Comparison of the yeast microbiota of different varieties of cool-climate grapes by PCR-RAPD

Abstract

The yeast microbiota occurring on different varieties of grapes grown in cool-climate is not completely researched. Therefore, its identification is important to research. On the other hand, yeasts occurring in these fruits can be potentially used as starter cultures to obtain particularly demanded features in the production of wine. In addition, rapid methods for yeast identification allow to eliminate the contamination with pathogenic yeasts, which could cause the loss of wine production. The aim of the study was to isolate and identify the yeasts occurring on the surface of the different varieties of white and red grapes, grown in cool-climate of Poland. Also, the aim was to compare the qualitative and quantitative composition of yeasts on the tested grapes. The 84 cultures of yeasts were isolated, that were initially macroscopic and microscopic analyzed and the purity of cultures was rated on the WL medium. Identification of yeasts by PCR-RAPD was carried using the M13 primer. In the PCR-RFLP method ITS1 and ITS4 primers, as well as restriction enzymes HhaI, HinfI, HaeIII, were used. Preliminary identification of yeasts by standard methods produced results very different from the results obtained by molecular methods. Among the isolated microorganisms yeasts were dominating, but bacteria and molds were also present. Using the PCR-RAPD method most strains of yeasts were identified. Yeast microflora of different varieties of white and red grapes was very similar as the same species of yeasts were identified. Yeasts of the genus Saccharomyces were present in all varieties of grapes. The Rhodotorula mucilaginosa, Saccharomyces cerevisiae, Metschnikowia pulcherrima, Rhodotorula minuta, Pichia kluyveri, Hanseniaspora uvarum and Rhodotorula mucilaginosa were identified by PCR-RAPD. 4 of the 33 tested strains of yeasts were identified by PCR-RFLP. By PCR-RAPD only Hanseniaspora uvarum was identified. The quantity and quality of microorganisms living on the surface of grape fruits is very important for the process of winemaking. Yeasts influence the course of alcoholic fermentation, the flavor, aroma, and thus the quality of the produced wine. To a large extent their presence depends on the condition of the surface of the fruit. Many researchers reported significant differences between yeast microflora in grapes of Mediterranean and cool-climate vineyards. As they are expected to affect the final wine properties precise researching of the microflora of cool-climate grapes may lead to the isolation of new species of yeasts and thus the wines with unique characteristics can be obtained.

Probiotic Yeasts and How to Find Them—Polish Wines of Spontaneous Fermentation as Source for Potentially Probiotic Yeasts

“…The forty-four strains examined belonged to six species: Hanseniaspora uvarum , Pichia kluyveri , Metschnikowia pulcherrima , Metschnikowia ziziphicola , Saccharomyces cerevisiae , and Starmerella bacillaris . Despite limited research on yeast biodiversity in Polish wines, our findings mostly align with the lists of species described by Drozdz et al and Cioch-Skoneczny et al [ 22 , 32 ], but include one species not mentioned by the authors— Starmerella bacillaris . The species, S. bacillaris , known also as Candida zemplinina , is frequently isolated from grapes and wines, and may affect the chemical composition of the musts and wines by its ability to produce various metabolites which influence their enological properties [ 48 , 49 , 50 ].…”

“…Over recent years, with the warming of Poland’s climate, there has been an increased interest in viticulture, resulting in the growth of vineyards by over 550 in the last decade [ 25 , 31 ]. At the time of publication preparation, only two publications characterizing native yeast strains of Polish wines and one metagenomic study were available, but none of them were dedicated to exploring the probiotic potential of the tested strains [ 22 , 25 , 32 ]. To the best of our knowledge, this is the first study focusing on the aspect of probiotic potential of yeast isolates derived from Polish wines and may provide some new perspectives on wine-derived yeasts besides the typical look inside their influence on wine’s enological properties, such as aroma, acidity, texture, etc.…”

Construction and Characterization of an Intergeneric Fusant That Degrades the Fungicides Chlorothalonil and Carbendazim

“…In existing studies, PCR-RAPD analysis has been proved to be a dependable and easy method to calculate the genetic distance between the parents and the fusion ( Savitha et al, 2010 ; Enan, 2011 ; Maharani and Dyah, 2012 ; Drożdż et al, 2015 ; Oluborode et al, 2018 ). The genomic DNA of the parents and the fusant-BD2 were amplified by PCR, and the results are shown in Figure 3 .…”

A new method for extracting DNA from the grape berry surface, beginning in the vineyard

“…Isolating DNA from the surface of a grape berry involves aggressive and disruptive actions, due to tight adhesion of microbes to the thick berry skin and cuticle, making it difficult to wash microbes off the surface using most commercial kits [1], with some exceptions [2, 3]. More commonly, researchers have used culture-based methods [4, 5, 6, 7, 8, 9] or sampled grape must [10, 11, 12] to conduct microbial ecology studies.…”