Skip to main content
Log in

Fungal endophytes of turmeric (Curcuma longa L.) and their biocontrol potential against pathogens Pythium aphanidermatum and Rhizoctonia solani

  • Original Paper
  • Published:
World Journal of Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Endophytic fungi have been isolated from the healthy turmeric (Curcuma longa L.) rhizomes from South India. Thirty-one endophytes were identified based on morphological and ITS–rDNA sequence analysis. The isolated endophytes were screened for antagonistic activity against Pythium aphanidermatum (Edson) Fitzp., and Rhizoctonia solani Kuhn., causing rhizome rot and leaf blight diseases in turmeric respectively. Results revealed that only six endophytes showed > 70% suppression of test pathogens in antagonistic dual culture assays. The endophyte T. harzianum TharDOB-31 showed significant in vitro mycelial growth inhibition of P. aphanidermatum (76.0%) and R. solani (76.9%) when tested by dual culture method. The SEM studies of interaction zone showed morphological abnormalities like parasitism, shriveling, breakage and lysis of hyphae of the pathogens by endophyte TharDOB-31. Selected endophytic isolates recorded multiple plant growth promoting traits in in vitro studies. The rhizome bacterization followed by soil application of endophyte TharDOB-31 showed lowest Percent Disease Incidence of rhizome rot and leaf blight, 13.8 and 11.6% respectively. The treatment of TharDOB-31 exhibited significant increase in plant height (85 cm) and fresh rhizome yield/plant (425 g) in comparison with untreated control under greenhouse condition. The confocal microscopy validates the colonization of the TharDOB-31 in turmeric rhizomes. The secondary metabolites in ethyl acetate extract of TharDOB-31 were found to contain higher number of antifungal compounds by high resolution liquid chromatograph mass spectrometer analysis. Thereby, endophyte T. harzianum isolate can be exploited as a potential biocontrol agent for suppressing rhizome rot and leaf blight diseases in turmeric.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • AbdSharad A, Usup G, Sahrani FK, Ahmad A (2016) Antimicrobial activity and determination of bioactive components from marine Alcaligenes faecalis extract against a sulfate-reducing bacteria. In AIP Conference Proceedings, vol 1784, No 1. AIP Publishing, p 020010

  • Abubakar MN, Majinda RR (2016) GC-MS analysis and preliminary antimicrobial activity of Albizia adianthifolia (Schumach) and Pterocarpus angolensis (DC). Medicines 3(1):3

    Article  Google Scholar 

  • Barnett HL, Hunter BB (1972) Illustrated genera of imperfect fungi, 3rd edn. Burgess Publishing Co., Minneapolis

    Google Scholar 

  • Bell DK, Wells HD, Markham CR (1982) In vitro antagonism of Trichoderma species against six fungal plant pathogens. Phytopathology 72(4):379–382

    Article  Google Scholar 

  • Benhamou N, Chet I (1996) Parasitism of sclerotia of Sclerotium rolfsii by Trichoderma harzianum: ultrastructural and cytochemical aspects of the interaction. Phytopathology 86(4):405–416

    Article  Google Scholar 

  • Borneman J, Hartin RJ (2000) PCR primers that amplify fungal rRNA genes from environmental samples. Appl Environ Microbiol 66(10):4356–4360

    Article  CAS  Google Scholar 

  • Buatong J, Phongpaichit S, Rukachaisirikul V, Sakayaroj J (2011) Antimicrobial activity of crude extracts from mangrove fungal endophytes. World J Microbiol Biotechnol 27(12):3005–3008

    Article  CAS  Google Scholar 

  • Chet I, Harman GE, Baker R (1981) Trichoderma hamatum, Its hyphal interactions with Rhizoctonia solani and Pythium sp. Microb Ecol 7:29–38

    Article  CAS  Google Scholar 

  • Cho JY, Moon JH, Seong KY, Park KH (1998) Antimicrobial activity of 4-hydroxybenzoic acid and trans 4-hydroxycinnamic acid isolated and identified from rice hull. Biosci Biotechnol Biochem 62(11):2273–2276

    Article  CAS  Google Scholar 

  • Choi JS, Park NH, Hwang SY, Sohn JH, Kwak I, Cho KK, Choi IS (2013) The antibacterial activity of various saturated and unsaturated fatty acids against several oral pathogens. J Environ Biol 34(4):673

    Google Scholar 

  • Colla G, Rouphael Y, Bonini P, Cardarelli M (2015) Coating seeds with endophytic fungi enhances growth, nutrient uptake, yield and grain quality of winter wheat. Int J Plant Prot 9:171–189

    Google Scholar 

  • Deshmukh S, Hückelhoven R, Schäfer 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 103(49):18450–18457

    Article  CAS  Google Scholar 

  • Domsch KH, Gams W, Anderson TH (1980) Compendium of soil fungi, vol 1. Academic Press, London

    Google Scholar 

  • Elsherbiny EA, El Khateeb AY, Azzaz NA (2016) Chemical composition and fungicidal effects of Ocimum basilicum essential oil on Bipolaris and Cochliobolus species. J Agric Sci Technol 18(4):1143–1152

    Google Scholar 

  • Evans HC, Holmes KA, Thomas SE (2003) Endophytes and mycoparasites associated with an indigenous forest tree, Theobroma gileri, in Ecuador and a preliminary assessment of their potential as biocontrol agents of cocoa diseases. Mycol Prog 2(2):149–160

    Article  Google Scholar 

  • Fisher PJ, Petrini O (1987) Location of fungal endophytes in tissue of Suaeda fruticosa: a preliminary study. Trans Br Mycol Soc 89:246–249

    Article  Google Scholar 

  • Fouda AH, Hassan SED, Eid AM, Ewais EED (2015) Biotechnological applications of fungal endophytes associated with medicinal plant Asclepias sinaica (Bioss.). Ann Agric Sci 60(1):95–104

    Google Scholar 

  • Gilma JC, Joseph C (1998) A manual of soil fungi. Daya Books, New Delhi

    Google Scholar 

  • Guo L, Wu JZ, Han T, Cao T, Rahman K, Qin LP (2008) Chemical composition, antifungal and antitumor properties of ether extracts of Scapania verrucosa Heeg. and its endophytic fungus Chaetomium fusiforme. Molecules 13(9):2114–2125

    Article  CAS  Google Scholar 

  • Gupta A (2016) Exploration of active metabolites from Aegle marmelos and Emblica officinalis for determining its immunosuppressive properties. J Prog Res Biol 3(1):119–126

    Google Scholar 

  • Gupta SC, Patchva S, Koh W, Aggarwal BB (2012a) Discovery of curcumin, a component of golden spice, and its miraculous biological activities. Clin Exp Pharmacol Physiol 39(3):283–299

    Article  CAS  Google Scholar 

  • Gupta P, Samant K, Sahu A (2012b) Isolation of cellulose-degrading bacteria and determination of their cellulolytic potential. Int J Microbiol. https://doi.org/10.1155/2012/578925

    Google Scholar 

  • Harman GE (2000) Myths and dogmas of biocontrol changes in perceptions derived from research on Trichoderma harzinum T-22. Plant Dis 84(4):377–393

    Article  Google Scholar 

  • Harman GE, Howell CR, Viterbo A, Chet I, Lorito M (2004) Trichoderma species—opportunistic, avirulent plant symbionts. Nat Rev Microbiol 2(1):43–56

    Article  CAS  Google Scholar 

  • Howell CR (2003) Mechanisms employed by Trichoderma species in the biological control of plant diseases, the history and evolution of current concepts. Plant Dis 87:4–10

    Article  Google Scholar 

  • Jeffers SN, Martin SB (1986) Comparison of two media selective for Phytophthora and Pythium species. Plant Dis 70:1038–1043

    Article  Google Scholar 

  • Johnston A, Booth C (1983) Plant pathologist’s pocketbook. Commonwealth Agricultural Bureaux, Slough

    Google Scholar 

  • Khan H, Khan Z, Amin S, Mabkhot YN, Mubarak MS, Hadda TB, Maione F (2017) Plant bioactive molecules bearing glycosides as lead compounds for the treatment of fungal infection: a review. Biomed Pharmacother 93:498–509

    Article  CAS  Google Scholar 

  • Kim HY, Choi GJ, Lee HB, Lee SW, Lim HK, Jang KS, Kim JC (2007) Some fungal endophytes from vegetable crops and their anti-oomycete activities against tomato late blight. Lett Appl Microbiol 44(3):332–337

    Article  Google Scholar 

  • Kruszewska JS, Perlinska-Lenart U, Gorka-Niec W, Orlowski J, Zembek P, Palamarczyk G (2008) Alterations in protein secretion caused by metabolic engineering of glycosylation pathways in fungi. Acta Biochim Pol 55(3):447–456

    CAS  Google Scholar 

  • Kruszewska H, Zareba T, Tyski S (2012) Examination of antimicrobial activity of selected non-antibiotic medicinal preparations. Act Pol Pharm Drug Res 69:1368–1371

    Google Scholar 

  • Kuete V, Ngameni B, Mbaveng AT, Ngadjui B, Meye JM, Lall N (2010) Evaluation of flavonoids from Dorstenia barteri for their antimycobacterial, antigonorrheal and anti-reverse transcriptase activities. Acta Trop 116(1):100–104

    Article  CAS  Google Scholar 

  • 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(1):137–151

    Article  Google Scholar 

  • Lahlali R, Hijri M (2010) Screening, identification and evaluation of potential biocontrol fungal endophytes against Rhizoctonia solani AG3 on potato plants. FEMS Microbiol Lett 311(2):152–159

    Article  CAS  Google Scholar 

  • Lahlali R, Serrhini MN, Friel D, Jijakli MH (2007) Predictive modelling of temperature and water activity (solutes) on the in vitro radial growth of Botrytis cinerea Pers. Int J Food Microbiol 114(1):1–9

    Article  Google Scholar 

  • Li E, Clark AM, Rotella DP, Hufford CD (1995) Microbial metabolites of ophiobolin A and antimicrobial evaluation of ophiobolins. J Nat Prod 58(1):74–81

    Article  CAS  Google Scholar 

  • Lledo S, Rodrigo S, Poblaciones MJ, Santamaria O (2016) Biomass yield, nutritive value and accumulation of minerals in Trifolium subterraneum L. as affected by fungal endophytes. Plant Soil 405(1–2):197–210

    Article  CAS  Google Scholar 

  • Medic-Saric M, Maysinger D, Movrin M, Dvoržak I (1980) Antibacterial and antifungal activities of nitroxoline Mannich bases. Chemotherapy 26(4):263–267

    Article  CAS  Google Scholar 

  • Mejia LC, Rojas EI, Maynard Z, Van Bael S, Arnold AE, Hebbar P, Herre EA (2008) Endophytic fungi as biocontrol agents of Theobroma cacao pathogens. Biol Control 46(1):4–14

    Article  Google Scholar 

  • Mishra VK (2010) In vitro antagonism of Trichoderma species against Pythium aphanidermatum. J Phytol 2(9):28–35

    Google Scholar 

  • Morandi MA, Sutton JC, Maffia LA (2000) Effects of host and microbial factors on development of Clonostachys rosea and control of Botrytis cinerea in rose. Eur J Plant Pathol 106(5):439–448

    Article  Google Scholar 

  • 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(5):1933–1944

    Article  CAS  Google Scholar 

  • Myrchiang P, Dkhar MS, Devi HR (2014) Studies on endophytic fungi associated with medicinally important aromatic plant Artemisia nilagirica (C.B. Clarke) Pamp. and their antagonistic activity against Phytophthora infestans. J Adv Lab Res Biol 4(4):112–119

    Google Scholar 

  • Nalini MS, Sunayana N, Prakash HS (2014) Endophytic fungal diversity in medicinal plants of Western Ghats, India. Int J Biodivers. https://doi.org/10.1155/2014/494213

    Google Scholar 

  • Naz S, Ahmad S, Rasool SA, Sayeed SA, Siddiqi R (2006) Antibacterial activity directed isolation of compounds from Onosma hispidum. Microbiol Res 161(1):43–48

    Article  CAS  Google Scholar 

  • 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 82:50–59

    Article  CAS  Google Scholar 

  • Olmedo GM, Cerioni L, González MM, Cabrerizo FM, Rapisarda VA, Volentini SI (2017) Antifungal activity of β-carbolines on Penicillium digitatum and Botrytis cinerea. Food Microbiol 62:9–14

    Article  CAS  Google Scholar 

  • Omero C, Inbar J, Rocha-Ramirez V, Herrera-Estrella A, Chet I, Horwitz BA (1999) G protein activators and cAMP promote mycoparasitic behaviour in Trichoderma harzianum. Mycol Res 103(12):1637–1642

    Article  CAS  Google Scholar 

  • Park M (1934) Report on the work of the Mycology Division. Admn Rep Dir Agric Ceylon, pp 126–133

  • Park ES, Moon WS, Song MJ, Kim MN, Chung KH, Yoon JS (2001) Antimicrobial activity of phenol and benzoic acid derivatives. Int Biodeterior Biodegradation 47(4):209–214

    Article  CAS  Google Scholar 

  • 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):14110–14123

    Article  CAS  Google Scholar 

  • Pujol I, Guarro J, Llop C, Soler L, Fernández-Ballart J (1996) Comparison study of broth macrodilution and microdilution antifungal susceptibility tests for the filamentous fungi. Antimicrob Agents Chem 40(9):2106–2110

    CAS  Google Scholar 

  • Rathaiah Y (1982) Rhizome rot of turmeric. Indian Phytopath 35(3):415–417

    Google Scholar 

  • Rinu K, Sati P, Pandey A (2014) Trichoderma gamsii (NFCCI 2177): a newly isolated endophytic, psychrotolerant, plant growth promoting, and antagonistic fungal strain. J Basic Microbiol 54(5):408–417

    Article  CAS  Google Scholar 

  • Roblin PM, Hammerschlag MR (1998) In vitro activity of a new ketolide antibiotic, HMR 3647, against Chlamydia pneumoniae. Antimicrob Agents Chem 42(6):1515–1516

    CAS  Google Scholar 

  • Rodriguez RJ, White JF Jr, Arnold AE, Redman ARA (2009) Fungal endophytes: diversity and functional roles. New phytol 182(2):314–330

    Article  CAS  Google Scholar 

  • Roy AK (1992) Severity of Rhizoctonia solani on the leaves of rice and turmeric. Indian Phytopath 45:344–347

    CAS  Google Scholar 

  • Rubini MR, Silva-Ribeiro RT, Pomella AW, Maki CS, Araújo WL, Dos Santos DR, Azevedo JL (2005) Diversity of endophytic fungal community of cacao (Theobroma cacao L.) and biological control of Crinipellis perniciosa, causal agent of Witches’ Broom Disease. Int J Biol Sci 1(1):24

    Article  CAS  Google Scholar 

  • Sanabria-Ríos DJ, Rivera-Torres Y, Maldonado-Domínguez G, Domínguez I, Ríos C, Díaz D, Montano N (2014) Antibacterial activity of 2-alkynoic fatty acids against multidrug-resistant bacteria. Chem Phys Lipids 178:84–91

    Article  Google Scholar 

  • 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 109:6241–6246

    Article  CAS  Google Scholar 

  • Shanmugam V, Kanoujia N (2011) Biological management of vascular wilt of tomato caused by Fusarium oxysporum f. sp. lycospersici by plant growth-promoting rhizobacterial mixture. Biol Control 57(2):85–93

    Article  Google Scholar 

  • Shanmugam V, Gupta S, Dohroo NP (2013) Selection of a compatible biocontrol strain mixture based on co-cultivation to control rhizome rot of ginger. Crop Prot 43:119–127

    Article  Google Scholar 

  • 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):30–34

    Article  CAS  Google Scholar 

  • Sriraj PP, Sundravadana S, Alice D (2014) Efficacy of fungicides, botanicals and bioagents against Rhizoctonia solani inciting leaf blight on turmeric (Curcuma longa L.). Afr J Microbiol Res 8(36):3284–3294

    Article  CAS  Google Scholar 

  • Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30(12):2725–2729

    Article  CAS  Google Scholar 

  • Thiripurasundari K, Selvarani K (2014) Production of turmeric in India: an analysis. Int J Bus Manag 2(9):229

    Google Scholar 

  • 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):4673–4680

    Article  CAS  Google Scholar 

  • 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):1147–1156

    Article  CAS  Google Scholar 

  • Ubaid JM, Hussein HM, Hameed IH (2016) Determination of bioactive chemical composition of Callosobruchus maculutus and investigation of its anti-fungal activity. Int J Pharmacogn Phytochem Res 8(8):1385–1397

    Google Scholar 

  • Ushamalini C, Nakkeeran P, Marimuthu T (2008) Induction of plant defense enzymes in turmeric plants by Trichoderma viride. Arch Phytopathol Plant Prot 41:79–93

    Article  CAS  Google Scholar 

  • Van der Plaats-Niterink AJ (1981) Monograph of the genus Pythium. Stud Mycol 21:1–24

    Google Scholar 

  • Vinale F, Ambrosio GD, Abadi K, Scala F, Marra R, Turrà D, Lorito M (2004) Application of Trichoderma harzianum (T22) and Trichoderma atroviride (P1) as plant growth promoters, and their compatibility with copper oxychloride. J Zhejiang Univ Sci 30(4):425–425

    Google Scholar 

  • Vinale F, Marra R, Scala F, Ghisalberti EL, Lorito M, Sivasithamparam K (2006) Major secondary metabolites produced by two commercial Trichoderma strains active against different phytopathogens. Lett Appl Microbiol 43(2):143–148

    Article  CAS  Google Scholar 

  • Vinale F, Ghisalberti EL, Sivasithamparam K, Marra R, Ritieni A, Ferracane R, Lorito M (2009) Factors affecting the production of Trichoderma harzianum secondary metabolites during the interaction with different plant pathogens. Lett Appl Microbiol 48(6):705–711

    CAS  Google Scholar 

  • Vinale F, Sivasithamparam K, Ghisalberti EL, Ruocco M, Woo S, Lorito M (2012) Trichoderma secondary metabolites that affect plant metabolism. Nat Prod Commun 7:1545–1550

    CAS  Google Scholar 

  • Vinale F, Sivasithamparam K, Ghisalberti EL, Woo SL, Nigro M, Marra R, Manganiello G (2014) Trichoderma secondary metabolites active on plants and fungal pathogens. Open Mycol J 8(1):127–139

    Article  Google Scholar 

  • Wahab S (2009) Biotechnological approaches in the management of plant pests, diseases and weeds for sustainable agriculture. J Biopest 2(2):115–134

    Google Scholar 

  • White TJ, Bruns T, Lee SJWT, Taylor JL (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR Protoc: Guide Methods Appl 18(1):315–322

    Google Scholar 

  • Yedidia I, Benhamou N, Chet I (1999) Induction of defense responses in cucumber plants (Cucumis sativus L.) by the biocontrol agent Trichoderma harzianum. Appl Environ Microbiol 65:1061–1070

    CAS  Google Scholar 

  • Youssef DA, Miller CW, El-Abbassi AM, Cutchins DC, Cutchins C, Grant WB, Peiris AN (2011) Antimicrobial implications of vitamin D. Dermatoendocrinol 3(4):220–229

    Article  CAS  Google Scholar 

  • Zhang L, Pornpattananangkul D, Hu CM, Huang CM (2010) Development of nanoparticles for antimicrobial drug delivery. Curr Med Chem 17(6):585–594

    Article  CAS  Google Scholar 

Download references

Acknowledgements

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. The authors extend thanks to Dr. K. Ramachandra Kini, Associate Professor, Department of Biotechnology, University of Mysore, Mysore for his help in Phylogenetic analysis of endophytes and Institute of Excellence (IOE), University of Mysore for providing the instrumentation facility.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to H. S. Prakash.

Ethics declarations

Conflict of interest

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

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Vinayarani, G., Prakash, H.S. Fungal endophytes of turmeric (Curcuma longa L.) and their biocontrol potential against pathogens Pythium aphanidermatum and Rhizoctonia solani. World J Microbiol Biotechnol 34, 49 (2018). https://doi.org/10.1007/s11274-018-2431-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s11274-018-2431-x

Keywords

Navigation