RESUMO
The aim of the present work was to evaluate the ability of the native yeast Rhodosporidium fluviale to control Botrytis cinerea on apple fruit and to study the possible mechanisms of action with the goal of improving the control of gray mold. For this, the influence of application time of the yeast was studied simulating preventive and curative effects. Also, the effect of nonviable cells of the yeast in the biocontrol was assessed. According to the results obtained, the following mechanisms of action of R. fluviale could be proposed: 1- competition for space, 2- direct interaction between antagonist and pathogen, 3- induction of ß-1,3-glucanase in apple tissue, 4- Probable production of glucanase in the apple wounds and 5- antifungal action of cellular components, probably chitin, present in the wall of yeast cells that could be the explanation for the activity of nonviable cells. SIGNIFICANCE AND IMPACT OF THE STUDY: Significance and Impact of the Study: Botrytis cinerea Pers: Fr, which causes gray mold of fruits and vegetables around the world, is difficult to control successfully because it is genetically variable and rapidly develops resistance to the chemicals commonly used for its control. This study is a contribution to the biocontrol of this phytopathogen fungus. The evaluation of the native yeast Rhodosporidium fluviale as biocontrol agent and the elucidation of possible mechanisms of action, including the participation of nonviable cells of this yeast, have not been reported up to date.
Assuntos
Antibiose/fisiologia , Antifúngicos/metabolismo , Basidiomycota/metabolismo , Agentes de Controle Biológico/metabolismo , Botrytis/crescimento & desenvolvimento , Malus/microbiologia , Argentina , Frutas/microbiologia , Doenças das Plantas/microbiologiaRESUMO
AIMS: The aim of the present study was to evaluate and compare freezing and freeze-drying treatments for conserving Rahnella aquatilis (BNM 0523) with the goal to achieve an adequate commercial formulation of this biocontrol agent. METHODS AND RESULTS: The effect of several protective agents, rehydration media and freezing temperatures on the viability and functional activity of the R. aquatilis was investigated. The storage stability at 3 months and 4 years was determined by checking the viability of the cells and their biocontrol capability against Botrytis cinerea by measuring the percentage of reduction of disease severity on apple. The best results were obtained by the freeze-drying of the cells using a mixture of skimmed nonfat milk 10%, yeast extract 0·5% and glucose 1% as the protecting and rehydrating medium, and a quickly freezing (-70°C) before the freeze-drying. In this case, the viability of the cells after 4 years was 98%, and their antagonistic ability showed a little decrease with respect fresh cells. CONCLUSIONS: The studies showed that R. aquatilis was resistant to freezing and freeze-drying when it was used a mixture of cryoprotectants and that it was possible to obtain inoculums with high viability and good effectiveness for reduction of decay caused by B. cinerea. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is probably the first report about the resistance of R. aquatilis to freezing and freeze-drying treatments and shows that these operations could be useful for obtaining a commercial formulation of this biocontrol agent.
Assuntos
Antibiose , Conservação de Alimentos/métodos , Rahnella/crescimento & desenvolvimento , Botrytis/crescimento & desenvolvimento , Botrytis/fisiologia , Temperatura Baixa , Hidratação , Liofilização/métodos , Congelamento , Humanos , Malus/microbiologia , Interações Microbianas , Rahnella/fisiologiaRESUMO
Yeasts produce hydroxamate-type siderophores (iron-binding compounds) in response to Fe-stress conditions. Because these siderophores are important to the biocontrol of postharvest diseases of apple and pears, a method for screening siderophore producer yeast was developed. The screening method was carried out in special Petri dishes with eight or nine wells (25-mm diameter). These wells were filled with siderophore production medium and seeded with yeasts isolated from epiphytic apple microflora. After yeasts grew (24-48 h), holes (2-mm diameter) were made in the agar of each well. Holes were filled with an acid solution of ferric perchlorate. After 10-15 min, reddish halos appeared in the bottom of the plate and their intensities were compared with standards. Standards were prepared in the same special dish with rhodotorulic acid solutions (concentrations between 0.05 and 1 g/l) plus 2% agar. When agar solidified into wells, holes were made and filled with ferric perchlorate solution. Color intensities of reddish halos were proportional to siderophore concentration and the detection limit was 0.1 g/l. It was possible to correlate the production of siderophore in solid medium with the results obtained in liquid medium. The methodology was also a useful tool for making a preliminary assessment of the influence of different factors on the siderophore production.
Assuntos
Meios de Cultura/química , Técnicas Microbiológicas , Sideróforos/biossíntese , Leveduras/metabolismo , Bactérias/crescimento & desenvolvimento , Bactérias/metabolismo , Ferro/farmacologia , Leveduras/crescimento & desenvolvimentoRESUMO
The production of rhodotorulic acid, a siderophore synthesized by Rhodotorula strains, was improved with the objective of achieving the biocontrol of phytopathogenic moulds. Rhodotorulic acid increased up to 60% in the presence of urea as a nitrogen source, pH near to 8 and a C:N ratio of 8:1. The siderophore-containing spent medium showed in vitro antifungal activity against important plant pathogens including Botrytis cinerea, which causes grey mould on a wide variety of host plants including numerous commercial crops. The antifungal activity was related to siderophore concentration.
Assuntos
Antifúngicos/metabolismo , Antifúngicos/farmacologia , Nitrogênio/farmacologia , Rhodotorula/efeitos dos fármacos , Rhodotorula/metabolismo , Sideróforos/biossíntese , Sideróforos/farmacologia , Aminoácidos/farmacologia , Fungos/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Nitrogênio/metabolismo , Controle Biológico de Vetores , Piperazinas/metabolismo , Piperazinas/farmacologia , Doenças das Plantas/microbiologia , Rhodotorula/classificação , Rhodotorula/crescimento & desenvolvimento , Ureia/metabolismo , Ureia/farmacologiaRESUMO
The recovery of lithium from spodumene (6.9% Li2O) by bioleaching was investigated. This process was carried out using heterotrophic micro-organisms previously isolated from the mineral. Penicillium purpurogenum, Aspergillus niger and Rhodotorula rubra were assayed separately. Two different media were used for bioleaching; one of them (M2 medium) was highly limited in Mg2+, Fe2+ and K+. The assays were carried out in 500 ml Erlenmeyer flasks with 1 g of ground mineral at 50-80 mesh and 150 ml of leaching medium. Lithium extracted and accumulated in biomass during 30 d of bioleaching with P. purpurogenum was 6.35 mg % dry weight (d.w.) in M1 medium and 10.8 mg % d.w. in M2 medium, while in the leach liquor, Li concentration was 1.06 ppm (M1 medium) and 1.26 ppm (M2 medium). Results of leaching on day 30 with R. rubra were 5.87 mg % d.w. and 16.7 mg % d.w. of lithium accumulated in biomass in M1 medium and M2 medium, respectively. In leach liquor, lithium was 0.5 ppm (M1 medium) and 1.53 ppm (M2 medium). Aspergillus niger was able to accumulate 1.60 mg % d.w. (M1 medium) and 5.1 mg % d.w. of lithium (M2 medium) in biomass. Lithium in leach liquor was 0.37 ppm (M1 medium) and 0.75 ppm (M2 medium). Chemical analysis of the leach liquor showed gluconic and citric acids. It was possible to detect capsular exopolymers in the yeast. These metabolic products seem to be related to leaching but a more important factor for enhancing this process may be microbial adaptation to a low nutrient environment.