Indian Journal of Experimental Biology Vol. 58, July 2020, pp. 461-473

Isolation and characterization of yeast strains from Badacsony, Hungary

Annamária Gerőcs1, Katalin Nemes-Barnás1, Sára Pál1, Barna Szőke2, János Májer2, Tibor Farkas1*# & Ferenc Olasz1*

1Agricultural Biotechnology Institute, National Agricultural Research and Innovation Centre (NARIC ABC), Szent-Györgyi Albert Street 4. H-2100 Gödöllő, Hungary 2Research Institute for Viticulture and Oenology, National Agricultural Research and Innovation Centre - (NARIC RIVE) Római Street 181. H-8261 Badacsonytomaj, Hungary

Supplementary Data

Suppl. Table S1 — Comparison of the literature data of carbohydrate utilization of yeast species to that of presented in this study Species Literature data of Data of carbohydrate assimilation (+) Literature data of carbohydrate carbohydrate assimilation (+) presented in Table 1 assimilation (-) Candida californica glucose, xylose, glycerol glucose, sucrose, maltose sucrose, melibiose, galactose, maltose, L-rhamnose, cellobiose, L/D-arabinose Candida zemplinina glucose, sucrose glucose, sucrose, xylose, maltose melibiose, galactose, maltose, (Starmerella bacillaris) cellobiose, L-rhamnose, xylose, L/D-arabinose Hanseniaspora uvarum glucose, cellobiose glucose, sucrose, galactose, xylose, maltose, sucrose, melibiose, galactose, glycerol maltose, L-rhamnose, xylose, L/D- arabinose Pichia fermentans glucose, xylose, glycerol* glucose, sucrose, galactose, glycerol, sucrose, melibiose, galactose, xylose,maltose, cellobiose maltose, cellobiose, L-rhamnose, L/D-arabinose Pichia kluyveri glucose, xylose, glycerol glucose, sucrose, galactose, maltose, sucrose, raffinose, melibiose, glycerol, xylose, L-arabinose, mannose, galactose, maltose, cellobiose, L- cellobiose, L- rhamnose rhamnose, L/D-arabinose, Saccharomyces glucose, sucrose*galactose, glucose, sucrose, galactose, maltose, melibiose, cellobiose, L-rhamnose, cerevisiae maltose* glycerol, melibiose* glycerol, mannose, xylose, xylose, L/D-arabinose melibiose,cellobiose, L- rhamnose Saccharomyves glucose, sucrose, galactose, glucose, sucrose, galactose, maltose, cellobiose, L-rhamnose, xylose, uvarum/bayanus maltose, glycerol, melibiose, glycerol, mannose L/D-arabinose, Saccharomyces glucose, sucrose, galactose, glucose, sucrose, galactose, maltose, melibiose, cellobiose, L-rhamnose, paradoxus maltose, glycerol glycerol, mannose xylose, L/D-arabinose, Saccharomyces glucose, sucrose, galactose, glucose, sucrose, galactose, maltose, melibiose, cellobiose, L-rhamnose, pastorianus maltose, glycerol glycerol, mannose xylose, L/D-arabinose,

Sign are the followings: Signs (+) and (-) in the header mean that the given species are able to utilize the indicated carbohydrate or not, respectively; yellow color: results in Table 1 corresponds to the literature data, blue color: indicates such sugars which were utilized according to our results, but data presented in the literature are different, green color: carbohydrates which were utilized according to the literature by the given species but data presented in Table 1 do not support this description. purple color: data are not available in the cited literature. Sign *: the cited literatures contradict each other The data in this table are based on the following literatures:

1. Kurtzman CP, Fell JW, Boekhout T & Robert V, Methods for isolation, phenotypic characterization and maintenance of yeasts. In: The Yeasts 5th Edition. (Ed. by C Kurtzman, JW Fell, T. Boekhout, Elsevier) 2011 https://doi.org/10.1016/B978-0-444-52149- 1.00007-0 2. Rib’ereau-Gayon P, Dubourdieu D, Don`eche B & Lonvaud A,Classification of yeast species, In: Handbook of Enology Volume 1The Microbiology of Wine and Vinifications 2nd Edition (Ed. by P Rib’ereau-Gayon, D Dubourdieu, B Don`eche & A Lonvaud, John Wiley &Sons, West Sussex, England) 2006 3. Vaughan-Martini A & Martini A, Saccharomyces Meyen ex Reess (1870), In: The Yeasts 5th Edition (Ed. by C Kurtzman, JW Fell, T. Boekhout, Elsevier) 2011 https://doi.org/10.1016/B978-0-444-52149-1.00061-6

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Suppl. Table S2 — Characteristics of CA isolates (species identity, identification number of carbohydrate (C) and nitrogen (N) group (see Tables 1 and 2), tolerance of 300 mg/l glucose, tolerance to10 and 15% ethanol, acid tolerance and acid production, Killer toxin production, H2S production) Isolates Species Group Group glucose EtOH EtOH acid acid Killer toxin H2S of C of N 300 mg/l 10% 15% tolerance production production production

CA1 P. fermentans 5 1 ++ +++ - - - + ++/+++ CA2 P. fermentans 5 1 ++ +++ - - - + ++++ CA3 P. kluyveri 3 1 ++ +++ - - - + ++/+++ CA4 P. kluyveri 3 1 ++ +++ - - - + ++/+++ CA5 P. kluyveri 6 1 ++ +++ - - - + +++ CA6 P. kluyveri 6 1 ++ +++ - - - + ++/+++ CA8 P. kluyveri 7 2 ++ + - +++ + - +++ CA9 P. kluyveri 2 1 ++ +++ - - - + +++ CA11 P. kluyveri 11 1 ++ +++ - - - + +++ CA12 P. fermentans 11 1 ++ +++ - - - + +++ CA17 P. kluyveri 12 4 +++ ++ - - - - +/++ CA27 P. kluyveri 37 1 ++ + - +++ + + + CA29 P. kluyveri 30 1 ++ + - +++ + + +++ CA30 P. kluyveri 30 1 ++ + - +++ + + +++ CA33 P. kluyveri 38 1 ++ + - +++ + + +++ CA34 P. kluyveri 13 1 ++ + - +++ + + +++ CA35 P. fermentans 13 7 ++ + - +++ + - +++ CA37 P. fermentans 29 1 +++ + - - - + +++ CA38 P. kluyveri 29 6 +++ + - - - + +++ CA40 P. fermentans 22 1 - - - + + + +++ CA7 C. zemplinina 7 3 ++ ++ - +++ + - +++ CA10 C. zemplinina 2 1 ++ + - +++ + - +++ CA13 C. zemplinina 8 6 ++ + - +++ + - +++ CA19 C. zemplinina 13 2 ++ + - +++ + + + CA20 C. zemplinina 13 8 ++ + - +++ + + +++ CA28 C. zemplinina 13 7 ++ + - +++ + + - CA31 C. zemplinina 7 2 ++ + - +++ + - +++ CA32 C. californica 7 7 + - - - - + + CA36 C. zemplinina 13 1 ++ + - +++ + - +++ CA18 S. cerevisiae 12 4 +++ ++ - - - + + CA21 S. cerevisiae 19 4 +++ ++ - - - - + CA22 S. cerevisiae 19 4 +++ +++ - - - + +++ CA23 S. uvarum 26 3 + + - - - - + CA24 S. uvarum 26 4 + ++ - - - - + CA25 S. cerevisiae 20 4 +++ ++ + - - + + CA26 S. cerevisiae 20 4 +++ ++ + + + + + CA39 S. cerevisiae 22 1 +++ ++ - - - - + CA14 H. uvarum 8 2 ++ + - +++ + - - CA15 H. uvarum 34 3 ++ + - - - - - CA16 H. uvarum 1 3 ++ ++ - - + + - [+: low, ++: middle, +++: high, -: no tolerance/production, respectively, underlined strains are suitable for wine making according to glucose tolerance, ethanol tolerance and hydrogen sulfide production]

GERŐCS et al.: ISOLATION AND CHARACTERIZATION OF YEAST STRAINS FROM, HUNGARY 3

Suppl. Table S3 — Characteristics of NA isolates (species identity, identification number of carbohydrate (C) and nitrogen (N) group (see Tables 1 and 2), tolerance of 300 mg/l glucose, tolerance to10 and 15% ethanol, acid tolerance and acid production, Killer toxin production, H2S production) Isolates Species Group Group glucose EtOH EtOH acid acid Killer toxin H2S of C of N 300 mg/l 10% 15% tolerance production production production NA2 S. cerevisiae 12 4 +++ ++ - - - + - NA3 S. cerevisiae 12 4 +++ ++ - - - - + NA4 S. cerevisiae/ 12 4 +++ ++ + - - + ++ paradoxus NA5 S. cerevisiae 19 4 +++ ++ - - - + ++ NA6 S. cerevisiae/ 12 4 +++ ++ - - - + ++ paradoxus NA8 S. cerevisiae 19 4 +++ +++ - - - - ++ NA9 S. cerevisiae 17 4 +++ ++ - - - ++ ++ NA10 S. cerevisiae 20 4 +++ ++ + - - + + NA11 S. cerevisiae 12 4 +++ ++ + - - + + NA12 S. cerevisiae 12 4 +++ ++ + - - + + NA13 S. cerevisiae 12 4 +++ ++ + - - + + NA14 S. cerevisiae/ 12 4 +++ ++ + - - + + paradoxus NA15 S. cerevisiae 12 4 +++ ++ + - - + + NA16 S. cerevisiae/ 12 1 ++ ++ + +++ + - +/++ paradoxus NA17 S. cerevisiae 12 4 +++ ++ - - - - + NA20 S. cerevisiae 12 4 +++ ++ - - - - ++ NA21 S. cerevisiae 12 4 +++ ++ - - - - + NA22 S. cerevisiae 12 4 +++ ++ - - - - + NA23 S. cerevisiae 12 4 +++ ++ - - - - + NA25 S. cerevisiae/ 12 4 +++ ++ + - - + + paradoxus NA26 S. paradoxus 12 4 ++ ++ - - - - + NA27 S. cerevisiae 12 4 + ++ - - - + + NA29 S. cerevisiae 12 4 ++ ++ ++ - - - + NA30 S. cerevisiae 12 4 +++ ++ ++ - - ++ + NA31 S. cerevisiae 12 4 +++ ++ ++ - - + + NA32 S. cerevisiae/ 12 4 +++ - - - - + + paradoxus NA33 S. cerevisiae 12 4 +++ ++ - - - + + NA34 S. cerevisiae 12 4 +++ ++ - - - + + NA35 S. cerevisiae/ 12 4 +++ ++ + - - + + paradoxus NA36 S. cerevisiae 12 4 +++ ++ - - - + + NA37 S. uvarum 26 4 + + - - - - + NA38 S. cerevisiae 12 4 +++ ++ ++ - - - + NA40 S. cerevisiae 12 4 +++ ++ ++ - - - ++ NA1 P. kluyveri 12 4 +++ ++ - - - + ++++ NA19 P. kluyveri 12 4 +++ ++ - - - + ++ NA24 P. kluyveri 12 4 +++ ++ + - - + ++ NA28 P. kluyveri 12 4 +++ ++ + - - + ++/+++ NA39 P. kluyveri 12 4 +++ ++ - - + + NA7 H. uvarum 19 4 +++ +++ - - - + - NA18 ND 12 4 +++ ++ - - - ND ND [ND: not determined, +: low, ++: middle, +++: high, ++++: very high, -: no tolerance/production, respectively, underlined strains are suitable for wine making according to glucose tolerance, ethanol tolerance and hydrogen sulfide production]

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Suppl. Fig. S1 — Phylogenetic tree of 15 isolated species from Badacsonybased on their ITS sequence.