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Effective biofertilizer Trichoderma spp. isolates with enzymatic activity and metabolites enhancing plant growth

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Trichoderma species have been widely recognized as biofertilizer fungi for their ability to produce phytohormones and enhance plant growth. In our current study, fifteen strains of Trichoderma spp. (T1–T15) were screened for their capacity to produce phytohormones and metabolites eliciting plant growth. The stains were previously isolated from olive rhizosphere soil in northern Algeria. Plant growth promoting (PGP) potential of Trichoderma spp. was evaluated in vitro through the production of phosphatases, siderophores, hydrogen cyanide (HCN), and ammonia (NH3). Besides, plant growth phytohormones such as gibberellic acid and indole-3-acetic acid (IAA) were assessed quantitatively by a colorimetric assay. Results showed an effective potential of Trichoderma spp. in plant growth-promoting biomolecule production. Importantly, qualitative estimation of phosphate solubilization indicates that T10 gave the highest phosphate solubilization on medium Pikovskaya’s with a solubilization index (SI) of 3, whereas, the high capacity nitrogen-fixing was related to T8. On the other hand, quantitative analysis of indole-3-acetic acid and gibberellic acid revealed a production varying between (1.30 μg mL−1 to 21.15 μg mL−1) and (0.53 μg mL−1 to 7.87 μg mL−1), respectively; the highest amount of both phytohormones was obtained by T11 isolate. Indeed, an analysis of ethyl acetate extracts of T11 isolate by high-performance liquid chromatography (HPLC) revealed a high amount (71.19 mg L−1) of IAA. Overall, the results showed clearly that isolate T11 has promising plant growth-promoting properties. Hence, this native Trichoderma isolate (T11) identified as Trichoderma harzianum strain (OL587563) could be used later as biofertilizer for sustainable olive crop agriculture.

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  • Abe CAL, Faria CB, De Castro FF, De Souza SR, Santos FCD, Da Silva CN, Barbosa-Tessmann IP (2015) Fungi isolated from maize (Zea mays L) grains and production of associated enzyme activities. Int J Mole Sci 16(7):15328–15346.

    Article  CAS  Google Scholar 

  • Ahemad M, Kibret M (2014) Mechanisms and Applications of Plant Growth Promoting Rhizobacteria: Current Perspective. J King Saud Univ Sci 26(1):1–20.

    Article  Google Scholar 

  • Akintokun AK, Akande GA, Akintokun PO, Popoola TOS, Babalola AO (2007) Solubilization on insoluble phosphate by organic acid-producing fungi isolated from Nigerian soil. Int J Soil Sci 2(4):301–307.

    Article  CAS  Google Scholar 

  • Anke H, Kinn J, Bergquist KE, Sterner O (1991) Production of siderophores by strains of the genus Trichoderma. Biol Met 4(3):176–180.

    Article  CAS  Google Scholar 

  • Bach E, dos Santos Seger GD, de Carvalho FG, Lisboa BB, Passaglia LMP (2016) Evaluation of biological control and rhizosphere competence of plant growth promoting bacteria. Appl Soil Ecol 99:141–149.

    Article  Google Scholar 

  • Bakker AW, Schippers B (1987) Microbial cyanide production in the rhizosphere in relation to potato yield reduction and Pseudomonas spp.-mediated plant growth-stimulation. Soil Biol Biochem 19:451–457.

    Article  CAS  Google Scholar 

  • Bric JM, Bostock RM, Silverstone SE (1991) Rapid in situ assay for indoleacetic acid production by bacteria immobilized on a nitrocellulose membrane. Appl Environ Microbiol, 57(2):535–538.0099–2240/91/020535–04$02.00/0

  • Brimecombe MJ, De Leij FA, Lynch JM (2001) The effect of root exudates on rhizosphere microbial populations. In: Pinton R, Varanini Z, Nanipieri P (eds) The rhizosphere. Marcel Dekker Inc, New York, pp 95–140

    Google Scholar 

  • Chanclud E, Morel JB (2016) Plant hormones: a fungal point of view. Mol Plant Pathol 17(8):1289–1297.

    Article  PubMed  PubMed Central  Google Scholar 

  • Cherkupally R, Amballa H, Reddy BN (2017) In vitro screening for enzymatic activity of Trichoderma species for biocontrol potential. Ann Plant Sci 6(11):1784–1789.

    Article  Google Scholar 

  • Chung KR, Shilts T, Ertürk Ü, Timmer LW, Ueng PP (2003) Indole derivatives produced by the fungus Colletotrichum acutatum causing lime anthracnose and postbloom fruit drop of citrus. FEMS Microbiol Lett 226(1):23–30.

    Article  CAS  PubMed  Google Scholar 

  • Dutta S, Kundu A, Chakraborty MR, Ojha S, Chakrabarti J, Chatterejee NC (2006) Production and optimization of Fe (III) specific ligand, the siderophore of soil inhabiting and wood rotting fungi as deterrent to plant pathogens. Acta Phytopathol Entomol Hung 41:237–248.

    Article  CAS  Google Scholar 

  • Ezzat AS, Ghoneem KM, Saber WIA, Alaskar AA (2015) Control of wilt, stalk and tuber rots diseases using Arbuscular mycorrhizal fungi, Trichoderma species and hydroquinone enhances yield quality and storability of Jerusalem artichoke (Helianthus tuberosus L.). Egypt J Biol Pest Control 25(1):11–22

    Google Scholar 

  • Fasim F, Ahmed N, Parson R, Gadd GM (2002) Solubilization of zinc salts by a bacterium isolated from air environment of a tannery. FEMS Microbiol Lett 213:1–6.

    Article  CAS  PubMed  Google Scholar 

  • Fayziev V, Jovlieva D, Juraeva U, Shavkiev J, Eshboev F (2020) Effects of PVXN-UZ 915 necrotic isolate of potato virus X on amount of pigments of Datura stramonium leaves. J Crit Rev 7(9):400–403.

    Article  Google Scholar 

  • Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791

    Article  Google Scholar 

  • Filiz, O, Takil E, Kayan N (2021) The role of plant growth promoting rhizobacteria (Pgpr) and phosphorus fertilization in improving phenology and physiology of bean (phaseolus vulgaris l.). Appl Ecol Environ Res 19(3):

  • Gardes M, Bruns TD (1993) ITS primers with enhanced specificity for basidiomycetes application to the identification of mycorrhizae and rusts. Mol Ecol 2(2):113–118.

    Article  CAS  PubMed  Google Scholar 

  • Ghosh SK, Banerjee S, Sengupta C (2017) Bioassay, characterization and estimation of siderophores from some important antagonistic fungi. J Biopest 10(2):105–112

    CAS  Google Scholar 

  • Gueye N, Dienaba Sall SY, Tahir A (2018) Strains isolated from agriculture field soil in Senegal. Key Words: Trichoderma, Enzymes.

    Article  Google Scholar 

  • Gupta R, Singal R, Shankar A, Kuhad RC, Saxena RK (1994) A modified plate assay for screening phosphate solubilizing microorganisms. J Gen Appl Microbiol 40(3):255–260

    Article  CAS  Google Scholar 

  • Hamayun M, Sumera A, Ilyas I, Bashir A, In-Jung L (2010) Isolation of a Gibberellin producing fungus (Penicillium sp. MH7) and growth promotion of crown daisy (Chrysanthemum coronarium). J Microbiol Biotechnol 20(1):202–207.

    Article  CAS  PubMed  Google Scholar 

  • Hankin L, Anagnostakis SL (1975) The use of solid media for detection of enzyme production by fungi. Mycologia 67(3):597–607.

    Article  Google Scholar 

  • Hassan MM, Daffalla HM, Modwi HI, Osman MG, Ahmed II, Gani MEA, Babiker AGE (2013) Effects of fungal strains on seeds germination of millet and Striga hermonthica. Univ J Agri Res 2(2):83–88.

    Article  Google Scholar 

  • Hemerly A (2016) Genetic controls of biomass increase in sugarcane by association with beneficial nitrogen-fixing bacteria. Plant and Animal Genome Conference XXIV. Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.

  • Holbrook AA, Edge WLW, Bailey F (1961) Spectrophotometric method for determination of gibberellic acid in gibberellins. ACS Washington, D.C. 159–167.

  • Hoyos-Carvajal L, Orduz S, Bissett J (2009) Growth stimulation in bean (Phaseolus vulgaris L) by Trichoderma. Biol Control 51(3):409–416.

    Article  Google Scholar 

  • Ismail I, Hamayun M, Sayyed A, Din IU, Gul H, Hussain A (2016) Gibberellin and indole acetic acid production capacity of endophytic fungi isolated from Zea mays L. Int J Biosci 8(3):35–43

    Article  CAS  Google Scholar 

  • Jaroszuk-Ściseł J, Kurek E, Trytek M (2014) Efficiency of indoleacetic acid, gibberellic acid and ethylene synthesized in vitro by Fusarium culmorum strains with different effects on cereal growth. Biologia 69(3):281–292.

    Article  CAS  Google Scholar 

  • Jaroszuk-Ściseł J, Tyśkiewicz R, Nowak A, Ozimek E, Majewska M, Hanaka A, Janusz G (2019) Phytohormones (auxin, gibberellin) and ACC deaminase in vitro synthesized by the mycoparasitic Trichoderma DEMTkZ3A0 strain and changes in the level of auxin and plant resistance markers in wheat seedlings inoculated with this strain conidia. Int J Mol Sci 20(19):4923

    Article  Google Scholar 

  • Johnson LF, Curl EA (1972) Methods for research on the ecology of soil-borne plant pathogens. Methods Res Ecol Soil-borne Plant Pathog

  • Jukes TH, Cantor CR (1969) Evolution of protein molecules. In: Munro HN (ed) Mammalian protein metabolism. Academic Press, New York, pp 21–132

    Chapter  Google Scholar 

  • Karadeniz A, Topcuoğlu ŞF, İnan S (2006) Auxin, gibberellin, cytokinin and abscisic acid production in some bacteria. World J Microbiol Biotechnol 22(10):1061–1064.

    Article  CAS  Google Scholar 

  • Kawaide H (2006) Biochemical and molecular analyses of gibberellin biosynthesis in fungi. Biosci Biotechnol Biochem 70(3):583–590

    Article  CAS  Google Scholar 

  • Lalngaihawmi BA (2019) Study on the different modes of action of potential Trichodermaspp. from banana rhizosphere against Fusarium oxysporum f. sp. cubense. Int J Curr Microbiol App Sci 8(01):1028–1040

    Article  CAS  Google Scholar 

  • Lanzuise S, Manganiello G, Lorito M (2014) Trichoderma secondary metabolites active on plants and fungal pathogens. Open Mycol J 8(Suppl-1, M5):127–139

    Google Scholar 

  • Li Z, Wang T, Luo X, Li X, Xia C, Zhao Y, Fan J (2018) Biocontrol potential of Myxococcus sp. strain BS against bacterial soft rot of calla lily caused by Pectobacterium carotovorum. Biol Control 126:36–44.

    Article  Google Scholar 

  • López AC, Alvarenga AE, Zapata PD, Luna MF, Villalba LL (2019) Trichoderma spp. from Misiones, Argentina: effective fungi to promote plant growth of the regional crop Ilex paraguariensis St. Hil Mycology 10(4):210–221

    Article  Google Scholar 

  • MacMillan J (2002) Erratum: occurrence of gibberellins in vascular plants, fungi, and bacteria. J Plant Growth Regul 21(3):242–243.

    Article  CAS  Google Scholar 

  • Meera T, Balabaskar P (2012) Isolation and characterization of Pseudomonas fluorescens from rice fields. Int J Food, Agri Veterinary Sci 2(1):113–120

    Google Scholar 

  • Mohiddin FA, Bashir I, Padder SA, Hamid B (2017) Evaluation of different substrates for mass multiplication of Trichoderma species. J Pharmacognosyand Phytochem 6(6):563–569

    CAS  Google Scholar 

  • Mohite B (2013) Isolation and characterization of indole acetic acid (IAA) producing bacteria from rhizospheric soil and its effect on plant growth. J Soil Sci Plant Nutr 13(3):638–649.

    Article  Google Scholar 

  • Nabi NG, Asghar M, Shah AH, Sheikh MA, Asad MJ (2003) Production of pectinase by Trichoderma harzianum in solid state fermentation of citrus peels. Pak J Agric Sci 40:193–201

    Google Scholar 

  • Napitupulu, TP, Kanti A, Sudiana IM (2019) Evaluation of the environmental factors modulating indole-3-acetic acid (IAA) production by Trichoderma harzianum InaCC F88. In IOP conference series: earth and environmental science (Vol. 308, No. 1, p. 012060). IOP Publishing.

  • Ng LC, Ngadin A, Azhari M, Zahari NA (2015) Potential of Trichoderma spp as biological control agents against bakanae pathogen (Fusarium fujikuroi) in rice. Asian J Plant Pathol 9(2):46–58

    Article  Google Scholar 

  • Noori MS, Saud HM (2012) Potential plant growth-promoting activity of Pseudomonas sp. isolated from paddy soil in Malaysia as biocontrol agent. J Plant Pathol Microbiol 3(2):1–4

    Google Scholar 

  • Pallardy S (2008) Plant hormones and other signaling molecules. Physiol Woody Plants :367–377

  • Premono ME, Moawad AM, Vlek PLG (1996) Effect of phosphate-solubilizing Pseudomonas putida on the growth of maize and its survival in the rhizosphere (No. REP-12113. CIMMYT.)

  • Reghmit A, Benzina-tihar F, López Escudero FJ, Halouane-Sahir F, Oukali Z, Bensmail S, Ghozali N (2021) Trichoderma spp. isolates from the rhizosphere of healthy olive trees in northern Algeria and their biocontrol potentials against the olive wilt pathogen, Verticillium dahliae. Org Agric 1–19.

  • Renuka N, Guldhe A, Prasanna R, Singh P, Bux F (2018) Microalgae as multi-functional options in modern agriculture: current trends, prospects and challenges. Biotechnol Adv 36(4):1255–1273.

    Article  CAS  PubMed  Google Scholar 

  • Rey M, Delgado-Jarana J, Rincón AM, Limón MDC, Benítez T (2000) Mejora de cepas de Trichoderma para su empleo como biofungicidas. Rev Iberoam Micol 17:31–36

    Google Scholar 

  • Rifai MA (1969) A revision of the genus Trichoderma. Mycol Pap 116:1–56

    Google Scholar 

  • Rijavec T, Lapanje A (2016) Hydrogen cyanide in the rhizosphere: not suppressing plant pathogens, but rather regulating availability of phosphate. Front Microbiol 7:1785

    Article  Google Scholar 

  • Saber WIA, Abd El-Hai KM, Ghoneem KM (2009) Synergistic effect of Trichoderma and Rhizobium on both biocontrol of chocolate spot disease and induction of nodulation, physiological activities and productivity of Vicia faba. Res J Microbiol 4(8):286–300

    Article  Google Scholar 

  • Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425

    CAS  PubMed  Google Scholar 

  • Samuels G, Dodd SL, Gams W, Castlebury LA, Petrini O (2002) Trichoderma species associated with the green mold epidemic of commercially grown Agaricus bisporus. Mycologia 94:146–170.

    Article  PubMed  Google Scholar 

  • Saravanakumar K, Arasu VS, Kathiresan K (2013) Effect of Trichoderma on soil phosphate solubilization and growth improvement of Avicennia marina. Aquat Bot 104:101–105.

    Article  CAS  Google Scholar 

  • Sharma S, Sharma A, Kaur M (2018) Extraction and evaluation of gibberellic acid from Pseudomonas sp: plant growth promoting rhizobacteria. J Pharmacognosy Phytochem 7(1):2790–2795

    CAS  Google Scholar 

  • Thakkar A, Saraf M (2015) Role of volatile metabolites from T citrinoviride in biocontrol of phytopathogens. Int J Res Chem Environ (IJRCE) 5(1):86–95

    Google Scholar 

  • Tsavkelova EA, Klimova SY, Cherdyntseva TA, Netrusov AI (2006) Microbial producers of plant growth stimulators and their practical use: a review. Appl Biochem Microbiol 42(2):117–126.

    Article  CAS  Google Scholar 

  • Ushamailini C, Nakkeeran S, Marimuthu T (2008) Development of biomanure for the management of turmeric rhizome rot. Arch Phytopathol Plant Protection 41(5):365–376.

    Article  Google Scholar 

  • Uthandi S, Karthikeyan S, Sabarinathan KG (2010) gibberellic acid production by Fusarium fujikuroi SG2. J Scient Ind Res 69:211–214

    CAS  Google Scholar 

  • Vinale F, Nigro M, Sivasithamparam K, Flematti G, Ghisalberti EL, Ruocco M, Varlese R, Marra R, Lanzuise S, Eid A, Woo SL, Lorito M (2013) Harzianic acid: a novel siderophore from Trichoderma harzianum. FEMS Microbiol Lett 347:123–129.

    Article  CAS  PubMed  Google Scholar 

  • Vinale F, Sivasithamparam K, Ghisalberti EL, Woo SL, Nigro M, Marra R, Lorito M (2014) Trichoderma secondary metabolites active on plants and fungal pathogens. The Open Mycology Journal 8(1)

  • Whipps JM (2001) Microbial interactions and biocontrol in the rhizosphere. J Exp Botany 52(suppl_1):487–511.

    Article  CAS  Google Scholar 

  • Zhang F, Chen C, Zhang F, Gao L, Liu G, Chen L, Fan X, Liu C, Zhang K, He Y, Chen C, Ji X (2017) Trichoderma harzianum containing 1-aminocyclopropane-1-carboxylate deaminase and chitinase improved growth and diminished adverse effect caused by Fusarium oxysporum in soybean. J Plant Physiol 210:84–94.

    Article  CAS  PubMed  Google Scholar 

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Partial financial support for this work was received from DGRST (General Delegation for Scientific and Technical Research) and socio economic project which are gratefully acknowledged.


This work was supported by DGRST (General Delegation for Scientific and Technical Research in Algeria) (grant number is not available).

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All the authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by Reghmit Abdenaceur, Benzina-tihar Farida, Djeziri Mourad, Hadjouti Rima, Oukali Zahia, and Sahir-Halouane Fatma. The first draft of the manuscript was written by Reghmit Abdenaceur and all the authors commented on the previous versions of the manuscript. All the authors read and approved the final manuscript.

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Correspondence to Reghmit Abdenaceur.

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This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the Ethics Committee of University of M’hamed Bougara, Department of Biology, Boumerdes 35000, Algeria (Date 10/02/2021/No number is not available).

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Abdenaceur, R., Farida, Bt., Mourad, D. et al. Effective biofertilizer Trichoderma spp. isolates with enzymatic activity and metabolites enhancing plant growth. Int Microbiol 25, 817–829 (2022).

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