New report of Trichoderma sp from Sub-Himalayan tea growing regions of West Bengal India

New report of Trichoderma sp from Sub-Himalayan tea growing regions of West Bengal India | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article New report of Trichoderma sp from Sub-Himalayan tea growing regions of West Bengal India Suvojeet Mukherjee, Saini Sultana, Vivek Chettri, Nahin Millat, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6903051/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Trichoderma species (Hypocreaceae) is one of the world’s most common widely used plant growth-promoting fungus and, ecologically and economically important genus. Three newly discovered species of the fungal genera Trichoderma , Trichoderma asperellum Trichoderma hamatum and Trichoderma yunnanense , isolated from the rhizosphere zone of tea of North Bengal region, India have been identified through morphology and phylogenetic study of sequences of nucleotide. The analysed DNA sequences encompassed the internal transcribed spacer (ITS) regions of the rDNA cluster, specifically ITS1 and ITS2.The analyses show that T. asperellum , T. yunnaense , T hamatum are grouped within the Hamatum clade. Biological sciences/Microbiology/Fungi Biological sciences/Microbiology/Fungi/Fungal ecology Morphology New species. Phylogenetic analysis Trichoderma yunnanense Figures Figure 1 Figure 2 Figure 3 Introduction Trichoderma spp. (Hypocreales, Pezizomycotina, and Ascomycota) is now recognized as a widely utilized biocontrol agent and biological fertilizer in agricultural industry. Their remarkable capacity to fight against plant pathogenic fungi results in the production of secondary metabolites that have bio-pesticide properties and bio-stimulant effects. 1 – 4 Trichoderma sp are cosmopolitan saprophytic in nature are naturally occurring, and are commonly found in various soils, decaying wood, organic matter, and as endophytes of plants. 5 . 6 , 7 Species within the genus Trichoderma are recognized because of their exceptional production of an assortment of secondary metabolites, which include polysaccharides, toxins, and antibiotics. 8 , 9 , 10 Moreover, various strains in this genus are extensively utilized for the biocontrol of soilborne plant pathogenic fungi. 11 Trichoderma sp used as both direct biological control techniques that control the targeted pathogens like mycoparasitism along with indirect approaches that enhance plant health, including the induction of systemic resistance, promotion of plant growth, and improvement of rhizosphere competence. Direct biocontrol strategies encompass competition for nutrients and space, the production of antimicrobial agents (antibiosis), and the formation of lytic enzymes. 4 Trichoderma has the ability to alter the biology of the rhizosphere, thereby minimizing the impact of pathogens. 12 Furthermore, certain species of Trichoderma show efficacy as bio-remediators for heavy metals. 13 , 14 The present study was carried out to distinguish species or strains of Trichoderma by cultural and morphological characters and further characterize the diversity of Trichoderma spp. isolated from rhizosphere soil of tea based on molecular characters. Material and Methods Study Area Soil sampling and Isolation Soil samples were collected from rhizosphere habitats of tea gardens of the North Bengal region, India, for isolating Trichoderma sp . The isolation of Trichoderma was carried out using a serial dilution method, 15 where each sample was diluted to 10 − 2 . Three replicates were made and one ml from each soil suspension were added to Trichoderma selected medium plates [TSM; rose Bengal 0.15 g/L, chloramphenicol 0.25 g/L, MgSO4.7H2O 0.2 g/L, KCl 0.15 g/L, K2HPO4 0.9 g/L, NH4NO3 1 g/L, glucose 3 g/L, agar 20 g/L with 1000 ml distilled water (dH2O), pH-6.5] 16 and incubated at 25 ± 1°C for 72 hours in the dark. The cultured petri plates were examined daily and Trichoderma sp transferred to another plate containing Rose Bengal Potato Dextrose media. Trichoderma sp Phenotypic characters. The morphological characteristics of five isolates of Trichoderma were examined in four distinct media: OMA, CMD, PDA, and TSM. 17 Mycelial discs (6 mm diameter) from actively growing cultures of each isolate were inoculated at the edges of the Petri plates containing the respective media and incubated at 28 ± 2°C for a week. The radius of the colony was assessed at 24, 48, and 72 hours. The daily growth of the isolates was recorded three times in triplicate and the observations of each isolate were summed up and recorded. Pigments, conidia colour, odor, and colony morphology of the isolate were also recorded. DNA Extraction Purified cultures have been grown in the potato dextrose broth for a duration of three days. A mycelial mat has been collected on filter paper, rinsed by distilled water 3–4 times, frozen, and then utilized for DNA extraction. DNA was extracted following the methodology given by. 18 The DNA was then suspended in 50 µl of TE buffer and estimated using ethidium bromide fluorescence. Molecular identification and Phylogenetic analysis To amplify the fragment of rDNA including ITS1 and ITS2 and 5.8S rDNA gene, were done by PCR by using combinations of primers SR6R and LR1 in a total volume of 50µl (White et al 1990).The PCR protocol consisted of the following steps: an initial denaturation at 95°C; followed by 30 cycles of denaturation at 95°C for 30 seconds, annealing at 52°C for 30 seconds, and a last extension at 72°C for 10 minute. 19 The amplified product of each gene was resolved using agarose gel of 1% and visible bands were excised and subsequently purified for the sequencing of analysis. Sequencing of purified product was carried out by Nelgen.Pvt.Ltd. Phylogenetic analysis Purified ITS sequence were analysed in comparison to the accessible ITS sequence through BLAST search in the NCBI GenBank records ( http://www.ncbi.nlm.nih.gov ). Alignment of multiple sequence was done by Clustal X (1.1). By the process of evolutionary distance has been calculated. 20 Phylogenetic tree was generated by bootstrap method of 1000 data set by using MEGA 12 (Molecular Evolutionary Genetic analysis). Results and Discussion A total of 03 Trichoderma sp were isolated from the tea gardens of Dooars and hilly region of West Bengal, India and their morphological and molecular characters were studied. Based on the observation of the conidia, phialides, colony texture, chlamydospore and conidiophore morphology. Table 1 Trichoderma sp isolated from the rhizosphere soil of tea gardens. Isolate Site Species Isolated NCBI Gene Bank Accession Number SUBH1 Subhasini T. E Trichoderma yunnanense Soil PV186710 MK1 Makaibari T. E Trichoderma asperellum Soil PV186709 S38 Diana T. E Trichoderma hamatum Soil PV186386 Table 2 Cultural and morphological characters (on PDA) of Trichoderma spp isolated from different tea gardens of North Bengal. Characters SUBH MK1 S38 Conidia (µm) 4.5-5.0×3.0–4.0 3.5-5.0× 2.7-3.0 3.0-4.5×2.0–3.0 Conidial shape Obovoid Oval to ellipsoidal Oblong-ellipsoidal Phialides in whorls 3–4 2–4 2–5 Phialides (µm) 6.0–10.0×4.0-5.5 5.5-6.0 × 2.0-4.5 6.0-7.5×2.5-4.0 Chlamydospores Abundant Abundant Abundant Pigments Green Green Yellowish green Green conidia + + + Colony appearance and Odor Green - dark green Dark green Yellowish -light green Colony radius (mm) A. 62–64 55–60 55 − 54 B. 55–65 48–54 55–58 C. 14–20 20–21 22–25 D. 80–82 65–72 68–70 A = PDA; B = CMA; C = TSM; D = OMA Phylogenetic analysis By analysing the ITS1 and ITS4 of our 03 isolated sequence phylogenetic tree were generated and the sequence of 11 Trichoderma sp were taken from NCBI Gene Bank showed in Fig. 1 . The ITS sequence was chosen for this analysis because it provides additional details about different regions the genus Trichoderma sp . 21 , 22 Our initial BLAST searches identified that sequences from these three new taxa were most closely related Hamatum Clade. 3 Conclusion To our knowledge only Trichoderma hamatum and Trichoderma asperellum have been reported from India. This is the first time Trichoderma yunnanense was reported from West Bengal and also from the rhizosphere soil of tea garden. More research has to be done to examine its potential as a plant growth stimulant for various plant species while also utilizing it for organic farming and its ability to biocontrol soil-borne diseases. Additionally, more research and surveys need to be done to ascertain the diversity of Trichoderma sp in new geographic locations. Declarations Funding: This work is supported by SERB-DST, Government of India, through project file nos. CRG/2022/007430. Conflict of interest: The authors declare there is no conflict of interest. Ethical Approval: Not applicable. Author Contribution SM: Writing—original draft, conceptualization, review & editing, investigation, validation, and visualization. SS: review & editing, VC: Sample collection & editing, NM: Visualization & collection, MS: Visualization & collection, SD: Supervision, conceptualization, validation, review & editing, and CG: Supervision, conceptualization, validation, correspondence, review & editing. Acknowledgement We sincerely acknowledge Department of Science and Technology, Government of India and Department of Tea Science, University of North Bengal, West Bengal, India for supporting with essential facilities for the work. SM is also thankful to Science and Engineering Research Board, Department of Science and Technology, Government of India, for providing fellowship. Data Availability All the DNA sequence generated during this study have been deposited to NCBI Gene Bank (https://www.ncbi.nlm.nih.gov) and Accession numbers of the same are given at Table 1. References Vinale, F. et al. Trichoderma–plant–pathogen interactions. Soil Biol. Biochem. 40 , 1–10 (2007). S, K. Mycofungicides and fungal biofertilizers. CiNii Research at (2009). https://cir.nii.ac.jp/crid/1572261549925129216 Druzhinina, I. S. et al. Trichoderma: the genomics of opportunistic success. Nat. Rev. Microbiol. 9 , 749–759 (2011). Sood, M. et al. Trichoderma: The Secrets of a multitalented biocontrol agent. Plants 9 , 762 (2020). Harman, G. E., Howell, C. R., Viterbo, A., Chet, I. & Lorito, M. Trichoderma species — opportunistic, avirulent plant symbionts. Nat. Rev. Microbiol. 2 , 43–56 (2004). Samuels, G. J. & Trichoderma Systematics, the Sexual State, and Ecology. Phytopathology 96 , 195–206 (2006). Lee, J. et al. The antagonistic properties of Trichoderma spp. inhabiting woods for potential biological control of wood-damaging fungi. Holzforschung 66 , 883–887 (2012). In CRC Press eBooks 17–48 (2002). 10.1201/9781482295320-8 Samuels, G. J. Trichoderma: a review of biology and systematics of the genus. Mycol. Res. 100 , 923–935 (1996). Bissett, J. A revision of the genus Trichoderma. II. Infrageneric classification. Can. J. Bot. 69 , 2357–2372 (1991). Bissett, J. A revision of the genus Trichoderma. III. Section Pachybasium. Can. J. Bot. 69 , 2373–2417 (1991). Asad, S. A. Mechanisms of action and biocontrol potential of Trichoderma against fungal plant diseases - A review. Ecol. Complex. 49 , 100978 (2021). Tripathi, P. et al. Trichoderma: a potential bioremediator for environmental clean up. Clean Technol. Environ. Policy . 15 , 541–550 (2013). Kumar, V. & Dwivedi, S. K. Hexavalent chromium stress response, reduction capability and bioremediation potential of Trichoderma sp. isolated from electroplating wastewater. Ecotoxicol. Environ. Saf. 185 , 109734 (2019). Askew, D. J. & Laing, M. D. An adapted selective medium for the quantitative isolation of Trichoderma species. Plant. Pathol. 42 , 686–690 (1993). Elad, Y., Chet, I. & Henis, Y. A selective medium for improving quantitative isolation ofTrichoderma spp. from soil. Phytoparasitica 9 , 59–67 (1981). Samuels, G. J., Dodd, S. L., Gams, W. & Castlebury, L. A. Petrini, O. Trichodermaspecies associated with the green mold epidemic of commercially grownAgaricus bisporus. Mycologia 94 , 146–170 (2002). Raeder, U. & Broda, P. Rapid preparation of DNA from filamentous fungi. Lett. Appl. Microbiol. 1 , 17–20 (1985). Fontana, P. D. et al. Isolation and genetic characterization of Acidovorax avenae from red stripe infected sugarcane in Northwestern Argentina. Eur. J. Plant Pathol. 137 , 525–534 (2013). Jukes, T. H. & Cantor, C. R. in Elsevier eBooks 21–132 (1969). 10.1016/b978-1-4832-3211-9.50009-7 Kuhls, K. et al. Molecular evidence that the asexual industrial fungus Trichoderma reesei is a clonal derivative of the ascomycete Hypocrea jecorina. Proceedings of the National Academy of Sciences 93, 7755–7760 (1996). Ospina-Giraldo, M. D., Royse, D. J., Thon, M. R., Chen, X. & Romaine, C. P. Phylogenetic relationships of Trichoderma harzianum causing mushroom green mold in Europe and North America to other species of Trichoderma from world-wide sources. Mycologia 90 , 76–81 (1998). Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6903051","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":487586430,"identity":"d6bf16e4-2fe0-4fc5-a27c-051d533706fd","order_by":0,"name":"Suvojeet Mukherjee","email":"","orcid":"","institution":"North Bengal University","correspondingAuthor":false,"prefix":"","firstName":"Suvojeet","middleName":"","lastName":"Mukherjee","suffix":""},{"id":487586431,"identity":"36aa7bab-f95b-4054-8d34-a9ff322368eb","order_by":1,"name":"Saini Sultana","email":"","orcid":"","institution":"North Bengal University","correspondingAuthor":false,"prefix":"","firstName":"Saini","middleName":"","lastName":"Sultana","suffix":""},{"id":487586432,"identity":"0eb3f5a0-b946-4eb9-ad54-bbe9ab475bd7","order_by":2,"name":"Vivek Chettri","email":"","orcid":"","institution":"North Bengal University","correspondingAuthor":false,"prefix":"","firstName":"Vivek","middleName":"","lastName":"Chettri","suffix":""},{"id":487586433,"identity":"5c553280-1908-4ef2-afd0-2d17bb64a988","order_by":3,"name":"Nahin Millat","email":"","orcid":"","institution":"North Bengal University","correspondingAuthor":false,"prefix":"","firstName":"Nahin","middleName":"","lastName":"Millat","suffix":""},{"id":487586434,"identity":"af5ee961-e82f-4459-8e83-2989dbc94ff0","order_by":4,"name":"Megha Saha","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Megha","middleName":"","lastName":"Saha","suffix":""},{"id":487586435,"identity":"64882c13-a042-44a1-b919-d4e996f505a9","order_by":5,"name":"Sukumar Debnath","email":"","orcid":"","institution":"North Bengal University","correspondingAuthor":false,"prefix":"","firstName":"Sukumar","middleName":"","lastName":"Debnath","suffix":""},{"id":487586436,"identity":"9ba70f4e-fcf2-4847-b6b9-afd570d976cc","order_by":6,"name":"Chandra Ghosh","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABAUlEQVRIiWNgGAWjYDACCSDmAWIDBgY2hoQKGxADJk6Mlgdn0kjUwviw7TBMC27AP7v52YO3e+zstrO3P3uQ2HY+z1wigfHDDwaLPJyW3DlmbjjnWXLyzp4z5gYJ524XW85IYJbsYZAoxqXFQCLBTJrnAHOywY0cNomEstuJG24kMEgDzUpswKkl/RtQSz1QS/oziQS2cyAtzL/xa8kB2XLYzuBGgplEQtsBkBY2vLZI3Mgpk5xz4HiCwZkzQC1nkhN39jxss+wxwK2Ff0b6Nok3B6rtDY63P5P8UWGXuJ09+fCNHxV1OLXAALICxgYGwrHDwGBPWMkoGAWjYBSMWAAAx69bSl6eLRYAAAAASUVORK5CYII=","orcid":"","institution":"North Bengal University","correspondingAuthor":true,"prefix":"","firstName":"Chandra","middleName":"","lastName":"Ghosh","suffix":""}],"badges":[],"createdAt":"2025-06-16 08:08:19","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6903051/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6903051/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":87184897,"identity":"be738c69-1c99-4c22-b980-ab0423a9aecf","added_by":"auto","created_at":"2025-07-21 10:19:07","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":299836,"visible":true,"origin":"","legend":"\u003cp\u003eShowing the study area.\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-6903051/v1/f3156a796cadc0724001e726.jpeg"},{"id":87184530,"identity":"abfa3ef6-f287-43cd-86a4-c1b82fbb2c4c","added_by":"auto","created_at":"2025-07-21 10:11:07","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":51353,"visible":true,"origin":"","legend":"\u003cp\u003eA:\u003cem\u003e Trichoderma hamatum;B: Trichoderma yunnanense\u003c/em\u003e; C:\u003cem\u003e Trichoderma asperellum\u003c/em\u003e;\u003c/p\u003e\n\u003cp\u003eD-F Conidia\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6903051/v1/0b9136e1a3e430bd1d302486.jpg"},{"id":87184898,"identity":"3be177d2-ec55-4780-8862-b38497483baf","added_by":"auto","created_at":"2025-07-21 10:19:07","extension":"jpeg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":288338,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic tree based on its ITS region of genomic rRNA gene of 03 isolates and 14 representative strains of \u003cem\u003eTrichoderma\u003c/em\u003e \u003cem\u003esp.\u003c/em\u003e\u003c/p\u003e","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-6903051/v1/7ce03c65a5a44e98b5ca84d9.jpeg"},{"id":91897450,"identity":"c4093e1b-9366-4a62-a4d3-1fa705842fb1","added_by":"auto","created_at":"2025-09-22 18:46:31","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1085983,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6903051/v1/e1a45cb9-dcdb-4d20-931e-bc4a7a42f822.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"New report of Trichoderma sp from Sub-Himalayan tea growing regions of West Bengal India","fulltext":[{"header":"Introduction","content":"\u003cp\u003eTrichoderma spp. (Hypocreales, Pezizomycotina, and Ascomycota) is now recognized as a widely utilized biocontrol agent and biological fertilizer in agricultural industry. Their remarkable capacity to fight against plant pathogenic fungi results in the production of secondary metabolites that have bio-pesticide properties and bio-stimulant effects.\u003csup\u003e\u003cspan additionalcitationids=\"CR2 CR3\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e Trichoderma sp are cosmopolitan saprophytic in nature are naturally occurring, and are commonly found in various soils, decaying wood, organic matter, and as endophytes of plants.\u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e.\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e,\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\u003cp\u003eSpecies within the genus Trichoderma are recognized because of their exceptional production of an assortment of secondary metabolites, which include polysaccharides, toxins, and antibiotics.\u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e,\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e,\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e Moreover, various strains in this genus are extensively utilized for the biocontrol of soilborne plant pathogenic fungi.\u003csup\u003e\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\u003cp\u003e\u003cem\u003eTrichoderma sp\u003c/em\u003e used as both direct biological control techniques that control the targeted pathogens like mycoparasitism along with indirect approaches that enhance plant health, including the induction of systemic resistance, promotion of plant growth, and improvement of rhizosphere competence. Direct biocontrol strategies encompass competition for nutrients and space, the production of antimicrobial agents (antibiosis), and the formation of lytic enzymes.\u003csup\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e Trichoderma has the ability to alter the biology of the rhizosphere, thereby minimizing the impact of pathogens.\u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e Furthermore, certain species of Trichoderma show efficacy as bio-remediators for heavy metals.\u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e,\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\u003cp\u003eThe present study was carried out to distinguish species or strains of Trichoderma by cultural and morphological characters and further characterize the diversity of Trichoderma spp. isolated from rhizosphere soil of tea based on molecular characters.\u003c/p\u003e"},{"header":"Material and Methods","content":"\u003cp\u003eStudy Area\u003c/p\u003e\n\u003cp\u003eSoil sampling and Isolation\u003c/p\u003e\n\u003cp\u003eSoil samples were collected from rhizosphere habitats of tea gardens of the North Bengal region, India, for isolating \u003cem\u003eTrichoderma sp\u003c/em\u003e. The isolation of \u003cem\u003eTrichoderma\u003c/em\u003e was carried out using a serial dilution method,\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e where each sample was diluted to 10\u003csup\u003e\u0026minus;\u0026thinsp;2\u003c/sup\u003e. Three replicates were made and one ml from each soil suspension were added to Trichoderma selected medium plates [TSM; rose Bengal 0.15 g/L, chloramphenicol 0.25 g/L, MgSO4.7H2O 0.2 g/L, KCl 0.15 g/L, K2HPO4 0.9 g/L, NH4NO3 1 g/L, glucose 3 g/L, agar 20 g/L with 1000 ml distilled water (dH2O), pH-6.5]\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e and incubated at 25\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C for 72 hours in the dark. The cultured petri plates were examined daily and Trichoderma sp transferred to another plate containing Rose Bengal Potato Dextrose media.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eTrichoderma sp\u003c/em\u003e Phenotypic characters.\u003c/p\u003e\n\u003cp\u003eThe morphological characteristics of five isolates of Trichoderma were examined in four distinct media: OMA, CMD, PDA, and TSM.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e Mycelial discs (6 mm diameter) from actively growing cultures of each isolate were inoculated at the edges of the Petri plates containing the respective media and incubated at 28\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C for a week. The radius of the colony was assessed at 24, 48, and 72 hours. The daily growth of the isolates was recorded three times in triplicate and the observations of each isolate were summed up and recorded. Pigments, conidia colour, odor, and colony morphology of the isolate were also recorded.\u003c/p\u003e\n\u003cp\u003eDNA Extraction\u003c/p\u003e\n\u003cp\u003ePurified cultures have been grown in the potato dextrose broth for a duration of three days. A mycelial mat has been collected on filter paper, rinsed by distilled water 3\u0026ndash;4 times, frozen, and then utilized for DNA extraction. DNA was extracted following the methodology given by.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003eThe DNA was then suspended in 50 \u0026micro;l of TE buffer and estimated using ethidium bromide fluorescence.\u003c/p\u003e\n\u003cp\u003eMolecular identification and Phylogenetic analysis\u003c/p\u003e\n\u003cp\u003eTo amplify the fragment of rDNA including ITS1 and ITS2 and 5.8S rDNA gene, were done by PCR by using combinations of primers SR6R and LR1 in a total volume of 50\u0026micro;l (White et al 1990).The PCR protocol consisted of the following steps: an initial denaturation at 95\u0026deg;C; followed by 30 cycles of denaturation at 95\u0026deg;C for 30 seconds, annealing at 52\u0026deg;C for 30 seconds, and a last extension at 72\u0026deg;C for 10 minute.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e The amplified product of each gene was resolved using agarose gel of 1% and visible bands were excised and subsequently purified for the sequencing of analysis. Sequencing of purified product was carried out by Nelgen.Pvt.Ltd.\u003c/p\u003e\n\u003cp\u003ePhylogenetic analysis\u003c/p\u003e\n\u003cp\u003ePurified ITS sequence were analysed in comparison to the accessible ITS sequence through BLAST search in the NCBI GenBank records (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.ncbi.nlm.nih.gov\u003c/span\u003e\u003c/span\u003e). Alignment of multiple sequence was done by Clustal X (1.1). By the process of evolutionary distance has been calculated.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e Phylogenetic tree was generated by bootstrap method of 1000 data set by using MEGA 12 (Molecular Evolutionary Genetic analysis).\u003c/p\u003e"},{"header":"Results and Discussion","content":"\u003cp\u003eA total of 03 Trichoderma sp were isolated from the tea gardens of Dooars and hilly region of West Bengal, India and their morphological and molecular characters were studied. Based on the observation of the conidia, phialides, colony texture, chlamydospore and conidiophore morphology.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n \u003ctable id=\"Tab1\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003e\u003cem\u003eTrichoderma sp\u003c/em\u003e isolated from the rhizosphere soil of tea gardens.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIsolate\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSite\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSpecies\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIsolated\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eNCBI Gene Bank Accession Number\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSUBH1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSubhasini T. E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eTrichoderma yunnanense\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSoil\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePV186710\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMK1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMakaibari T. E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eTrichoderma asperellum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSoil\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePV186709\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eS38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eDiana T. E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eTrichoderma hamatum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSoil\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePV186386\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cdiv class=\"gridtable\"\u003e\n \u003cdiv align=\"left\" class=\"colspec\"\u003e\u003cbr\u003e\u003c/div\u003e\n \u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eCultural and morphological characters (on PDA) of \u003cem\u003eTrichoderma spp\u003c/em\u003e isolated from different tea gardens of North Bengal.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eCharacters\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSUBH\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMK1\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eS38\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eConidia (\u0026micro;m)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.5-5.0\u0026times;3.0\u0026ndash;4.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.5-5.0\u0026times; 2.7-3.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.0-4.5\u0026times;2.0\u0026ndash;3.0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eConidial shape\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eObovoid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOval to ellipsoidal\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOblong-ellipsoidal\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePhialides in whorls\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u0026ndash;4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u0026ndash;4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u0026ndash;5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePhialides (\u0026micro;m)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.0\u0026ndash;10.0\u0026times;4.0-5.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.5-6.0 \u0026times; 2.0-4.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.0-7.5\u0026times;2.5-4.0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eChlamydospores\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAbundant\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAbundant\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAbundant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePigments\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGreen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGreen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eYellowish green\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGreen conidia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eColony appearance and Odor\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGreen - dark green\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eDark green\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eYellowish -light green\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003eColony radius (mm)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e62\u0026ndash;64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e55\u0026ndash;60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e55\u0026thinsp;\u0026minus;\u0026thinsp;54\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eB.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e55\u0026ndash;65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e48\u0026ndash;54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e55\u0026ndash;58\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eC.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14\u0026ndash;20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u0026ndash;21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e22\u0026ndash;25\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eD.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e80\u0026ndash;82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e65\u0026ndash;72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e68\u0026ndash;70\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;A = PDA; B = CMA; C = TSM; D = OMA\u003c/p\u003e\n\u003cp\u003ePhylogenetic analysis\u003c/p\u003e\n\u003cp\u003eBy analysing the ITS1 and ITS4 of our 03 isolated sequence phylogenetic tree were generated and the sequence of 11 \u003cem\u003eTrichoderma sp\u003c/em\u003e were taken from NCBI Gene Bank showed in Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e. The ITS sequence was chosen for this analysis because it provides additional details about different regions the genus \u003cem\u003eTrichoderma sp\u003c/em\u003e.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e21\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e Our initial BLAST searches identified that sequences from these three new taxa were most closely related Hamatum Clade.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eTo our knowledge only \u003cem\u003eTrichoderma hamatum\u003c/em\u003e and \u003cem\u003eTrichoderma asperellum\u003c/em\u003e have been reported from India. This is the first time \u003cem\u003eTrichoderma yunnanense\u003c/em\u003e was reported from West Bengal and also from the rhizosphere soil of tea garden. More research has to be done to examine its potential as a plant growth stimulant for various plant species while also utilizing it for organic farming and its ability to biocontrol soil-borne diseases. Additionally, more research and surveys need to be done to ascertain the diversity of \u003cem\u003eTrichoderma\u003c/em\u003e sp in new geographic locations.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eFunding:\u003c/h2\u003e\u003cp\u003eThis work is supported by SERB-DST, Government of India, through project file nos. CRG/2022/007430.\u003c/p\u003e\u003cp\u003eConflict of interest: The authors declare there is no conflict of interest.\u003c/p\u003e\u003cp\u003eEthical Approval: Not applicable.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eSM: Writing\u0026mdash;original draft, conceptualization, review \u0026amp; editing, investigation, validation, and visualization. SS: review \u0026amp; editing, VC: Sample collection \u0026amp; editing, NM: Visualization \u0026amp; collection, MS: Visualization \u0026amp; collection, SD: Supervision, conceptualization, validation, review \u0026amp; editing, and CG: Supervision, conceptualization, validation, correspondence, review \u0026amp; editing.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eWe sincerely acknowledge Department of Science and Technology, Government of India and Department of Tea Science, University of North Bengal, West Bengal, India for supporting with essential facilities for the work. SM is also thankful to Science and Engineering Research Board, Department of Science and Technology, Government of India, for providing fellowship.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eAll the DNA sequence generated during this study have been deposited to NCBI Gene Bank (https://www.ncbi.nlm.nih.gov) and Accession numbers of the same are given at Table 1.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eVinale, F. et al. Trichoderma\u0026ndash;plant\u0026ndash;pathogen interactions. \u003cem\u003eSoil Biol. Biochem.\u003c/em\u003e \u003cb\u003e40\u003c/b\u003e, 1\u0026ndash;10 (2007).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eS, K. 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Phylogenetic relationships of Trichoderma harzianum causing mushroom green mold in Europe and North America to other species of Trichoderma from world-wide sources. \u003cem\u003eMycologia\u003c/em\u003e \u003cb\u003e90\u003c/b\u003e, 76\u0026ndash;81 (1998).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Morphology, New species. Phylogenetic analysis, Trichoderma yunnanense","lastPublishedDoi":"10.21203/rs.3.rs-6903051/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6903051/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cem\u003eTrichoderma\u003c/em\u003e species (Hypocreaceae) is one of the world\u0026rsquo;s most common widely used plant growth-promoting fungus and, ecologically and economically important genus. Three newly discovered species of the fungal genera \u003cem\u003eTrichoderma\u003c/em\u003e, \u003cem\u003eTrichoderma asperellum Trichoderma hamatum\u003c/em\u003e and \u003cem\u003eTrichoderma yunnanense\u003c/em\u003e, isolated from the rhizosphere zone of tea of North Bengal region, India have been identified through morphology and phylogenetic study of sequences of nucleotide. The analysed DNA sequences encompassed the internal transcribed spacer (ITS) regions of the rDNA cluster, specifically ITS1 and ITS2.The analyses show that \u003cem\u003eT. asperellum\u003c/em\u003e ,\u003cem\u003eT. yunnaense\u003c/em\u003e, \u003cem\u003eT hamatum\u003c/em\u003e are grouped within the Hamatum clade.\u003c/p\u003e","manuscriptTitle":"New report of Trichoderma sp from Sub-Himalayan tea growing regions of West Bengal India","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-21 10:11:03","doi":"10.21203/rs.3.rs-6903051/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"5a548069-09c1-4805-9e39-3307e7971b3a","owner":[],"postedDate":"July 21st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":51769474,"name":"Biological sciences/Microbiology/Fungi"},{"id":51769475,"name":"Biological sciences/Microbiology/Fungi/Fungal ecology"}],"tags":[],"updatedAt":"2025-09-22T18:38:25+00:00","versionOfRecord":[],"versionCreatedAt":"2025-07-21 10:11:03","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6903051","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6903051","identity":"rs-6903051","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}