Evaluation of Indigenous Fluorescent Pseudomonads for the Management of Newly Emerging Wilt of Pomegranate Caused by Ceratocystis fimbriata

  • Gururaj Sunkad
  • T. H. Shruthi
  • Raja


Pomegranate (Punica granatum L.) is a commercial fruit crop in India and regarded a vital cash crop in many states including Karnataka. Wilt disease (Ceratocystis fimbriata Elli. and Halst.) is a new devastating disease on pomegranate that caused losses estimated up to 30% and becoming a major threat leading to the destruction of several pomegranate orchards. Although suitable wilt management practices including cultural, sanitation, and chemical have been developed, there is still a need to develop more economical, feasible, and effective environmentally friendly alternative control methods. Recently, there is an increased interest for use of plant growth-promoting rhizobacteria (PGPR) for sustainable pomegranate cultivation. Thus, the present investigation is intended to evaluate indigenous fluorescent pseudomonad isolates for biocontrol, PGPR traits, and plant growth promotion. In the present investigation, 35 native isolates of fluorescent pseudomonads from wilt-affected pomegranate orchards were isolated, purified, and characterized. These were screened for their in vitro efficacy using standard dual culture plate culture technique for the inhibition of the growth of C. fimbriata. Out of 35 isolates, 11 isolates (PFP-1–PFP-11) significantly reduced the vegetative growth of C. fimbriata. These isolates were tested for PGPR traits such as the production of indoleacetic acid (IAA), hydrogen cyanide (HCN), hydrogen sulfide, and siderophore. All the 11 isolates showed bright fluorescence under UV light, and morphological and biochemical tests proved them as Pseudomonas fluorescens. The isolates were positive for siderophore production and plant growth-promoting activities. Two isolates PFP-11 and PFP-10 showed remarkable antifungal activity against C. fimbriata with the inhibition of 72.04% and 71.20%, respectively. These isolates showed significant producers of HCN, IAA, siderophore, and enzymes. Overall, our results suggest that these fluorescent pseudomonad isolates can be estimated as efficient bioagents for the sustainable management of newly emerging wilt of pomegranate caused by C. fimbriata and warrant further evaluations.


Fluorescent pseudomonads PGPR Biocontrol Wilt of pomegranate Ceratocystis fimbriata 



The authors are grateful to the vice chancellor and the University of Agricultural Sciences, Raichur-584101, Karnataka, India, for their support.


  1. Blazevic DJ, Ederer GM (1975) Principles of biochemical tests in diagnostic microbiology. Wiley, New York, pp 13–45Google Scholar
  2. Cappuccino JC, Sherman N (1992) Microbiology. A laboratory manual. Benjamin/Cummings Pub. Co, New York, pp 125–179Google Scholar
  3. Cartwright DK, Benson DM (1995) Comparison of Pseudomonas species and application techniques for biocontrol of Rhizoctonia stem rot of poinsettia. Plant Dis 79:309–313CrossRefGoogle Scholar
  4. Eckford MQ (1927) Thermophilic bacteria in milk. Am J Hyg 7:200–202Google Scholar
  5. Garrity GM, Brenner DJ, Krieg NR, Staley JT (2005) Bergey’s manual of systematic bacteriology, 2nd edn. Springer, New York, pp 323–359CrossRefGoogle Scholar
  6. Gupta K, Sahm DF, Mayfield D, Stamm WE (2001) Antimicrobial resistance among uropathogens that cause community-acquired urinary tract infections in women: a nationwide analysis. Clin Infect Dis 33:89–94CrossRefGoogle Scholar
  7. He SY, Huang HC, Collmer A (1993) Pseudomonas syringae happiness: a protein that is secreted via the Hrp pathway and elicits the hypersensitive response in plants. Cell 73:1255–1266CrossRefGoogle Scholar
  8. Laha GS, Verma JP (1998) Role of fluorescent pseudomonads in the suppression of root rot and damping off of cotton. Indian Phytopathol 51:275–278Google Scholar
  9. Liu L, Kloepper JW, Tuzun S (1995) Induction of systemic resistance in cucumber against bacterial leaf spot by plant growth promoting rhizobacteria. Phytopathology 85:843–847CrossRefGoogle Scholar
  10. Maurhofer M, Reimmann C, Sacherer SP, Heeb S, Haas D, Defago G (1998) Salicylic acid biosynthetic genes expressed in Pseudomonas fl fluorescent strain P3 improve the induction of systemic resistance in tobacco against tobacco necrosis virus. Phytopathology 88:678–684CrossRefGoogle Scholar
  11. Mayer AM, Harel E, Shaul RB (1965) Assay of catechol oxidase: a critical comparison of methods. Phytochemistry 5:783–789CrossRefGoogle Scholar
  12. O’Sullivan DJ, O’Gara F (1988) Delivery system for one-step creation of in vivo lac gene fusions in Pseudomonas spp. involved in biological control. Appl Environ Microbiol 54:2877–2880Google Scholar
  13. Radjacommare R, Kandan A, Nandakumar R, Samiyappan R (2004) Association of the hydrolytic enzyme chitinase against Rhizoctonia solani in rhizobacteria-treated rice plants. J Phytopathol 152:365–370CrossRefGoogle Scholar
  14. Raja (2017) Investigations on wilt of pomegranate caused by ceratocystis fimbriata Ell. and Halst. In Karnataka. University of Agricultural Sciences, Ph. D. thesis, Raichur, Karnataka, pp 102–105Google Scholar
  15. Ramette A, Loy M, Defago G (2006) Genetic diversity and biocontrol protection of fluorescens pseudomonas producing phloroglucinols and hydrogen cyanide from swiss soils naturally suppressive or conducive to Thieviopsis basicola mediated black rot of tobacco. FEMS Microbial Ecol 55(3):369–381CrossRefGoogle Scholar
  16. Saranraj P, Sivasakthivelan P, Sakthi SS (2013) Prevalence and production of plant growth promoting substance by Pseudomonas fluorescens isolates from Paddy rhizosphere soil of Cuddalore District, Tamil Nadu, India. Afr J Basic Appl Sci 5(2):95–101Google Scholar
  17. Schaad NW (1992) Laboratory guide for identification of plant pathogenic bacteria. In: Schaad NW (ed) The American Phytopathological Society, Minneapolis, pp 89–94Google Scholar
  18. Sharma KK, Sharma J, Jadhav VT (2008) Prevalence and etiology of wilt of pomegranate. Indian Phytopathol 61:381Google Scholar
  19. Sharma KK (2009) Vascular wilt of pomegranate caused by Ceratocystis fimbriata Ellis and Halsted and its control. In: 5th International conference on plant pathology in the globalized Era, Nov, at IARI, New Delhi, p 240Google Scholar
  20. Shivangi S, Dutta U, Bhat AK, Gupta S, Vikas G, Jamwal S (2017) Morpho-cultural and biochemical identification of Pseudomonas sp. isolated from the rhizosphere of different vegetable crops and study its efficacy on Solanum melongena (Brinjal). J Pharmacol Phytochem 6(2):22–28Google Scholar
  21. Somasekhara YM (1999) New record of Ceratocystis fimbriata causing wilt of pomegranate in India. Plant Dis 83:406CrossRefGoogle Scholar
  22. Sonyal S, Giri MS, Mahesha HS, Palanna KB (2015) Effect of Botanicals and bioagents on the growth of Ceratocystis fimbriata ELL. and Halst. Causing wilt in pomegranate. Int J Pure Appl Biosci 3(4):42–48Google Scholar
  23. Tziros GT, Tzavella-Klonari K (2008) First report of verticillium wilt of pomegranate caused by Verticillium dahlia in Greece. J Plant Pathol 90:589Google Scholar
  24. Vinay JU, Naik MK, Rameshwaram R, Chennappa G, Sohel S, Shaikh RZS (2016) Detection of antimicrobial traits in fluorescent pseudomonads and molecular characterization of an antibiotic pyoluteorin. 3Biotech 6:227Google Scholar
  25. Vincent JM (1947) Distortion of fungal hyphae in presence of certain inhibitors. Nature 159:850CrossRefGoogle Scholar

Copyright information

© Springer Nature Singapore Pte Ltd. 2019

Authors and Affiliations

  • Gururaj Sunkad
    • 1
  • T. H. Shruthi
    • 1
  • Raja
    • 1
  1. 1.Department of Plant Pathology, Agriculture CollegeUniversity of Agricultural SciencesBangaloreIndia

Personalised recommendations