Mejda Daami-Remadi

Endophytic Bacteria from Datura stramonium for Fusarium WiltSuppression and Tomato Growth Promotion

Ten nonpathogenic bacterial isolates, recovered from Datura stramonium organs, and successfully colonizing the internal stem tissues of tomato cv. Rio Grande were screened for their ability to suppress tomato Fusarium wilt, caused by Fusaium oxysporum f. sp. lycopersici (FOL), and to enhance tomato growth. S37 and S40 isolates were found to be the most effective in decreasing leaf yellowing by 94-88% and the vascular browning extent by 96-95%, respectively, as compared to FOL-inoculated and untreated control. A significant enhancement of growth parameters was recorded on tomato plants inoculated or not with the pathogen. The two bioactive isolates were morphologically and biochemically characterized and identified using 16S rDNA sequencing genes as Stenotrophomonas maltophilia str. S37 and Bacillus mojavensis str. S40. Screened in vitro for their antifungal activity toward FOL, these strains led to 43.8 and 39% decrease in pathogen radial growth and to the formation of an inhibition zone of about 11.37 and 12.12 mm in diameter, respectively. S. maltophilia str. S37 and B. mojavensis str. S40 were found to be chitinase-, protease- and pectinase-producing strains but only S. maltophilia str. S37 was able to produce the volatile metabolite hydrogen cyanide. Indole-3-acetic acid production, phosphate solubilizing ability and pectinase activity were investigated for elucidating their plant growth promoting traits and their endophytic colonization ability. To our knowledge, this is the first report on endophytic bacteria from D. stramonium exhibiting Fusarium wilt suppression potential and plant growth-promoting ability on tomato.

Bio-suppression of Sclerotinia Stem Rot of Tomato and Biostimulation ofPlant Growth Using Tomato-associated Rhizobacteria

A collection of 25 rhizobacterial strains, recovered from rhizospheric soils around healthy tomato plants grown in Rhizoctonia-infested fields, belonging to Bacillus amyloliquefaciens, B. thuringiensis, B. megaterium, B. subtilis, Enterobacter cloacae, Chryseobacterium jejuense, and Klebsiella pneumoniae was screened for its suppressive effects of Sclerotinia Stem Rot of tomato caused by Sclerotinia sclerotiorum and plant growth-promoting ability. The inhibitory effects of diffusible and volatile metabolites from these rhizobacteria against pathogen mycelial growth depended significantly upon strains tested. Growth inhibition caused by diffusible and volatile compounds was of about 37-57% and 24-54%, respectively. All strains tested had totally suppressed myceliogenic germination of sclerotia and improved germination of bacterized tomato seeds as compared to the untreated controls. The screening of their disease-suppressive and plant growth-promoting abilities revealed 72-100% decrease in Sclerotinia Stem Rot severity and significant increments in plant height by 52-67%, roots fresh weight by about 66-88% and aerial part weight by 47-75%, compared to S. sclerotiorum-inoculated and untreated control. The most promising strains combining disease-suppressive and growth-promoting abilities were B. subtilis B10 (KT921327) and B14 (KU161090), B. thuringiensis B2 (KU158884), B. amyloliquefaciens B13 (KT951658) and B15 (KT923051), and E. cloacae B16 (KT921429).

Endophytic bacteria from Datura metel for plant growth promotion and bioprotection against Fusarium wilt in tomato

ABSTRACT Nine non-pathogenic bacterial isolates, recovered from Datura metel organs and able to colonise the internal stem tissues of tomato cultivar Rio Grande, were screened for their ability to suppress tomato Fusarium wilt disease caused by Fusarium oxysporum f. sp. lycopersici (FOL), and to enhance plant growth. S33 and S85 isolates tested were found to be the most effective in decreasing Fusarium wilt severity by 94–95% compared to FOL-inoculated and untreated control. A significant enhancement of growth parameters was recorded on tomato plants inoculated or not with FOL. Both isolates were characterised and identified using 16S rDNA sequencing genes as Stenotrophomonas sp. str. S33 (KR818084) and Pseudomonas sp. str. S85 (KR818087). Screened in vitro for their antifungal activity towards FOL, these isolates led to 38.7% and 22.5% decrease in pathogen radial growth and to the formation of an inhibition zone of 12.75 and 8.37 mm respectively. Stenotrophomonas sp. str. S33 and Pseudomonas sp. str. S85 were found to be chitinase-, protease- and pectinase-producing strains but unable to produce hydrogen cyanide. Production of indole-3-acetic acid-like compounds, phosphate solubilising ability and pectinase activity were investigated for elucidating their plant growth-promoting traits and their endophytic colonisation ability.

Efficacy of Bacillus subtilis V26 as a biological control agent against Rhizoctonia solani on potato.

The aim of this study is to evaluate the efficacy of the strain Bacillus subtilis V26, a local isolate from the Tunisian soil, to control potato black scurf caused by Rhizoctonia solani. The in vitro antifungal activity of V26 significantly inhibited R. solani growth compared to the untreated control. Microscopic observations revealed that V26 caused considerable morphological deformations of the fungal hyphae such as vacuolation, protoplast leakage and mycelia crack. The most effective control was achieved when strain V26 was applied 24h prior to inoculation (protective activity) in potato slices. The antagonistic bacterium V26 induced significant suppression of root canker and black scurf tuber colonization compared to untreated controls with a decrease in incidence disease of 63% and 81%, respectively, and promoted plant growth under greenhouse conditions on potato plants. Therefore, B. subtilis V26 has a great potential to be commercialized as a biocontrol agent against R. solani on potato crops.

Variation in Chitosan and Salicylic Acid Efficacy Towards Soil-borne and Air-borne Fungi and Their Suppressive Effect of Tomato Wilt Severity

Two resistance inducers (RIs), chitosan and salicylic acid (SA), were assessed in vitro for their antifungal activity against ten tomato phytopathogenic fungi i.e. Fusarium oxysporum f. sp. lycopersici, F. oxysporum f. sp. radicis-lycopersici, F. solani, Verticillium dahliae, Rhizoctonia solani, Colletotrichum coccodes, Pythium aphanidermatum, Sclerotinia sclerotiorum, Botrytis cinerea, and Alternaria solani. The impact of these RIs, applied as soil drench, on Verticillium wilt, Fusarium wilt, and Fusarium Crown and Root Rot severity and on growth parameters of tomato cv. Rio Grande plants were also investigated. Chitosan (0.5-4 mg/ml) and SA (1-25 mM) inhibited mycelial growth of all pathogens in Potato Dextrose Agar (PDA) medium in a concentration-dependent manner, with the greatest inhibition achieved using the highest chitosan and SA concentrations. Inter specific variations in sensitivity to chitosan and SA were detected. P. aphanidermatum and S. Sclerotiorum were the most sensitive to both RIs. Single treatments with chitosan (4 mg/ml) and SA (10 mM) resulted in varied degree of protection against wilt diseases. Chitosan-and SA-based treatments resulted in 42.1-73.68, 60.86-78.26 and 45- 50% reductions in wilt severity, as compared to VD-, FOL- and FORL-inoculated and untreated controls, respectively. All growth parameters noted were enhanced using RIs compared to pathogen-inoculated controls. In fact, SA-based treatment had significantly increased plant height, root and aerial part fresh weights by 17.94, 52.17 and 33.33%, by 23.01, 55.40 and 29.72%, and by 17.72, 50 and 46.84%,respectively, while compared to VD-, FOL- and FORL-inoculated and untreated plants. Chitosan-treated plants showed increment in their height, root and aerial part fresh weights by 13.81, 62.16 and 38.97%, respectively, compared to FORL-inoculated and untreated control. Results from this investigation showed that SA and chitosan may be used as potential inducers of systemic acquired resistance for successfully controlling fungal tomato diseases in Tunisia.

Composition and Antimicrobial Activities of Essential Oils From the Aerial Parts and Flowers of Thymus hirtus W. Growing in Tunisia

Abstract The chemical composition of essential oils isolated from fresh flowers and aerial parts of Thymus hirtus Willd was analyzed using GC and GC/MS. Forty-six components were identified in the oil of each organ. Camphor occurred as the major constituent of the aerial part oil, while α-pinene was the most notably dominating compound in the flower oil. Biological activities of the indicated oils were evaluated against eight bacteria, nine phytopathogenic, and 15 human pathogenic fungi.

Synthesis and antimicrobial activity of novel coumarin derivatives from 4-methylumbelliferone

AbstractConsidering the potential interest of heterocyclic compounds, the aim of the present study is to synthesize new coumarin derivatives, to provide their full chemical characterization and to evaluate their antimicrobial activities. The reaction of ethyl 2-(4-methyl-2-oxo-2H-chromen-7-yloxy) acetate 2 with sodium hydroxide afforded the corresponding 2-(2-oxo-4-methyl-2H-chromen-7-yloxy) acetic acid 3 which was esterified using a series of alcohols in the presence of iodine to yield a new series of coumarin esters 4a–j. On the other hand, treatment of the key intermediate 2 with an aqueous solution of hydrazine in ethanol at reflux gave the corresponding hydrazide 5 which further converted into coumarin derivatives 6a–f and 7a–c by condensation with a series of aromatic aldehydes and cyclic anhydrides, respectively. The synthesized compounds were completely characterized by 1H NMR, 13C NMR, IR and HRMS. The antibacterial and antifungal activities of the new synthesized compounds were evaluated using the disc diffusion method and seemed to be significant.

Biocontrol of Rhizoctonia Root Rot in Tomato and Enhancement of PlantGrowth using Rhizobacteria Naturally associated to Tomato

In the present study, 25 rhizobacterial isolates, obtained from rhizosphere of healthy tomato plants collected from various tomato-growing sites in Tunisia, were tested in vitro and in vivo against Rhizoctonia solani. This bacterial collection, composed of isolates belonging to Bacillus spp., Enterobacter cloacae, Chryseobacterium jejuense, and Klebsiella pneumoniae, was assessed for its antifungal potential against R. solani the causative agent of Rhizoctonia Root Rot disease in various crops including tomato. Antifungal activity of diffusible and volatile metabolites derived from these isolates was tested against target pathogen using dual and distance culture bioassays, respectively. Growth inhibition rates, recorded after 5 days of incubation at 25°C, depended significantly upon tested bacterial isolates and screening methods and reached 34-59% and 18-45% for diffusible and volatile metabolites, respectively. The screening of disease-suppressive and plant growth-promoting abilities of these tomato-associated rhizobacteria showed 47-100% decrease in disease severity and significant increments in plant height by 62-76%, roots fresh weight by 53-86%, and aerial parts fresh weight by 34-67% compared to pathogen-inoculated and untreated control. B. thuringiensis B2 (KU158884), B. subtilis B10 (KT921327) and E. cloacae B16 (KT921429) were found to be the most efficient isolates in decreasing R. solani radial growth, suppressing disease severity, and enhancing plant growth.

Isolation of Endophytic Bacteria from Withania Somnifera and Assessment of their Ability to Suppress Fusarium Wilt Disease in Tomato and to Promote Plant Growth

Four nonpathogenic and putative endophytic bacterial isolates, recovered from Withania somnifera fruits (S7, S8 and S9) and stems (S15), were evaluated for their in vivo and in vitro antifungal activity against Fusarium oxysporum f. sp. lycopersici (FOL), and their plant-growth promoting ability. Tomato plants challenged and/or not with FOL and treated using these bacterial isolates exhibited a significant increment in their growth parameters (plant height, aerial part fresh weight, maximum root length, and root fresh weight). The strong suppressive effect against Fusarium wilt was achieved using two isolates (namely S15 and S8) leading to 92-96% lower disease severity compared to pathogen-inoculated and untreated control. Both isolates were characterized and only the isolate S8 was identified as Alcaligenes faecalis subsp. faecalis str. S8 (KR818077) using 16S rDNA gene sequencing. The unidentified bacterial isolate S15 had improved germination of bacterized tomato seeds relative to the untreated ones. Tested using streak and sealed plates methods, diffusible and volatile compounds from S15 and S8 isolates inhibited FOL by 10.7-16.8% and 53.8-20.7%, respectively. Moreover, an inhibition zone (8.5-8.25 mm) was formed around FOL colonies using the disc diffusion method. Alcaligenes faecalis subsp. faecalis str. S8 and the unidentified bacterium str. S15 were shown able to produce chitinolytic, proteolytic and pectinolytic enzymes and hydrogen cyanide. Production of indole-3-acetic acid and phosphate solubilizing ability were also investigated for elucidation of their plant growth-promoting traits.

Antifungal activity of aqueous and organic extracts from Withania somnifera L. against Fusarium oxysporum f. sp. radicis-lycopersici

The aim of this study was to evaluate the in vitro antifungal activity of aqueous and organic extracts from native Withania somnifera L. leaves, stems, and fruits against Fusarium oxysporum f. sp. radicis-lycopersici (FORL), the causal agent of Fusarium Crown and Root Rot disease in tomato. Aqueous and organic extracts (used at 1, 2, 3 and 4%) were added to molten Potato Dextrose Agar (PDA) medium. After pathogen challenge, cultures were incubated at 25°C for 5 days. All extracts tested, whatever the concentrations used, showed a strong antifungal activity toward targeted pathogen. FORL response to the different extracts assessed using the poisoned food technique, varied depending on plant organs, concentrations tested and organic solvent used for extraction. For aqueous extracts, fruit extract used at 2% exhibited the highest antifungal potential where FORL growth was decreased by 56.27%, relative to the untreated control, compared to 52 and 45.34% achieved using stem and leaf extracts at 3%, respectively. The highest antifungal activity of organic extracts was registred at the highest concentration used (4%). FORL was found to be more sensitive to fruit extracts than those from leaves and stems. Among the three organic extracts tested, butanolic fractions were the most active against FORL growth. The highest antifungal potential expressed by 62.03% decrease in pathogen radial growth was displayed by butanolic stem extracts applied at 4%. These results indicate that native W. somnifera plants may be exploited as potential source of allelochemicals biologically active against FORL.