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Expression analysis on mycoparasitism related genes during antagonism of Trichoderma with Colletotrichum falcatum causing red rot in sugarcane

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Abstract

Present study was aimed to select a suitable Trichoderma isolate as candidate antagonist based on its efficacy in producing cell wall degrading enzymes (CWDEs), its mycoparasitism activity and expression of related genes against the red rot pathogen caused by Colletotrichum falcatum in sugarcane. For which, six different isolates of Trichoderma selected from our earlier studies (T. harzianum, T. asperullum) were evaluated based on their capability in releasing cell wall degrading enzymes individually and during antagonism with C. falcatum in dual plate. Amongst T. harzianum (T20) exhibited the greatest mycoparasitic potential against the C. falcatum, by producing higher concentration of  CWDEs viz., chitinase and β-1, 3-glucanase, slightly lower amounts of cellulase and protease with significant reduction in polygalacturonase produced by pathogen. Further microscopic observation on interaction of C. falcatum with the selected isolate of T. harzianum (T20) exhibited the mycoparasitic activity of antagonist over pathogen in dual culture and inhibition of C. falcatum pathogenesis in detached sugarcane leaves. In addition, expression pattern of eight genes coding various enzymes involved in mycoparasitism by T. harzianum over C. falcatum were analyzed using qRT-PCR in vitro and on sugarcane leaves. In in vitro interactions, five genes of  cell wall degrading enzymes viz., chitinase (chit33), endochitinase (endo42), β-1, 3-glucanase (glu), exochitinase 1 (exc1), exochitinase 2 (exc2), were upregulated during and after contact as compared to before contact, while three genes related with proteases such as alkaline proteinase (prb1), trypsin-like protease (Pra1), subtilin-like serine protease (ssp), genes were upregulated during the contact with C. falcatum and slightly down regulated after contact. In detached leaves, seven genes were potentially upregulated except subtilin-like serine protease, which was down regulated during interaction of C. falcatum and T. harzianum as compared to T. harzianum inoculation alone. All these biochemical and molecular results confirm the efficacy of T. harzianum (T20) against C. falcatum and justify the right selection of candidate antagonist for our further studies on identification of antifungal genes/proteins against C. falcatum in sugarcane.

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References

  • Anson ML (1938) The estimation of pepsin, trypsin, papain and cathepsin with hemoglobin. J Gen Physiol 22:79–89

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Boller T, Mauch F (1988) Colorimetric assay of chitinase. Methods Enzymol 161:431–435

    Google Scholar 

  • Dotaniya ML, Datta SC, Biswas DR, Dotaniya CK, Meena BL, Rajendiran S, Regar KL, Lata M (2016) Use of sugarcane industrial by-products for improving sugarcane productivity and soil health. Int J Recycl Org Waste Agric 5:185–194

    Article  Google Scholar 

  • Elad Y, Kapat A (1999) The role of Trichoderma harzianum protease in the biocontrol of Botrytis cinerea. Eur J Plant Pathol 105:177–189

    Article  CAS  Google Scholar 

  • Elamathi E, Malathi P, Viswanathan R, Ramesh Sunder A (2017a) Molecular characterization of antagonists in sugarcane ecosystem and their potential to suppress Colletotrichum falcatum and express antifungal proteins. J Sugarcane Res 7(2)

  • Elamathi E, Malathi P, Viswanathan R, Ramesh Sundar A (2017b) Molecular analysis of the suppression of Colletotrichum falcatum by Trichoderma harzianum in sugarcane. J Plant Pathol 99:211–218

    Google Scholar 

  • Emani C, Garcia JM, Lopata-Finch E, Pozo MJ, Uribe P, Kim DJ, Sunilkumar G, Cook DR, Kenerley CM, Rathore KS (2003) Enhanced fungal resistance in transgenic cotton expressing an endochitinase gene from Trichoderma virens. Plant Biotechnol J 1:321–336

    Article  PubMed  CAS  Google Scholar 

  • Fan H, Liu Z, Zhang R, Wang N, Dou K, Mijiti G, Diao G, Wang Z (2014) Functional analysis of a subtilisin-like serine protease gene from biocontrol fungus Trichoderma harzianum. J Microbiol 52:129–138

    Article  PubMed  CAS  Google Scholar 

  • Gajera HP, Vakharia DN (2010) Molecular and biochemical characterization of Trichoderma isolates inhibiting a phytopathogenic fungi Aspergillus niger van tieghem. Physiol Mol Plant Pathol 74:274–282

    Article  CAS  Google Scholar 

  • Gajera HP, Vakharia DN (2012) Production of lytic enzymes by Trichoderma isolates during in vitro antagonism with Aspergillus niger, the causal agent of collar rot of peanut. Braz J Microbiol 43:43–52

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Geremia RA, Goldman GH, Jacobs D, Aviles W, Vila SB, Van Montagu M, Herrera-Estrella A (1993) Molecular characterization of the proteinase encoding gene, prb1, related to mycoparasitism by Trichoderma harzianum. Mol Microbiol 8:603–613

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  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 

  • Jayakumar V, Tsushima S, Malathi P, Viswanathan R (2012) Elution, biological activity and characterization of phytotoxin produced by sugarcane red rot pathogen C. falcatum. Funct Plant Sci Biotechnol 6:149–163

    Google Scholar 

  • Kapat A, Zimand Y, Elad Y (1998) Effect of two isolates of Trichoderma harzianum on the activity of hydrolytic enzymes produced by Botrytis cinerea. Physiol Mol Plant Pathol 52:127–137

    Article  CAS  Google Scholar 

  • Lopez-Mondejar R, Ros M, Pascual JA (2011) Mycoparasitism-related genes expression of Trichoderma harzianum isolates to evaluate their efficacy as biological control agent. Biol Control 56:59–66

    Article  CAS  Google Scholar 

  • Malathi P, Padmanaban P, Viswanathan R, Mohanraj D (2008) Interaction between Colletotrichum falcatum pathotypes and biocontrol agents. Arch Phytopathol Plant Prot 41(5):311–317

    Article  Google Scholar 

  • Malathi P, Viswanathan R, Padmanaban P (2012) Identification of antifungal proteins from fungal and bacterial antagonists against Colletotrichum falcatum causing sugarcane red rot. J Biol Control 26(1):49–54

    Google Scholar 

  • Mamarabadi M, Jensen B, Jensen DF, Lubeck M (2008) Real-time RT-PCR expression analysis of chitinase and endoglucanase genes in the three-way interaction between the biocontrol strain Clonostachys rosea IK726, Botrytis cinerea and strawberry. FEMS Microbiol Lett 285:101–110

    Article  PubMed  CAS  Google Scholar 

  • Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Annal Chem 31:426–428

    Article  CAS  Google Scholar 

  • Papavizas GC (1985) Trichoderma and Gliocladium: biology, ecology, and potential for biocontrol. Annu Rev Phytopathol 23:23–54

    Article  Google Scholar 

  • Pozo MJ, Baek JM, García JM et al (2004) Functional analysis of tvsp1, a serine protease-encoding gene in the biocontrol agent Trichoderma virens. Fungal Genet Biol 41:336–348

    Article  PubMed  CAS  Google Scholar 

  • Rao GP, Viswanathan R, Singh SB (2002) Current situation of sugarcane diseases in India. In: Singh SB, Rao GP, Easwaramoorthy S (eds) Sugarcane crop management, vol 734. SCI Tech Publishing LLC, Houstan

    Google Scholar 

  • Roatti B, Perazzolli M, Gessler C, Pertot I (2013) Abiotic stresses affect Trichoderma harzianum T39-induced resistance to downy mildew in grapevine. Phytopathology 103(12):1227–1234

    Article  PubMed  CAS  Google Scholar 

  • Sadasivam S, Manickam K (1992) Biochemical method for agricultural sciences. Wiley, New Delhi

    Google Scholar 

  • Shah MR, Mukherjee PK, Eapen S (2010) Expression of a fungal endochitinase gene in transgenic tomato and tobacco results in enhanced tolerance to fungal pathogens. Physiol Mol Biol Plants 16:39–51

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Sharon E, Bar-Eyal M, Chet I, Herrera-Estrella A, Kleifeld O, Spiegel Y (2001) Biological control of the root-knot nematode meloidogyne javanica by T. harzianum. Phytopathology 91(7):687–693

    Article  PubMed  CAS  Google Scholar 

  • Steyaert JM, Stewart A, Jaspers MV, Carpenter M, Ridgway HJ (2004) Co-expression of two genes, a chitinase (chit42) and proteinase (prb1), implicated in mycoparasitism by Trichoderma hamatum. Mycologia 96(6):1245–1252

    Article  PubMed  CAS  Google Scholar 

  • Suarez B, Rey M, Castillo P, Monte E, Llobell A (2004) Isolation and characterizarion of PRA1, a tripsin- like protease from the biocontrol agent Trichoderma harzianum CECT 2413 displaying nematicidal activity. Appl Microbiol Biotechnol 65:46–55

    Article  PubMed  CAS  Google Scholar 

  • Verma M, Brar SK, Tyagi RD, Surampalli RY, Valero JR (2007) Antagonistic fungi Trichoderma spp.: panoply of biological control. Biochem Eng J 37:1–20

    Article  Google Scholar 

  • Vieira PM, Guedes Coelho AS, Steindorff AS, Siqueira SL, Silvaand RN, Ulhoa CJ (2013) Identification of differentially expressed genes from Trichoderma harzianum during growth on cell wall of Fusarium solani as a tool for biotechnological application. BMC Genom 14:1–11

    Article  CAS  Google Scholar 

  • Vinalea F, Sivasithamparam K, Ghisalbertic EL, Marra R, Woo SL, Loritoa M (2008) Trichoderma-plant-pathogen interaction. Soil Biol Biochem 40:1–10

    Article  CAS  Google Scholar 

  • Viswanathan R (2010) Plant disease: red rot of sugarcane. Anmol Publishers, New Delhi

    Google Scholar 

  • Viswanathan R, Rao GP (2011) Disease scenario and management of major sugarcane diseases in India. Sugar Tech 13(4):336–353

    Article  CAS  Google Scholar 

  • Viswanathan R, Ramesh Sunder A, Premkumari SM (2003) Mycolytic enzymes of extracellular enzymes of antagonistic microbes to Colletotrichum falcatum, red rot pathogen of sugarcane. World J Microbiol Biotechnol 19:953–959

    Article  CAS  Google Scholar 

  • Viswanathan R, Merina Premkumari S, Ramesh Sundar A, Kathiresan T (2006) Cloning partial endochitinase cDNA of Trichoderma harzianum antagonistic to Colletotrichum falcatum causing red rot of sugarcane. Curr Sci 91:7–10

    Google Scholar 

  • Viterbo A, Ramot O, Chemin L, Chet I (2002) Significance of lytic enzymes from Trichoderma spp. in the biocontrol of fungal plant pathogens. Antonie Van Leeuwenhoek 81(1–4):549–556

    Article  PubMed  CAS  Google Scholar 

  • Yang HH, Yang SL, Peng KC, Lo CT, Liu SY (2008) Induced proteome of Trichoderma harzianaum by Botrytis cinerea. Mycol Res 113:924–932

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors are grateful to the Director, ICAR-Sugarcane Breeding Institute for providing facilities and constant encouragement. The financial support received from Sugarcane Development Fund, Ministry of Consumer Affairs, Food & Public, New Delhi is duly acknowledged.

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Correspondence to Palaniyandi Malathi.

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Elamathi, E., Malathi, P., Viswanathan, R. et al. Expression analysis on mycoparasitism related genes during antagonism of Trichoderma with Colletotrichum falcatum causing red rot in sugarcane. J. Plant Biochem. Biotechnol. 27, 351–361 (2018). https://doi.org/10.1007/s13562-018-0444-z

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  • DOI: https://doi.org/10.1007/s13562-018-0444-z

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