Cyclic lipopeptides (cLPs) of the surfactin, iturin and fengycin families synthesized by plant-associated Bacilli represent an important class of antibiotics as they may be tightly involved in the protective effect of selected strains against phytopathogens. However, their production by Bacillus cells developing on roots under rhizosphere conditions is still poorly understood. In this work, we combined electrospray and imaging mass spectrometry-based approaches to determine the detailed pattern of surfactins, iturins and fengycins produced in planta by Bacillus amyloliquefaciens S499. Very different production rates were observed for the three cLPs families. Whereas surfactin accumulated in significant amounts, much lower quantities of iturins and fengycins were detected in the environment of colonized roots in comparison with laboratory medium. In addition, the surfactin pattern produced by strain S499 evolving on roots is enriched in homologues with long fatty acid chains (C15) compared with the chains typically secreted under in vitro conditions. Additional experiments revealed that lipopeptide production by root-associated S499 cells is qualitatively and quantitatively dictated by the specific nutritional context of the rhizosphere (exudates enriched in organic acids, oxygen limitation) but also by the formation of biofilm-related structures around root hairs. As surfactins, iturins and fengycins retain specific functions and bioactivities, the biological relevance of their differential production observed in planta is discussed in the context of biocontrol of plant diseases.

Dried ground leaves of Psidium guajava L. (guava) were extracted by water and aqueous ethyl alcohol 50% (1:10) ratio, and the total phenolic content in the extracts was determined spectrophotometrically according to Folin-Ciocalteu's phenol method and calculated as gallic acid equivalent (GAE). Remarkably high total phenolic content 575.3 ±15.5 and 511.6±6.2 mg of GAE/g of dried weight material (for ethanol guava leaf extracts and water guava leaf extracts, respectively) were obtained. The antioxidant activity of lyophilized extracts was determined at ambient temperature by means of a 2,2-diphenyl-1-picrylhydryzyl (DPPH˙) colorimetry with detection scheme at 515 nm. The activity was evaluated by the decrease in absorbance as the result of DPPH˙ color change from purple to yellow. The higher the sample concentration used, the stronger was the free radical-scavenging effect. The results obtained showed that ascorbic acid was a substantially more powerful antioxidant than the extracts from guava leaf. On the other hand, the commercial guava leaf extracts and ethanol guava leaf extracts showed almost the same antioxidant power whereas water guava leaf extracts showed lower antioxidant activity. The parameter EC 50 and the time needed to reach the steady state to EC 50 concentration (50 EC T) affected the antiradical capacity of the sample. The antioxidant efficiency (AE) has been shown to be a more adequate parameter for selecting antioxidants than the widely used EC 50. This study revealed that guava leaf extracts comprise effective potential source of natural antioxidants.

Cyclic lipopeptides and particularly surfactins produced by Bacillus species retain antibacterial, antiviral, biofilm-forming and plant resistance-inducing activities. In most cases, their role in biological control of plant diseases was evoked on the basis of in vitro assays or by using non-producing/overproducing mutants but there is a need for more direct evidence of an efficient lipopeptide biosynthesis in the rhizosphere. In this work, we coupled LC-MS quantification of the lipopeptides secreted by cells colonizing tomato plants with the use of psrfA-lacZ reporter system integrated within the BGS3 chromosome to study the expression of the surfactin operon in planta. Results showed that a higher level of psrfA induction was observed upon the establishment of a stable BGS3 population on roots and surfactins extracted from the rhizosphere were produced in biologically significant quantities. Our results also demonstrate that BGS3 efficiently utilizes the main substrates from plant exudates to produce surfactins. This synthesis is also efficient in cells forming colonies and the production may be favoured in bacteria developing slowly in the rhizosphere. This provides a first understanding of how environmental factors may influence lipopeptide production by beneficial Bacillus strains.

A novel and convenient method based on native polyacrylamide gel electrophoresis (PAGE) and homogenization extraction was used for the purification of xylanase from crude enzymes. Two xylanases were purified by this method from the crude enzyme preparation from the selected strain of Bacillus subtilis. Subsequent analysis with thin layer chromatography and high pressure liquid chromatography (HPLC) indicated that these two xylanases were endo-acting enzymes, designated xyl I and xyl II. Both enzymes showed their activities in the pH range from 5.0 to 9.0 at 50 8C and had similar optimum activities at pH 7.0 and at 50 8C. Mn 2+ ions enhanced their xylanolytic activities to 2.7-fold whereas Fe 3+ completely inhibited them. The action of endoxylanase xyl II on wheat bran insoluble dietary fibre was also studied. The hydrolysis products were shown to contain feruloyl oligosaccharides by paper chromatography. #