A total of 30 fluorescent pseudomonads (FPs) were showed significant antagonistic activity against different fungal phytopathogens with different level of the zone of inhibition (ZOI) for Rhizoctonia solani (5mm-34mm), Macrophomina phaseolina (9mm-37mm), Scleotium rolfsii (4mm-36mm), Helminthusporium solani (5mm-27mm), Fusarium oxysporum (2mm-25mm) and Fusarium oxysporum RACE (4mm-31mm) compared to control. The maximum growth of our selected isolate VSMKU4036 was observed in King'B Broth (KBB), pH 7.0 and at 37°C. The VSMKU4036 isolate has been recognized as Pseudomonas sp, based on the morphological, biological, and different functional characteristics. Antagonistic rhizobacterium Pseudomonas sp VSMKU4036 produced antimicrobial traits, such as plant growth promotion and various functional characters like siderophores, hydrogen cyanide (HCN), phosphate solubilization, indole acetic acid (IAA), biofilms formation, protease, gelatinase, amylase, and pectinase. Our superior biocontrol isolate VSMKU4036 was high resistance to tetracycline, streptomycin and nalidixic acid, however, it was sensitive to ampicillin and rifamycin. Pseudomonas sp VSMKU4036 showed maximum resistance to cadmium, nickel chloride, copper sulphate, magnesium sulphate, zinc chloride and ferric chloride where as highly sensitive to mercuric chloride, and selenium dioxide compared to control.
Microbial bio-products are becoming an appealing and viable alternative to chemical pesticides for effective management of crop diseases. These bio-products are known to have potential to minimize agrochemical applications without losing crop yield and also restore soil fertility and productivity. In this study, the inhibitory efficacy of 2,4-diacetylphloroglucinol (DAPG) produced by Pseudomonas fluorescens VSMKU3054 against Ralstonia solanacearum was assessed. Biochemical and functional characterization study revealed that P. fluorescens produced hydrogen cyanide (HCN), siderophore, indole acetic acid (IAA) and hydrolytic enzymes such as amylase, protease, cellulase and chitinase, and had the ability to solubilize phosphate. The presence of the key antimicrobial encoding gene in the biosynthesis of 2,4-diacetylphloroglucinol (DAPG) was identified by PCR. The maximum growth and antimicrobial activity of P. fluorescens was observed in king’s B medium at pH 7, 37 °C and 36 h of growth. Glucose and tryptone were found to be the most suitable carbon and nitrogen sources, respectively. DAPG was separated by silica column chromatography and identified by various methods such as UV-Vis, FT-IR, GC-MS and NMR spectroscopy. When R. solanacearum cells were exposed to DAPG at 90 µg/mL, the cell viability was decreased, reactive oxygen species (ROS) were increased and chromosomal DNA was damaged. Application of P. fluorescens and DAPG significantly reduced the bacterial wilt incidence. In addition, P. fluorescens was also found effective in promoting the growth of tomato seedlings. It is concluded that the indigenous isolate P. fluorescens VSMKU3054 could be used as a suitable biocontrol agent against bacterial wilt disease of tomato.
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