Journal of Plant Pathology

, Volume 101, Issue 4, pp 965–980 | Cite as

Induction of systemic resistance in turmeric by rhizospheric isolate Trichoderma asperellum against rhizome rot disease

  • Govindegowda Vinayarani
  • Kallahally Nagendra Madhusudhan
  • Harishchandra Sripathi PrakashEmail author
Original Article


The rhizospheric fungal isolates were screened for growth promotion and induced systemic resistance against rhizome rot disease in turmeric caused by Pythium aphanidermatum (Edson) Fitzp. Thirty isolates from the turmeric rhizosphere were identified by morphological characteristics and using internal transcribed spacer sequence homology. In in vitro antagonistic activity, out of 30 isolates tested only five isolates showed >70% suppression of P. aphanidermatum. The rhizospheric isolates viz., T. viride PGPFDOB-V6, Chaetomium sp. PGPFDOB-V13, T. asperellum PGPFDOB-V11, T. harzianum PGPFDOB-V22 and T. asperellum PGPFDOB-V36 also showed multiple plant growth promoting traits under in vitro studies. In greenhouse studies, rhizome treatment followed by soil application of PGPFDOB-V36 significantly increased the plant height to 87.60 cm and fresh rhizome yield/plant to 430 g. This isolate reduced the percent disease incidence (PDI) of rhizome rot to 14.2% when compared to the control 78.70%. The rhizome colonization of PGPFDOB-V36 was observed by using the confocal microscope. The selected PGPF isolates were tested for their ability to induce production of defense-related enzymes in plants. Temporal expression pattern of defense-related enzymes such as peroxidase (PO), phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO) and PR-protein β-1,3 glucanase in turmeric plants pretreated with PGPF isolates followed by challenge inoculation with the pathogen was studied. The defense enzymes were increased by two to three folds compared to uninoculated control. The accumulation of phenolics was higher in plants pre-treated with the PGPF isolates. The study revealed that the rhizosphere isolate PGPFDOB-V36 has promising plant growth promoting and rhizome rot suppression ability in turmeric.


Antagonism Biocontrol Defense enzymes Pythium aphanidermatum Plant growth promotion 



This work was carried out with the financial assistance from the Department of Science and Technology (DST), Government of India, New Dehli, under the Women Scientist Scheme (DST-WOS A) awarded to Mrs. Vinaya Rani. G (DST sanction No.SR/WOS-A/LS-104/2013 (G) dated 22.04.2014. We also thank the Institution of Excellence (IOE) at the University of Mysore for providing instrumentation facility.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interests regarding the publication of this paper.


  1. Ahmed AS, Ezziyyani M, Sanchez CP, Candela ME (2003) Effect of chitin on biological control activity of Bacillus spp. and Trichoderma harzianum against root rot disease in pepper (Capsicum annuum) plants. Eur J Plant Pathol 109(6):633–637Google Scholar
  2. Benhamou N, Garand C, Goulet A (2002) Ability of nonpathogenic fusarium oxysporum strain Fo47 to induce resistance against Pythium ultimum infection in cucumber. Appl Environ Microbiol 68(8):4044–4060PubMedPubMedCentralGoogle Scholar
  3. Benítez T, Rincon AM, Limon MC, Codon AC (2004) Biocontrol mechanisms of Trichoderma strains. Int Microbiol 7(4):249–260PubMedGoogle Scholar
  4. Boominathan U, Sivakumaar PK (2012) Induction of systemic resistance by mixtures of rhizobacterial isolates against Pythium aphanidermatum. Res J Biotechnol 7(4):192–197Google Scholar
  5. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72(1–2):248–254PubMedPubMedCentralGoogle Scholar
  6. Chen C, Belanger RR, Benhamou N, Paulitz TC (2000) Defense enzymes induced in cucumber roots by treatment with plant growth-promoting rhizobacteria (PGPR) and Pythium aphanidermatum. Physiol Mol Plant Pathol 56(1):13–23Google Scholar
  7. Chet I (1987) Trichoderma, application, mode of action, and potential as biocontrol agent of soilborne plant pathogenic fungi. Innovative approaches to plant disease control, pp 137–160Google Scholar
  8. Chet I, Chernin L (2003) Biocontrol, microbial agents in soil. Encyclopedia of Environmental Microbiology. Willey, New York, pp 450–465Google Scholar
  9. Datnoff LE, Nemec S, Pernezny K (1995) Biological control of fusarium crown and root rot of tomato in Florida using Trichoderma harzianum and Glomus intraradices. Biol Control 5(3):427–431Google Scholar
  10. De Meyer G, Bigirimana J, Elad Y, Höfte M (1998) Induced systemic resistance in Trichoderma harzianum T39 biocontrol of Botrytis cinerea. Eur J Plant Pathol 104(3):279–286Google Scholar
  11. Deshmukh S, Hückelhoven R, Schafer P, Imani J, Sharma M, Weiss M, Kogel KH (2006) The root endophytic fungus Piriformospora indica requires host cell death for proliferation during mutualistic symbiosis with barley. Proc Natl Acad Sci U S A 103(49):18450–18457PubMedPubMedCentralGoogle Scholar
  12. Dickerson DP, Pascholati SF, Hagerman AE, Butler LG, Nicholson RL (1984) Phenylalanine ammonia-lyase and hydroxycinnamate, CoA ligase in maize mesocotyls inoculated with Helminthosporium maydis or Helminthosporium carbonum. Physiol Plant Pathol 25(2):111–123Google Scholar
  13. Djonovic S, Pozo MJ, Dangott LJ, Howell CR, Kenerley CM (2006) Sm1, a proteinaceous elicitor secreted by the biocontrol fungus Trichoderma virens induces plant defense responses and systemic resistance. Mol Plant-Microbe Interact 19(8):838–853PubMedGoogle Scholar
  14. Gaigole AH, Wagh GN, Khadse AC (2011) Antifungal activity of Trichoderma species against soil borne pathogen. Asiatic J Biot Resour 4:461–465Google Scholar
  15. Gowtham HG, Murali M, Brijesh Singh S, Lakshmeesha TR, Murthy KN, Amruthesh KN, Niranjana SR (2018) Plant growth promoting rhizobacteria Bacillus amyloliquefaciens improves plant growth and induces resistance in chilli against anthracnose disease. Biol Control 126:209–217Google Scholar
  16. Gupta P, Samant K, Sahu A (2012) Isolation of cellulose-degrading bacteria and determination of their cellulolytic potential. Int J Microbiol:1–5Google Scholar
  17. Hammerschmidt R, Nuckles EM, Kuc J (1982) Association of enhanced peroxidase activity with induced systemic resistance of cucumber to Colletotrichum lagenarium. Physiol Plant Pathol 20(1):73–82Google Scholar
  18. Hariprasad P, Chandrashekar S, Singh SB, Niranjana SR (2014) Mechanisms of plant growth promotion and disease suppression by Pseudomonas aeruginosa strain 2apa. J Basic Microbiol 54(8):792–801PubMedGoogle Scholar
  19. Harman GE, Howell CR, Viterbo A, Chet I, Lorito M (2004) Trichoderma species opportunistic, avirulent plant symbionts. Nat Rev Microbiol 2(1):43–56Google Scholar
  20. Hjeljord LG, Tronsmo A (2003) Effect of germination initiation on competitive capacity of Trichoderma atroviride P1 conidia. Phytopathology 93(12):1593–1598PubMedGoogle Scholar
  21. Howell CR, Hanson LE, Stipanovic RD, Puckhaber LS (2000) Induction of terpenoid synthesis in cotton roots and control of Rhizoctonia solani by seed treatment with Trichoderma virens. Phytopathology 90(3):248–252PubMedGoogle Scholar
  22. Jeyaseelan EC, Tharmila S, Niranjan K (2012) Antagonistic activity of Trichoderma spp. and Bacillus spp. against Pythium aphanidermatum isolated from tomato damping off. Arch Appl Sci Res 4(4):1623–1627Google Scholar
  23. John RP, Tyagi RD, Prevost D, Brar SK, Pouleur S, Surampalli RY (2010) Mycoparasitic Trichoderma viride as a biocontrol agent against Fusarium oxysporum f. sp. adzuki and Pythium arrhenomanes and as a growth promoter of soybean. Crop Protect 29(12):1452–1459Google Scholar
  24. Kavitha K, Mathiyazhagan S, Senthilvel V, Nakkeeran S, Chandrasekar G (2005) Development of bioformulations of antagonistic bacteria for the management of damping off of Chilli (Capsicum annuum L.). Arch Phytopathol Plant Protect 38(1):19–30Google Scholar
  25. Kavitha K, Nakkeeran S, Chandrasekar G (2012) Rhizobacterial-mediated induction of defense enzymes to enhance the resistance of turmeric (Curcuma longa L) to Pythium aphanidermatum causing rhizome rot. Arch Phytopathol Plant Protect 45(2):199–219Google Scholar
  26. Koike N, Hyakumachi M, Kageyama K, Tsuyumu S, Doke N (2001) Induction of systemic resistance in cucumber against several diseases by plant growth-promoting fungi, lignification and superoxide generation. Eur J Plant Pathol 107(5):523–533Google Scholar
  27. Kovach J, Petzoldt R, Harman GE (2000) Use of honey bees and bumble bees to disseminate Trichoderma harzianum 1295-22 to strawberries for Botrytis control. Biol Control 18(3):235–242Google Scholar
  28. Kumakura K, Watanabe S, Toyoshima J, Makino T, Ichikawa T, Lyozumi H, Nagayama K (2003) Effect Trichoderma sp. SKT-1 on suppression of six different seedborne disease of rice (Oryzae sativa). Jpn J Phytopathol 69:384–392Google Scholar
  29. Le Floch G, Rey P, Benizri E, Benhamou N, Tirilly Y (2003) Impact of auxin-compounds produced by the antagonistic fungus Pythium oligandrum or the minor pathogen Pythium group F on plant growth. Plant Soil 257(2):459–470Google Scholar
  30. Lugtenberg B, Kamilova F (2009) Plant-growth-promoting rhizobacteria. Ann Rev Microbiol 63:541–556Google Scholar
  31. Mayer AM, Harel E, Ben-Shaul R (1966) Assay of catechol oxidase-a critical comparison of methods. Phytochemistry 5(4):783–789Google Scholar
  32. Meera MS, Shivanna MB, Kageyama K, Hyakumachi M (1995) Persistence of induced systemic resistance in cucumber in relation to root colonization by plant growth promoting fungal isolates. Crop Prot 14(2):123–130Google Scholar
  33. Mishra VK (2010) In vitro antagonism of Trichoderma species against Pythium aphanidermatum. J Phytol 2(9):28–35Google Scholar
  34. Murali M, Amruthesh KN (2015) Plant growth promoting fungus Penicillium oxalicum enhances plant growth and induces resistance in pearl millet against downy mildew disease. J Phytopathol 163(9):743–754Google Scholar
  35. Murali M, Sudisha J, Amruthesh KN, Ito SI, Shetty HS (2013) Rhizosphere fungus Penicillium chrysogenum promotes growth and induces defence-related genes and downy mildew disease resistance in pearl millet. Plant Biol 15(1):111–118PubMedGoogle Scholar
  36. Muthukumar A, Eswaran A, Sangeetha G (2011) Induction of systemic resistance by mixtures of fungal and endophytic bacterial isolates against Pythium aphanidermatum. Acta Physiol Plant 33:1933–1944Google Scholar
  37. Pan SQ, Ye XS, Kuc J (1991) A technique for detection of chitinase, beta-1, 3-glucanase, and protein patterns after a single separation using polyacrylamide gel electrophoresis or isoelectrofocusing. Phytopathology 81(9):970–974Google Scholar
  38. Pereira SIA, Castro PML (2014) Diversity and characterization of culturable bacterial endophytes from Zea mays and their potential as plant growth-promoting agents in metal-degraded soils. Environ Sci Pollut Res 21(24):14110–14123Google Scholar
  39. Pieterse CM, Zamioudis C, Berendsen RL, Weller DM, Van Wees SC, Bakker PA (2014) Induced systemic resistance by beneficial microbes. Annu Rev Phytopathol 52:347–375PubMedGoogle Scholar
  40. Pikovskaya RI (1948) Mobilization of phosphorus in soil in connection with vital activity of some microbial species. Mikrobiologiya 17:362–370Google Scholar
  41. Ravindran PN, Babu KN, Sivaraman K (2007) Turmeric: the genus Curcuma. CRC pressGoogle Scholar
  42. Rohini GHG, Hariprasad P, Singh SB, Niranjana SR (2016) Biological control of Phomopsis leaf blight of brinjal (Solanum melongena L.) with combining phylloplane and rhizosphere colonizing beneficial bacteria. Biol Control 101:123–129Google Scholar
  43. Rojo FG, Reynoso MM, Ferez M, Chulze SN, Torres AM (2007) Biological control by Trichoderma species of Fusarium solani causing peanut brown root rot under field conditions. Crop Prot 26(4):549–555Google Scholar
  44. Schwyn B, Neilands JB (1987) Universal chemical assay for the detection and determination of siderophores. Anal Biochem 160(1):47–56PubMedGoogle Scholar
  45. Shanmugam V, Thakur H, Kaur J, Gupta S, Rajkumar S, Dohroo NP (2013) Genetic diversity of Fusarium spp. inciting rhizome rot of ginger and its management by PGPR consortium in the western Himalayas. Biol Control 66(1):1–7Google Scholar
  46. Shoresh M, Harman GE, Mastouri F (2010) Induced systemic resistance and plant responses to fungal biocontrol agents. Annu Rev Phytopathol 48:21–43PubMedGoogle Scholar
  47. Singh H, Reddy MS (2011) Effect of inoculation with phosphate solubilizing fungus on growth and nutrient uptake of wheat and maize plants fertilized with rock phosphate in alkaline soils. Eur J Soil Biol 47(1):30–34Google Scholar
  48. Singh PP, Shin YC, Park CS, Chung YR (1999) Biological control of Fusarium wilt of cucumber by chitinolytic bacteria. Phytopathology 89:92–99PubMedGoogle Scholar
  49. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5, molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28(10):2731–2739PubMedPubMedCentralGoogle Scholar
  50. Thanavelu R, Sundararaju P, Sathiamoorthy S, Reghuchander T, Velazhahan R, Nakkeeran S, Palanisamy A (2001) Status of Fusarium wilt of banana in India. In International Workshop on the Banana Fusarium Wilt Disease, Genting Highlands Resort (Malaysia), 18-20 Oct 1999Google Scholar
  51. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W, improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22(22):4673–4680PubMedPubMedCentralGoogle Scholar
  52. Triveni S, Prasanna R, Shukla L, Saxena AK (2013) Evaluating the biochemical traits of novel Trichoderma-based biofilms for use as plant growth-promoting inoculants. Ann Microbiol 63(3):1147–1156Google Scholar
  53. Ushamalini C, Nakkeeran P, Marimuthu T (2008) Induction of plant defense enzymes in turmeric plants by Trichoderma viride. Arch Phytopathology Plant Prot 41(2):79–93Google Scholar
  54. Van Wees SC, Van der Ent S, Pieterse CM (2008) Plant immune responses triggered by beneficial microbes. Curr Opin Plant Biol 11(4):443–448PubMedGoogle Scholar
  55. Vinayarani G, Prakash HS (2018) Fungal endophytes of turmeric (Curcuma longa L.) and their biocontrol potential against pathogens Pythium aphanidermatum and Rhizoctonia solani. World J Microbiol Biotechnol 34(3):49PubMedGoogle Scholar
  56. Viterbo ADA, Chet I (2006) TasHyd1, a new hydrophobin gene from the biocontrol agent Trichoderma asperellum, is involved in plant root colonization. Mol Plant Pathol 7(4):249–258PubMedGoogle Scholar
  57. Whipps JM, Lumsden RD (1991) Biological control of Pythium species. Biocontrol Sci Tech 1(2):75–90Google Scholar
  58. White TJ, Bruns T, Lee SJWT, Taylor JL (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR protocols: a guide to methods and applications 18(1):315–322Google Scholar
  59. Windham MT (1986) A mechanism for increased plant growth induced by Trichoderma spp. Phytopathology 76:518–521Google Scholar
  60. Woo SL, Scala F, Ruocco M, Lorito M (2006) The molecular biology of the interactions between Trichoderma spp., phytopathogenic fungi, and plants. Phytopathology 96(2):181–185PubMedGoogle Scholar
  61. Yedidia L, Benhamou N, Chet I (1999a) Induction of defense responses in cucumber plants (Cucumis sativus L.) by the biocontrol agent Trichoderma harzianum. Appl Environ Microbiol 65(3):1061–1070PubMedPubMedCentralGoogle Scholar
  62. Yedidia L, Benhamou N, Chet I (1999b) Induction of defense responses in cucumber plants (Cucumis sativus L.) in the biocontrol agent Trichoderma harzianum. Appl Environ Microbiol 65:929–935Google Scholar
  63. Yedidia I, Shoresh M, Kerem Z, Benhamou N, Kapulnik Y, Chet I (2003) Concomitant induction of systemic resistance to Pseudomonas syringae pv. lachrymans in cucumber by Trichoderma asperellum (T-203) and accumulation of phytoalexins. Appl Environ Microbiol (12):7343–7353PubMedPubMedCentralGoogle Scholar
  64. Yoshioka Y, Ichikawa H, Naznin HA, Kogure A, Hyakumachi M (2012) Systemic resistance induced in Arabidopsis thaliana by Trichoderma asperellum SKT-1, a microbial pesticide of seed-borne diseases of rice. Pest Manag Sci 68(1):60–66PubMedGoogle Scholar
  65. Zieslin N, Ben-zaken R (1993) Peroxidase activity and presence of phenolic substances in peduncles of rose flowers. Plant Physiol Biochem 31(3):333–339Google Scholar

Copyright information

© Società Italiana di Patologia Vegetale (S.I.Pa.V.) 2019

Authors and Affiliations

  • Govindegowda Vinayarani
    • 1
  • Kallahally Nagendra Madhusudhan
    • 2
  • Harishchandra Sripathi Prakash
    • 1
    Email author
  1. 1.Department of Studies in BiotechnologyUniversity of MysoreMysuruIndia
  2. 2.Molecular Biology LaboratoryCentral Sericultural Research and Training InstituteMysuruIndia

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