Ability of endophytic fungi associated with Withania somnifera L. to control Fusarium Crown and Root Rot and to promote growth in tomato

  • Ahlem NefziEmail author
  • Rania Aydi Ben Abdallah
  • Hayfa Jabnoun-Khiareddine
  • Nawaim Ammar
  • Mejda Daami-Remadi
Environmental Microbiology - Research Paper


Fusarium crown and root rot (FCRR), caused by Fusarium oxysporum f. sp. radicis-lycopersici (FORL), is a soilborne tomato disease of increased importance worldwide. In this study, Withania somnifera was used as a potential source of biological control and growth-promoting agents. Seven fungal isolates naturally associated with W. somnifera were able to colonize tomato seedlings. They were applied as conidial suspensions or a cell-free culture filtrate. All isolates enhanced treated tomato growth parameters by 21.5–90.3% over FORL-free control and by 27.6–93.5% over pathogen-inoculated control. All tested isolates significantly decreased by 28.5–86.4% disease severity over FORL-inoculated control. The highest disease suppression, by 86.4–92.8% over control and by 81.3–88.8% over hymexazol-treated control, was achieved by the I6 isolate. FORL radial growth was suppressed by 58.5–82.3% versus control when dual cultured with tested isolates and by 61.8–83.2% using their cell-free culture filtrates. The most active agent was identified as Fusarium sp. I6 (MG835371), which displayed chitinolytic, proteolytic, and amylase activities. This has been the first report on the potential use of fungi naturally associated with W. somnifera for FCRR suppression and for tomato growth promotion. Further investigations are required in regard to mechanisms of action involved in disease suppression and plant growth promotion.


Antifungal activity Associated fungi Fusarium oxysporum f. sp. radicis-lycopersici W. somnifera Tomato growth 


Funding information

This work was funded by the Ministry of Higher Education and Scientific Research of Tunisia through the funding allocated to the research unit UR13AGR09-Integrated Horticultural Production in the Tunisian Centre-East and by IRESA through the funding attributed to the multidisciplinary and multi-institutional project CleProD.

Compliance with ethical standards

All the experiments undertaken in this study comply with the current law of the country where they were performed.

Conflict of interest

The authors declare that they have no conflicts of interest.


  1. 1.
    Zhang L, Khabbaz SE, Wang A, Li H, Abbasi PA (2015) Detection and characterization of broad-spectrum antipathogen activity of novel rhizobacterial isolates and suppression of Fusarium crown and root rot disease of tomato. J Appl Microbiol 118:685–703CrossRefGoogle Scholar
  2. 2.
    Lin F, Liu N, Lai D, Kang XH, Pang NW, Jiang HK, Xu HH (2017) A formulation of neem cake seeded with Bacillus sp. provides control over tomato Fusarium crown and root rot. Biocontrol Sci Tech 27:393–407CrossRefGoogle Scholar
  3. 3.
    Hibar K, Daami-Remadi M, El Mahjoub M (2007) Induction of resistance in tomato plants against Fusarium oxysporum f. sp. radicis-lycopersici by Trichoderma spp. Tunis J Plant Prot 2:47–58Google Scholar
  4. 4.
    Rowe RC, Farley JD (1977) New greenhouse tomato disease can be controlled. Ohio Dis Reporter 62:41–43Google Scholar
  5. 5.
    Ozbay N, Newman SE, Bashan CW, Brown WM (2004) Biological control of Fusarium crown and root rot of tomato with Trichoderma harzianum. Pak J Biol Sci 7:478–484CrossRefGoogle Scholar
  6. 6.
    Ajilogba CF, Babalola OO (2013) Integrated management strategies for tomato Fusarium wilt. Biocontrol Sci 18:117–127CrossRefGoogle Scholar
  7. 7.
    Madbouly AK, Abdelbacki AM (2017) Biocontrol of certain soilborne disease and promotion of growth of capsicum annum using biofungicides. Pak J Bot 49:371–378Google Scholar
  8. 8.
    Zeng Q, Wu S, Sukumaran J, Rodrigo A (2017) Models of microbiome evolution incorporating host and microbial selection. Microbiome. 5:127–136CrossRefGoogle Scholar
  9. 9.
    Rodriguez RJ, White JJF, Arnold AE, Redman ARA (2009) Fungal enodophytes: diversity and functional roles. New Phytol 182:314–330CrossRefGoogle Scholar
  10. 10.
    Kusari P, Kusari S, Spiteller M, Kayser O (2013) Endophytic fungi harbored in Cannabis sativa L.: diversity and potential as biocontrol agents against host plant-specific phytopathogens. Fungal Divers 60:137–151CrossRefGoogle Scholar
  11. 11.
    Mei C, Flinn BS (2010) The use of beneficial microbial endophytes for plant biomass and stress tolerance improvement. Recent Pat Biotechnol 4:81–95CrossRefGoogle Scholar
  12. 12.
    Le Cocq K, Gurr SJ, Hirsch PR, Mauchline TH (2017) Exploitation of endophytes for sustainable agricultural intensification. Mol Plant Pathol 18:469–473CrossRefGoogle Scholar
  13. 13.
    Patel JK, Archana G (2017) Diverse culturable diazotrophic endophytic bacteria from Poaceae plants show cross-colonization and plant growth promotion in wheat. Plant Soil 417:99–116CrossRefGoogle Scholar
  14. 14.
    Garne Brahim B, Ouhdouch Y (2017) Management of tomato foot and root rot (TFRR) by biocontrol agents with emphasis on factors affecting its effectiveness. In: Kumar V, Kumar M, Sharma S, Prasad R (eds) Probiotics and plant health. Springer, Singapore, pp 1–9Google Scholar
  15. 15.
    Kavroulakis N, Ntougias S, Zervakis GI, Ehaliotis C, Haralampidis K, Papadopoulou KK (2007) Role of ethylene in the production of tomato plants against soil-borne fungal pathogens conferred by an endophytic Fusarium solani strain. J Exp Bot 58:3853–3864CrossRefGoogle Scholar
  16. 16.
    Horinouchi H, Muslim A, Suzuki T, Hyakumachi M (2007) Fusarium equiseti GF191 as an effective biocontrol agent against Fusarium crown and root rot of tomato in rock wool systems. Crop Prot 26:1514–1523CrossRefGoogle Scholar
  17. 17.
    Fakhro A, Andrade-Linares DR, Von Bargen S, Bandte M, Büttner C, Grosch R, Schwarz D, Franken P (2010) Impact of Piriformospora indica on tomato growth and on interaction with fungal and viral pathogens. Mycorrhiza. 20:191–200CrossRefGoogle Scholar
  18. 18.
    Qin Y, Pan X, Kubicek C, Druzhinina I, Chenthamara K, Labbé JL, Yuan Z (2017) Diverse plant-associated pleosporalean fungi from saline areas: ecological tolerance and nitrogen-status dependent effects on plant growth. Front Microbiol 8:158–169Google Scholar
  19. 19.
    Nisa H, Kamili AN, Nawchoo IA, Shafi S, Shameem N, Bandh SA (2015) Fungal endophytes as prolific source of phytochemicals and other bioactive natural products: a review. Microb Pathog 82:50–59CrossRefGoogle Scholar
  20. 20.
    Bilal L, Asaf S, Hamayun M, Gul H, Iqbal A, Ullah I, Hussain A (2018) Plant growth promoting endophytic fungi Asprgillus fumigatus TS1 and Fusarium proliferatum BRL1 produce gibberellins and regulates plant endogenous hormones. Symbiosis. 2:1–11Google Scholar
  21. 21.
    Mishra A, Singh SP, Mahfooz S, Singh SP, Bhattacharya A, Mishra N, Nautiyal CS (2018) Endophyte-mediated modulation of defense-related genes and systemic resistance in Withania somnifera (L.) Dunal under Alternaria alternata stress. J Appl Environ Microbiol 84:2817–2845CrossRefGoogle Scholar
  22. 22.
    Aswani P, Tijith K, George Jisha MS (2018) Characterization of bioactive metabolites of endophytic Fusarium solani isolated from Withania somnifera. J Biol Active Products Nat 7:411–426Google Scholar
  23. 23.
    Singh AK, Rathod V, Dattu Singh JM, Kulkarni P (2016) Effect of silver nanoparticles (AgNps) produced by an endophytic fungus Fusarium Semitectum isolated from a medicinal plant Withania Somnifera (Ashwagandha) on seed germination. Int J Res Stud Agric Sci 2:6–12Google Scholar
  24. 24.
    Bensaci OA, Harzallah D, Gouaref K (2015) Endophytic mycoflora of Cedrus atlantica: diversity patterns and determinism of the phytosanitary situation of Atlas cedar forests in Belezma massif (Algeria). For Sci Technol 11:36–43Google Scholar
  25. 25.
    Luna MF, Aprea J, Crespo JM, Boiardi JL (2012) Colonization and yield promotion of tomato by Gluconacetobacter diazotrophicus. Appl Soil Ecol 61:225–229CrossRefGoogle Scholar
  26. 26.
    Cao L, Qiu Z, You J, Tan H, Zhou S (2004) Isolation and characterization of endophytic Streptomyces strains from surface-sterilized tomato (Lycopersicon esculentum) roots. J Appl Microbiol 39:425–430CrossRefGoogle Scholar
  27. 27.
    Szűcs Z, Plaszkó T, Cziáky Z, Kiss-Szikszai A, Emri T, Bertóti R, Gonda S (2018) Endophytic fungi from the roots of horseradish (Armoracia rusticana) and their interactions with the defensive metabolites of the glucosinolate-myrosinase-isothiocyanate system. Plant Biol 18:85–96Google Scholar
  28. 28.
    Potshangbam M, Devi SI, Sahoo D, Strobel GA (2017) Functional characterization of endophytic fungal community associated with Oryza sativa L. and Zea mays L. Front Microbiol 8:325–336CrossRefGoogle Scholar
  29. 29.
    Nath A, Chattopadhyay A, Joshi SR (2015) Biological activity of endophytic fungi of Rauwolfia serpentina Benth: an ethnomedicinal plant used in folk medicines in Northeast India. Biol Sci 85:233–240Google Scholar
  30. 30.
    Yuan H, Lei Z, Rondon SI, Gao Y (2017) Potential of a strain of Beauveria bassiana (Hypocreales: Cordycipitaceae) for the control of the potato tuberworm, Phthorimaea operculella (Zeller). Int J Pest Manag 63:352–354CrossRefGoogle Scholar
  31. 31.
    Hallmann J, Berg G, Schulz B (2006) Isolation procedures for endophytic microorganisms. In: Schulz BJE, Boyle CJC, Sieber TN (eds) Microbial root endophytes. Springer, Berlin, pp 299–319CrossRefGoogle Scholar
  32. 32.
    Kumaresan V, Suryanarayanan TS, Johnson JA (1998) Foliar fungal endophytes from two species of the mangrove Rhizophora. Can J Microbiol 44:1003–1006CrossRefGoogle Scholar
  33. 33.
    Vakalounakis DJ, Fragkiadakis GA (1999) Genetic diversity of Fusarium oxysporum isolates for cucumber: differentiation by pathogenicity, vegetative compatibility and RAPD fingerprinting. Phytopathol. 89:161–168CrossRefGoogle Scholar
  34. 34.
    Campanile G, Ruscelli A, Luisi N (2007) Antagonistic activity of endophytic fungi towards Diplodia corticola assessed by in vitro and in planta tests. Eur J Plant Pathol 117:237–246CrossRefGoogle Scholar
  35. 35.
    Kumar A, Shukla R, Singh P, Prasad CS, Dubey NK (2008) Assessment of Thymus vulgaris L. essential oil as a safe botanical preservative against post harvest fungal infestation of food commodities. Innov Food Sci Emerg Technol 9:575–580CrossRefGoogle Scholar
  36. 36.
    Xiao Y, Li HX, Li C, Wang JX, Li J, Wang MH, Ye YH (2013) Antifungal screening of endophytic fungi from Ginkgo biloba for discovery of potent anti-phytopathogenic fungicides. Fed Eur Microbiol Soc 339:130–136CrossRefGoogle Scholar
  37. 37.
    Schoch CL, Seifert KA, Huhndorf S, Robert V, Spouge JL, Levesque CA, Miller AN (2012) Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi. Proc Natl Acad Sci 109:6241–6246CrossRefGoogle Scholar
  38. 38.
    Saleem A, Ebrahim MK (2014) Production of amylase by fungi isolated from legume seeds collected in Almadinah Almunawwarah, Saudi Arabia. J Taibah Univ Sci 8:90–97CrossRefGoogle Scholar
  39. 39.
    Kotogán A, Németh B, Vágvölgyi C, Papp T, Takó M (2014) Screening for extracellular lipase enzymes with transesterification capacity in Mucoromycotina strains. Food Technol Biotechnol 52:73–82Google Scholar
  40. 40.
    Choudhary V, Jain PC (2012) Screening of alkaline protease production by fungal isolates from different habitats of Sagar and Jabalpur district (MP). J Acad Ind Res 1:215–220Google Scholar
  41. 41.
    Okay S, Tefon BE, Ozkan M, Ozcengiz G (2008) Expression of chitinase A (chiA) gene from a local isolate of Serratia marcescens in Coleoptera-specific Bacillus thuringiensis. J Appl Microbiol 104:16–170Google Scholar
  42. 42.
    Bogner CW, Kariuki GM, Elashry A, Sichtermann G, Buch AK, Schouten A (2016) Fungal root endophytes of tomato from Kenya and their nematode biocontrol potential. Mycol Prog 15:30–39CrossRefGoogle Scholar
  43. 43.
    Kumar DSS, Hyde KD (2004) Biodiversity and tissue-recurrence of endophytic fungi in Tripterygium wilfordii. Fungal Divers 17:69–90Google Scholar
  44. 44.
    Sun X, Guo LD, Hyde KD (2011) Community composition of endophytic fungi in Acer truncatum and their role in decomposition. Fungal Divers 47:85–95CrossRefGoogle Scholar
  45. 45.
    Gazis R, Chaverri P (2010) Diversity of fungal endophytes in leaves and stems of wild rubber trees (Hevea brasiliensis) in Peru. Fungal Ecol 3:240–254CrossRefGoogle Scholar
  46. 46.
    Khan R, Shahzad S, Choudhary MI, Khan SA, Ahmad A (2010) Communities of endophytic fungi in medicinal plant Withania somnifera. Pak J Bot 42:1281–1287Google Scholar
  47. 47.
    Qadri M, Johri S, Shah BA, Khajuria A, Sidiq T, Lattoo SK, Riyaz-Ul-Hassan S (2013) Identification and bioactive potential of endophytic fungi isolated from selected plants of the Western Himalayas. SpringerPlus. 2:8–12CrossRefGoogle Scholar
  48. 48.
    Kuldau GA, Yates IE (2000) Evidence for Fusarium endophytes in cultivated and wild plants. Microbial Endophytes 5:85–117Google Scholar
  49. 49.
    Ratnaweera PB, De Silva ED, Williams DE, Andersen RJ (2015) Antimicrobial activities of endophytic fungi obtained from the arid zone invasive plant Opuntia dillenii and the isolation of equisetin, from endophytic Fusarium sp. BMC Complement Altern Med 15:220–229CrossRefGoogle Scholar
  50. 50.
    Vinodhini D, Agastian P (2013) Berberine production by endophytic fungus Fusarium solani from Coscinium fenestratum. Int J Biol Pharm Res 4:1239–1245Google Scholar
  51. 51.
    Wang XJ, Min CL, Ge M, Zuo RH (2014) An endophytic sanguinarine-producing fungus from Macleaya cordata, Fusarium proliferatum BLH51. Curr Microbiol 68:336–341CrossRefGoogle Scholar
  52. 52.
    Khan AR, Ullah I, Waqas M, Shahzad R, Hong SJ, Park GS, Shin JH (2015) Plant growth-promoting potential of endophytic fungi isolated from Solanum nigrum leaves. World J Microbiol Biotechnol 31:1461–1466CrossRefGoogle Scholar
  53. 53.
    Patle PN, Navnage NP, Ramteke PR (2018) Endophytes in plant system: roles in growth promotion, mechanism and their potentiality in achieving agriculture sustainability. Int J Chem Stud 6:270–274Google Scholar
  54. 54.
    Saldajeno MGB, Hyakumachi M (2011) The plant growth-promoting fungus Fusarium equiseti and the arbuscular mycorrhizal fungus Glomus mosseae stimulate plant growth and reduce severity of anthracnose and damping-off diseases in cucumber (Cucumis sativus) seedlings. Ann Appl Biol 159:28–40CrossRefGoogle Scholar
  55. 55.
    Yong YH, Dai CC, Gao FK, Yang QY, Zhao M (2009) Effects of endophytic fungi on growth and two kinds of terpenoids for Euphorbia pekinensis. Chin Tradit Herbal Drugs 40:1136–1139Google Scholar
  56. 56.
    Elsharkawy MM, Shimizu M, Takahashi H, Hyakumachi M (2012) The plant growth-promoting fungus Fusarium equiseti and the arbuscular mycorrhizal fungus Glomus mosseae induce systemic resistance against Cucumber mosaic virus in cucumber plants. Plant Soil 361:397–409CrossRefGoogle Scholar
  57. 57.
    Šišić A, Baćanović J, Finckh MR (2017) Endophytic Fusarium equiseti stimulates plant growth and reduces root rot disease of pea (Pisum sativum L.) caused by Fusarium avenaceum and Peyronellaea pinodella. Eur J Plant Pathol 148:271–282CrossRefGoogle Scholar
  58. 58.
    Patil NN, Waghmode MS, Gaikwad PS, Gajbhiye MH, Gunjal AB, Nawani NN, Kapadnis BP (2014) Potential of Microbispora sp. V2 as biocontrol agent against Sclerotium rolfsii, the causative agent of southern blight of Zea mays L. (Baby corn)–in vitro studies. Indian J Exp Biol 52:1147–1151Google Scholar
  59. 59.
    Yang VW, Clausen CA (2005) Determining the suitability of Lactobacilli antifungal metabolites for inhibiting mould growth. World J Microbiol Biotechnol 21:977–981CrossRefGoogle Scholar
  60. 60.
    Li XJ, Zhang Q, Zhang AL, Gao JM (2012) Metabolites from Aspergillus fumigatus, an endophytic fungus associated with Melia azedarach, and their antifungal, antifeedant, and toxic activities. J Agric Food Chem 60:3424–3431CrossRefGoogle Scholar
  61. 61.
    Tayung K, Barik BP, Jha DK (2010) Antifungal activity and biocontrol potential of metabolite produced by an endophytic Fusarium (MTCC-9622) against some postharvest pathogens. J Agric Technol 6:409–419Google Scholar
  62. 62.
    Shen WY, Bai R, Wang AR, He JY, Wang H, Zhang Y, Dong JY (2016) Two new polyhydroxysterols produced by Fusarium solani, an endophytic fungus from Chloranthus multistachys. Nat Prod Res 30:2173–2182CrossRefGoogle Scholar

Copyright information

© Sociedade Brasileira de Microbiologia 2019

Authors and Affiliations

  • Ahlem Nefzi
    • 1
    Email author
  • Rania Aydi Ben Abdallah
    • 2
  • Hayfa Jabnoun-Khiareddine
    • 2
  • Nawaim Ammar
    • 2
  • Mejda Daami-Remadi
    • 2
  1. 1.Faculty of Sciences of BizerteZarzounaTunisia
  2. 2.UR13AGR09-Integrated Horticultural Production in the Tunisian Centre-East, Regional Center of Research on Horticulture and Organic AgricultureUniversity of SousseChott-MariemTunisia

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