The hidden treasures in endophytic fungi: a comprehensive review on the diversity of fungal bioactive metabolites, usual analytical methodologies, and applications.

This review provides a comprehensive overview of the key aspects of the natural metabolite production by endophytic fungi, which has attracted significant attention due to its diverse biological activities and wide range of applications. Synthesized by various fungal species, these metabolites encompass compounds with therapeutic, agricultural, and commercial significance. We delved into strategies and advancements aimed at optimizing fungal metabolite production. Fungal cultivation, especially by Aspergillus, Penicillium, and Fusarium, plays a pivotal role in metabolite biosynthesis, and researchers have explored both submerged and solid-state cultivation processes to harness the full potential of fungal species. Nutrient optimization, pH, and temperature control are critical factors in ensuring high yields of the targeted bioactive metabolites especially for scaling up processes. Analytical methods that includes High-Performance Liquid Chromatography (HPLC), Liquid Chromatography-Mass Spectrometry (LC-MS), Gas Chromatography-Mass Spectrometry (GC-MS), Nuclear Magnetic Resonance (NMR), and Mass Spectrometry (MS), are indispensable for the identification and quantification of the compounds. Moreover, genetic engineering and metabolic pathway manipulation have emerged as powerful tools to enhance metabolite production and develop novel fungal strains with increased yields. Regulation and control mechanisms at the genetic, epigenetic, and metabolic levels are explored to fine-tune the biosynthesis of fungal metabolites. Ongoing research aims to overcome the complexity of the steps involved to ensure the efficient production and utilization of fungal metabolites.

Methods used for the study of endophytic fungi: a review on methodologies and challenges, and associated tips.

Endophytic fungi are microorganisms that colonize the interior of plant tissues (e.g. leaves, seeds, stem, trunk, roots, fruits, flowers) in intracellular and/or extracellular spaces without causing symptoms of disease in host plants. These microorganisms have been isolated from plant species in a wide variety of habitats worldwide, and it is estimated that all terrestrial plants are colonized by one or more species of endophytic fungus. In addition, these microorganisms have been drawing the attention of researchers because of their ability to synthesize a wide range of bioactive molecules with potential for applications in agriculture, medicine and biotechnology. However, several obstacles come up when studying the diversity and chemical potential of endophytic fungi. For example, the usage of an inappropriate surface disinfection method for plant tissue may not eliminate the epiphytic microbiota or may end up interfering with the endophytic mycobiota, which consequently generates erroneous results. Moreover, the composition of the culture medium and the culture conditions can favor the growth of certain species and inhibit others, which generates underestimated results. Other inconsistencies can arise from the fungus misidentification and consequent exploration of its chemical potential. Based on the methodological biases that may occur at all stages of studies dealing with endophytic fungi, the objective of this review is to discuss the main methods employed in these studies as well as highlight the challenges derived from the different approaches. We also report associated tips to help future studies on endophytic fungi as a contribution.

Insights into taxonomic diversity and bioprospecting potential of Cerrado endophytic fungi: a review exploring an unique Brazilian biome and methodological limitations.

Cerrado is the second largest biome in Brazil, and it is known for harboring a wide variety of endemic plant and microbial species, among which are endophytic fungi. Endophytic fungi are microorganisms capable of colonizing the interior of plant tissues without causing disease in host plants. Especially in the Cerrado biome, this group of microorganisms is still poorly studied and information on species estimation, ecological and evolutionary importance is not accurate and remains unknown. Also, it is extremely important to emphasize that great part of studies available on Cerrado endophytic fungi are national literature, including master's dissertations, course conclusion works or unpublished doctoral theses. The majority of these studies has highlighted that the endemic plant species are an important habitat for fungal endophytes, and new species have increasingly been described. Due to the lack of international literature on Cerrado endophytic fungi, the present review brings a bibliographic survey on taxonomic diversity and bioprospecting potential of fungal endophytes from a unique environment. This review also emphasizes the importance of studying Brazilian endophytic fungi from Cerrado as a source of new technologies (biofertilizer and biocontroller), since they are secondary metabolite-producing organisms with different biological activities for biotechnological, agricultural and pharmaceutical applications.

Brief history of biofertilizers in Brazil: from conventional approaches to new biotechnological solutions.

Brazil has a long history of research with rhizobia and plant growth-promoting rhizobacteria (PGPR). Currently, the use of bio-based products in Brazil, containing microorganisms that are effective in promoting plant growth through various mechanisms, is already a consolidated reality for the cultivation of several crops of agricultural interest. This is due to the excellent results obtained over many years of research, which contributed to reinforce the use of rhizobia and PGPR by farmers. The high quality of the products offered, containing elite strains, allows the reduction and prevention in the use of mineral fertilization, contributing to low-cost and sustainable agriculture. Currently, research has turned its efforts in the search for new products that further increase the efficiency of those already available on the market and for new formulations or inoculation strategies that contribute to greater productivity and efficiency of these products. In this review, the history of biological products for main crops of agricultural interest and the new biotechnologies and research available in the agricultural market are discussed.

Isolation, Identification, and Screening of Lactic Acid Bacteria with Probiotic Potential in Silage of Different Species of Forage Plants, Cocoa Beans, and Artisanal Salami.

The objective of this study was to isolate and characterize lactic acid bacteria with probiotic potential in silages of different species of forage plants, cocoa beans, and artisanal salami. The obtained isolates were submitted to the following evaluations: (i) screening for tolerance to pH 2 and bile salts, (ii) genotypic identification of isolates, (iii) survival in simulated gastric and pancreatic conditions, (iv) antimicrobial activity, (v) antibiotic susceptibility and safety, and (vi) properties associated with adhesion capacity. A total of 82 isolates were obtained and were screened for pH 2.0 tolerance and capacity to growth in the presence of bile salts (1.0 and 2.0%). Only 19 strains simultaneously presented tolerance to pH 2.0 and bile salts. These 19 strains were evaluated for genetic profile by Box-PCR. Subsequently, the selected strains were subjected to partial sequencing of the 16S rRNA gene. The species Lactobacillus plantarum was prevalent. The identified strains were evaluated for survival under simulated gastric and pancreatic conditions. Some strains have shown tolerance in both conditions. Different strains showed variations in antimicrobial activity, susceptibility to antibiotics, and properties associated with adhesion (hydrophobicity, autoaggregation, coaggregation, and adhesion to CaCo2 cells). All strains were negative for hemolysis, DNase, gelatinase, and biogenic amine synthesis activity. The L. plantarum SBR64.7 strain can be considered the most promising for it presented the lowest viability reduction when exposed to gastric and pancreatic juices.

Correction to: Identification of soybean Bradyrhizobium strains used in commercial inoculants in Brazil by MALDI-TOF mass spectrometry.

The original version of this article unfortunately contained a mistake. The presentation of Fig. 1was incorrect. The correct version is given below.

Identification of soybean Bradyrhizobium strains used in commercial inoculants in Brazil by MALDI-TOF mass spectrometry.

Biological nitrogen fixation (BNF) with the soybean crop probably represents the major sustainable technology worldwide, saving billions of dollars in N fertilizers and decreasing water pollution and the emission of greenhouse gases. Accordingly, the identification of strains occupying nodules under field conditions represents a critical step in studies that are aimed at guaranteeing increased BNF contribution. Current methods of identification are mostly based on serology, or on DNA profiles. However, the production of antibodies is restricted to few laboratories, and to obtain DNA profiles of hundreds of isolates is costly and time-consuming. Conversely, the matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS technique might represent a golden opportunity for replacing serological and DNA-based methods. However, MALDI-TOF databases of environmental microorganisms are still limited, and, most importantly, there are concerns about the discrimination of protein profiles at the strain level. In this study, we investigated four soybean rhizobial strains carried in commercial inoculants used in over 35 million hectares in Brazil and also in other countries of South America and Africa. A supplementary MALDI-TOF database with the protein profiles of these rhizobial strains was built and allowed the identification of unique profiles statistically supported by multivariate analysis and neural networks. To test this new database, the nodule occupancy by Bradyrhizobium strains in symbiosis with soybean was characterized in a field experiment and the results were compared with serotyping of bacteria by immuno-agglutination. The results obtained by both techniques were highly correlated and confirmed the viability of using the MALDI-TOF MS technique to effectively distinguish bacteria at the strain level.