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Invitro Screening and Molecular Characterization of Trichoderma Harzianum Against Phytophthora Palmivora (Butl.,) Causing Root Rot Disease in Caricapapaya L.

Authors(4) :-M. Mahadevi, A. Thavaselvi, V. Latha, A. Panneerselvam

The present investigation showed Trichoderma. harzianum exhibited promising biocontrol ability against P. palmivora (Butl.) by dual culture experiment and poisoned food technique. T. harzianum was found to be the most effective antagonists against P. palmivora (Butl.). GC-MS analysis of ethyl acetate extract of the filtrate of T. harzianum revealed the presence of 17 compounds by representing 17 prominent peaks. The major phytocompound of T. harzianumwas Diethyl Phthalate (RT= 4.105 min) with 29.994 % of peak area.The ITS region gene sequence of potential biocontrol fungus obtained in this study were deposited in GenBank under the accession number KY346985. The sequence of potential biocontrol fungus showed the maximum homology with T. harzianum (GenBank Accession Number KC33021) by BLAST homology analysis. Therefore potential biocontrol fungus was further confirmed as T. harzianum by molecular analysis. The secondary structure of ITS region gene of T. harzianum showed 45 stems, 28 bulge loops and 12 hairpin like structure respectively. The free energy of ITS region gene of T. harzianum secondary structure was – 291.0 kkal/mol. The total restriction enzyme sites of ITS region gene of T. harzianum were 35. The GC and AT content of ITS region gene of T. harzianum were found to be 56 and 44% respectively. These exacting results suggested that fungicide play an important role in controlling the phytopathogenic fungi.
M. Mahadevi, A. Thavaselvi, V. Latha, A. Panneerselvam
Fungicides, T. harzianum, GC-MS, Biocontrol.
  1. Al-Askar, A. A,. 2012. Invitro antifungal activity of three Saudi plant extracts against some phytopathogenic fungi. J. Pl. Prot. Res., 52(4): 458-462.
  2. Al-Rahmah A. N., Mostafa A. A., Abdel-Megeed A., Yakout S. M. and Hussein S. A., 2013. Fungicidal activities of certain methanolic plant extracts against tomato phytopathogenic fungi. Afr. J. Microbiol. Res., 7(6): 517-524.
  3. Bae, S. J., Mohanta, T. K., Chung, J. Y., Ryu, M., Park, G., Shim, S., Hong, S. B., Seo, H., Bae, D. W., Bae, I., Kim, J. J. and Bae, H., 2016. Trichoderma metabolites as biological control agents against Phytophthora pathogens. Biol. Cont., 92: 128–138.
  4. Dickinson, C.H. and Broadman, F., 1971. Physiological studies of some fungi isolated from peat. Trans. Br. Mycol, Soc., 55:293 -305.
  5. Elshahawya, I. E., Haggagb, K. H. E. and Abd-El-Khaira, H., 2016. Compatibility of Trichoderma spp. with seven chemical fungicides used in the control of soil borne plant pathogens. Res. J. Pharm. Biol. Chem. Sci., 7(1): 1772 – 1785.
  6. Felsenstein, J., 1985. Confidence limits on phylogenies: An approach using the bootstrap. Evolution,39:783-791.
  7. Gherbawy, Y.A., Hussein, N.A. and Al- Qurashi, A. A., 2014. Molecular Characterization of Trichoderma populations isolated from soil of Taif City, Saudi Arabia. Int. J. Curr. Microbiol. App. Sci., 3(9)1059-1071.
  8. Grover, R.K. and Moore, J.D., 1962. Toximetric studies of fungicides against brown rot organism. Sclerotina fruticola. Phytopathol., 52:876-880.
  9. Hadizadeh, I., Peivastegan, B. and Kolahi, M., 2009. Antifungal activity of nettle (Urticad ioica L.), colocynth (Citrullus colocynthis L. Schrad), oleander (Nerium oleander L.) and konar (Ziziphus spina-christi L.) extracts on plants pathogenic fungi. Pakistan J. Biol. Sci., 12: 58-63.
  10. Hamed. Eman R , Hassan M. Awad , Eman A. Ghazi , Nadia G. El-Gamal and Heba S. Shehata 2015. Trichoderma asperellum isolated from salinity soil using rice straw waste as biocontrol agent for cowpea plant pathogens. J of Appl Pharmaceuti Sci 5 (Suppl 2); 091-098.
  11. Higgins, D.G., Bleasby, A.T. and Fuchs, R., 1992. Clustal W: Improved software for multiple sequence alignment. CABIOS, 8:189 – 191.
  12. Hung, P. M., Wattanachai, P., Kasem, S. and Poaim, S., 2015. Biological control of Phytophthora palmivora causing root rot of pomelo using Chaetomium sp. Mycobiol., 43(1): 63-70.
  13. Kelepertzis, E., 2014. Accumulation of heavy metals in agricultural soils of Mediterranean: insights from Argolida basin, Peloponnese, Greece. Geoderma 221-222, 82-90.
  14. Khan, K., Lu, Y., Khan, H., Ishtiaq, M., Sardar, K., Waqas, M., Luo, W and Wang, T., 2013. Heavy metals in agricultural soils and crops and their health risks in Swat District, northern Pakistan. Food Chem. Toxicol. 58, 449-458.
  15. Kubicek, C.P., Bissett, J., Druzhinina, I., Kullnig Gradinger, C. and Szakacs, G., 2003. Genetic and metabolic diversity of Trichoderma: a case study on South East Asian isolates. Fungal Genet. Biol., 38: 310–319.
  16. Lalitha, V., Kiran, B. and Raveesha, K.A., 2012. Antifungal activity of Trichoderma koningii oudm. against seed borne fungal species of paddy. Int. J. Innov. Bio Sci.., 2 (4): 176-180.
  17. Mpika, J., Kébé, I. B., Issali, A. E., Guessan, F.K.N., Druzhinina, S., Komon-Zélazowska, M., Kubicek, C. P. and Aké, S., 2009. Antagonist potential of Trichoderma indigenous isolates for biological control of Phytophthora palmivora the causative agent of black pod disease on cocoa (Theobroma cacao L.) in Côte d’Ivoire. Afr. J. Biotechnol., 8 (20): 5280-5293.
  18. Perez, C., Pauli, M. and Bazerque, P., 1990. An antibiotic assay by the well agar method. Acta. Biol. Med. Exper., 15: 113–115.
  19. Porter, C. L., 1924. Concerning the characters of certain fungi as exhibited by their growth in the presence of other fungi. Am. J. Bot., 11:168 - 188.
  20. Romão-Dumaresq, A.S., de Araújo, W.L., Talbot, N.J. and Thornton, C.R., 2012. RNA interference of endochitinases in the sugarcane endophyte Trichoderma virens 223 reduces its fitness as a biocontrol agent of pineapple disease. PLoS One, 7: e47888.
  21. Saitou, N. and Nei, M., 1987. The neighbor joining method: A new method for reconstructing phylogenetic trees. Mol. Biol. Evolution.,4:406-425.
  22. Senthilkumar G., Madhanraj P and Panneerselvam A. 2011. Studies on DNA extraction, molecular identification and genetic evolution of Trichoderma harzianum. Asian J. Research Chem. 4(8): August, 2011; Page 1225-1230.
  23. Senthilkumar, G., Madhanraj, P. and Panneerselvam A., 2011. Studies on DNA extraction, molecular identification and genetic evolution of Trichoderma harzianum. Asian J. Res. Chem., 4(8): 1225 – 1230.
  24. Sette, L. D., Passarini, M. R. Z., Delarmelina, C., Salati, F. and Duarte, M. C. T., 2006. Molecular characterization and antimicrobial activity of endophytic fungi from coffee plants. World J. Microbiol. Biotechnol., 22:1185-1195.
  25. Siddiquee, S., Cheong, B. E., Taslima, K., Kausar, H. and Hasan, M. M., 2012. Separation and identification of volatile compounds from liquid cultures of Trichoderma harzianum by GC-MS using three different capillary columns. J. Chromatogr. Sci., 50: 358-367.
  26. Siddiquee, S., Soon Guan, F. A. T. and Aziz, E. R., 2007. Phylogenetic relationships of Trichoderma harzianum based on the sequence analysis of the internal transcribed spacer region -1 of the rDNA.J. Appl. Sci. Res., 3(9): 896-903.
  27. Siddiquee, S., Yusuf, U.K., Hossain, K. and Jahan, S. 2009. In vitro studies on the potential Trichoderma harzianum for antagonistic properties against Ganoderma boninense. J. Food Agric. Environ., 7(3&4): 970-976.
  28. Singh, P. and Srivastava, D. 2013. Phytochemical screening and in vitro antifungal investigation of Parthenium hysterophorus extracts against A. alternata. Int. Res. J. Pharm., 4: 190-193.
  29. Siva, N., Ganesan , S., Banumathy, N. and Muthuchelian, 2008. Antifungal effect of leaf extract of some medicinal plants against Fusarium oxysporum causing wilt disease of Solanum melogena L. Ethnobot. Leaflets, 12: 156-163.
  30. Skidmore, A. M. and Dickinson, C.M., 1976. Colony interactions and hyphal interferences between Septoria nodorum and phylloplane fungi. Trans. Br. Mycol, Soc., 66:57 -64.
  31. Srivastav, R., Singh, R. and Prasad, R., 2011. Relative antagonistic effect of different isolates of Trichoderma viride and T. harzianum against Phytophthora capsici - a bell pepper pathogen. J. Biol. Control., 25: 239-241.
  32. Tamura, K., Dudley, J., Nei, M. and Kumar, S., 2007. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Molecular Biol. Evolution24:1596-1599.
  33. Tapwal, A., Nisha, Garg, S., Guatam, N. and Kumar, R. 2011. In vitro antifungal potency of plant extracts against five phytopathogens. Braz. Arch. Biol. Technol., 54(6): 1093- 1098.
  34. Thakur, M.K. and Pandey, S. 2016. GC-MS analysis of phytochemical constituents in leaf extract Vitex negundo (L.). World J. Pharm. Pharm. Sci., 5(8): 672-675.
  35. Vawdrey. L.L, M. Male and K.R.E. Grice 2015. Field and laboratory evaluation of fungicides for the control of Phytophthora fruit rot of papaya in far north Queensland, Australia. Crop Protection 67,116-120.
  36. Vazquez-Angulo, J.C., Mendez-Trujillo, V., González-Mendoza, D., Morales-Trejo, A., Grimaldo-Juarez, O. and Cervantes-Díaz, L., 2012. A rapid and inexpensive method for isolation of total DNA from Trichoderma sp. (Hypocreaceae). Gen. Mol. Res., 11 (2): 1379-1384.
  37. Vinale, F., Marra, R., Scala, F., Lorito, M., Ghisalberti, E. L. and Sivasithampram, K., 2005. Secondary metabolites produced by two commercial strains of Trichoderma harzianum.J. Plant Pathol., 87(4):267–309.
  38. Vinale, F., Sivasithamparam, K., Ghisalberti, L. E., Marra, R., Woo, L. S. and Lorito, M., 2008b. Trichoderma-plant-pathogen interactions. Soil. Biol. Biochem., 40: 1-10.
Publication Details
  Published in : Volume 4 | Issue 5 | March-April 2018
  Date of Publication : 2018-02-28
License:  This work is licensed under a Creative Commons Attribution 4.0 International License.
Page(s) : 156-168
Manuscript Number : IJSRST1841274
Publisher : Technoscience Academy
PRINT ISSN : 2395-6011
ONLINE ISSN : 2395-602X
Cite This Article :
M. Mahadevi, A. Thavaselvi, V. Latha, A. Panneerselvam, "Invitro Screening and Molecular Characterization of Trichoderma Harzianum Against Phytophthora Palmivora (Butl.,) Causing Root Rot Disease in Caricapapaya L.", International Journal of Scientific Research in Science and Technology(IJSRST), Print ISSN : 2395-6011, Online ISSN : 2395-602X, Volume 4, Issue 5, pp.156-168, March-April-2018.
Journal URL : http://ijsrst.com/IJSRST1841274

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