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volume 38 issue 3 (june 2015) : 375-381,   Doi: 10.5958/0976-0571.2015.00121.6

Screening of antagonistic potential and plant growth promotion activities of Trichoderma spp. and fluorescent Pseudomonas spp. isolates against Sclerotinia sclerotiorum causing stem rot of French bean

R
Ruchita Dixit*
R
Ram Bahadur Singh
H
Harikesh Bahadur Singh
1Department of Mycology and Plant Pathology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi-221 005, India.
Cite article:- Dixit* Ruchita, Singh Bahadur Ram, Singh Bahadur Harikesh (2025). Screening of antagonistic potential and plant growth promotion activities of Trichoderma spp. and fluorescent Pseudomonas spp. isolates against Sclerotinia sclerotiorum causing stem rot of French bean. Legume Research. 38(3): 375-381. doi: 10.5958/0976-0571.2015.00121.6.

ABSTRACT

Sclerotinia sclerotiorum have been a major plant pathogen with wide host range in agriculture. Annually, 60% crop losses due to this pathogen have been reported worldwide. Biological management have been sought to be an alternative method for controlling the disease. Trichoderma and Pseudomonas are well known for their biocontrol and plant growth promotion activities. Isolation was done from nearby and within Varanasi. Among all isolates 20 isolates of Trichoderma and 11 isolates of fluorescent Pseudomonas were selected on the basis of dual culture assay against S. sclerotiorum. They were further characterised on the basis of IAA, siderophore, phosphate solubilization, effect of volatile and crude extract, HCN production, ammonia production, chitinase and ß 1, 3–glucanase enzymatic assay. Among the Trichoderma isolates, maximum plant growth promotion activity and mycolytic enzyme activity was observed by isolate T9. Among fluorescent Pseudomonas isolates the best plant promotion activity as well efficient antagonistic activity was observed in isolate Pf11.

REFERENCES

  1. Abdullah, M.T., Ali, N.Y., and Suleman, P. (2008). Biological control of Sclerotinia sclerotiorum (Lib.) de Bary with Trichoderma harzianum and Bacillus amyloliquefaciens isolates. Crop. Prot. 27: 1354-1359.
  2. Ahmad, F., Ahmad, I., and Khan, M.S. (2004). Indole Acetic acid production by the indigenous isolates of Azotobacter and fluorescent Pseudomonas in the presence and absence of tryptophan. Turk. J. Biol. 29: 29-34.
  3. Amin, F., Razdan, V.K., Mohiddin, F.A., Bhat, K.A., and Banday, S. (2010). Potential of Trichoderma species as bioconttrol agents of soil borne fungal propagules. J. Phytol. 2: 38-41.
  4. Bakker, A.W., and Schipper, B. (1987). Microbial cyanide production in the rhizosphere in relation to potato yield reduction and Pseudomonas sp. mediated plant growth stimulation. Soil. Biochem., 19: 451-457.
  5. Bell, D.K., Wells, H.D., and Markham, C.R. (1982). In vitro antagonism of Trichoderma spp. against six fungal plant pathogens. Phytopatholgy. 72: 379–382.
  6. Benhamou, N., and Chet, I. (1993). Hyphal interactions between Trichoderma harzianum and Rhizoctonia solani: Ultrastructure and gold cytochemistry of the mycoparasitic process. Phytopathology. 83: 1062-1071.
  7. Castric, P.A. (1975). Hydrogen cyanide a secondary metabolite of Pesudomonas aeruginosa. Can. J. Microbial. 21: 613-618. De Marco, J.L., and Felix, C.R. (2007). Purification and Characterization of a â-Glucanase produced by Trichoderma harzianum showing biocontrol potential. Braz. Arch. Biol. Technol. 50: 21-29.
  8. De Souza, J.T., Bailey, B.A., Pomella, A.W.V., Erbe, E.F., Murphy, C.A., Bae, H., and Hebbar, P.K. (2008). Colonization of cocoa seedlings by Trichoderma stromaticum, a mycoparasite of the witches’ broom pathogen, and its influence on plant growth and resistance. Biol. Control. 46: 36-45.
  9. Dennis, C., and Webster, J. (1971). Antagonistic properties of species groups of Trichoderma II. Production of volatile antibiotics. Trans. Br. Mycol. Soc. 57: 41–48.
  10. Diby, P., Saju, K.A., Jisha, P.J., Sarma, Y.R., Kumar, A., and Anandaraj, M. (2005). Mycolytic enzymes produced by Pseudomonas fluorescens and Trichoderma spp. against Phytophthora capsici, the foot rot pathogen of black pepper (Pipernigrum L.). Ann. Microbiol. 55: 129-133.
  11. Elad, Y., Chet, I., and Henis, Y. (1982). Degradation of plant pathogenic fungi by Trichoderma harzianum. Can. J. Microbiol. 28: 719-725.
  12. El-Katatny, M.H., Gudelj, M., Robra, K.H., Elnaghy, M.A., and Gubitz, G.M. (2001). Charaterization of a chitinase and an endo-b-1,3-glucanase from T. harzianum Rifai T-24 involved in control of the phytopathogen Sclerotium rolfsii. Appl. Microbiol. Biot. 56: 137-143.
  13. Fernando, W.D.G., Nakkeeran, S., Zhang, Y., and Savchuk, S. (2007). Biological control of Sclerotinia sclerotiorum (Lib.) de Bary by Pseudomonas and Bacillus species on canola petals. Crop. Prot. 26: 100-107.
  14. Gravel, V., Antoun, H., and Tweddell, R.J. (2007). Growth stimulation and fruit yield improvement of greenhouse tomato plants by inoculation with Pseudomonas putida or Trichoderma atroviride: possible role of indole acetic acid (IAA). Soil. Biol. and Biochem. 39: 1968-1977.
  15. Harman, G.E., Chet, I., and Baker, R. (1980). Trichoderma hamatum effects on seed and seedlings disease induced in radish and pea by Pythium spp. or Rhizoctonia solani. Phytopathology. 70: 1167-1172.
  16. Hassanein, W.A., Awny, N.M., El-Mougith, A.A., and Salah El-Dien, S.H. (2009). The antagonistic activities of some metabolites produced by Pseudomonas aeruginosa Sha8. J. Appl. Sc. Res. 5: 404-414.
  17. Henri, F., Laurette, N.N., Annette, D., John, Q., Wolfgang, M., Francois-Xavier, E., and Dieudonne, N. (2008). Solubilization of inorganic phosphates and plant growth promotion by strains of Pseudomonas fluorescens isolated from acidic soils of Cameroon. Af. J. Microbiol. Res. 2: 171-178.
  18. Hernandez, C.F.D., Berlanga, P.A.M., Gallegos, M.G., and Cepeda, S.M. (2011). In vitro antagonist action of Trichoderma strains against Sclerotinia sclerotiorum and Sclerotium cepivorum. Am. J. Agric. Biol. Sc. 6: 410-417. Hoyos-Carvajal, L., Orduz, S., and Bissett, J. (2009). Growth stimulation in bean (Phaseolus vulgaris L.) by Trichoderma. Biol. Control. 51: 409-416.
  19. Huang, H.C., and Haes, J.A. (1976). Penetration and infection of Sclerotinia sclerotiorum by Coniothyrium minitans. Can. J. Bot. 54: 406-410.
  20. Johnson, L.A. (1957). Effect of antibiotics on the number of bacteria and fungi isolated from soil by dilution plate method. Phytopathology. 47: 21-22.
  21. Johnson, D.A., and Atallah, Z.K. (2006). Timing fungicide applications for managing Sclerotinia stem rot of potato. Plan. Dis. 90: 755-758.
  22. Kapri, A., and Tewari, L. (2010). Phosphate solubilization potential and phosphatise activity of rhizospheric Trichoderma spp. Brazil. J. Microbiol. 41: 787-795.
  23. Little, T.M., and Hills, F.J. (1978). Agricultural experimentation: design and analysis. John Wiley and Sons Inc, Indianapolis Ind Mehta, S., and Nautiyal, C.S. (2001). An efficient method for qualitative screening of phosphate-solubilizing bacteria. Curr. Microbiol. 43: 51–56.
  24. Miller, G.L. (1959). Use of dinitrosalicylic acid reagent for the determination of reducing sugar. Anal. Chem. 31: 426- 428.
  25. Pareek, R.P., and Gaur, A.C. (1973). Release of phosphorus from tricalcium phosphate and rock phosphate by organic acids. Curr. Sci. 43: 278–279.
  26. Patten, C.L., and Glick, B.R. (2002). Role of Pseudomonas putida indole acetic acid in development of the host plant root system. App. Environ. Microb. 68: 3795-3801.
  27. Rini, C.R., and Sulochana, K.K. (2007). Usefullness of Trichoderma and Pseudomonas against Rhizoctonia solani and Fusarium oxysporum infecting tomato. J. Trop. Agric. 45: 21-28.
  28. Rini, C.R., and Sulochana, K.K. (2006). Management of seedling rot of chilli (Capsicum annuum L.) using Trichoderma spp. and fluorescent pseudomonads (Pseudomonas fluorescens). J. Trop. Agric. 44: 79-82.
  29. Rudresh, D.L., Shivaprakash, M.K., and Prasad, R.D. (2005). Tricalcium phosphate solubilizing abilities of Trichoderma spp. in relation to P uptake and growth and yield parameters of chickpea (Cicer arietinum L.). Can. J. Microbiol. 51: 217–222.
  30. Schwyn, B., and Neilands, J.B. (1987). Universal chemical assay for the detection and determination of siderophores. Anal. Biochem. 160: 47-56.
  31. Sandhya, C., Adapa, L.K., Nampoothiri, M., Binod, P., Szakacs, G., and Pandey, A. (2004). Extracellular chitinase production by Trichoderma harzianum in submerged fermentation. J. Basic. Microbiol. 44: 49-58.
  32. Shaigan, S., Seraji, A., and Moghaddam, S.A.M. (2008). Identification and investigation on antagonistic effect of Trichoderma spp. on tea seedlings white foot and root rot (Sclerotium rolfsii Sacc) in vitro condition. Pakistan. J. Biol. Sci. 11: 3246-3250.
  33. Vincent, J.H. (1947). Distortion of fungal hyphae in the presence of certain inhibitors. Nature. 15: 850. Viterbo, A., Ramot, O., Chernin, L., and Chet, I. (2002). Significance of lytic enzymes from Trichoderma spp. in the biocontrol of fungal plant pathogens. Van. Leeuw. J. Microb. 81: 549-556.
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    volume 38 issue 3 (june 2015) : 375-381,   Doi: 10.5958/0976-0571.2015.00121.6

    Screening of antagonistic potential and plant growth promotion activities of Trichoderma spp. and fluorescent Pseudomonas spp. isolates against Sclerotinia sclerotiorum causing stem rot of French bean

    R
    Ruchita Dixit*
    R
    Ram Bahadur Singh
    H
    Harikesh Bahadur Singh
    1Department of Mycology and Plant Pathology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi-221 005, India.
    Cite article:- Dixit* Ruchita, Singh Bahadur Ram, Singh Bahadur Harikesh (2025). Screening of antagonistic potential and plant growth promotion activities of Trichoderma spp. and fluorescent Pseudomonas spp. isolates against Sclerotinia sclerotiorum causing stem rot of French bean. Legume Research. 38(3): 375-381. doi: 10.5958/0976-0571.2015.00121.6.

    ABSTRACT

    Sclerotinia sclerotiorum have been a major plant pathogen with wide host range in agriculture. Annually, 60% crop losses due to this pathogen have been reported worldwide. Biological management have been sought to be an alternative method for controlling the disease. Trichoderma and Pseudomonas are well known for their biocontrol and plant growth promotion activities. Isolation was done from nearby and within Varanasi. Among all isolates 20 isolates of Trichoderma and 11 isolates of fluorescent Pseudomonas were selected on the basis of dual culture assay against S. sclerotiorum. They were further characterised on the basis of IAA, siderophore, phosphate solubilization, effect of volatile and crude extract, HCN production, ammonia production, chitinase and ß 1, 3–glucanase enzymatic assay. Among the Trichoderma isolates, maximum plant growth promotion activity and mycolytic enzyme activity was observed by isolate T9. Among fluorescent Pseudomonas isolates the best plant promotion activity as well efficient antagonistic activity was observed in isolate Pf11.

    REFERENCES

    1. Abdullah, M.T., Ali, N.Y., and Suleman, P. (2008). Biological control of Sclerotinia sclerotiorum (Lib.) de Bary with Trichoderma harzianum and Bacillus amyloliquefaciens isolates. Crop. Prot. 27: 1354-1359.
    2. Ahmad, F., Ahmad, I., and Khan, M.S. (2004). Indole Acetic acid production by the indigenous isolates of Azotobacter and fluorescent Pseudomonas in the presence and absence of tryptophan. Turk. J. Biol. 29: 29-34.
    3. Amin, F., Razdan, V.K., Mohiddin, F.A., Bhat, K.A., and Banday, S. (2010). Potential of Trichoderma species as bioconttrol agents of soil borne fungal propagules. J. Phytol. 2: 38-41.
    4. Bakker, A.W., and Schipper, B. (1987). Microbial cyanide production in the rhizosphere in relation to potato yield reduction and Pseudomonas sp. mediated plant growth stimulation. Soil. Biochem., 19: 451-457.
    5. Bell, D.K., Wells, H.D., and Markham, C.R. (1982). In vitro antagonism of Trichoderma spp. against six fungal plant pathogens. Phytopatholgy. 72: 379–382.
    6. Benhamou, N., and Chet, I. (1993). Hyphal interactions between Trichoderma harzianum and Rhizoctonia solani: Ultrastructure and gold cytochemistry of the mycoparasitic process. Phytopathology. 83: 1062-1071.
    7. Castric, P.A. (1975). Hydrogen cyanide a secondary metabolite of Pesudomonas aeruginosa. Can. J. Microbial. 21: 613-618. De Marco, J.L., and Felix, C.R. (2007). Purification and Characterization of a â-Glucanase produced by Trichoderma harzianum showing biocontrol potential. Braz. Arch. Biol. Technol. 50: 21-29.
    8. De Souza, J.T., Bailey, B.A., Pomella, A.W.V., Erbe, E.F., Murphy, C.A., Bae, H., and Hebbar, P.K. (2008). Colonization of cocoa seedlings by Trichoderma stromaticum, a mycoparasite of the witches’ broom pathogen, and its influence on plant growth and resistance. Biol. Control. 46: 36-45.
    9. Dennis, C., and Webster, J. (1971). Antagonistic properties of species groups of Trichoderma II. Production of volatile antibiotics. Trans. Br. Mycol. Soc. 57: 41–48.
    10. Diby, P., Saju, K.A., Jisha, P.J., Sarma, Y.R., Kumar, A., and Anandaraj, M. (2005). Mycolytic enzymes produced by Pseudomonas fluorescens and Trichoderma spp. against Phytophthora capsici, the foot rot pathogen of black pepper (Pipernigrum L.). Ann. Microbiol. 55: 129-133.
    11. Elad, Y., Chet, I., and Henis, Y. (1982). Degradation of plant pathogenic fungi by Trichoderma harzianum. Can. J. Microbiol. 28: 719-725.
    12. El-Katatny, M.H., Gudelj, M., Robra, K.H., Elnaghy, M.A., and Gubitz, G.M. (2001). Charaterization of a chitinase and an endo-b-1,3-glucanase from T. harzianum Rifai T-24 involved in control of the phytopathogen Sclerotium rolfsii. Appl. Microbiol. Biot. 56: 137-143.
    13. Fernando, W.D.G., Nakkeeran, S., Zhang, Y., and Savchuk, S. (2007). Biological control of Sclerotinia sclerotiorum (Lib.) de Bary by Pseudomonas and Bacillus species on canola petals. Crop. Prot. 26: 100-107.
    14. Gravel, V., Antoun, H., and Tweddell, R.J. (2007). Growth stimulation and fruit yield improvement of greenhouse tomato plants by inoculation with Pseudomonas putida or Trichoderma atroviride: possible role of indole acetic acid (IAA). Soil. Biol. and Biochem. 39: 1968-1977.
    15. Harman, G.E., Chet, I., and Baker, R. (1980). Trichoderma hamatum effects on seed and seedlings disease induced in radish and pea by Pythium spp. or Rhizoctonia solani. Phytopathology. 70: 1167-1172.
    16. Hassanein, W.A., Awny, N.M., El-Mougith, A.A., and Salah El-Dien, S.H. (2009). The antagonistic activities of some metabolites produced by Pseudomonas aeruginosa Sha8. J. Appl. Sc. Res. 5: 404-414.
    17. Henri, F., Laurette, N.N., Annette, D., John, Q., Wolfgang, M., Francois-Xavier, E., and Dieudonne, N. (2008). Solubilization of inorganic phosphates and plant growth promotion by strains of Pseudomonas fluorescens isolated from acidic soils of Cameroon. Af. J. Microbiol. Res. 2: 171-178.
    18. Hernandez, C.F.D., Berlanga, P.A.M., Gallegos, M.G., and Cepeda, S.M. (2011). In vitro antagonist action of Trichoderma strains against Sclerotinia sclerotiorum and Sclerotium cepivorum. Am. J. Agric. Biol. Sc. 6: 410-417. Hoyos-Carvajal, L., Orduz, S., and Bissett, J. (2009). Growth stimulation in bean (Phaseolus vulgaris L.) by Trichoderma. Biol. Control. 51: 409-416.
    19. Huang, H.C., and Haes, J.A. (1976). Penetration and infection of Sclerotinia sclerotiorum by Coniothyrium minitans. Can. J. Bot. 54: 406-410.
    20. Johnson, L.A. (1957). Effect of antibiotics on the number of bacteria and fungi isolated from soil by dilution plate method. Phytopathology. 47: 21-22.
    21. Johnson, D.A., and Atallah, Z.K. (2006). Timing fungicide applications for managing Sclerotinia stem rot of potato. Plan. Dis. 90: 755-758.
    22. Kapri, A., and Tewari, L. (2010). Phosphate solubilization potential and phosphatise activity of rhizospheric Trichoderma spp. Brazil. J. Microbiol. 41: 787-795.
    23. Little, T.M., and Hills, F.J. (1978). Agricultural experimentation: design and analysis. John Wiley and Sons Inc, Indianapolis Ind Mehta, S., and Nautiyal, C.S. (2001). An efficient method for qualitative screening of phosphate-solubilizing bacteria. Curr. Microbiol. 43: 51–56.
    24. Miller, G.L. (1959). Use of dinitrosalicylic acid reagent for the determination of reducing sugar. Anal. Chem. 31: 426- 428.
    25. Pareek, R.P., and Gaur, A.C. (1973). Release of phosphorus from tricalcium phosphate and rock phosphate by organic acids. Curr. Sci. 43: 278–279.
    26. Patten, C.L., and Glick, B.R. (2002). Role of Pseudomonas putida indole acetic acid in development of the host plant root system. App. Environ. Microb. 68: 3795-3801.
    27. Rini, C.R., and Sulochana, K.K. (2007). Usefullness of Trichoderma and Pseudomonas against Rhizoctonia solani and Fusarium oxysporum infecting tomato. J. Trop. Agric. 45: 21-28.
    28. Rini, C.R., and Sulochana, K.K. (2006). Management of seedling rot of chilli (Capsicum annuum L.) using Trichoderma spp. and fluorescent pseudomonads (Pseudomonas fluorescens). J. Trop. Agric. 44: 79-82.
    29. Rudresh, D.L., Shivaprakash, M.K., and Prasad, R.D. (2005). Tricalcium phosphate solubilizing abilities of Trichoderma spp. in relation to P uptake and growth and yield parameters of chickpea (Cicer arietinum L.). Can. J. Microbiol. 51: 217–222.
    30. Schwyn, B., and Neilands, J.B. (1987). Universal chemical assay for the detection and determination of siderophores. Anal. Biochem. 160: 47-56.
    31. Sandhya, C., Adapa, L.K., Nampoothiri, M., Binod, P., Szakacs, G., and Pandey, A. (2004). Extracellular chitinase production by Trichoderma harzianum in submerged fermentation. J. Basic. Microbiol. 44: 49-58.
    32. Shaigan, S., Seraji, A., and Moghaddam, S.A.M. (2008). Identification and investigation on antagonistic effect of Trichoderma spp. on tea seedlings white foot and root rot (Sclerotium rolfsii Sacc) in vitro condition. Pakistan. J. Biol. Sci. 11: 3246-3250.
    33. Vincent, J.H. (1947). Distortion of fungal hyphae in the presence of certain inhibitors. Nature. 15: 850. Viterbo, A., Ramot, O., Chernin, L., and Chet, I. (2002). Significance of lytic enzymes from Trichoderma spp. in the biocontrol of fungal plant pathogens. Van. Leeuw. J. Microb. 81: 549-556.
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