Nitrogen fixing bacteria in the family Acetobacteraceae and their role in agriculture.

For centuries, the Acetobacteraceae is known as a family that harbors many species of organisms of biotechnological importance for industry. Nonetheless, since 1988 representatives of this family have also been described as nitrogen fixing bacteria able to plant growth promotion by a variety of mechanisms. Nitrogen fixation is a biological process that guarantees that the atmospheric N2 is incorporated into organic matter by several bacterial groups. Most representatives of this group, also known as diazotrophic, are generally associated with soil rhizosphere of many plants and also establishing a more specific association living inside roots, leaves, and others plants tissues as endophyte. Their roles as plant growth-promoting microorganisms are generally related to increase in plant biomass, phosphate and other mineral solubilization, and plant pathogen control. Here, we report many of these plant growth-promoting processes related to nitrogen fixing species already described in Acetobacteraceae family, especially Gluconacetobacter diazotrophicus and their importance to agriculture. In addition, a brief review of the state of art of the phylogenetics, main physiological and biochemical characteristics, molecular and functional genomic data of this group of Acetobacteraceae is presented.

Validation of a Tn5 transposon mutagenesis system for Gluconacetobacter diazotrophicus through characterization of a flagellar mutant.

Gluconacetobacter diazotrophicus is a nitrogen-fixing bacterium, which was originally isolated from the interior of sugarcane plants. The genome of strain PAL5 of G. diazotrophicus has been completely sequenced and a next step is the functional characterization of its genes. The aim of this study was to establish an efficient mutagenesis method, using the commercial Tn5 transposon EZ::Tn5Tnp Transposome (Epicentre). Up to 1 x 10(6) mutants per microgram of transposome were generated in a single electroporation experiment. Insertion-site flanking sequences were amplified by inverse PCR and sequenced for 31 mutants. For ten of these mutants, both insertion flanks could be identified, confirming the 9 bp duplication that is typical for Tn5 transposition. Insertions occurred in a random fashion and were genetically stable for at least 50 generations. One mutant had an insertion in a homolog of the flagellar gene flgA, and was therefore predicted to be affected in flagella-dependent traits and used to validate the applied mutagenesis methodology. This mutant lacked flagella and was non-motile on soft agar. Interestingly, it was also strongly affected in the ability to form biofilm on glass wool.

Manufacture of a beverage from cheese whey using a "tea fungus" fermentation.

Kombucha is a sour beverage reported to have potential health effects prepared from the fermentation of black tea and sugar with a "tea fungus", a symbiotic culture of acetic acid bacteria and yeasts. Although black tea is the preferred substrate for Kombucha fermentation, other beverages have also been tested as substrates with fair results. Cheese whey is a by-product with a good amount of fermentable lactose that has been used before in the production of beverages, so the objective of this study was to test three types of whey (fresh sweet, fresh acid and reconstituted sweet) in the elaboration of a fermented beverage using a kombucha culture as inoculum. The isolation and identification of bacteria and yeasts from the fermented tea and wheys was done along with the study of the rates of change in sugar consumption, acid production and pH decrease. Several species of acetic acid bacteria (Acetobacter aceti subsp. aceti, Gluconobacter oxydans subsp. industrius, subsp. oxydans and Gluconoacetobacter xylinus) were isolated from the different kombuchas along with the yeasts Saccharomyces cerevisiae, Kluyveromyces marxianus, and Brettanomyces bruxelensis. The main metabolic products in the fermented wheys included ethanol, lactic and acetic acids. A good growth was obtained in both sweet wheys in which a pH of 3.3 and a total acid content (mainly lactic and acetic acids) of 0.07 mol/l was reached after 96 h. The sweet whey fermented beverages contained a relatively low lactose concentration (< 12 g/l). The final ethanol content was low (5 g/l) in all the fermented wheys. The whey products were strongly sour and salty non sparkling beverages.