An insight into the role of the organic acids produced by Enterobacter sp. strain 15S in solubilizing tricalcium phosphate: in situ study on cucumber.

BACKGROUND: The release of organic acids (OAs) is considered the main mechanism used by phosphate-solubilizing bacteria (PSB) to dissolve inorganic phosphate in soil. Nevertheless, little is known about the effect of individual OAs produced by a particular PSB in a soil-plant system. For these reasons, the present work aimed at investigating the effect of Enterobacter sp. strain 15S and the exogenous application of its OAs on (i) the solubilization of tricalcium phosphate (TCP), (ii) plant growth and (iii) P nutrition of cucumber. To this purpose two independent experiments have been performed. RESULTS: In the first experiment, carried out in vitro, the phosphate solubilizing activity of Enterobacter 15S was associated with the release of citric, fumaric, ketoglutaric, malic, and oxalic acids. In the second experiment, cucumber plants were grown in a Leonard jar system consisting of a nutrient solution supplemented with the OAs previously identified in Enterobacter 15S (jar's base) and a substrate supplemented with the insoluble TCP where cucumber plants were grown (jar's top). The use of Enterobacter 15S and its secreted OAs proved to be efficient in the in situ TCP solubilization. In particular, the enhancement of the morpho-physiological traits of P-starved cucumber plants was evident when treated with Enterobacter 15S, oxalate, or citrate. The highest accumulation of P in roots and shoots induced by such treatments further corroborated this hypothesis. CONCLUSION: In our study, the results presented suggest that organic acids released by Enterobacter 15S as well as the bacterium itself can enhance the P-acquisition by cucumber plants.

Diversity and antimicrobial potential of the culturable rhizobacteria from medicinal plant Baccharis trimera Less D.C.

Plant microbiota is usually enriched with bacteria producers of secondary metabolites and represents a valuable source of novel species and compounds. Here, we analyzed the diversity of culturable root-associated bacteria of the medicinal native plant Baccharis trimera (Carqueja) and screened promising isolates for their antimicrobial properties. The rhizobacteria were isolated from the endosphere and rhizosphere of B. trimera from Ponta Grossa and Ortigueira localities and identified by sequencing and restriction analysis of the 16S rDNA. The most promising isolates were screened for antifungal activities and the production of siderophores and biosurfactants. B. trimera presented a diverse community of rhizobacteria, constituted of 26 families and 41 genera, with a predominance of Streptomyces and Bacillus genera, followed by Paenibacillus, Staphylococcus, Methylobacterium, Rhizobium, Tardiphaga, Paraburkholderia, Burkholderia, and Pseudomonas. The more abundant genera were represented by different species, showing a high diversity of the microbiota associated to B. trimera. Some of these isolates potentially represent novel species and deserve further examination. The communities were influenced by both the edaphic properties of the sampling locations and the plant niches. Approximately one-third of the rhizobacteria exhibited antifungal activity against Sclerotinia sclerotiorum and Colletotrichum gloeosporioides, and a high proportion of isolates produced siderophores (25%) and biosurfactants (42%). The most promising isolates were members of the Streptomyces genus. The survey of B. trimera returned a diverse community of culturable rhizobacteria and identified potential candidates for the development of plant growth-promoting and protection products, reinforcing the need for more comprehensive investigations of the microbiota of Brazilian native plants and habitats.

Plant growth-promoting bacteria improve leaf antioxidant metabolism of drought-stressed Neotropical trees.

MAIN CONCLUSION: Plant growth-promoting bacteria association improved the enzymatic and non-enzymatic antioxidant pathways in Neotropical trees under drought, which led to lower oxidative damage and enhanced drought tolerance in these trees. Water deficit is associated with oxidative stress in plant cells and may, thus, negatively affect the establishment of tree seedlings in reforestation areas. The association with plant growth-promoting bacteria (PGPB) is known to enhance the antioxidant response of crops, but this strategy has not been tested in seedlings of Neotropical trees. We evaluated the effects of inoculation with two PGPB (Azospirillum brasilense and Bacillus sp.) on the antioxidant metabolism of Cecropia pachystachya and Cariniana estrellensis seedlings submitted to drought. We measured the activity of antioxidant enzymes and the content of non-enzymatic antioxidants in leaves, and biometrical parameters of the seedlings. In both tree species, drought decreased the activity of antioxidant enzymes and the content of non-enzymatic antioxidant compounds. For C. pachystachya, the enzymatic and non-enzymatic pathways were mostly influenced by A. brasilense inoculation, which enhanced ascorbate peroxidase (APX) and superoxide dismutase activities and positively affected the level of non-enzymatic antioxidant compounds. In C. estrellensis, A. brasilense inoculation enhanced APX activity. However, A. brasilense and Bacillus sp. inoculation had more influence on the non-enzymatic pathway, as both bacteria induced a greater accumulation of secondary compounds (such as chlorogenic acid, gallic acid, rutin and synapic acid) compared to that in non-inoculated plants under drought. For both species, PGPB improved biometrical parameters related to drought tolerance, as specific leaf area and leaf-area ratio. Our results demonstrate that PGPB induced antioxidant mechanisms in drought-stressed Neotropical trees, increasing drought tolerance. Thus, PGPB inoculation provides a biotechnological alternative to improve the success of reforestation programmes.

Isolation and Identification of Aspergillus Section Nigri, and Genotype Associated with Ochratoxin A and Fumonisin B2 Production in Garlic Marketed in Brazil.

The garlic contains sulfur bioactive compounds responsible for medicinal properties. The decrease of these compounds due to inadequate storage conditions reduces the beneficial properties and favors infection by microorganisms. Several studies have shown high frequency of garlic infected with Aspergillus section Nigri that potentially produce mycotoxin. Garlic samples were collected in markets of Brazil and a total of 32 samples (of 36) had the fungal infection with predominant genus Aspergillus (50.3%), Penicillium (34.7%), and Fusarium (11%). A total of 63% (649/1031) of infection with Aspergillus section Nigri, of which 60 isolates were selected for analysis of genetic variability that resulted in 4 clusters. Representatives of clusters were identified by the calmodulin gene. Isolates from cluster I were subdivided into A-I and identified as A. niger (16 isolates) and the isolates of clusters B-I, II, and III were identified as A. welwitschiae (43 isolates). Besides, an isolate of the IV-cluster was identified by A. luchuensis. Further, we used the multiplex PCR to verify genotypes of 59 isolates, and none of these had OTA production-associated genotype. Moreover, 19 A. welwitschiae and 15 A. niger were FB2 production-associated genotype. Our study is the first report to the incidence of garlic infection in Brazil and to show that A. welwitschiae causes most of these infections.

Enhanced drought tolerance in seedlings of Neotropical tree species inoculated with plant growth-promoting bacteria.

The inoculation of tree species with plant growth-promoting bacteria (PGPB) has emerged as an important strategy for the acclimation of seedlings by improving plant tolerance to biotic and abiotic stresses. This study aimed to evaluate the effects of inoculation with bacterial species (Azospirillum brasilense - Ab-V5, Bacillus sp., Azomonas sp. and Azorhizophillus sp.) on the growth and physiology of the Neotropical tree species Trema micrantha and Cariniana estrellensis under drought conditions. When associated with Ab-V5 and Azomonas sp., T. micrantha showed increased protein in the leaves, starch in the leaves and roots, photosynthesis, instantaneous carboxylation efficiency and root and shoot dry mass. Moreover, there were reductions in hydrogen peroxide, lipid peroxidation, water potential and proline. In C. estrellensis associated with Ab-V5, higher values of photosynthesis and instantaneous carboxylation efficiency were observed, in addition to higher starch content in the leaves and roots and higher protein content in the leaves; lower hydrogen peroxide and lipid peroxidation contents were also observed. The associations of T. micrantha with Ab-V5 and Azomonas sp. and C. estrellensis with Ab-V5 favored the activation of metabolic processes under drought, leading to greater drought tolerance. This work demonstrates the effects of compatible associations of Neotropical tree and PGPB species and suggests that the identification of compatible PGPB strains can result in tree seedlings with increased tolerance to abiotic stresses, such as drought.

Formulations of polymeric biodegradable low-cost foam by melt extrusion to deliver plant growth-promoting bacteria in agricultural systems.

The extrusion technology of blends formed by compounds with different physicochemical properties often results in new materials that present properties distinctive from its original individual constituents. Here, we report the use of melt extrusion of blends made from low-cost materials to produce a biodegradable foam suitable for use as an inoculant carrier of plant growth-promoting bacteria (PGPB). Six formulations were prepared with variable proportions of the raw materials; the resulting physicochemical and structural properties are described, as well as formulation performance in the maintenance of bacterial viability during 120 days of storage. Differences in blend composition influenced foam density, porosity, expansion index, and water absorption. Additionally, differences in the capability of sustaining bacterial viability for long periods of time were more related to the foam composition than to the resulting physicochemical characteristics. Microscopic analyses showed that the inoculant bacteria had firmly attached to the extruded material by forming biofilms. Inoculation assays using maize plants demonstrated that the bacteria attached to the extruded foams could survive in the soil for up to 10 days before maize sowing, without diminishing its ability to promote plant growth. The results presented demonstrate the viability of the new matrix as a biotechnological material for bacterial delivery not only in agriculture but also in other biotechnological applications, according to the selected bacterial strains.