scholarly journals Aislamiento e Identificación Molecular de Levaduras Autóctonas en Viñedo de la Variedad Malvasía Blanca

Author(s):  
María Berradre ◽  
Luzmila Meza ◽  
Braulio Zarzoso ◽  
Jorge Ortega ◽  
Betzabé Sulbarán ◽  
...  

Se realizó el aislamiento e identificación de levaduras nativas de un viñedo de la especie Vitis vinifera variedad Malvasía blanca, en la Región Zuliana. Se muestrearon aséptica y aleatoriamente bayas y partes de la planta (hojas, raquis, corteza y suelo), de un total de 123 plantas, correspondientes a un viñedo de la variedad de uva Malvasía. A aproximadamente a 500m del viñedo, se encuentra la bodega. La identificación de las levaduras aisladas en el viñedo se realizó por técnicas moleculares mediante PCR-RFLP, sometiendo los productos amplificados a un análisis de restricción con las enzimas Hinf I, Hae III, CfoI y DdeI.  La distribución de las levaduras en los diversos sustratos fue en el suelo 60% Hanseniaspora guillermondii y 40% Hanseniaspora uvarum, en la corteza 90% Candida sake y 10% Hanseniaspora uvarum, en hojas 100% por Rhodotorula mucilagenosa, en raquis 100% por Aureobasidium pullulans y en las bayas 96% Rhodotorula mucilagenosa y 4% Aureobasidium pullulans.  En el viñedo están ampliamente difundidos los géneros Ascomycetos Hanseniaspora, Candida y Aureobasidium y el género Basdiomycetos Rhodotorula, siendo las levaduras oxidativas Aureobasidium y Rhodotorula las de mayor difusión en el mismo, sin embargo, cabe destacar la presencia de levaduras fermentativas como los géneros Hanseniaspora y Candida, importantes levaduras con reconocido potencial enológico, que podrán ser utilizadas en futuras fermentaciones alcohólicas para obtener vinos con calidad única por ser fermentados con levaduras autóctonas adaptadas a clima tropical.  Palabras clave: levaduras autóctonas, variedad Malvasía, PCR-RFLP 5,8S.    ABSTRACT The isolation and identification of native yeasts from a vineyard of the Vitis vinifera white variety Malvasia was carried out in Zulia, Venezuela. Aseptically and randomly, berries and parts of the plant (leaves, rachis, bark and soil) were sampled from a total of 123 plants, corresponding to a vineyard of the Malvasia grape variety. A winery is located at approximately 500 m from the vineyard. The identification of the yeasts isolated in the vineyard was carried out by molecular techniques by PCR-RFLP, subjecting the amplified products to a restriction analysis with the enzymes Hinf I, Hae III, CfoI and DdeI. The distribution of the yeasts in the different substrates was in the soil 60% Hanseniaspora guillermondii and 40% Hanseniaspora uvarum, in the bark 90% Candida sake and 10% Hanseniaspora uvarum, in leaves 100% by Rhodotorula mucilagenosa, in rachis 100% by Aureobasidium pullulans and in berries 96% Rhodotorula mucilagenosa and 4% Aureobasidium pullulans. In the vineyard, the Ascomycetos type Hanseniaspora, Candida and Aureobasidium and the genus Basdiomycetos Rhodotorula are widely spread, with the oxidative yeasts Aureobasidium and Rhodotorula being the most widespread, however, the presence of fermentative yeasts such as the Hanseniaspora  and Candida genera, important yeasts with recognized oenological potential, which can be used in future alcoholic fermentations to obtain wines with unique quality by being fermented with native yeasts adapted to tropical climate. Key words: native yeasts, Malvasía variety, PCR-RFLP 5.8S.

2021 ◽  
Vol 285 ◽  
pp. 05020
Author(s):  
Elena Lobodina ◽  
Ivan Suprun ◽  
Natalya Ageeva ◽  
Ekaterina Al-Nakib

The studies present the results of morphological, cultural and genetic analysis of the ITS1-ITS4 region of the autochthonous yeast strains genome by using the HaeIII restriction enzyme. On the red and white grapes varieties, based on the morphology of the cells, autochthonous strains belonging to the genus Saccharomyces prevail – 83.3%, what is confirmed by genetic analysis of rDNA ITS region. Restriction analysis showed that all strains of the genus Saccharomyces belong to the species Saccharomyces cerevisiae / S. paradoxus. The percentage of Saccharomyces isolated on the Pervenets Magaracha variety is 86.7%, Krasnostop Anapsky - 80%. The non-Saccharomyces yeast had a product size of 750 bp, presumably of the species Hanseniaspora uvarum.


Fermentation ◽  
2020 ◽  
Vol 6 (2) ◽  
pp. 41
Author(s):  
José Juan Mateo ◽  
Patricia Garcerà ◽  
Sergi Maicas

There a lot of studies including the use of non-Saccharomyces yeasts in the process of wine fermentation. The attention is focused on the first steps of fermentation. However, the processes and changes that the non-Saccharomyces yeast populations may have suffered during the different stages of grape berry ripening, caused by several environmental factors, including antifungal treatments, have not been considered in depth. In our study, we have monitored the population dynamics of non-Saccharomyces yeasts during the ripening process, both with biochemical identification systems (API 20C AUX and API ID 32C), molecular techniques (RFLP-PCR) and enzymatic analyses. Some unusual non-Saccharomyces yeasts have been identified (Metschnikowia pulcherrima, Aureobasidium pullulans, Cryptococcus sp. and Rhodotorula mucilaginosa). These yeasts could be affected by antifungal treatments used in wineries, and this fact could explain the novelty involved in their isolation and identification. These yeasts can be a novel source for novel biotechnological uses to be explored in future work.


2001 ◽  
Vol 22 (5) ◽  
pp. 294-298 ◽  
Author(s):  
Thomas A. Wichelhaus ◽  
Klaus-Peter Hunfeld ◽  
Boris Böddinghaus ◽  
Peter Kraiczy ◽  
Volker Schàfer ◽  
...  

AbstractObjective:To establish a new, rapid, and reliable genotypic fingerprinting technique for methicillin-resistantStaphylococcus aureus(MRSA) typing in routine epidemiological surveillance.Design:The method is based on polymerase chain reaction (PCR) restriction fragment-length polymorphism (RFLP) followingHaeII digestion of simultaneously amplified parts of the protein A gene, the coagulase gene, and the hypervariable region adjacent tomecA. A total of 46 MRSA initial isolates were analyzed, including 14 isolates from five countries; the six German epidemic strains; 16 isolates from the Frankfurt metropolitan area, which were known to be heterogeneous by pulsed-field gel electrophoresis (PFGE); and 10 isolates obtained during three epidemics, all of which displayed an identical genotype.Results:Restriction analysis by PCR-RFLP permitted discrimination of 10 of 14 international isolates, all six German epidemic strains, and 15 of 16 national isolates. It also confirmed the homogeneous character of the 10 outbreak isolates.Conclusions:This new and rapid PCR-RFLP typing method is an attractive tool in routine epidemiological surveillance. Its impressive characteristics are ease of performance and interpretation, while at the same time guaranteeing good discriminatory power, reproducibility, and typeability.


2004 ◽  
Vol 54 (5) ◽  
pp. 1653-1667 ◽  
Author(s):  
Mark F. Schinsky ◽  
Roger E. Morey ◽  
Arnold G. Steigerwalt ◽  
Michael P. Douglas ◽  
Rebecca W. Wilson ◽  
...  

The Mycobacterium fortuitum third biovariant complex (sorbitol-negative and sorbitol-positive) contains unnamed taxa first characterized in 1991. These organisms can cause respiratory infections, a spectrum of soft tissue and skeletal infections, bacteraemia and disseminated disease. To evaluate this group of organisms, clinical reference isolates and the type strains of M. fortuitum third biovariant complex sorbitol-negative (n=21), M. fortuitum third biovariant complex sorbitol-positive (n=3), M. fortuitum (n=3), Mycobacterium peregrinum (pipemidic acid-susceptible) (n=1), Mycobacterium porcinum (n=1), Mycobacterium senegalense (n=2) and Mycobacterium septicum (n=1) were characterized by using conventional phenotypic (morphological, physiological and antimicrobial susceptibilities), chemotaxonomic (HPLC and cellular fatty acids) and genotypic [RFLP of the rRNA gene (ribotyping), PCR-RFLP of a 439 bp segment of the 65 kDa hsp gene (PCR restriction analysis) and 16S rRNA gene sequence] analysis, DNA G+C content and DNA–DNA relatedness analyses. The results of these studies indicated that the strains comprised M. porcinum (n=13), M. septicum (n=1) and four novel closely related genetic groups within the M. fortuitum third biovariant complex: Mycobacterium boenickei sp. nov. (n=6), Mycobacterium houstonense sp. nov. (n=2), Mycobacterium neworleansense sp. nov. (n=1) and Mycobacterium brisbanense sp. nov. (n=1), with type strains ATCC 49935T (=W5998T=DSM 44677T), ATCC 49403T (=W5198T=DSM 44676T) ATCC 49404T (=W6705T=DSM 44679T) and ATCC 49938T (=W6743T=DSM 44680T), respectively.


2012 ◽  
Vol 58 (12) ◽  
pp. 1344-1352 ◽  
Author(s):  
Renáta Vadkertiová ◽  
Jana Molnárová ◽  
Dana Vránová ◽  
Elena Sláviková

Yeasts are common inhabitants of the phyllosphere, but our knowledge of their diversity in various plant organs is still limited. This study focused on the diversity of yeasts and yeast-like organisms associated with matured fruits and fully open blossoms of apple, plum, and pear trees, during 2 consecutive years at 3 localities in southwest Slovakia. The occurrence of yeasts and yeast-like organisms in fruit samples was 2½ times higher and the yeast community more diverse than that in blossom samples. Only 2 species (Aureobasidium pullulans and Metschnikowia pulcherrima) occurred regularly in the blossom samples, whereas Galactomyces candidus, Hanseniaspora guilliermondii, Hanseniaspora uvarum, M. pulcherrima, Pichia kluyveri, Pichia kudriavzevii, and Saccharomyces cerevisiae were the most frequently isolated species from the fruit samples. The ratio of the number of samples where only individual species were present to the number of samples where 2 or more species were found (consortium) was counted. The occurrence of individual species in comparison with consortia was much higher in blossom samples than in fruit samples. In the latter, consortia predominated. Aureobasidium pullulans, M. pulcherrima, and S. cerevisiae, isolated from both the fruits and blossoms, can be considered as resident yeast species of various fruit tree species cultivated in southwest Slovakia localities.


2000 ◽  
Vol 66 (9) ◽  
pp. 3987-3997 ◽  
Author(s):  
Florence Doignon-Bourcier ◽  
Anne Willems ◽  
Renata Coopman ◽  
Gisele Laguerre ◽  
Monique Gillis ◽  
...  

ABSTRACT We examined the genotypic diversity of 64Bradyrhizobium strains isolated from nodules from 27 native leguminous plant species in Senegal (West Africa) belonging to the genera Abrus, Alysicarpus,Bryaspis, Chamaecrista, Cassia,Crotalaria, Desmodium, Eriosema,Indigofera, Moghania, Rhynchosia,Sesbania, Tephrosia, and Zornia, which play an ecological role and have agronomic potential in arid regions. The strains were characterized by intergenic spacer (between 16S and 23S rRNA genes) PCR and restriction fragment length polymorphism (IGS PCR-RFLP) and amplified fragment length polymorphism (AFLP) fingerprinting analyses. Fifty-three reference strains of the different Bradyrhizobium species and described groups were included for comparison. The strains were diverse and formed 27 groups by AFLP and 16 groups by IGS PCR-RFLP. The sizes of the IGS PCR products from the Bradyrhizobium strains that were studied varied from 780 to 1,038 bp and were correlated with the IGS PCR-RFLP results. The grouping of strains was consistent by the three methods AFLP, IGS PCR-RFLP, and previously reported 16S amplified ribosomal DNA restriction analysis. For investigating the whole genome, AFLP was the most discriminative technique, thus being of particular interest for future taxonomic studies in Bradyrhizobium, for which DNA is difficult to obtain in quantity and quality to perform extensive DNA:DNA hybridizations.


2005 ◽  
Vol 55 (1) ◽  
pp. 197-207 ◽  
Author(s):  
A. M. Rodas ◽  
S. Ferrer ◽  
I. Pardo

One hundred and seventy-eight lactobacilli isolated from wine were characterized by a polyphasic approach. Strains were phenotypically identified at genus and species level by classical tests including the analysis of cell morphology, homo/heterofermentative character, sugar fermentation patterns, growth at different temperatures and the optical nature of the isomer of lactic acid produced from glucose. Molecular techniques such as random amplification of polymorphic DNA (RAPD), amplified 16S rDNA restriction analysis (16S-ARDRA), PFGE-RFLP and ribotyping were used to characterize strains, and their potential for identification and/or typing was evaluated. The information obtained with these techniques was processed with the BioNumerics software in order to analyse relationships existing between isolated strains and various reference species of the genus. Then, taxonomic dendrograms were obtained, and this information allowed the proposal of molecular procedures suitable for the identification and typing of these wine micro-organisms. The techniques useful for both identification and typing were RAPD and ribotyping, while 16S-ARDRA was only useful for identification and PFGE-RFLP only for typing purposes. The wine strains were identified as Lactobacillus brevis (19 strains), Lactobacillus collinoides (2 strains), Lactobacillus hilgardii (71 strains), Lactobacillus paracasei (13 strains), Lactobacillus pentosus (2 strains), Lactobacillus plantarum (34 strains) and Lactobacillus mali (10 strains).


2020 ◽  
Vol 3 (6) ◽  
pp. 23-32
Author(s):  
Karol Bazurto-Vera ◽  
María Cevallos-Cedeño ◽  
Anlly Vilcacundo-Alcívar

El presente trabajo se fundamenta en el proceso de deshidratación de la uva (Vitis vinifera L.), puesto que el proceso de secado es de gran importancia en diversos procesos químicos industriales, ya que permite alargar la vida útil de cualquier alimento, obteniéndose productos con mayor valor agregado. El objetivo de esta investigación fue evaluar la cinética de deshidratación de la uva a partir de la curva de velocidad contra la humedad media y la curva de humedad total contra el tiempo. El contenido de humedad en la uva se determinó por el método de secado por estufa de aire caliente forzado, basándose en la pérdida de peso de la uva en relación al tiempo, la muestra desde el tiempo cero se pesó y cada 5 minutos durante 1 hora y 25 minutos, se realizó el procedimiento a una temperatura de 60°C. Al obtener los datos de cada peso, siendo un total de 15 datos, se realizó los procedimientos para calcular la humedad total, humedad media, humedad libre y la velocidad crítica de secado. El contenido de agua en la uva tuvo un valor final de 7,02x10-5 kg de agua, con una velocidad de secado de 0,095951332 Kg/h y una humedad media de 0,0001016 KgH2O/Kg sólido seco. Se concluye que la pérdida de peso de la uva es directamente proporcional al tiempo y a la temperatura a la que fue sometida la muestra. Palabras clave: deshidratación, uva, tiempo, humedad, temperatura.


Agriculture ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 1250
Author(s):  
Markéta Kulišová ◽  
Maria Vrublevskaya ◽  
Petra Lovecká ◽  
Blanka Vrchotová ◽  
Milena Stránská ◽  
...  

Endophytes are microorganisms that live asymptomatically inside plant tissues. They are beneficial to their host in many aspects, especially as a defense against foreign phytopathogens through the production of a variety of metabolites. These substances can serve as sources of new natural products for medicinal, agricultural, and industrial purposes. This article is focused on endophytic fungi from Vitis vinifera. The purpose of the research was their isolation and identification during the Vitis vinifera growing season. Subsequently, the isolates were tested for the production of biotechnologically interesting metabolites (siderophores, antioxidants, and antifungal compounds). In total, 24 endophytic fungi were isolated, the most represented genus was Cladosporium sp. The results of the test for antioxidant and antifungal properties, as well as siderophore production, have shown that the population of Vitis vinifera endophytic microscopic fungi could serve as a promising source of metabolites with a wide range of applications.


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