Phenotypic and molecular diagnosis of Trichoderma spp. isolated from the local soil of Nineveh Governorate, Iraq
Article Main
Abstract
Trichoderma spp. is a common soil fungus that penetrates plant roots and engages with various microorganisms and plants. The present study aimed to determine the phylogenetic relationships of rhizosphere isolates of Trichoderma spp. obtained from the local soil in Mosul in the north of Iraq utilizing the sequence of the internal transcribed spacer-1 (ITS-1) region of the ribosomal DNA of eight Trichoderma spp.. Approximately 192 samples were collected between March and October 2023 from the rhizospheric soil of Potato )Solanum tuberosum), Tomato (Solanum lycopersicum), Wheat (Triticum aestivum), Barley (Hordeum vulgare), Eucalyptus (Eucalyptus globulus) and Citrus (Citrus limon (. Only 5.8% of the rhizosphere samples belonged to Trichoderma spp. The samples were identified based on morphological characteristics and molecular techniques. All the selected isolates amplified a similar band (660 bp). The nucleotide sequences were then analyzed, and a phylogenetic tree was constructed to compare the present isolates with the closest isolates submitted to the National Center for Biotechnology Information (NCBI). Isolates were submitted to NCBI as new strains under the accession numbers: Trichoderma harzianum (PP968131), T. longibrachiatum (PP977534), T. harzianum (PP977576), T. longibrachiatum (PP989798), Trichoderma longibrachiatum (PP989800), T. longibrachiatum (PP989803), T. longibrachiatum (PP989804), and T. harzianum (PP989805). This study expands the NCBI database with novel Trichoderma strains, enhancing fungal genetic resources globally. These new entries provide unique genomic data, potentially revealing valuable traits, resolving taxonomic ambiguities within Trichoderma, and improving the reliability of the NCBI database for future studies.
Article Details
Article Details
Keywords
Transcribed spacer-1, Trichoderma longibrachiatum, Trichoderma harzianum, ITS1, ITS4
References
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Allaga, H., Zhumakayev, A., Büchner, R., Kocsubé, S., Szűcs, A., Vágvölgyi, C., Kredics, L. & Hatvani, L., 2021. Members of the Trichoderma harzianum species complex with mushroom pathogenic potential. Agronomy, 11(12), p.2434. https://www.mdpi.com/2073-4395/11/12/2434
Allawi, Y. & Hmoshi, R.M., 2022. Isolation and Identification of Penicillium rubens from the Local Strain in Mosul, Iraq, and Investigation of Potassium Phosphate Effect on its Growth. Archives of Razi Institute, 77(1), p.421. doi: 10.22092/ARI.2021.356684.1896
Almashhadani, M.A. & Qassim, W.S., 2025. Bioactivity of silver nanoparticles produced by the aqueous extract of local Trichoderma longibrachiatum isolates against some types of MDR bacteria. Regulatory Mechanisms in Biosystems, 16(1), pp.e25037-e25037. https://doi.org/10.1542 1/0225037
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Alwadai, A.S., Al Wahibi, M.S., Alsayed, M.F., Alshaikh, N.A., Perveen, K. & Elsayim, R., 2024. Molecular characterization of plant growth-promoting Trichoderma from Saudi Arabia. Scientific Reports, 14(1), p.23236. https://doi.org/10.1038/s41598-024-73762
Bensons, A.E.B., 2005. Microbiology Applications” 9thed., Laboratory Manual in general, New York.
Cai, F. & Druzhinina, I.S., 2021. In honor of John Bissett: authoritative guidelines on molecular identification of Trichoderma. Fungal diversity, 107(1), pp.1-69. https://doi.org/10.1007/s13225-020-00464-4
Calleros, L., Panzera, F., Bargues, M.D., Monteiro, F.A., Klisiowicz, D.R., Zuriaga, M.A., Mas-Coma, S. & Pérez, R., 2010. Systematics of Mepraia (Hemiptera-Reduviidae): cytogenetic and molecular variation. Infection, Genetics and Evolution, journal of molecular epidemiology and evolutionary genetics in infectious diseases, 10(2), 221–228. https://doi.org/10.1016/j.meegid.2009.1 2.002
Carro-Huerga, G., Mayo-Prieto, S., Rodríguez-González, Á., Cardoza, R.E., Gutiérrez, S. & Casquero, P.A., 2023. Vineyard management and physicochemical parameters of soil affect native Trichoderma populations, sources of biocontrol agents against Phaeoacremonium minimum. Plants, 12(4), p.887. https://doi.org/10.3390/plants12040887
Damgaard, J., Andersen, N.M. & Meier, R., 2005. Combining molecular and morphological analyses of water strider phylogeny (Hemiptera–Heteroptera, Gerromorpha): effects of alignment and taxon sampling. Systematic Entomology, 30(2), pp.289-309. https://doi.org/10.1111/j.1365-3113.2004.00275.x
Druzhinina, I.S., Kopchinskiy, A.G. & Kubicek, C.P., 2006. The first 100 Trichoderma species characterized by molecular data. Mycoscience, 47(2), pp.55-64. https://doi.org/10.1007/S10267-006-0279-7
Eman, A.A.F., Mohamed, I.A., Allah, S.F., Shams, A.H. & Elsokkary, I.H., 2023. Trichoderma species: An overview of current status and potential applications for sustainable agriculture. Indian Journal of Agricultural Research, 57(3), pp.273-282.http://dx.doi.org/10.18805/IJARe.AF-751
Haque, Z., Iqbal, M.S., Ahmad, A., Khan, M.S., Singh, S.P. & Prakash, J., 2020. Explorations of Tolerant Trichoderma spp. as Plant Growth Promoter and Biocontrol Agent against Colletotrichum falcatum. Journal of Pure & Applied Microbiology, 14(1). https://doi.org/10.22207/JPAM.14.1.34
Harnelly, E., Kusuma, H.I., Thomy, Z. & Samingan, S., 2022. Internal Transcribed Spacer (Its) Gene As An Accurate Dna Barcode For Identification Of Macroscopic Fungus In Aceh. Biodiversitas Journal Of Biological Diversity, 23(5). https://doi.org/10.13057/biodiv/d230514
Hastuti, U.S. & Rahmawati, I., 2016. The antagonism mechanism of Trichoderma spp. towards Fusarium solani mold. Interaction. https://doi.org/10.21776/ub.jpacr.2016.005.03.260
Hillis, D.M., Moritz, C., Porter, C.A. & Baker, R.J., 1991. Evidence for biased gene conversion in concerted evolution of ribosomal DNA. Science, 251(4991), pp.308-310. https://doi.org/10.1126/science.1987647
Kale, G.J., Rewale, K.A., Sahane, S.P. & Magar, S.J., 2018. Isolation of Trichoderma spp. from the rhizospheric soils of tomato crop grown in Marathwada region. Journal of Pharmacognosy and Phytochemistry, 7(3), pp.3360-3362.
Kindermann, J., El-Ayouti, Y., Samuels, G.J. & Kubicek, C.P., 1998. Phylogeny of the genus Trichoderma based on sequence analysis of the internal transcribed spacer region 1 of the rDNA cluster. Fungal Genetics and Biology, 24(3), pp.298-309. https://doi.org/10.1006/fgbi.1998.1049
Kredics, L., Naeimi, S., Hatvani, L., Vágvölgyi, C., Cai, F., Druzhinina, I.S. & Manczinger, L., 2021. ‘The Good, the Bad and the Ugly’in the shades of green: the genus Trichoderma in the spotlight. Indian Phytopathology, 74(2), pp.403-411. https://doi.org/10.1007/s42360-021-00352-0
Kubiak, A., Wolna-Maruwka, A., Pilarska, A.A., Niewiadomska, A. & Piotrowska-Cyplik, A., 2023. Fungi of the Trichoderma genus: future perspectives of benefits in sustainable agriculture. Applied Sciences, 13(11), p.6434. https://doi.org/10.3390/app13116434
Kubicek, C.P., Steindorff, A.S., Chenthamara, K., Manganiello, G., Henrissat, B., Zhang, J., Cai, F., Kopchinskiy, A.G., Kubicek, E.M., Kuo, A. & Baroncelli, R., 2019. Evolution and comparative genomics of the most common Trichoderma species. BMC genomics, 20, pp.1-24.https://doi.org/10.1186/s12864-019-5680-7
Li, X., Liao, Q., Zeng, S., Wang, Y. & Liu, J., 2025. The use of Trichoderma species for the biocontrol of postharvest fungal decay in fruits and vegetables: Challenges and opportunities. Postharvest Biology and Technology, 219, p.113236.
https://doi.org/10.1016/j.postharvbio.2024.113236
Marcilla, A., Bargues, M.D., Ramsey, J.M., Magallon-Gastelum, E., Salazar-Schettino, P.M., Abad-Franch, F., Dujardin, J.P., Schofield, C.J. & Mas-Coma, S., 2001. The ITS-2 of the nuclear rDNA as a molecular marker for populations, species, and phylogenetic relationships in Triatominae (Hemiptera: Reduviidae), vectors of Chagas disease. Molecular Phylogenetics and Evolution, 18(1), pp.136-142. https://doi.org/10.1006/mpev.2000.0864
Matei, G.M. & Matei, S., 2024. The Influence Of Tomato Root Exudates On Structure And Diversity of Rhizosphere Communities of Bacteria And Fungi. Scientific Papers. Series B. Horticulture, 68(1). http://dx.doi.org/10.13140/RG.2.2.34643.39202
Mohammed, M. & Al-Taie, B., 2024. Isolation and identification of aquatic fungi from some water samples in Mosul City. Egyptian Journal of Aquatic Biology and Fisheries, 28(5), pp.793-804. https://dx.doi.org/10.21608/ejabf.2024.380430
Pokhrel, A., Adhikari, A., Oli, D., Paudel, B., Pandit, S., GC, B. & Tharu, B.R.R., 2022. Biocontrol potential and mode of action of Trichoderma against fungal plant diseases. Acta Scientific AGRICULTURE (ISSN: 2581-365X), 6(10). http://actascientific.com/ASAG/pdf/ASAG-06-1184.pdf
Prahl, R.E., Khan, S. & Deo, R.C., 2021. The role of internal transcribed spacer 2 secondary structures in classifying mycoparasitic Ampelomyces. PloS One, 16(6), p.e0253772.https://doi.org/10.1371/journal.pone.0253772
Prameeladevi, T., Prabhakaran, N., Kamil, D., Toppo, R.S. & Tyagi, A., 2018. Trichoderma pseudokoningii identified based on morphology was re-identified as T. longibrachiatum through molecular characterization. Indian phytopathology, 71, pp.579-587. https://doi.org/10.1007/s42360-018-0067-2
Qassim, W.S., Mohamed, A.H. & Hamdoon, Z.K., 2024. Biological control of root rot fungi in cowpea. SABRAO J. Breed. Genet, 56(1), pp.302-309. http://doi.org/10.54910/sabrao2024.56.1.27.
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Phenotypic and molecular diagnosis of Trichoderma spp. isolated from the local soil of Nineveh Governorate, Iraq. (2025). Journal of Applied and Natural Science, 17(2), 934-942. https://doi.org/10.31018/jans.v17i2.6595
