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volume 36 issue 6 (december 2013) : 569-575

ANTAGONISTIC EFFECT OF PSEUDOMONAS FLUORESCENS AGAINST FUSRIUM OXYSPORUM F.SP. CICERI CAUSING WILT IN CHICKPEA

U
U.K. Kandoliya*
D
D.N. Vakharia
1Department of Biotechnology, College of Agriculture, Junagadh Agricultural University, Junagadh- 362 001, India

  • Submitted|

  • First Online |

  • doi

Cite article:- Kandoliya* U.K., Vakharia D.N. (2025). ANTAGONISTIC EFFECT OF PSEUDOMONAS FLUORESCENS AGAINST FUSRIUM OXYSPORUM F.SP. CICERI CAUSING WILT IN CHICKPEA. Legume Research. 36(6): 569-575. doi: .

ABSTRACT

Biocontrol agent, Pseudomonas fluorescens  which inhibit the growth of fungal pathogen Fusarium oxysporum f. sp. ciceri  in vitro on potato dextrose agar + Kings’B  (1:1) medium, were evaluated for their potential to produce various extra cellular enzymes at 6 days after inoculation (DAI). The highest extra cellular activities of cellulase and polygalacturonase (PG) were seen in the culture medium of F. oxysporum compared to culture media of sole F. oxysporum  and  P.  fluorescens treatments. Among the ten isolates of biocontrol agents, Pf-3 showed highest per cent inhibition of growth of the pathogen on culture medium. The per cent inhibition was positively correlated with extra cellular specific activities of protease, chitinase and b-1, 3-glucanase. The possible involvements of these enzymes in the antagonistic interaction between P. fluorescens and F. oxysporum f. sp. ciceri were observed. This suggested that isolates of P. fluorescens was suitable to be used in biological control of plant pathogen F. oxysporum f. sp. ciceri. Moreover, the substance like salicylic acid, siderophore and IAA production by Pf-3 exhibited its efficacy to work as biocontrol agent as well as PGPR.

REFERENCES

  1. Boller, T. and Mauch, F. (1988). Colorimetric assay of chitinase. Meth Enzymol 161:431-5.
  2. Buyer, J.S. and Leong, J. (1986). Iron trans- port mediated antagonism between plant growth promoting and plant deleterious Pseu- domonas strains. J Biol Chemist., 261: 791-794.
  3. Collmer, A., Reid, J. L. and Mount, M. S. (1988). Assay methods for pectic enzymes. Meth Enzymol 161:329-35.
  4. Dave, B. P. and Dube, H. (2000). Detection and chemical characterisation of siderophores of rhizobacterial fluorescent pseudomonads. Indian. Phytopathol., 53: 97-98.
  5. DeMeyer, G. and Hofte, M. (1997). Salicylic acid produced by the rhizobacterium P. auregenosa 7 NSK 2 induces resistance to leaf infection by Botrytis cinerea on bean. Phytopathol., 87: 588-593.
  6. Emmert, E.A.B., and Handelsman, J. (1999). Biocontrol of plant disease: a (Gram) positive perspective. FEMS Microbiology letters, 171: 1-9.
  7. Inam-ul-Haq, M., Javed, N., Ahmad, R. and Rehman, A. (2003). Evaluation of different strain of pseudomonas fluorescens for the control of fusarium wilt of chickpea. Pak. J. Pl. Patholo., 2(a): 65-74.
  8. Gupta, S. C. and Sahu, S. (2012). Response of chickpea to micronutrients and biofertilizers in vertisol. Legume Res., 35 (3): 248-251.
  9. Jagdees, K. S., Kulkarni, J. H. and Krishnaraj, P. U. (2001). Evaluation of the role of fluorescent siderophore in the, biological control of bacterial wilt in tomato using Tn5 mutants of fluorescent Pseudomonas sp. Curr. Sci., 81(8): 56-59.
  10. Kauffman, S., Legrand, M., Geoffory, P. and Fritig, B. (1987). Biological functions of ‘pathogenesis related’ proteins four PR proteins of tobacco have â-1,3-glucanase activity. EMBO J. 6: 3209-12.
  11. Kaur, R., Kaur, J., Singh, R. S. and Alabouvette, C. (2007). Biological control of Fusarium oxysorum f.sp. ciceri by nonpathogenic Fusarium and Fluorescent Pseudomonas. Int. J. of Bot. 3(1): 114-113.
  12. Kishore, G. K., Pande, S. and Podile, A. R. (2006). Pseudomonas aeruginosa GSE 18 inhibits the cell wall degrading enzymes of Aspergillus niger and activates defence-related enzymes of groundnut in control of collar rot disease. Aus. Pl. Patho., 35: 259-263.
  13. Malick C P, and Singh M. B. (1980). Plant Enzymology and Histo enzymology. Ludhiana: Kalyani Publishers, Ludhiana. pp 54-56, 71-72.
  14. Maurhofer, M., Reimmann, C., Schmidti-Sacherer, P., Heeb, S., Haas, D. and Defago, G. (1998). Salicylic acid biosynthesis genes expressed in Pseudomonas fluorescens strain P3 improve the induction of systemic resistance in tobacco against tobacco mosaic virus. Phytopath., 88: 678-684.
  15. Mazumdar, T., Goswami, C. and Talukdar, N. C. (2007). Characterization and screening of beneficial bacteria obtained on King’s B agar from tea rhizosphere. Indian. J. Biotech., 6: 490-494.
  16. Meyer, J.M., Ajelvandre, P. and Georges, C. (1992). Iron metabolism in Pseudomonas: Salicylic acid, a siderophore of Pseudomonas fluorescens CHA0. Biofactors, 4: 23-27.
  17. Nandakumar, R., Babu, S., Raguchander, T. and Samiyappan, R. (2007). Chitinolytic Activity of Native Pseudomonas fluorescens Strains. J. Agric. Sci. Techno., 9: 61-68.
  18. Nene, Y. L., Sheila, V. K. and Sharma, S. B. A. (1996). A world list of chickpea and pigeonpea pathogens, ICRISAT, Patencheru, 5: 27-29.
  19. Nielsen, M. N. and Sorensen, J. (1999). Chitinolytic activity of Pseudomonas fluorescens isolates from barley and sugar beet rhizosphere. FEMS microbio. Eco., 30(3): 217-227.
  20. Pal, K. K., Dey, R., Bhatt, D. M. and Chauhan, S. M. (1999). Enhancement of groundnut growth and yield by plant growth promoting rhizobacteria, Int. Arch. News L., 19: 51-53.
  21. Ramnathan, G., Banupriya, S. and Birami, D. A. (2009). Production and optimization of cellulose from Fusarium oxysporum by submerged fermentation. J. Sci & Ind. Res., 69: 454-459.
  22. Reddy, B. P., Reddy, K. R. N., Rao, M. S. and Rao, K. S. (2008). Efficacy of Antimicrobial Metabolites of Pseudomonas fluorescens Against Rice Fungal Pathogens. Curr. Tre. in Biotech. and Pharma., 2: 178-182.
  23. Reeves, M., Pine, L., Neilands, J.B. and Bullows, A. (1983). Absence of siderophore activity in Legionella spp. grown in iron deficient media. J Bacteriology., 154: 324-329.
  24. Reissig J L , Strominger J L, and Lefloir L F (1955). A modified colorimetric method for the estimation of N-acetyl amino sugars. J Biol Chem 217:959-66.
  25. Sadasivam S, and Manickam K. (1992). Biochemical Method for Agricultural Sciences. Wiley Estern Limited, Coimbatore. pp.116-117.
  26. Saikia, R., Singh, B. P., Kumar, R. and Arora, D. K. (2005). Detection of pathogenesis related proteins-chitinase and â- 1, 3-glucanase in induced chickpea. Curr. Sci., 89: 659-663.
  27. Saikia, R., Singh, T., Kumar, R., Srivastava, J., Srivastava, A. K., Singh, K. and Arora, D. K. (2003). Role of salicylic acid in systemic resistance induced by Pseudomonas fluorescens against Fusarium oxysporum f. sp. ciceri in chickpea. Microbio. Res., 158: 203-213.
  28. Simon, A. and Ridge, E. H. (1974). The use of ampicillin in a simplified selective medium for the isolation of fluorescent Pseudomonas. J. Appl. Bacterio., 37: 459-460.
  29. Singh, U. P., Sarma, B. K., and Singh, D. P. (2003). Effect of plant growth promoting rhizobacteria and culture filtrate of Sclerotium rolfsii on phenolic and salicylic acid contents in chickpea (Cicer arietinum). Curr. Microbio., 46: 131-140.
  30. Subramanian, C.V. (1954). Studies on south India Fusarium III. Fusarium isolated from some crop plants. J. Madras. Univ., 24:21-46.
  31. Sullivan, D.J. and Gara, F. (1992). Traits of fluorescent pseudomonads involved in suppres- sion of plant root pathogens. Microbiological Review., 56: 662-676.
  32. Velzahan, R., Samiyappan, R. and Vidyasekaran, P. (1999). Relationship between antagonistic activities of P. fluorescens isolates against R. solani and their production of lytic enzymes. J Pl. Dis. Prot., 106: 244-250.
  33. Vidhyasekaran, P. and Muthalian, M. (1995). Development of formulations of Pseudomonas fluorescens for control of chickpea wilt. Pl. Dis., 79(8): 782-786.
  34. Viswanathan, R. and Samiyappan, R. (2001). Antifungal activity of chitinase produced by Fluorescent Pseudomonads against Colletotrichum falcaturn Went causing red root disease in sugarcane. Microbio. Res., 155: 305-314.
  35. Vlassak, K., Van holm, L., Duchateau, L., Vanderleyden, J. and De Mot, R. (1992). Isolation and characterization of fluorescent Pseudomonas associated with the roots of rice and banana grown in Sri Lanka. Pl. Soil, 145: 51-63.
  36. Yasmin, F., Radziah, O., Sijam, K. and Saad, M. S., (2009). Characterization of beneficial properties of plant growth- promoting rhizobacteria isolated from sweet potato rhizosphere, Afri. J. of Microbio. Res., 3(11): 637-640.
  37. Zarrin F, Saleemi, M., Zia, M., Sultan, T., Aslam, M. , Rehman, R. and Chaudhary, M.F. (2009). Antifungal activity of plant growth-promoting rhizobacteria isolates against Rhizoctonia solani in wheat. Afri. J. Biotech., 8(2): 219–225.
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    volume 36 issue 6 (december 2013) : 569-575

    ANTAGONISTIC EFFECT OF PSEUDOMONAS FLUORESCENS AGAINST FUSRIUM OXYSPORUM F.SP. CICERI CAUSING WILT IN CHICKPEA

    U
    U.K. Kandoliya*
    D
    D.N. Vakharia
    1Department of Biotechnology, College of Agriculture, Junagadh Agricultural University, Junagadh- 362 001, India

    • Submitted|

    • First Online |

    • doi

    Cite article:- Kandoliya* U.K., Vakharia D.N. (2025). ANTAGONISTIC EFFECT OF PSEUDOMONAS FLUORESCENS AGAINST FUSRIUM OXYSPORUM F.SP. CICERI CAUSING WILT IN CHICKPEA. Legume Research. 36(6): 569-575. doi: .

    ABSTRACT

    Biocontrol agent, Pseudomonas fluorescens  which inhibit the growth of fungal pathogen Fusarium oxysporum f. sp. ciceri  in vitro on potato dextrose agar + Kings’B  (1:1) medium, were evaluated for their potential to produce various extra cellular enzymes at 6 days after inoculation (DAI). The highest extra cellular activities of cellulase and polygalacturonase (PG) were seen in the culture medium of F. oxysporum compared to culture media of sole F. oxysporum  and  P.  fluorescens treatments. Among the ten isolates of biocontrol agents, Pf-3 showed highest per cent inhibition of growth of the pathogen on culture medium. The per cent inhibition was positively correlated with extra cellular specific activities of protease, chitinase and b-1, 3-glucanase. The possible involvements of these enzymes in the antagonistic interaction between P. fluorescens and F. oxysporum f. sp. ciceri were observed. This suggested that isolates of P. fluorescens was suitable to be used in biological control of plant pathogen F. oxysporum f. sp. ciceri. Moreover, the substance like salicylic acid, siderophore and IAA production by Pf-3 exhibited its efficacy to work as biocontrol agent as well as PGPR.

    REFERENCES

    1. Boller, T. and Mauch, F. (1988). Colorimetric assay of chitinase. Meth Enzymol 161:431-5.
    2. Buyer, J.S. and Leong, J. (1986). Iron trans- port mediated antagonism between plant growth promoting and plant deleterious Pseu- domonas strains. J Biol Chemist., 261: 791-794.
    3. Collmer, A., Reid, J. L. and Mount, M. S. (1988). Assay methods for pectic enzymes. Meth Enzymol 161:329-35.
    4. Dave, B. P. and Dube, H. (2000). Detection and chemical characterisation of siderophores of rhizobacterial fluorescent pseudomonads. Indian. Phytopathol., 53: 97-98.
    5. DeMeyer, G. and Hofte, M. (1997). Salicylic acid produced by the rhizobacterium P. auregenosa 7 NSK 2 induces resistance to leaf infection by Botrytis cinerea on bean. Phytopathol., 87: 588-593.
    6. Emmert, E.A.B., and Handelsman, J. (1999). Biocontrol of plant disease: a (Gram) positive perspective. FEMS Microbiology letters, 171: 1-9.
    7. Inam-ul-Haq, M., Javed, N., Ahmad, R. and Rehman, A. (2003). Evaluation of different strain of pseudomonas fluorescens for the control of fusarium wilt of chickpea. Pak. J. Pl. Patholo., 2(a): 65-74.
    8. Gupta, S. C. and Sahu, S. (2012). Response of chickpea to micronutrients and biofertilizers in vertisol. Legume Res., 35 (3): 248-251.
    9. Jagdees, K. S., Kulkarni, J. H. and Krishnaraj, P. U. (2001). Evaluation of the role of fluorescent siderophore in the, biological control of bacterial wilt in tomato using Tn5 mutants of fluorescent Pseudomonas sp. Curr. Sci., 81(8): 56-59.
    10. Kauffman, S., Legrand, M., Geoffory, P. and Fritig, B. (1987). Biological functions of ‘pathogenesis related’ proteins four PR proteins of tobacco have â-1,3-glucanase activity. EMBO J. 6: 3209-12.
    11. Kaur, R., Kaur, J., Singh, R. S. and Alabouvette, C. (2007). Biological control of Fusarium oxysorum f.sp. ciceri by nonpathogenic Fusarium and Fluorescent Pseudomonas. Int. J. of Bot. 3(1): 114-113.
    12. Kishore, G. K., Pande, S. and Podile, A. R. (2006). Pseudomonas aeruginosa GSE 18 inhibits the cell wall degrading enzymes of Aspergillus niger and activates defence-related enzymes of groundnut in control of collar rot disease. Aus. Pl. Patho., 35: 259-263.
    13. Malick C P, and Singh M. B. (1980). Plant Enzymology and Histo enzymology. Ludhiana: Kalyani Publishers, Ludhiana. pp 54-56, 71-72.
    14. Maurhofer, M., Reimmann, C., Schmidti-Sacherer, P., Heeb, S., Haas, D. and Defago, G. (1998). Salicylic acid biosynthesis genes expressed in Pseudomonas fluorescens strain P3 improve the induction of systemic resistance in tobacco against tobacco mosaic virus. Phytopath., 88: 678-684.
    15. Mazumdar, T., Goswami, C. and Talukdar, N. C. (2007). Characterization and screening of beneficial bacteria obtained on King’s B agar from tea rhizosphere. Indian. J. Biotech., 6: 490-494.
    16. Meyer, J.M., Ajelvandre, P. and Georges, C. (1992). Iron metabolism in Pseudomonas: Salicylic acid, a siderophore of Pseudomonas fluorescens CHA0. Biofactors, 4: 23-27.
    17. Nandakumar, R., Babu, S., Raguchander, T. and Samiyappan, R. (2007). Chitinolytic Activity of Native Pseudomonas fluorescens Strains. J. Agric. Sci. Techno., 9: 61-68.
    18. Nene, Y. L., Sheila, V. K. and Sharma, S. B. A. (1996). A world list of chickpea and pigeonpea pathogens, ICRISAT, Patencheru, 5: 27-29.
    19. Nielsen, M. N. and Sorensen, J. (1999). Chitinolytic activity of Pseudomonas fluorescens isolates from barley and sugar beet rhizosphere. FEMS microbio. Eco., 30(3): 217-227.
    20. Pal, K. K., Dey, R., Bhatt, D. M. and Chauhan, S. M. (1999). Enhancement of groundnut growth and yield by plant growth promoting rhizobacteria, Int. Arch. News L., 19: 51-53.
    21. Ramnathan, G., Banupriya, S. and Birami, D. A. (2009). Production and optimization of cellulose from Fusarium oxysporum by submerged fermentation. J. Sci & Ind. Res., 69: 454-459.
    22. Reddy, B. P., Reddy, K. R. N., Rao, M. S. and Rao, K. S. (2008). Efficacy of Antimicrobial Metabolites of Pseudomonas fluorescens Against Rice Fungal Pathogens. Curr. Tre. in Biotech. and Pharma., 2: 178-182.
    23. Reeves, M., Pine, L., Neilands, J.B. and Bullows, A. (1983). Absence of siderophore activity in Legionella spp. grown in iron deficient media. J Bacteriology., 154: 324-329.
    24. Reissig J L , Strominger J L, and Lefloir L F (1955). A modified colorimetric method for the estimation of N-acetyl amino sugars. J Biol Chem 217:959-66.
    25. Sadasivam S, and Manickam K. (1992). Biochemical Method for Agricultural Sciences. Wiley Estern Limited, Coimbatore. pp.116-117.
    26. Saikia, R., Singh, B. P., Kumar, R. and Arora, D. K. (2005). Detection of pathogenesis related proteins-chitinase and â- 1, 3-glucanase in induced chickpea. Curr. Sci., 89: 659-663.
    27. Saikia, R., Singh, T., Kumar, R., Srivastava, J., Srivastava, A. K., Singh, K. and Arora, D. K. (2003). Role of salicylic acid in systemic resistance induced by Pseudomonas fluorescens against Fusarium oxysporum f. sp. ciceri in chickpea. Microbio. Res., 158: 203-213.
    28. Simon, A. and Ridge, E. H. (1974). The use of ampicillin in a simplified selective medium for the isolation of fluorescent Pseudomonas. J. Appl. Bacterio., 37: 459-460.
    29. Singh, U. P., Sarma, B. K., and Singh, D. P. (2003). Effect of plant growth promoting rhizobacteria and culture filtrate of Sclerotium rolfsii on phenolic and salicylic acid contents in chickpea (Cicer arietinum). Curr. Microbio., 46: 131-140.
    30. Subramanian, C.V. (1954). Studies on south India Fusarium III. Fusarium isolated from some crop plants. J. Madras. Univ., 24:21-46.
    31. Sullivan, D.J. and Gara, F. (1992). Traits of fluorescent pseudomonads involved in suppres- sion of plant root pathogens. Microbiological Review., 56: 662-676.
    32. Velzahan, R., Samiyappan, R. and Vidyasekaran, P. (1999). Relationship between antagonistic activities of P. fluorescens isolates against R. solani and their production of lytic enzymes. J Pl. Dis. Prot., 106: 244-250.
    33. Vidhyasekaran, P. and Muthalian, M. (1995). Development of formulations of Pseudomonas fluorescens for control of chickpea wilt. Pl. Dis., 79(8): 782-786.
    34. Viswanathan, R. and Samiyappan, R. (2001). Antifungal activity of chitinase produced by Fluorescent Pseudomonads against Colletotrichum falcaturn Went causing red root disease in sugarcane. Microbio. Res., 155: 305-314.
    35. Vlassak, K., Van holm, L., Duchateau, L., Vanderleyden, J. and De Mot, R. (1992). Isolation and characterization of fluorescent Pseudomonas associated with the roots of rice and banana grown in Sri Lanka. Pl. Soil, 145: 51-63.
    36. Yasmin, F., Radziah, O., Sijam, K. and Saad, M. S., (2009). Characterization of beneficial properties of plant growth- promoting rhizobacteria isolated from sweet potato rhizosphere, Afri. J. of Microbio. Res., 3(11): 637-640.
    37. Zarrin F, Saleemi, M., Zia, M., Sultan, T., Aslam, M. , Rehman, R. and Chaudhary, M.F. (2009). Antifungal activity of plant growth-promoting rhizobacteria isolates against Rhizoctonia solani in wheat. Afri. J. Biotech., 8(2): 219–225.
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