLETIN OF THE IRAQ NATURAL HISTORY MUSEUM Al Anbagi and Al-Khesraji Bull. Iraq nat. Hist. Mus. (2022) 17 (1): 89-101. https://doi.org/10.26842/binhm.7.2022.17.1.0089 ORIGINAL ARTICLE MORCHELLA CONICA PERS., 1818 (PEZIZALES, MORCHELLACEAE): A NEW RECORD FROM IRAQ Rajaa Abdulrazzaq Al Anbagi*♦ and Talib Owaid Al-Khesraji** *Department of Genetic Engineering, College of Biotechnology, Al-Qasim Green University, Babylon, Iraq. **Department of Biology, College of Education for Pure Sciences, Tikrit University, Tikrit, Iraq. ♦Corresponding author E-mail: dr.rajaa.a.a@gmail.com Received Date: 28 February 2022, Accepted Date: 08 May 2022, Published Date: 20 June 2022 This work is licensed under a Creative Commons Attribution 4.0 International License ABSTRACT The present study reports Morchella conica Pers.1818, which belongs to the family, Morchellaceae as a new record of Iraqi macromycota based on the morphological and molecular methods. During their short and often sporadic fruiting season, this fungal species was found in mixed forest unburned areas in Branan ranges (Suliamaniya Province, Northeast Iraq). Currently, M. conica is the second Morchella species reported from Iraq. The current study aimed to introduce this new record, which is poorly studied in the Middle East. M. conica is morphologically described and phylogenetically confirmed. The relationship between this species and other species within the genus was studied using the nrDNA ITS sequences from different species and diverse geographical regions. Maximum likelihood (ML) analyses were also conducted to build the molecular phylogeny of this species. The results of the presented species are essential for assessing the genus geographic distribution and developing information about species of this highly prized edible, industrial medicinal fungus. Keywords: Bioinformatic, Iraq, Morchella conica, Morchellaceae, Phylogenetics, rRNA. INTRODUCTION True morels (species of Morchella Dill. ex Pers.: Fr.) are edible ascomycetous fungi belonging to the Pezizales Morchellaceae (Hibbett et al., 2007). The wild mushroom fruits sometimes exist prolifically in various forest types throughout western North America. Morels are amongst the most highly prized and valuable edible fungi in the world (Olfati et al., 2009; Nitha et al., 2017; Tietel and Masaphy, 2018). Morels have considerably gained attention and are widely in demand due to their nutritional and medicinal values as well as the diversity of bioactive ingredients as being radioprotective for mitochondria and DNA and anti-inflammatory, immunostimulants (Tietel and Masaphy, 2018; Yang et al., 2019; Nitha et al., 2020). True morels are also known for their broad ecological plasticity due to being colonized a wide range of habitats (burned BULLETIN OF THE IRAQ NATURAL HISTORY MUSEUM Iraq Natural History Research Center & Museum, University of Baghdad https://jnhm.uobaghdad.edu.iq/index.php/BINHM/Home Online ISSN: 2311-9799 Print ISSN: 1017-8678 https://doi.org/10.26842/binhm.7.2022.17.1.0089 https://orcid.org/0000-0001-8873-9448 https://orcid.org/0000-0001-8378-3319 https://creativecommons.org/licenses/by/4.0/ https://jnhm.uobaghdad.edu.iq/index.php/BINHM/Home 90 BULLETIN OF THE IRAQ NATURAL HISTORY MUSEUM Morchella conica Pers., 1818 (Pezizales, Morchellaceae) areas, coastal dunes, orchards, grasslands, and forests) with a wide spectrum of trophic strategies. Through secreting enzymes into the external substrates and absorbing the released nutrients, the taxon obtains its nourishment mostly as saprotrophs, slight parasitism and mycorrhiza-like interactions have also been reported (Du et al., 2012). Although Morchella is an imperative genus, the genus still has large gaps in our knowledge about its taxonomy, biology, ecology, and diversity. Taxonomically, Morchella is a complex genus and its taxonomic classification system of fungi is still unsolved (Wurtz et al., 2005; Masaphy, 2011; Richard et al., 2014; Loizides et al., 2016). It has been proposed the genus needs in-depth morphological studies and considers species with unique habitats for moving more research into an ecosystem perspective. The species of this genus have been mostly reported and morphologically described from Europe, with only few species described in Asia and USA (Du et al., 2012). Based on the classical morphological taxonomy and molecular phylogenetic and biogeographic studies, the genus has been divided into three main distinct groups or clades, Esculenta clade (yellow morels, e.g., M. esculenta, M. deliciosa, and M. crassipes (Vent.) Pers., Elata clade (black morels, e.g., M. angusticeps Peck, M. conica and M. elata Fr.), including semifree capped morels deeply nested. The third group was blushing morels presented an early diverging basal lineage containing M. rufobrunnea, and M. rigidoide, distributed in the tropics or subtropics (O’Donnell et al., 2011; Du et al., 2012). The recent molecular phylogenetic studies revealed that the occurrence of at least 60 species of Morchella worldwide (Kuo et al., 2012; Du et al., 2012; Baroni et al., 2018; Petrzelova and Sochor, 2019; Clowez et al., 2020). Only a part of the total macrofangal wealth has been subjected to scientific investigation and, mycologists remain to unravel the unexplored and hidden Iraqi macrofungi (e.g. Abdulla et al., 1989; Al-Khesraji, 2016). Despite its biogeographic significance, Iraq is still unexplored from macrofungal point of view (Suliaman et al., 2017). Reports on Pezizales including Morchellaceae from the country are very limited (Al Anbagi, 2014; Al-Khesraji, 2016, 2018; Al- Khesraji and Al-Hayawi, 2019; Al-Khesraji and Suliaman, 2019). With increasing interest in the genus Morchella worldwide, only M. esculenta has so far been reported from northeast Iraq (Al-Khesraji, 2016). The current study presents the results of morphological and molecular identifications which were confirmed using a molecular phylogeny of a newly recorded black morel, Morchella conica Pers., 1818 from Iraq. MATERIALS AND METHODS Specimens' collection and morphological features Fresh specimens of morels were collected during the investigation on macrofungi in April-May 2019 on Baranan Mountain (elevation 1200-1400 m) north Darbandikhan City (35.116258°N 45.686245°E) of Suliamaniya Province, Eastern North of Iraq, as a part of Iraqi Kurdistan Region. Data related to a natural habitat, site coordinates, a soil type, and vegetation were recorded at the field. The specimens were photographed in 91 BULLETIN OF THE IRAQ NATURAL HISTORY MUSEUM Al Anbagi and Al-Khesraji their natural habitats as well as in the laboratory after being somewhat dried, and transferred to the laboratory for morphological and molecular analyses. Later, the fruit bodies were air-dried and deposited in the Biology Department, College of Education for Pure Sciences, Tikrit University, Iraq. The macromorphological features were described for fresh fruiting bodies. Measurements of the microscopic features were also performed on the fresh materials; these specimens were described and photographed under a light microscope. The classical technique for fungal identification was concluded according to the relevant literature and keys (Negi, 2006; Watanabe, 2010; Lakhanpal et al., 2010; Kuo et al., 2012). Later, the morphological identification was confirmed based on amplification of the ribosomal DNA internal transcribed spacer (rDNA-ITS) for two isolates and phylogenetic analyses (White et al., 1990). Molecular and DNA sequencing techniques The identity of isolated strains was confirmed using DNA amplifications and sequencings of the fungal gene ITS region. Genomic DNA was extracted from fresh fruiting body specimens using the ZR Plant/Seed DNA MiniPrep kit (Zymo Research), following the manufacturer's instructions. The extracted DNA was stored at -20 °C for further analysis. The rDNA-ITS was amplified using the ITS1 and ITS4 primers (White et al., 1990). The PCR amplifications were performed in a total volume of 25 µl. This consisted of 1.5 µl DNA, 5 µl Taq PCR PreMix (Intron, Korea), and 1µl of each primer (10 pmol). The deionized distilled water was finally added to complete the total volume. The thermal cycling conditions were as follows: 94 °C for 3 min, 35 cycles of 94 °C for 45 s, 52°C for 1 min, and 72 °C for 1min, followed by 72 °C for 7 min using a Thermal Cycler (Gene Amp, PCR system 9700; Applied Biosystem). Amplicons were loaded and visualized on 2 % agarose gels stained with Red safe (Intron Korea) under ultraviolet light (309 nm) for quantity determination (Al- Khesraji et al., 2021). The successful DNA amplifications were sent to Macrogen Inc./ South Korea for sequencing . Bioinformatic and molecular phylogenetic analyses Obtained sequence data of fungal isolates were trimmed, and low-quality edges were removed. The individual sequences were assembled to contigs with the Geneious program (Kearse et al., 2012). These sequences were blasted against the ITS database in the NCBI’s Gene Bank database for species identification based on the query sequence similarity. The generated sequences in the present study were deposited in GenBank under the accession number MW291450. The results of BLASTN search with the sequences having 99% and above similarity were used for further analyses. Phylogenetic analyses were performed to assess the fungal sequence based on the result of the BLAST search in GenBank and sequences of close related Morchella species as recommended (Du et al., 2012). The DNA sequences were aligned using MAFFT v7.309 (Katoh and Standley, 2013), and the aligned sequences were manually modified as necessary. Maximum likelihood (ML) analyses were conducted using RAxML V7.2.8 (Stamatakis, 2014). Bootstrap algorithm was realized on the dataset for 1000 replicates in Geneious 92 BULLETIN OF THE IRAQ NATURAL HISTORY MUSEUM Morchella conica Pers., 1818 (Pezizales, Morchellaceae) version 9.1.8. All characters were preserved as unordered and equally weighted. The alignment gaps were treated as missing data. The resulting tree was visualized in Geneious version 9.1.8. RESULTS AND DISCUSSION Habitat and distribution Solitary on soil under mixed forest unburned sites during April and May in 2019. The fruiting bodies were found in the wet ground between rocky and brown soil covered with decayed plant litter and woody debris. Samples were collected under diverse tree species including Quercus sp., Populus sp., and Nerium sp. However, some specimens were found away from the trees. Several fruiting bodies of M. conica so far are known only from northern Iraq in the current study. The fungus was collected from various localities on the Baranan Mountain in Darbandikhan area (10 Km north Darbandikhan Lake), in Suliamaniya. Morphological features The Cap of M. conica has 2-6 cm high and 1.5-3 cm wide, conical with acute apex, attached to the stipe, yellowish when young and brownish or black at age, with vertical ribs connected by transverse ribs, forming a series of rectangular or square pits. Stipe 3-8 cm long and 1-3 cm wide, cylindrical with swollen base, hollow, white to yellowish- white, surface with fine granules, rarely bald (Pl. 1A-C). The Asci was cylindrical, hyaline, with a base rounded, usually 8-spored 300-350 × 18-22 µm (Pl. 1D). Ascospores were elliptical, smooth-walled, hyaline, homogenous with no oil droplets, inside sacs, some external oil droplets occur adjacent to each end of the spores 20-22 × 10-13 µm (Pl. 1E). Paraphyses cylindrical with rounded apex, septate, hyaline, 220-300 × 8 -10 µm. In the present paper, M. conica was reported as a new addition to Iraqi macrofungi and a second Morchella species described from the country. Macro-and microscopic features of M. conica are in agreement with previously related descriptions (Negi, 2006; Watanabe, 2010; Lakhanpal et al., 2010). However, some morphological variations within and between Morchella species may be expected due to the plasticity of macro- and micromorphological traits (Phanpadith et al., 2019; Ali et al., 2021). In the study area, morphological differences were not observed between fruiting bodies of M. conica collected from different substrates/ hosts in unburned sites. The fruiting bodies of current species were found under diverse tree species including Quercus sp., Populus sp. and Nerium sp. presumably to survive using diverse niches. However, it has been reported that 70% of the Elata clade species were found in a coniferous forest with a few species found within temperate deciduous forests (Du et al., 2012). Regarding the trophic status, M. conica and other true morels were considered saprotrophic, mycorrhizal, or even facultative (Keefer, 2005; Du et al., 2012; Kuo, 2012). In this study, the presence of M. conica under decomposed plant debris in places away from trees may suggest that the fungus acts as a saprotroph rather than mycorrhizal. True 93 BULLETIN OF THE IRAQ NATURAL HISTORY MUSEUM Al Anbagi and Al-Khesraji morels including M. conica are known to fruit after a forest fire (Wurtz et al., 2005; Negi, 2006; Masaphy, 2011; Du et al., 2012; Larson et al., 2016; Miller et al., 2017) presumably supporting their saprotrophic mode. Li et al (2013) mentioned that black morels like M. conica were saprotrophic and those with yellow caps like M. esculenta were mycorrhizal. However, switching lifestyles, for example, from symbiotic to saprotrophic interactions and vice versa in macrofungal species have been suggested and recorded to access organically restricted nutrients (Bahnmann et al., 2018; Al Anbagi, 2020). This aspect in Morchella has not been resolved yet and needs further attention in the future studies. Molecular identification and phylogenetic analysis Amplification of the ITS rRNA region of morels resulted in size product 663 bp. A BLASTN query of investigated sequence in GenBank revealed that they had more than 99% similarity with M. conica (accession numbers AJ544195 and EF080999). However, the Iraqi morel sequence had more than 99% sequence similarity with M. elata Fr. (accession numbers MN462953 and MN462952), M. importuna M. Kuo, O‘Donnell & T.J. Volk, and M. esculenta (L.) Pers. (accession numbers MF170632 and GU373504 respectively) and Morchella sp. (MK955411, Diag. 1). The size of the cap and stipe of M. elata and M. importuna were wider. However, compared with their asci, the size of asci of M. conica were wider with a broadly rounded base containing smaller ascospores with taller and narrower paraphyses (Ali et al., 2021). The insufficiently identified sequences and detected named sequences in GenBank have been reported. Being has a high percentage similarity with other species in the current study; it may be potentially suggested misidentifications of Morchella sequences in GenBank. The ITS gene alone was able to identify 77.4% of the known phylospecies in Morchella using ITS1 and ITS2. At least 66% of the named Morchella sequences in GenBank have been found misidentified such as those with binomials (Du et al., 2012; Petrželová and Sochor, 2019). Some species of this genus such as M. esculenta, M. crassipes, and M. elata included different phylogenetic species. That is because of the near absence of type studies and informational databases, including references to voucher specimens and/or cultures (Kanwal et al., 2011; Du et al., 2012) for aiding DNA sequence-based identifications of true morels. Additionally, predominant cryptic speciation has been discovered between macrofungi including true morels. With only 2.6 and 4.5% of the estimated fungal species discovered (Hyde et al., 2020), detecting a new species in Morchella is highly raised especially with reporting the most novel species in North America, Turkey, and Asia as well as Europe (Du et al., 2012; Loizides et al., 2016). In the present study, the phylogenetic analyses of Morchella sequences confirmed the species identification after being a BLAST query and reconstructed some related black Morchella sequences similar to other researchers who have used a molecular phylogeny to confirm the species identification (Du et al., 2012). The phylogenetic tree of black morels was divided into conica groups as well as other clustered species. M. conica from Iraq interestingly clustered into one clade with M. conica from Germany while the 94 BULLETIN OF THE IRAQ NATURAL HISTORY MUSEUM Morchella conica Pers., 1818 (Pezizales, Morchellaceae) specimens belong to this species from different countries such as China were separated into other clades. These relations may suggest that the disjunct distributions of species have been produced through distinct processes during their evolutionary relationships. The current results agree with other findings that geographically isolated populations of the M. conica may have more variation than those between two putatively different species (Bunyard et al., 1994; Li et al., 2013; Ali et al., 2021). The sequences of M. conica grouped nearest from M. costata, M. esculenta, and M. elata similar to the outcomes of other scientists (Kanwal et al., 2011; El-Wakil and Al-Gifri, 2020). However, some species within the Elata clade still have unsolved evolutionary relationships (Due et al., 2012). Plate (1): Morchella conica; (A) Fruiting body at the natural habitat, (B) Fruiting bodies in different age of the development, (C) Air-dried fruiting bodies, (D) Ascus with 8 ascospores, (E) Ascospores. 95 BULLETIN OF THE IRAQ NATURAL HISTORY MUSEUM Al Anbagi and Al-Khesraji Diagram (1): The maximum likelihood tree based on ITS/5.8S rRNA gene sequences represented the position of M. conica from Iraq, resulting from Rapid Bootstrapping algorithm and a ML search in RAxML. The number within parentheses indicates the GenBank accession number. The bootstrap probability value is shown at the nodes. CONCLUSIONS The present study reported the first black morel M. conica; this species was found in mixed forest unburned area in Branan ranges, northeast Iraq. Previously, yellow morel, M. esculenta, has been collected from northern Iraq, but the identification was entirely 96 BULLETIN OF THE IRAQ NATURAL HISTORY MUSEUM Morchella conica Pers., 1818 (Pezizales, Morchellaceae) based on the morphological features. The current results were based on the morphological description and molecular analyses. The phylogenetic analysis clustered the Iraqi M. conica sequence closest to other specimens from different geological regions. Further surveys on this group of fungi are urgently needed to detect the distribution of this genus as well as other macrofungi. Detecting, describing, and identifying the first record of fruiting bodies of M. conica, Elata clade, in Iraq with yearly high temperature and low rainfall, and complex topography conceivably offered useful information about environmental tolerance, especially with being considered Asia or China as the center of species diversification and distribution of the modern Morchella. CONFLICT OF INTEREST STATEMENT We are the authors of the submitting manuscript, declare and verify there are no significant financial supporters for the current research from any providers. LITERATURE CITED Abdullah, S. K., Al-Issa, A., Ewaz, J. O. and Al-Bader, S. M. 1989. Taxonomy of edible hypogenous ascomycotina of Iraq. International Journal of Mycology and Lichenology, 4: 9-21. Al Anbagi, R. A. 2020. A comparative taxonomic and diversity study of litter-associated fungi in Northwest Arkansas forests. Ph.D. thesis in Cell and Molecular biology, the Fulbright College of Arts and Sciences, Arkansas University, Arkansas, USA, 214pp. Al Anbagi, R. A. 2014. Histological study of the discomycetes fungus Cheilymina theleboloides. Journal of Babylon University/Pure and Applied Sciences, 22 (2): 769-778. [Click here] AL-Khesraji, T. O. 2016. Seven new records of ascomycetous macrofungi from Suliamaniya province (Northeast of Iraq). Journal of Biology, Agriculture and Healthcare, 6 (16): 94-107. [Click here] AL-Khesraji, T. O. 2018. Two Peziza taxa (Peziza proteana f. proteana (Boud.) Seaver and Peziza proteana f. sparassoides (Boud.) Korf) new to Iraq and bordering countries. African Journal of Plant Science, 12(2): 24-27. [CrossRef] Al- Khesraji, T. O. and Al-Hayawi, A. Y. 2019. Five new taxa records for the macromycota of Iraq from Suliamaniya and Tikrit provinces (Iraq). Plant Archives, 19(2): 4067-4072. [Click here] Al-Khesraji, T. O. and Suliaman, S. 2019. New taxa records for macromycota of Iraq from Salahadin Governorate. The Journal of Research on the Lepidoptera, 50 (3): 125-135. [CrossRef] https://www.iasj.net/iasj/download/d69258d7a6cecf7b https://www.iiste.org/Journals/index.php/JBAH/article/view/32523/33415 https://doi.org/10.5897/AJPS2017.1613 http://plantarchives.org/19-2/4067-4072%20(5933).pdf http://dx.doi.org/10.36872/LEPI/V50I3/201032 97 BULLETIN OF THE IRAQ NATURAL HISTORY MUSEUM Al Anbagi and Al-Khesraji Ali, S. Imran, A. Fiaz, M. Khalid, A. N. and Khan, S. M. 2021. Molecular identification of true morels (Morchella spp.) from the Hindu Kush temperate forests leads to three new records from Pakistan. Gene Reports, 23:101125. [CrossRef] Bahnmann, B., Mašínová, T., Halvorsen, R., Davey, M. L., Sedlák, P., Tomšovský, M. and Baldrian, P. 2018. Effects of oak, beech and spruce on the distribution and community structure of fungi in litter and soils across a temperate forest. Soil Biology and Biochemistry, 119: 162-173. [CrossRef] Bunyard, B. A., Nicholson, M. S and Royse, D. J. 1994. A systematic assessment of Morchella using RFLP-analysis of the 28S ribosomal RNA gene. Mycologia, 86 (6): 762-772. [CrossRef] Clowez, P., Marcos Martinez, J., Sanjaume, R., Marques, G., Bellanger, J.-M. and Moreau, P.-A. 2020. A survey of half-free morels in Spain reveals a new species: Morchella iberica sp. nov. (Ascomycota, Pezizales). Ascomycete.org., 12 (1): 11- 18. [CrossRef] Du, X.- H., Zhao, Q., Yang, Z. L., Hansen, K., Taşkın, H., Büyükalaca, S., Dewsbury, D., Moncalvo, J.-M., Douhan, G. W., Robert, V. A. R. G., Crous, P. W., Rehner, S. A., Rooney, A. P., Sink, S. and O’Donnell, K. 2012. How well do ITS rDNA sequences differentiate species of true morels (Morchella)? Mycologia, 104 (6): 1351-1368. [CrossRef] El-Wakil, D. A. and Al-Gifri, A. N. 2020. Identification of wild macrofungi from Southern West of Saudi Arabia. International Journal of Current Research in Bioscinces and Plant Biology, 7(1): 17-22. [CrossRef] Hibbett, D. S., Binder, M., Bischoff, J. F., Blackwell, M., Cannon, P. F., Eriksson, O. E., Huhndorf, S., James, T., Kirk, P. M., Lücking, R., Lumbsch, H. T., Lutzoni, F., Matheny, P. B., McLaughlin, D. J., Powell, M. J., Redhead, S., Schoch, C. L., Spatafora, J. W., Stalpers, J. A., Vilgalys, R., Aime, M. C., Aptroot, A., Bauer, R., Begerow, D., Benny, G. L., Castlebury, L. A., Crous, P. W., Dai, Y.-C., Gams, W., Geiser, D. M., Griffith, G. W., Gueidan, C., Hawksworth, D. L., Hestmark, G. Hosaka, K., Humber, R. A., Hyde, K. D., Ironside, J. E., Kõljalg, U., Kurtzman, C. P., Larsson, K.-H., Lichtwardt, R., Longcore, J., Miądlikowska, J., Miller, A., Moncalvo, J.-M., Mozley-Standridge, S., Oberwinkler, F., Parmastoa, E., Reeb, V., Rogers, J. D., Roux, C., Ryvarden, L., Sampaio, J. P., Schüßler, A., Sugiyama, J., Thorn, R. G., Tibell, L., Untereiner, W. A., Walker, C., Wang, Z., Weir, A., Weiss, M., White, M. M., Winka, K., Yao, Y.-J. and Zhang, N. 2007. A higher-level phylogenetic classification of the fungi. Mycological Research, 111: 509-547. [CrossRef] https://doi.org/10.1016/j.genrep.2021.101125 https://doi.org/10.1016/j.soilbio.2018.01.021 https://doi.org/10.2307/3760589 https://doi.org/10.25664/ART-0291 https://doi.org/10.3852/12-056 https://doi.org/10.20546/ijcrbp.2020.701.002 https://doi.org/10.1016/j.mycres.2007.03.004 98 BULLETIN OF THE IRAQ NATURAL HISTORY MUSEUM Morchella conica Pers., 1818 (Pezizales, Morchellaceae) Hyde, K. D., Jeewon, R., Chen, Y. J. Bhunjun, C. S., Calabon, M. S., Jiang, H. B, Lin, C. G., Norphanphoun, C., Sysouphanthong, P., Pem, D. and Tibpromma, S. 2020. The numbers of fungi: is the descriptive curve flattening? Fungal Diversity, 103 (1): 219-271. [CrossRef] Kanwal, H. K., Acharya, K. Ramesh, G. and Reddy, M. S. 2011. Molecular characterization of Morchella species from the Western Himalayan region of India. Current Microbiology, 62 (4): 1245-1252. [CrossRef] Katoh, K. and Standley, D. M. 2013. MAFFT multiple sequence alignment soft- ware version 7: improvements in performance and usability. Molecular Biology and Evolution, 4 (30):772-780. [CrossRef] Kearse, M., Moir, R., Wilson, A., Stones-Havas, S., Cheung, M., Sturrock, S., Buxton, S., Cooper, A., Markowitz, S., Duran, C., Thierer, T., Ashton, B., Meintjes, P. and Drummond, A. 2012. Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics, 28 (12): 1647-1649. [CrossRef] Keefer, M. E. 2005.The ecology and economy of morels in British Columbiaʼs east Kootenay. MSc. Thesis, Royal Roads University, Canada, 99 pp. Kuo, M., Dewsbury, D. R., O'Donnell, K., Carter, M. C., Rehner, S. A., Moore, J. D., Moncalvo, J. M., Canfield, S. A., Stephenson, S. L., Methven, A. S. and Volk, T. J. 2012. Taxonomic revision of true morels (Morchella) in Canada and the United States. Mycologia, 104 (5):1159-1177. [CrossRef] Kuo, M. 2012. The Morchellaceae: True morels and verpas. [Click here] Lakhanpal, T. N. 2010. Biology of Indian morels. IK International Pvt Ltd, 245 pp. Larson, A. J., Cansler, C. A., Cowdery, S. G. S., Hiebert, S., Furniss, T. J., Swanson, M. E. and Lutz, J. A. 2016. Post-fire morel (Morchella) mushroom abundance, spatial structure, and harvest sustainability. Forest Ecology and Management, 377: 16–25. [CrossRef] Li, Q. L. Ding, C. and Fan, L. 2013. Trophic manner of morels analyzed by using stable carbon isotopes. Mycosystema, 32: 213-223 . Loizides, M., Bellanger, J.-M., Clowez, P., Richard, P., Moreau, P.-A. 2016. Combined phylogenetic and morphological studies of true morels (Pezizales, Ascomycota) in Cyprus reveal significant diversity, including Morchella arbutiphila and M. disparilis spp. nov. Mycological Progress, 15: 1-28. [CrossRef] https://doi.org/10.1007/s13225-020-00458-2 https://doi.org/10.1007/s00284-010-9849-1 https://doi.org/10.1093/molbev/mst010 https://doi.org/10.1093/bioinformatics/bts199 https://doi.org/10.3852/11-375 http://www.mushroomexpert.com/morchellaceae.html https://doi.org/10.1016/j.foreco.2016.06.038 https://doi.org/10.1007/s11557-016-1180-1 99 BULLETIN OF THE IRAQ NATURAL HISTORY MUSEUM Al Anbagi and Al-Khesraji Masaphy, S. 2011. Diversity of fruiting patterns of wild black morel mushroom. In: Proceedings of the 7th International Conference on Mushroom Biology and Mushroom Products; 2011 Oct 4–7; Arcachon, France. Villenave d’Ornon: Institut National de la Recherche Agronomique, p. 165-169. Miller, A. N., Raudabaugh, D. B., Iturriaga, T. Matheny, P. B., Petersen, R. H., Hughes, K. W., Gube, M., Rob, A., Powers, R. A., James, T. Y and O’Donnell, K. 2017. First report of the post-fire morel Morchella exuberans in eastern North America. Mycologia, 109 (5):710-714. [CrossRef] Negi, C. S. 2006. Morels (Morchella spp.) in Kumaun Himalaya. Natural Products Radiance, 5(4): 306-310. Nitha, B., Smina, T. P. and Janardhanan, K. K. 2017. Chemopreventive effect of Morchella esculenta against DMBA induced skin papilloma in mice. European Journal of Pharmaceutical and Medical Research, 4 (4): 658-662 . Nitha, B., De, S., Devasagayam, T. P. A. and Janardhanan, K. K. 2020. Edible mushroom Morchella esculenta (L.) Pers. mycelium protects DNA and mitochondria from radiation induced damages. Indian Journal of Experimental Biology, 58 (12): 842-847. [Click here] O’Donnell, K. Rooney, A. P., Mills, G. L., Kuo, M., Weber, N. S. and Rehner, S. A. 2011. Phylogeny and historical biogeography of true morels (Morchella) reveals an early Cretaceous origin and high continental endemism and provincialism in the Holarctic. Fungal Genetics and Biology, 48: 252-265. [CrossRef] Olfati, J., Peyvast, G. and Mami, Y. 2009. Identification and chemical properties of popular wild edible mushrooms from northern Iran. Journal of Horticulture and Forestry, 1 (3): 048-051. Petrzelova, I. and Sochor, M. 2019. How useful is the current species recognition concept for the determination of true morels? Insights from the Czech Republic. MycoKeys, 52: 17-43. [CrossRef] Phanpadith, P., Yu, Z. and Li, T. 2019. High diversity of Morchella and a novel lineage of the esculenta clade from the North Qinling Mountains revealed by GCPSR- based study. Scientific Reports, 9 (1): 1-10. [CrossRef] Richard, F., Sauve, M., Bellanger, J. M., Clowez, P., Hansen, K., O’Donnell, K., Urban, A., Courtecuisse, R. and Moreau, P. A. 2014. True morels (Morchella, Pezizales) of Europe and North America: evolutionary relationships inferred from multilocus data and a unified taxonomy. Mycologia, 107 (2): 359-382. [CrossRef] https://doi.org/10.1080/00275514.2017.1408294 http://nopr.niscair.res.in/bitstream/123456789/55807/1/IJEB%2058%2812%29%20842-847.pdf https://doi.org/10.1016/j.fgb.2010.09.006 https://doi.org/10.3897/mycokeys.52.32335 https://doi.org/10.1038/s41598-019-56321-1 https://doi.org/10.3852/14-166 100 BULLETIN OF THE IRAQ NATURAL HISTORY MUSEUM Morchella conica Pers., 1818 (Pezizales, Morchellaceae) Stamatakis, A. 2014. RAxML version 8: a tool for phylogenetic analysis and post- analysis of large phylogenies. Bioinformatics, 30 (9): 1312-1313. [CrossRef] Al- Khesraji, T. O., Suliaman, S. Q. and Al-Hayawi, A. Y. 2021. Morphological and molecular characterization of Bjerkandera adusta (Meruliaceae), a new addition to macromycota of Iraq. Annals of the Romanian Society for Cell Biology, 25 (4): 11968-11975. [Click here] Tietel, Z. and Masaphy, S. 2018. True morels (Morchella)-nutritional and phytochemical composition, health benefits and flavor: a review. Critical Reviews in Food Science and Nutrition, 58 (11): 1888-1901. [CrossRef] Watanabe, T. 2010. Pictorial atlas of soil and seed fungi: Morphologies of cultured fungi and key to species. CRC Press, New York, USA, 426 pp. White, T. J., Bruns, T., Lee, S. and Taylor, J. W. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis, M. A., Gelfand, D. H., Sninsk, M. J. J. and White, T. J. (eds.), PCR protocols: A guide to methods and applications. Academic Press, New York, p. 315-322. [CrossRef] Wurtz, T. L., Wiita, A. L., Weber, N. S. and Pilz, D. 2005. Harvesting morels after wildfire in Alaska. Res. Note PNW-RN-546. Portland, OR: US Department of Agriculture, Forest Service, Pacific Northwest Research Station, 31 pp. [CrossRef] Yang, C., Zhou, X., Meng, Q., Wang, M., Zhang, Y. and Fu, S. 2019. Secondary metabolites and antiradical activity of liquid fermentation of Morchella sp. isolated from Southwest China. Molecules, 24: 1706. [CrossRef] https://doi.org/10.1093/bioinformatics/btu033 https://www.annalsofrscb.ro/index.php/journal/article/view/4052 https://doi.org/10.1080/10408398.2017.1285269 https://doi.org/10.1016/B978-0-12-372180-8.5004 https://doi.org/10.2737/PNW-RN-546 https://doi.org/10.3390/molecules24091706 101 BULLETIN OF THE IRAQ NATURAL HISTORY MUSEUM Al Anbagi and Al-Khesraji Bull. Iraq nat. Hist. Mus. (2022) 17 (1): 89 -101. Morchella Conica Pers., 1818تسجيل جديد للفطر (Pezizales, Morchellaceaeمن العراق ) رجاء عبد الرزاق العنبكي* و طالب عويد الخزرجي** * قسم الهندسة الوراثية، كليه التقانات األحيائية، جامعه القاسم الخضراء، بابل، العراق. ** قسم علوم الحياة، كليه التربية للعلوم الصرفة، جامعه تكريت، تكريت، العراق. 20/06/2022، تأريخ النشر: 08/05/2022، تأريخ القبول: 28/02/2022تأريخ االستالم: الخالصة العائد للعائلة Morchella conica Pers.1818جلت الدراسة الحاليه النوع س Morchellaceae كتسجيل جديد ضمن الفطريات الكبيرة العراقية، اعتمادا على الطرق املورفولوجية والجزيئية. خالل موسم اإلثمار القصير واملتقطع في كثير من األحيان، عثر لغابات املختلطة غير املحترقة ضمن منطقه بروانه على هذا النوع الفطري في مناطق ا )محافظة السليمانية، شمال شرق العراق(. كان الهدف من الدراسة هو تقديم التسجيل ، الذي قلما درس في الشرق األوسط. حاليا، النوع املدروس هو ثاني نوع لهذا النوعالجديد تم تسجيله من العراق. Morchellaمن أنواع الجنس و اكد الوصف من الناحية الجزيئية، كما درست العالقة بين M. conicaصف و ً مظهريا من nrDNA ITSهذا النوع واألنواع األخرى ضمن الجنس باستخدام تسلسالت املنطقة أنواع مختلفة ومناطق جغرافية متنوعة. أيضا، تم أجراء تحليالت االحتمال األقص ى (MLلبناء األصل التطوري الجزيئي ) .لهذا النوع تعتبر نتائج النوع املسجل الحالي اساسيه لتقييم التوزيع الجغرافي للجنس وتوفير املعلومات حول أنواع هذا الفطر عالي القيمة الغذائية، الصناعية والطبية.