Starter cultures as biocontrol strategy to prevent Brettanomyces bruxellensis proliferation in wine

Brettanomyces bruxellensis is a common and significant wine spoilage microorganism. B. bruxellensis strains generally detain the molecular basis to produce compounds that are detrimental for the organoleptic quality of the wine, including some classes of volatile phenols that derive from the sequential bioconversion of specific hydroxycinnamic acids such as ferulate and p-coumarate. Although B. bruxellensis can be detected at any stage of the winemaking process, it is typically isolated at the end of the alcoholic fermentation (AF), before the staring of the spontaneous malolactic fermentation (MLF) or during barrel aging. For this reason, the endemic diffusion of B. bruxellensis leads to consistent economic losses in the wine industry. Considering the interest in reducing sulfur dioxide use during winemaking, in recent years, biological alternatives, such as the use of tailored selected yeast and bacterial strains inoculated to promote AF and MLF, are actively sought as biocontrol agents to avoid the “Bretta” character in wines. Here, we review the importance of dedicated characterization and selection of starter cultures for AF and MLF in wine, in order to reduce or prevent both growth of B. bruxellensis and its production of volatile phenols in the matrix.

     

A Nested Polymerase Chain Reaction Protocol for in planta Detection of Fusicladium eriobotryae,Causal Agent of Loquat Scab

Scab caused by the fungus Fusicladium eriobotryae is the most serious disease affecting loquat in Spain. Isolation of F. eriobotryae from infected tissue on culture media can be difficult due to its slow growth. A polymerase chain reaction (PCR)‐based protocol was developed for F. eriobotryae‐specific identification from pure culture or infected loquat tissues. The primer set was designed in the elongation factor 1‐α gene (EF1‐α), and specificity and sensitivity for single and nested PCR were validated. The nested PCR assay resulted in 100% positive detection of F. eriobotryae in naturally and artificially infected tissues. This protocol can be useful for routine diagnosis, disease monitoring programmes and epidemiological research.     

Saccharomyces cerevisiae and Oenococcus oeni immobilized in different layers of a cellulose/starch gel composite for simultaneous alcoholic and malolactic wine fermentations

The production of a two-layer composite biocatalyst for immobilization of two different microorganisms for simultaneous alcoholic and malolactic fermentation (MLF) of wine in the same bioreactor is reported. The biocatalyst consisted of a tubular delignified cellulosic material (DCM) with entrapped Oenococcus oeni cells, covered with starch gel containing the alcohol resistant and cryotolerant strain Saccharomyces cerevisiae AXAZ-1. The biocatalyst was found effective for simultaneous low temperature alcoholic fermentation resulting to conversion of malic acid to lactic acid in 5 days at 10 °C. Improvement of wine quality compared with wine fermented with S. cerevisiae AXAZ-1 immobilized on DCM was attributed to MLF as well as to increased ester formation and lower higher alcohols produced at low fermentation temperatures (10 °C) as shown by GC and headspace SPME GC/MS analysis. Scanning electron microscopy showed that the preparation of a three-layer composite biocatalyst is also possible. The significance of such composite biocatalysts is the feasibility of two or three bioprocesses in the same bioreactor, thus reducing production cost in the food industry     

Characterization of Fusarium circinatum from Pinus spp. in northern Spain

Pitch canker caused by Fusarium circinatum was recently reported on Pinus spp. in Spain. In this study, a collection of 157 isolates of F. circinatum obtained from different geographical origins and hosts in northern Spain were identified and characterized by cultural and morphological features, PCR-RFLPs of the histone H3 gene, IGS region, and the translation elongation factor 1-alpha gene (TEF). Mating types were determined by multiplex PCR and sexual compatibility was performed under laboratory conditions. Both mating types were present in Spain and were able to form the teleomorph Gibberella circinata. Morphological differences between mating types, not previously reported, were observed: MAT-1 isolates showed clear, coiled, sterile hyphae characteristic of F. circinatum, whereas MAT-2 isolates presented sterile hyphae but not coiled. Virulence of representative isolates was tested on seven to eight-month-old P. nigra, P. pinaster and P. sylvestris seedlings. All isolates tested were pathogenic to these pine species, MAT-1 isolates being more virulent than MAT-2 isolates.