Search Results (9)

This study characterised the diversity of filamentous fungi and yeasts during Malbec grape fermentation in the Southern Oasis of Mendoza (Argentina) winegrowing region, under different sanitary conditions and SO₂ treatments, using morphological and ITS-RFLP-based molecular methods. Alternaria, Cladosporium and Penicillium were present in both sound and damaged grapes, while Aspergillus and Botrytis were primarily found in damaged grapes. The predominant yeast species in both sound and damaged grape must, at lower and higher maturity levels, were Aureobasidium pullulans and Hanseniaspora spp. At higher grape ripening levels species diversity increased, with Hanseniaspora vineae, Metschnikowia pulcherrima and Candida membranifaciens dominating, and others such as Pichia kudriavzevii and Issatchenkia terricola appearing. A. pullulans and M. pulcherrima were highly tolerant to SO₂. Notably, the species Meyerozyma guilliermondii, Zygoascus hellenicus and Hanseniaspora uvarum were exclusively present in damaged grape must, while Zygosaccharomyces bailii was also found in sound grape must. Hanseniaspora spp. and P. kudriavzevii predominated at mid-fermentation and persisted at the end of the process, highlighting their resistance to wine conditions and their potential to influence post-fermentative dynamics. These findings emphasise the significant influence of grape sanitary status on mycobiota composition, with important implications for fermentation behaviour and final wine quality. Full article

Dual-frequency and moderate thermosonication (TS, 300 + 300 W, 20 + 40 kHz, 25~60 °C) was employed to inactivate Pichia membranifaciens in soybean paste. The aim was to evaluate the effect of TS on the inactivation of P. membranaefaciens and on the quality of soybean paste. The Weibull model fitted the survival data of P. membranaefaciens in thermosonicated soybean paste well and a decrease of 5 log of P. membranaefaciens in soybean paste was obtained at TS_50°C, TS_55°C, TS_60°C, and T_65°C for 15.41, 7.49, 2.27, and 18.61 min. Scanning electron microscope observation revealed TS_50°C damaged the cell structure, leading to the leakage of intracellular contents. The physicochemical properties of soybean paste treated by TS were more retained than in paste treated by heat. The GC-MS analysis indicated that the flavor components had increased after TS treatment, especially at TS_50°C. In conclusion, TS can inactive P. membranaefaciens in soybean paste without causing significant changes in its physicochemical and flavor qualities. Full article

The increasing demand for craft beer is driving the search for novel ale yeast cultures from brewing-related wild environments. The focus of bioprospecting for craft cultures is to identify feral yeasts suitable to imprint unique sensorial attributes onto the final product. Here, we integrated phylogenetic, genotypic, genetic, and metabolomic techniques to demonstrate that sour beer during aging in wooden barrels is a source of suitable craft ale yeast candidates. In contrast to the traditional lambic beer maturation phase, during the aging of sour-matured production-style beer, different biotypes of Saccharomyces cerevisiae dominated the cultivable in-house mycobiota, which were followed by Pichia membranifaciens, Brettanomyces bruxellensis, and Brettanomyces anomalus. In addition, three putative S. cerevisiae × Saccharomyces uvarum hybrids were identified. S. cerevisiae feral strains sporulated, produced viable monosporic progenies, and had the STA1 gene downstream as a full-length promoter. During hopped wort fermentation, four S. cerevisiae strains and the S. cerevisiae × S. uvarum hybrid WY213 exceeded non-Saccharomyces strains in fermentative rate and ethanol production except for P. membranifaciens WY122. This strain consumed maltose after a long lag phase, in contrast to the phenotypic profile described for the species. According to the STA1+ genotype, S. cerevisiae partially consumed dextrin. Among the volatile organic compounds (VOCs) produced by S. cerevisiae and the S. cerevisiae × S. uvarum hybrid, phenylethyl alcohol, which has a fruit-like aroma, was the most prevalent. In conclusion, the strains characterized here have relevant brewing properties and are exploitable as indigenous craft beer starters. Full article

Fungal endophytes are not widely known for their role in bioactive metabolite production and salinity stress alleviation in different crop plants. Presently, we investigated the salt stress (NaCl, KCl, and H₂SO₄) mitigation capabilities of fungal endophyte Candida membranifaciens (FH15) isolated from Euphorbia milii L. The pure culture filtrate (CF) of C. membranifaciens revealed siderophore production and solubilization of phosphate, with high levels of indoleacetic acid (IAA: 35.8µg/mL), phenolics (70 µg/mL), and flavonoids (50 µg/mL) by using a UV spectrophotometer. The LC/MS analysis of the CF showed different phenols and flavonoids that were identified as Salicylic acid, Baicalein, Aconitic acid, Feruloylquinic acid, Coniferyl aldehyde hexoside, Pentose, Chlorogenic acid, Myricetin, Propoxyphene, and Amino-flunitrazepam. Inoculation of maize seedlings with C. membranifaciens significantly (p = 0.05) enhanced the fresh and dry biomass, carotenoid, and chlorophyll contents under 100 mM salt stress conditions. Similarly, the catalase, peroxidase activity, phenols, proline flavonoids and relative water contents (RWC) of the maize plants were enhanced. More interestingly, the inoculation of C. membranifaciens on maize revealed a higher endogenous IAA level as compared to non-inoculated control plants. Endophyte C. membranifaciens inoculation on maize seedlings under salt stress revealed a 20.87% and 16.60% increase in fresh and dry biomass, as well as significantly enhanced root shoot length and allied growth attributes, in addition to an alleviation of the adverse effects of salinity stress. Conclusively, endophytic C. membranifaciens significantly enhanced the growth attributes of maize and mitigated the adverse effects of salinity stress. Such endophytic fungal strain could be used for further field trails to enhance agricultural productivity and facilitate sustainable agricultural practices. Full article

A new method was proposed to produce alginate bio-films containing Pichia membranifaciens and Wickerhamomyces anomalus killer yeast to control the post-harvest fungal decay in organic apples caused by Botrytis cinerea and Penicillium italicum. Coatings with W. anomalus killer yeast effectively controlled the growth of P. italicum during storage at 22 °C. W. anomalus killer yeast incorporated in alginate reduced the P. italicum incidence from 90% (control) to 35% after 14 days of storage at 22 °C. Alginate biofilms with W. anomalus or P. membranifaciens also limited the incidence of the fungal decay of apples inoculated with B. cinerea compared with the control fruits, although the antagonistic capability against B. cinerea was lower than against P. italicum. The survival of W. anomalus cells in alginate coating was higher than P. membranifaciens. The incorporation of killer yeasts into alginate had no significant effect on the mechanical properties (tensile strength, percent elongation at break) of alginate coating, however, they increased the thickness of the biofilm. The bioactive coating reduced the fruit weight loss and had no significant effects on the fruit firmness during storage at 2 °C. As organic apples, produced without any synthetic fungicides, are especially prone to fungal decay during storage, the proposed alginate biofilms containing killer yeast seem to be a very promising solution by offering non-chemical, biological control of post-harvest pathogens. Full article

The surfaces of grapes are covered by different yeast species that are important in the first stages of the fermentation process. In recent years, non-Saccharomyces yeasts such as Torulaspora delbrueckii, Lachancea thermotolerans, Metschnikowia pulcherrima, and Pichia kluyveri have become popular with regard to winemaking and improved wine quality. For that reason, several manufacturers started to offer commercially available strains of these non-Saccharomyces species. P. kluyveri stands out, mainly due to its contribution to wine aroma, glycerol, ethanol yield, and killer factor. The metabolism of the yeast allows it to increase volatile molecules such as esters and varietal thiols (aroma-active compounds), which increase the quality of specific varietal wines or neutral ones. It is considered a low- or non-fermentative yeast, so subsequent inoculation of a more fermentative yeast such as Saccharomyces cerevisiae is indispensable to achieve a proper fermented alcohol. The impact of P. kluyveri is not limited to the grape wine industry; it has also been successfully employed in beer, cider, durian, and tequila fermentation, among others, acting as a promising tool in those fermentation processes. Although no Pichia species other than P. kluyveri is available in the regular market, several recent scientific studies show interesting improvements in some wine quality parameters such as aroma, polysaccharides, acid management, and color stability. This could motivate yeast manufacturers to develop products based on those species in the near future. Full article

The aim of the present study was to assess the microecosystem of 13 homemade spontaneously fermented wheat sourdoughs from different regions of Greece, through the combined use of culture-dependent (classical approach; clustering by Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) and identification by PCR species-specific for Lactiplantibacillus plantarum, and sequencing of the 16S-rRNA and 26S-rRNA gene, for Lactic Acid Bacteria (LAB) and yeasts, respectively) and independent approaches [DNA- and RNA-based PCR-Denaturing Gradient Gel Electrophoresis (DGGE)]. The pH and Total Titratable Acidity (TTA) values ranged from 3.64–5.05 and from 0.50–1.59% lactic acid, respectively. Yeast and lactic acid bacteria populations ranged within 4.60–6.32 and 6.28–9.20 log CFU/g, respectively. The yeast: LAB ratio varied from 1:23–1:10,000. A total of 207 bacterial and 195 yeast isolates were obtained and a culture-dependent assessment of their taxonomic affiliation revealed dominance of Lb. plantarum in three sourdoughs, Levilactobacillus brevis in four sourdoughs and co-dominance of these species in two sourdoughs. In addition, Companilactobacillusparalimentarius dominated in two sourdoughs and Fructilactobacillussanfranciscensis and Latilactobacillus sakei in one sourdough each. Lactococcus lactis, Lb. curvatus, Leuconostoc citreum, Ln. mesenteroides and Lb. zymae were also recovered from some samples. Regarding the yeast microbiota, it was dominated by Saccharomyces cerevisiae in 11 sourdoughs and Pichia membranifaciens and P. fermentans in one sourdough each. Wickerhamomyces anomalus and Kazachstania humilis were also recovered from one sample. RNA-based PCR-DGGE provided with nearly identical results with DNA-based one; in only one sample the latter provided an additional band. In general, the limitations of this approach, namely co-migration of amplicons from different species to the same electrophoretic position and multiband profile of specific isolates, greatly reduced resolution capacity, which resulted in only partial verification of the microbial ecology detected by culture-dependent approach in the majority of sourdough samples. Our knowledge regarding the microecosystem of spontaneously fermented Greek wheat-based sourdoughs was expanded, through the study of sourdoughs originating from regions of Greece that were not previously assessed. Full article

Fusarium species coexist as toxigenic, systemic pathogens in sweet corn seed production in southwestern Idaho, USA. We hypothesized that fungal antagonists of seedborne Fusarium would differentially alter production of Fusarium mycotoxins directly and/or systemically. We challenged the Fusarium complex by in vitro antagonism trials and in situ silk and seed inoculations with fungal antagonists. Fungal antagonists reduced growth and sporulation of Fusarium species in vitro from 40.5% to as much as 100%. Pichia membranifaciens and Penicillium griseolum reduced fumonisin production by F. verticillioides by 73% and 49%, respectively, while P. membranifaciens and a novel Penicillium sp. (WPT) reduced fumonisins by F. proliferatum 56% and 78%, respectively. In situ, pre-planting inoculation of seeds with Penicillium WPT systemically increased fumonisins in the resulting crop. Morchella snyderi applied to silks of an F1 cross systemically reduced deoxynivalenol by 47% in mature seeds of the F2. Antagonists failed to suppress Fusarium in mature kernels following silk inoculations, although the ratio of F. verticillioides to total Fusarium double with some inoculants. Fusarium mycotoxin concentrations in sweet corn seed change systemically, as well as locally, in response to the presence of fungal antagonists, although in Fusarium presence in situ was not changed. Full article

The killer phenomenon is defined as the ability of some yeast to secrete toxins that are lethal to other sensitive yeasts and filamentous fungi. Since the discovery of strains of Saccharomyces cerevisiae capable of secreting killer toxins, much information has been gained regarding killer toxins and this fact has substantially contributed knowledge on fundamental aspects of cell biology and yeast genetics. The killer phenomenon has been studied in Pichia membranifaciens for several years, during which two toxins have been described. PMKT and PMKT2 are proteins of low molecular mass that bind to primary receptors located in the cell wall structure of sensitive yeast cells, linear (1→6)-β-d-glucans and mannoproteins for PMKT and PMKT2, respectively. Cwp2p also acts as a secondary receptor for PMKT. Killing of sensitive cells by PMKT is characterized by ionic movements across plasma membrane and an acidification of the intracellular pH triggering an activation of the High Osmolarity Glycerol (HOG) pathway. On the contrary, our investigations showed a mechanism of killing in which cells are arrested at an early S-phase by high concentrations of PMKT2. However, we concluded that induced mortality at low PMKT2 doses and also PMKT is indeed of an apoptotic nature. Killer yeasts and their toxins have found potential applications in several fields: in food and beverage production, as biocontrol agents, in yeast bio-typing, and as novel antimycotic agents. Accordingly, several applications have been found for P. membranifaciens killer toxins, ranging from pre- and post-harvest biocontrol of plant pathogens to applications during wine fermentation and ageing (inhibition of Botrytis cinerea, Brettanomyces bruxellensis, etc.). Full article