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We have shown, the outcome of antifungal activity of phenazine derivatives which is produced by fluorescent pseudomonads (FPs) for the control of sheath blight of rice. A total of 50 fluorescent pseudomonads (FPs) were isolated from rice rhizosphere. Off which, 36 FPs exhibited antagonistic activity against Rhizoctonia solani , Macrophomina phaseolina , Fusarium oxysporum , Alternaria alternata and Sclerotium rolfsii up to 70–80% compared to control by dual culture method. BOX-PCR analyses of antagonistic isolates indicated that two phylogenetic group, where group I consisted of 28 isolates and eight isolates belongs to group II. Among 36 FPs, a total of 10 FPs revealed that the presence of phenazine derivatives on thin layer chromatography (TLC), which is coincided with that of authentic phenazine with R f value 0.57. Similar to TLC analysis, antibiotic encoding gene phenazine-1-carboxamide (PCN) was detected in 10 FPs by PCR analysis with respective primer. Among, PCN detected isolates of FPs, a significant biocontrol potential possessing isolate designated as VSMKU1 and it was showed prominent antifungal activity against R. solani and other tested fungal pathogens. Hence, the isolate VSMKU1 was selected for further studies. The selected isolate VSMKU1 was identified as Pseudomonas aeruginosa by 16S rDNA sequence analysis. The antifungal metabolite phenazine like compound produced by VSMKU1 was confirmed by UV, FT-IR and HPLC analysis. The phenazine compound from VSMKU1 significantly arrest the growth of R. solani compared to carbendazim by well diffusion method. The detached leaf assay showed remarkable inhibition of lesion height 80 to 85% by the treatments of culture (VSMKU1), cell free culure filtrate and phenazine like compound compared to control and other treatments was observed in detached leaves of rice. These results emphasized that VSMKU1 isolate can be used as an alternative potential biocontrol agent against sheath blight of rice, instead of using commercial fungicide such as validamycin and carbendazim which cause environmental pollution and health hazards. [ABSTRACT FROM AUTHOR]

Soil samples were collected from different rhizosphere plants grown in SEKEM farm, Bilbis, El-Sharkyia governorate, Egypt. Four fluorescent pseudomonads isolates, out of seventy one, were selected according to their efficiency to produce IAA, cyanide, antagonistic effect and high ability to solubilize potassium and phosphorus. These isolates were identified using 16S rRNA gene sequencing technique to be different strains of Pseudomonas otitidis . Growth curve and growth kinetics were determined on king’s broth medium. Ps. otitidis SE8 gave the highest specific growth rate, multiplication rate and number of generation being 0.287 h −1 , 0.4147 h −1 and 4.98 h −1 , respectively. This strain also showed the lowest of doubling time. Ps. otitidis SE8 and OL2 had higher activity to solubilize phosphorus than other tested strains. Bacterial strain SE8 also gave a considerable amount of soluble potassium as compared with other strains being 80.7 ppm. The maximum IAA and gibberellic acid production was also recorded by Pseudomonas SE8 and OL2. A remarkable quantity of siderophores was detected in the case of Ps. otitidis SE8 being 28.20 mM DFOM. Application of these isolates as inoculants for tomato plants in green house was performed. The results showed that inoculation of tomato seedling with Pseudomonas strains led to suppress the soil-borne pathogen, increased of NPK uptake and supported tomato plant growth. [ABSTRACT FROM AUTHOR]

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Abstract: Charcoal rot of sorghum caused by Macrophomina phaseolina is a disease of economic importance for which a high level of genetic resistance is not available. Therefore, we made an effort to manage this disease with fluorescent pseudomonads associated with field-grown sorghum crops. One hundred and twenty-six fluorescent Pseudomonas spp. from different sorghum growing regions of India were isolated, selected and evaluated for in vitro antifungal activities and three selected strains were further tested for management of charcoal rot under field conditions. Antifungal activities (inhibition of growth, biomass, microsclerotia production, spore germination) of the secondary metabolites and the cell-free culture filtrates of the selected fluorescent pseudomonad strains (SRB129, SRB288 and Pseudomonas chlororaphis SRB127) were studied in detail. SRB127, SRB129 and SRB288 inhibited mycelial growth of M. phaseolina ranging from 30.5 to 76.5% in dual culture assay. The cell-free culture filtrates of these strains at 20% (v/v) concentration significantly reduced the formation and germination of microsclerotia of M. phaseolina. In the field, P. chlororaphis SRB127 emerged as an effective biocontrol agent of charcoal rot of sorghum. The bacterium, when applied as seed treatment, reduced the charcoal rot incidence by >40%, crop-lodging by >20%, and increased grain mass. P. chlororaphis SRB127, when grown in a gnotobiotic sand system, effectively colonized the sorghum root and formed microcolony-like cell-aggregates in some parts of the root. Under glasshouse conditions the bacterium survived in the sorghum rhizosphere without a significant reduction in population. We conclude that the use of a selection of effective strains of bacteria can be a useful component of integrated management of charcoal rot in sorghum. [Copyright &y& Elsevier]

A total of 87 FPs have been isolated from tomato (Lycopersicon esculentum) rhizosphere and evaluated by dual plate assay method for the antimicrobial activity of Ralstonia solanacearum. Of which, 30 FPs showed antagonistic activity against R. solanacearum with the zone of inhibitions (ZOI) ranging from 19 mm to 28 mm. Similarly, antagonistic FPs significantly controlled various fungal phytopathogens such as Rhizoctonia solani (6.3 mm−20 mm) , Macrophomina phaseolina (8.7 mm−19.3 mm), Fusarium oxysporum (7.3 mm−30.7 mm), and Sclerotium rolfsii (5.3 mm−21 mm). The cell-free culture filtrate of Pseudomonas sp VSMKU3054 significantly suppressed the fungal pathogens and increased the root and shoot length of the tomato seedlings compared to the control. Furthermore, the antagonistic FPs effectively produced lytic enzymes and antimicrobial traits such as amylase (6), protease (30), cellulase (28), pectinase (12), chitinase (22), gelatinase (27), siderophore (28), hydrogen cyanide (25), phosphate solubilization (30), and Indole acetic acid (IAA) (23). Genetic diversity of FPs was assessed by BOX-PCR with specific primers revealed that two distinct clusters were observed, whereas, RFLP analyses were showed that 5 to 16 bands ranged from 75 bp to 1.2 bp with two major clusters using restriction enzyme Hae III. The antibiotic encoding genes were detected from FPs, of which 10 FPs were positive for 2,4-diacetylphloroglucinol (DAPG), 9 FPs were positive for pyoleuterin (PLT), 10 FPs were positive for hydrogen cyanide (HCN) and none of them obtained pyrrolnitrin (PRN) with respective primers. Based on the superior antagonistic isolates against R. solanacearum and phytofungal pathogens and other antimicrobial traits, the isolate VSMKU3054 was selected for further studies. Based on the morphological, physiological and 98% of 16 s rDNA sequence similarity, the selected isolate VSMKU3054 was identified as Pseudomonas fluorescens. Further, the potential isolate P. fluorescens VSMKU3054 and cell free culture filtrate remarkably enhance tomato seedling growth such as root and shoot length, fresh and dry weight and vigor index compared to control. This study indicated that rhizospheric isolates of FPs have more potential for plant growth promotion and plant protection from plant pathogenic microbes. [ABSTRACT FROM AUTHOR]