Agrobacterium tumefaciens is considered a prominent phytopathogen, though most isolates are nonpathogenic. Agrobacteria can inhabit plant tissues interacting with other microorganisms. Yeasts are likewise part of these communities. We analyzed the quorum sensing (QS) systems of A. tumefaciens strain 6N2, and its relevance for the interaction with the yeast Meyerozyma guilliermondii , both sugarcane endophytes. We show that strain 6N2 is nonpathogenic, produces OHC8-HSL, OHC10-HSL, OC12-HSL and OHC12-HSL as QS signals, and possesses a complex QS architecture, with one truncated, two complete systems, and three additional QS-signal receptors. A proteomic approach showed differences in QS-regulated proteins between pure (64 proteins) and dual (33 proteins) cultures. Seven proteins were consistently regulated by quorum sensing in pure and dual cultures. M. guilliermondii proteins influenced by QS activity were also evaluated. Several up- and down- regulated proteins differed depending on the bacterial QS. These results show the importance of the QS regulation in the bacteria-yeast interactions. Highlights: The avirulent A. tumefaciens 6N2 has two replicons and a complex QS architecture The profile of QS-regulated proteins is modified in dual cultures with Pa. laurentii The bacterial QS activity alters the proteome of the yeast Pa. laurentii

Vinasse is a by-product with a key role in the circular economy. In this work, we analyze sugarcane vinasse as culture medium for obtaining single and mixed inoculants. Trichoderma harzianum was cultured in single and sequential co-culture with Pseudomonas capeferrum or Rhizobium sp. Fungal biomass was higher in vinasse than in a laboratory medium. Residual vinasses presented almost neutral pH and lower conductivities and toxicity than raw vinasse. Fertigation with residual vinasses improves characteristics of soil evidenced in the total N, cation exchange capacity, urease and acid phosphatase, and the microbial metabolic diversity, in comparison to raw vinasse. The evaluation of the treatment indicates that vinasse is suitable for the production of inoculants containing T. harzianum and that the co-culture with P. capeferrum improves the characteristics of the residual vinasse in comparison to Rhizobium sp. Obtaining this valuable biomass in vinasse is relevant for the circular and green economy.

Agrobacterium tumefaciens is considered a prominent phytopathogen, though most isolates are nonpathogenic. Agrobacteria can inhabit plant tissues interacting with other microorganisms. Yeasts are likewise part of these communities. We analyzed the quorum sensing (QS) systems of A. tumefaciens strain 6N2, and its relevance for the interaction with the yeast Meyerozyma guilliermondii, both sugarcane endophytes. We show that strain 6N2 is nonpathogenic, produces OHC8-HSL, OHC10-HSL, OC12-HSL and OHC12-HSL as QS signals, and possesses a complex QS architecture, with one truncated, two complete systems, and three additional QS-signal receptors. A proteomic approach showed differences in QS-regulated proteins between pure (64 proteins) and dual (33 proteins) cultures. Seven proteins were consistently regulated by quorum sensing in pure and dual cultures. M. guilliermondii proteins influenced by QS activity were also evaluated. Several up- and down- regulated proteins differed depending on the bacterial QS. These results show the QS regulation in the bacteria-yeast interactions.

After a time away from the classrooms and laboratories due to the global pandemic, the return to the teaching activities during the semester represented a challenge to both teachers and students. Our particular situation in a Microbial Physiology course was the necessity of imparting in a shorter time, laboratory practices that usually take longer. This article describes a two-week long laboratory exercise that covers several concepts in an interrelated way: conjugation as a gene transfer mechanism, regulation of microbial physiology, production of secondary metabolites, degradation of macromolecules and biofilm formation. Utilizing a Quorum Quenching (QQ) strategy, the Quorum Sensing (QS) system of Pseudomonas aeruginosa is first attenuated. Then, phenotypes regulated by QS are evidenced. QS is a regulatory mechanism of the microbial physiology that relays on signal molecules. QS is related in P. aeruginosa to several virulence factors, some of which are exploited in the laboratory practices presented in this work. QQ is phenomenon by which QS is interrupted or attenuated. We utilized a QQ approach based on the enzymatic degradation of the P. aeruginosa QS signals in order to put in evidence QS-regulated traits that are relevant for our Microbial Physiology course.

In the environment, communication among microorganisms is a key aspect of their lives. Through communication, single cells of a microbial population can coordinate their gene expression and, in consequence, their physiology. Quorum sensing systems (QS) are one of these mechanisms that allow this communication through the production and detection of signal molecules whose concentration is related to the density of the signaling cells. In several Gram-negative bacteria, these QS signals belongs to the N-acyl homoserine lactone (AHL) family, which can be detected in the laboratory by means of simple bioassays. In this report, we present for undergraduate students a detailed protocol that allows us to evidence QS activity in environmental samples. First, with a selective culture medium, Gram-negative bacteria are isolated from samples provided by the students. Then, isolates are screened with two biosensor strains for the detection of AHL production. Positive results are visualized by the induction of violacein production in the biosensors. These experiments evidence the presence of communicating microorganisms in the analyzed samples. This simple protocol allows the students to analyze a particular microbial activity in samples provided by them, which enhance their learning experiences.