Isolation and screening of Pseudomonas fluorescens isolates against Fusarium oxysporum f. sp. radicis-lycopersici and their effects on seedling growth of Paraserianthes falcataria




Abstract. Yusran Y. 2023. Isolation and screening of Pseudomonas fluorescens isolates against Fusarium oxysporum f.sp. radicis-lycopersici and their effects on seedling growth of Paraserianthes falcataria. Biodiversitas 24: 2294-2301. Pseudomonas fluorescens plays a major role in biological control of pathogens and plant growth promotion as well with various mechanisms. The aim of the present study was to determine the biochemical characteristics of several Pseudomonas spp. isolates and to screen them against root pathogenic fungus Fusarium oxysporum f.sp radicis-lycopersici (FORL) and to evaluate their effect on the growth enhancement of Paraserianthes falcataria (L.) I.C. Nielsen seedlings. About 28 Pseudomonas fluorescens isolates were isolated from the rhizosphere of several plants and identified based on their physiological and morphological characters as well as their biochemical reactions. All isolates were tested for their ability to suppress FORL growth in vitro. All isolates inhibited FORL growth with varying inhibition zones. Six P. fluorescens isolates showed highest inhibition zones, namely TMTP4, TMTP5, TMTP6, SCPA1, SCPB3 and Proradix. The results showed that eight Pseudomonas isolates, namely TMTP4, TMTP5, TMTP6, SCPA1, SCPB3, SMPB3, TMTA2 and Proradix had a significantly different effect compared to the control treatment in increasing the growth of Paraserianthes seedlings. Based on our findings, it was confirmed that some P. fluorescens isolates could be used as potential bio-inoculants for controlling plant diseases caused by FORL and as a biological fertilizer to increase plant growth.


Ahmad F, Ahmad I, Khan MS. 2008. Screening of free living rhizospheric bacteria for their multiple plant growth promoting activities. Microbiol Res 163: 173–181.
Alori ET, Glick BR, Babalola OO. 2017. Microbial Phosphorus Solubilization and Its Potential for Use in Sustainable Agriculture. Front. Microbiol. 8:971. doi: 10.3389/fmicb.2017.00971
Baker KF, Cook RJ. 1982. Examples of biological control. In: Biological control of plant pathogens. The American Phytopathological Society, St. Paul, MN. 61-106.
Bjeli? D, Marinkovi? J, Tintor B, Mrkova?ki N. 2018. Antifungal and plant growth promoting activities of indigenous rhizobacteria isolated from maize (Zea mays L.) rhizosphere. Commun Soil Sci Plant Anal 49: 88–98.
Cook D, Dreyer D, Bonnet D, Howell M, Nony E, VandenBosch K. 1995. Transient induction of a peroxidase gene in Medicago truncatula precedes infection by Rhizobium meliloti. Plant Cell 7, 43–55. doi: 10.1105/tpc.7.1.43.
Cumming JR, Zawaski C, Desai S, Collart FR. 2015. Phosphorus disequilibrium in the tripartite plant-ectomycorrhiza-plant growth promoting rhizobacterial association. J. Soil Sci. Plant Nutr. 15, 464–485. doi: 10.4067/s0718-95162015005000040.
Damiri N, Mulawarman, Effendi RS, 2019. Antagonism of Pseudomonas fluorescens from plant roots to Rigidoporus lignosus pathogen of rubber white roots in vitro. Biodiversitas. 20(6): 1549-1554. DOI: 10.13057/biodiv/d200609
David BV, Chandrasehar G, Selvam PN, 2018. Chapter 10 - Pseudomonas fluorescens: A Plant-Growth-Promoting Rhizobacterium (PGPR) With Potential Role in Biocontrol of Pests of Crops. Crop Improvement Through Microbial Biotechnology. New and Future Developments in Microbial Biotechnology and Bioengineering. 221-243.
Defago G, Berling CH, Burger U, Haas D, Kahr G, Keel C, Voisard C, Wirthner P, Wurthrich B. 1990. Suppression of black root rot of tobacco and other root diseases by strain of Pseudomonas fluorescens: Potential application and mechanisms. In : Biological control of soilborne pathogens. Hornby D, ed. CAB International,Walling Ford, UK, 93 – 108.
De La Fuente L, Tomashow L, Weller D, Bajsa N, Quagliotto L, Chermin L, Arias A. 2004. Pseudomonas fluorescens UP61 isolated from birdsfoot trefoil rhizosphere produces multiple antibiotics extracts and exerts a broad spectrum of biocontrol activity. European J. of Plant Pathol. 110: 671 – 681.
Ghadamgahi F, Tarighi S, Taheri P, Saripella GV, Anzalone A, Kalyandurg PB, Catara V, Ortiz R, Vetukuri RR. 2022. Plant GrowthPromoting Activity of Pseudomonas aeruginosa FG106 and Its Ability to Act as a Biocontrol Agent against Potato, Tomato and Taro Pathogens. Biology. 11, 140. https://
Giles CD, Hsu PC, Richardson AE, Hurst M H, Hill JE. 2014. Plant assimilation of phosphorus from an insoluble organic form is improved by addition of an organic anion producing Pseudomonas sp. Soil Biol. Biochem. 68, 263–269. doi: 10.1016/j.soilbio.2013.09.026.
Gusmiaty, Restu M, Bachtiar B, Larekeng SH. 2019. Gibberellin And IAA Production by Rhizobacteria From Various Private Forest. IOP Conf. Series: Earth and Environmental Science 270.012018 doi:10.1088/1755-1315/270/1/012018
Habibi S, Djedidi S, Prongjunthuek K, Mortuza MF, Ohkama-Ohtsu N, Sekimoto H, Yokoyoma T. 2014. Physiological and genetic characterization of rice nitrogen fixer PGPR isolated from rhizosphere soils of different crops. Plant Soil 379, 51–66. doi: 10.1007/s11104-014-2035-7
Hmouni A, Hajlaoui MR, Mlaiki A. 1996. Résistance de Botrytis cinerea aux benzimidazoles et aux dicarboximides dans les cultures abritées de tomate en Tunisie. OEPP/EPPO Bull 26:697–705
Holt JG, Krieg NR, Sneath PHA, Staley JT, Williams ST. 1994. Bergey’s Manual of Determinative Bacteriology. 9????? ed. Williams and Williams. Baltimore. p. 566
Höfte M. 2019. The Use of Pseudomonas spp. as Bacterial Biocontrol Agents to Control Plant Disease. In Microbial Bioprotectants for Plant Disease Management; Burleigh Dodds Science Publishing: Cambridge, UK. p. 75.
Höfte M. 2021. The use of Pseudomonas spp. as bacterial biocontrol agents to control plant diseases. Chapter taken from: Köhl, J. and Ravensberg, W. (ed.), Microbial bioprotectants for plant disease management, Burleigh Dodds Science Publishing, Cambridge, UK.
Howell CR, Stipanovic RD. 1980. Suppression of Phytium ultimum-induced damping-off of cotton seedlings by Pseudomonas fluorescens and its antibiotic pyoluterin. Phytopathology. 70:712 – 715.
Jarvis WR, Shoemaker RA. 1978. Taxonomic status of Fusarium oxysporum causing foot and root rot of tomato. Phytopathology 68: 1679–1680.
Keel C, Schneider U, Maurhofer M, Voisard C, Laville J, Burger U, Wirthner P, Haas D, Defago G. 1992. Suppression of root diseases by Pseudomonas fluorescens CHAO: importance of the bacterial secondary metabolite 2,4-diacetylphloroglucinol. Mol. Plant-Microbe Interact. 5: 4-13.
King EO, Ward MK, Raney DE. 1954. Two simple media for the demonstration of pyocyanin and fluorescin. J. Lab. Clin. Med. 44, 301-307.
Kloepper JW, Leong J, Teintze T, Schroth MN. 1980. Pseudomonas siderophores: a mechanism explaining disease-suppressive soils. Curr. Microbiol. 4. 317-320.
Kurek J, Kirk JL, Muir DC, Wang X, Evans MS, Smol JP. 2013. Legacy of a half century of Athabasca oil sands development recorded by lake ecosystems. Proc. Natl. Acad. Sci. U.S.A. 110, 1761–1766. doi: 10.1073/pnas.1217675110
Kumari NV, Vickram AS, Sridharan TB. 2020. Plant Growth Promoting Rhizobacteria (PGPR) Pseudomonas stutzeri from forest soil: A Review. European Journal of Molecular & Clinical Medicine. 7(2): 5721-5738.
Lambers H, Mougel C, Jaillard B, Hinsinger P. 2009. Plant-microbesoil interactions in the rhizosphere: an evolutionary perspective. Plant Soil 321, 83–115. doi: 10.1007/s11104-009-0042-x
Li X, Guo H, Qi Y, Liu H, Zhang X, Ma P, Liang Z, Dong J. 2016. Salicylic acid-induced cytosolic acidification increases the accumulation of phenolic acids in Salvia miltiorrhiza cells. Plant Cell Tissue Organ Cult. (PCTOC). 126: 333–341
Liu XX, Jiang XX, He XY, Zhao W. 2019. Phosphate-solubilizing Pseudomonas sp. strain P34-L promotes wheat growth by colonizing the wheat rhizosphere and improving the wheat root system and soil phosphorus nutritional status. J Plant Growth Regul 38: 1314–1324.
Lemaire JM, Lot M, Biancard D, Leong H. 1988. Disease in protected crops-relations with integrated crop protection. EUR-report, EUR-9386E, 134 – 137.
Lugtenberg B. 2015. Life of microbes in the rhizosphere. In: B. Lugtenberg ed. Principles of Plant-Microbe Interactions. Springer International Publishing Switzerland, Heidelberg, pp. 7-15
Marathe RJ, Phatake YB, Shaikh AC, Shinde BP, Gajbhiye MH. 2017. Effect of IAA produced by Pseudomonas aeruginosa 6A (BC4) on seed germination and plant growth of Glycin max. Journal of Experimental Biology and Agricultural Sciences. 5(3): 352-358.
Malmierca MG, Cardosa RE, Alexander NJ, McComick SP, Hermosa R, Monte E, Gutierrez S. 2012. Involvement of Trichoderma trichothecenesin the biocontrol activity and in the induction of plant defense-related genes. Appl Environ Microb.78(14): 456-486.
Maurhofer M, Keel C, Haas D, Defago G. 1994. Pyoluterin production by Pseudomonas fluorescens strain CHAO is involved in the suppression of Phytium damping-off of cress but not cucumber. Eur. J. Plant Pathol. 100: 221 – 232.
McKellar RC. 1992. Factors influencing the production of extracellular proteinase by Pseudomonas fluorescens. Journal of Applied Bacteriology. 53: 305-316.
Meyer JM. 2000. Pyoverdines: pigments, siderophores and potential taxonomic markers of fluorescent Pseudomonas species. Arch. Microbiol. 174(3):135–142. https://doi .org/10.1007/s002030000188.
Millar RL, Higgins VJ. 1970. Association with infection of birdsfoot trefoil by Stemphylium loti. Phytopathology. 60: 104 – 110.
Morgan JL, Darling AE, Eisen JA. 2010. Metagenomic sequencing of an in vitro-simulated microbial community. PLoS One 5:e10209. doi: 10.1371/journal.pone.0010209.
Nagarajkumar M, Bhaskaran R, Velazhahan R. 2004. Involvement of secondary metabolites and extracellular lytic enzymes produced by Pseudomonas fluorescens in inhibition of Rhizoctonia solani, the rice sheath blight pathogen. Microbiol Res 159: 73–81.
Nerek E, Soko?owska B. 2022. Pseudomonas spp. in biological plant protection and growth promotion. AIMS Environmental Science. 9(4): 493–504. DOI: 10.3934/environsci.2022029
Oni FE, Esmaeel Q, Onyeka JT, Adeleke R, Jacquard C, Clement C, Gross H, Barka EA, Höfte M. 2022. Pseudomonas Lipopeptide-Mediated Biocontrol: Chemotaxonomy and Biological Activity. Molecules 2022, 27, 372. molecules27020372
Panth M, Hassler SC, Baysal-Gurel F. 2020. Methods for Management of Soilborne Diseases in Crop Production. Agriculture, 10(1), 16. https://
Pastor N, Rosas S, Luna, V, Rovera M. 2014. Inoculation with Pseudomonas putida PCI2, a phosphate solubilizing rhizobacterium, stimulates the growth of tomato plants. Symbiosis 62, 157–167. doi: 10.1007/s13199-014-0281-3
Peix A, Ramírez-Bahena MH, Velázquez E. 2009. Historical evolution and current status of the taxonomy of genus Pseudomonas. Infect. Genet. Evol. 9(6), 1132–1147.
Pothiraj GA, Kamalakkannan V, Amirthalingam, Balamurugan A. 2018. Screening of Pseudomonas fluorescens against Dry Root Rot Pathogen Macrophomina phaseolina in Black Gram. Int. J. Curr. Microbiol. App. Sci. 7(09): 3300-3307. doi:
Press CM, Wilson M, Tuzun S, Kloepper JW. 1997. Salicylic acid produced by Serratia marcescens 90-166 is not the primary determinant of induced systemic resistance in Cucumber or Tobacco. Mol. Plant Microbe Interact. 10: 761–768.
Radhapriya P, Ramachandran A, Anandham R, Mahalingam S. 2015. Pseudomonas aeruginosa RRALC3 Enhances the Biomass, Nutrient and Carbon Contents of Pongamia pinnata Seedlings in Degraded Forest Soil. PLoS ONE 10(10): e0139881. doi:10.1371/journal.pone.0139881
Rathore R, Vakharia DN, Rathore DS. 2020. In vitro screening of different Pseudomonas fluorescens isolates to study lytic enzyme production and growth inhibition during antagonism of Fusarium oxysporum f. sp. cumini, wilt causing pathogen of cumin. Egyptian Journal of Biological Pest Control. 30:57
Saha M, Maurya BR, Meena VS, Bahadur I, Kumar A. 2016. Identification and characterization of potassium solubilizing bacteria (KSB) from Indo-Gangetic Plains of India. Biocatal Agric Biotechnol 7: 202–209.
Saraf M, Jha CK, Patel D. 2010. Plant Growth and Health Promoting Bacteria. Springer, Berlin. 365–385.
Saravanan VS, Subramoniam SR, Raj SA. 2003. Assessing in vitro solubilization potential of different zinc solubilizing bacterial (ZSB) isolates. Braz J Microbiol 34: 121–125.
Szczechura W, Staniaszek M, Habdas H. 2013. Fusarium oxysporum f. sp. radicis-lycopersici – The cause of Fusarium Crown and Root Rot in Tomato cultivation. J of. Plant Protection Research. 53(2): 172-176. DOI: 10.2478/jppr-2013-0026
Sulochana MB, Jayachandra SY, Kumar SA, Parameshwar AB, Reddy KM, Dayanand A. 2014. Siderophore as a Potential Plant Growth-Promoting Agent Produced by Pseudomonas aeruginosa JAS-25. Appl Biochem Biotechnol. 174: 297–308. DOI 10.1007/s12010-014-1039-3
Thomashow LS, Weller DM. 1988. Role of a phenazine antibiotic from Pseudomonas fluorescens in biological control of Gaeumannomyces graminis var. tritici. Journal of Bacteriology.170: 3499 – 3508.
Velazhahan R, Samiyappan R, Vidhyasekaran P. 1999. Relationship between antagonistic activities of Pseudomonas fluorescens isolates against Rhizoctonia solani and their production of lytic enzymes. J Plant Dis Prot 106: 244–250.
VDLUFA. 2007. Methodenbuch Bd. 1: Die Untersuchung von Böden. VDLUFA-Verlag, Darmstadt.
Wang B, Jeffers SN. 2000. Fusarium root and crown rot: a disease of container-grown hosts. Plant Dis. 84:980 – 988.
Weller DM. 2007. Pseudomonas Biocontrol Agents of Soilborne Pathogens: Looking Back Over 30 Years. Phytopathology. 97:250–256
Whipps JM. 2001. Microbial interactions and biocontrol in the rhizosphere. J.of. Experimental Botany. 52: 487 – 451.
Wood WA. 1988. Lignin, pectin and chitin. In Methods in Enzymology, vol.161. Academic Press.
Yusran Y, Roemheld V, Mueller T. 2009. Effects of Plant Growth-Promoting Rhizobacteria and Rhizobium on Mycorrhizal Development and Growth of Paraserianthes falcataria (L.) Nielsen Seedlings in Two Types of Soils with Contrasting levels of pH. UC Davis The Proceedings of the International Plant Nutrition Colloquium XVI.
Zhao LF, Xu YJ, Ma ZQ, Deng ZS, Shan CJ, Wei GH. 2013. Colonization and plant growth promoting characterization of endophytic Pseudomonas chlororaphis strain Zong1 isolated from Sophoraalopecuroides root nodules, Braz. J. Microbiol., Vol. 44. No. 2, pp. 629-637
Zhu F, Qu L, Hong X, Sun X. 2011. Isolation and Characterization of a Phosphate-Solubilizing Halophilic Bacterium Kushneria sp. YCWA18 from Daqiao Saltern on the Coast of Yellow Sea of China. Evid. Based Complementary Altern. Med. 615032. doi:10.1155/2011/615032.