RESUMO
Bacterial genome sequencing has revealed a vast number of novel biosynthetic gene clusters (BGC) with potential to produce bioactive natural products. However, the biosynthesis of secondary metabolites by bacteria is often silenced under laboratory conditions, limiting the controlled expression of natural products. Here we describe an integrated methodology for the construction and screening of an elicited and pre-fractionated library of marine bacteria. In this pilot study, chemical elicitors were evaluated to mimic the natural environment and to induce the expression of cryptic BGCs in deep-sea bacteria. By integrating high-resolution untargeted metabolomics with cheminformatics analyses, it was possible to visualize, mine, identify and map the chemical and biological space of the elicited bacterial metabolites. The results show that elicited bacterial metabolites correspond to ~45% of the compounds produced under laboratory conditions. In addition, the elicited chemical space is novel (~70% of the elicited compounds) or concentrated in the chemical space of drugs. Fractionation of the crude extracts further evidenced minor compounds (~90% of the collection) and the detection of biological activity. This pilot work pinpoints strategies for constructing and evaluating chemically diverse bacterial natural product libraries towards the identification of novel bacterial metabolites in natural product-based drug discovery pipelines.
RESUMO
As a result of the capability of fungi to respond to culture conditions, we aimed to explore and compare the antibacterial activity and chemical diversity of two endophytic fungi isolated from Hyptis dilatata and cultured under different conditions by the addition of chemical elicitors, changes in the pH, and different incubation temperatures. Seventeen extracts were obtained from both Pestalotiopsis mangiferae (man-1 to man-17) and Pestalotiopsis microspora (mic-1 to mic-17) and were tested against a panel of pathogenic bacteria. Seven extracts from P. mangiferae and four extracts from P. microspora showed antibacterial activity; while some of these extracts displayed a high-level of selectivity and a broad-spectrum of activity, Pseudomonas aeruginosa was the most inhibited microorganism and was selected to determine the minimal inhibitory concentration (MIC). The MIC was determined for extracts man-6 (0.11 µg/mL) and mic-9 (0.56 µg/mL). Three active extracts obtained from P. mangiferae were analyzed by Liquid Chromatography-Electrospray Ionization-Quadrupole-Time of Flight-Mass Spectrometry (LC-ESI-Q-TOF-MS) to explore the chemical diversity and the variations in the composition. This allows us to propose structures for some of the determined molecular formulas, including the previously reported mangiferaelactone (1), an antibacterial compound.
RESUMO
Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae) biotype B is a key pest of Solanum lycopersicum L. (Solanaceae) throughout the world. In this study, we examined the induction of resistance on tomato plants treated with SA, BABA, and Trichoderma either individually or in combination against B. tabaci biotype B through the assessment of some biological and behavioral aspects of this insect pest. Also, to understand the mode of action of these inducers, we correlated and analyzed the biochemical basis of plant resistance, by measuring levels of polyphenol oxidase (PPO), peroxidase (POD), phenylalanine ammonia lyase (PAL), and phenolic content in leaves of treated tomato plants. The longest development time of whitefly immature stages was recorded for plants treated with root ß-aminobutyric acid application (RBABA) + root Trichoderma application (RT), root salicylic acid application (RSA) + RT, and RT. In a free-choice assay, B. tabaci adults showed a significantly lower preference for settling and oviposition in RBABA + RT, RSA + RT, and RT in comparison with control. In a no-choice assay, B. tabaci females laid significantly fewer eggs on treatments than those in control, with better results observed in RBABA + RT. Plants responded to different treatments and showed higher induction of PPO, POD, and PAL activities, besides the higher accumulation of phenols in RBABA + RT, RSA + RT, and RT treatments. These results suggest that RBABA + RT, RSA + RT, and RT could be utilized for the induction of effective plant defense against B. tabaci.