Non-Conventional Grape Varieties and Yeast Starters for First and Second Fermentation in Sparkling Wine Production Using the Traditional Method

Abstract

Sparkling wine production using the traditional method involves a second fermentation of still wines in bottles, followed by prolonged aging on lees. The key factors affecting the organoleptic profiles of these wines are the grape varieties, the chemical and sensory attributes of the base wines elaborated, the yeast strains used for first and second fermentation, and the winery practices. While Chardonnay and Pinot noir are gold standard grape varieties in sparkling wine production, other valuable grape cultivars are used worldwide to elaborate highly reputable sparkling wines. Fundamental research on the chemical and sensory profiles of innovative sparkling wines produced by the traditional method, using non-conventional grape varieties and novel yeast strains for first and/or second fermentation, is accompanying their market diversification. In this review, we summarize relevant aspects of sparkling wine production using the traditional method and non-conventional grape varieties and yeast starters.

Full text

fermentation
Review
Non-Conventional Grape Varieties and Yeast Starters for First
and Second Fermentation in Sparkling Wine Production Using
the Traditional Method
María Laura Raymond Eder 1and Alberto Luis Rosa 1, 2, *


Citation: Raymond Eder, M.L.; Rosa,
A.L. Non-Conventional Grape
Varieties and Yeast Starters for First
and Second Fermentation in
Sparkling Wine Production Using the
Traditional Method. Fermentation
2021,7, 321. https://doi.org/
10.3390/fermentation7040321
Academic Editor:
Jean-Marie Sablayrolles
Received: 11 November 2021
Accepted: 16 December 2021
Published: 20 December 2021
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Attribution (CC BY) license (https://
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4.0/).
1Laboratorio de Genética y Biología Molecular, IRNASUS-CONICET, Facultad de Ciencias Químicas,
Universidad Católica de Córdoba, Córdoba X5016DHK, Argentina; mlraymondeder@ucc.edu.ar
2Bodega Finca las Acacias, Córdoba X5194DHK, Argentina
*Correspondence: alrosa@ucc.edu.ar; Tel.: +54-351-4938000 (ext. 609); Fax: +54-351-4938061
Abstract:
Sparkling wine production using the traditional method involves a second fermentation of
still wines in bottles, followed by prolonged aging on lees. The key factors affecting the organoleptic
profiles of these wines are the grape varieties, the chemical and sensory attributes of the base
wines elaborated, the yeast strains used for first and second fermentation, and the winery practices.
While Chardonnay and Pinot noir are gold standard grape varieties in sparkling wine production,
other valuable grape cultivars are used worldwide to elaborate highly reputable sparkling wines.
Fundamental research on the chemical and sensory profiles of innovative sparkling wines produced
by the traditional method, using non-conventional grape varieties and novel yeast strains for first
and/or second fermentation, is accompanying their market diversification. In this review, we
summarize relevant aspects of sparkling wine production using the traditional method and non-
conventional grape varieties and yeast starters.
Keywords: sparkling wine; non-conventional grape varieties; second fermentation; yeast
1. Introduction
The traditional method for sparkling wine production (i.e., méthode champenoise) con-
sists of the fermentation of a base wine following the addition of a liqueur de tirage (i.e., wine,
sucrose, ethanol-adapted yeast cells, nutrients, and a clarifying agent), in characteristic
bottles [
1
] (Figure 1). The development of this second fermentation in sealed bottles greatly
contributes to the sensory complexity of the resulting wine [
2
,
3
]. Among other factors, the
in bottle fermentation method contributes to a refined development of foam and bubbles,
which are key properties identifying champenoise sparkling wines [3,4].
Conventional grape varieties used in this industry are linked to prestigious sparkling
wines produced in France (e.g., Chardonnay, Pinot noir, and Pinot meunier), Italy (e.g.,
Chardonnay, Pinot nero, Pinot bianco, and Lambrusco), and Spain (e.g., Macabeo, Xarel.lo,
Parellada, Pinot noir, and Chardonnay) (Table 1). Each country has different officially
authorized grape varieties for their production. These varieties are linked to various
Protected Designations of Origin [3,4] (Table 1).
Accompanying the global expansion of sparkling wine production, emerging grape va-
rieties for the elaboration of fine quality sparkling wines are being explored worldwide [
3
,
4
]
(Table 2). Non-conventional grape varieties challenge the wine industry to recognize and
market products with novel sensory profiles, and to anticipate potential consumer accep-
tance [
2
,
5
]. Adding complexity to the scenario of innovation in sparkling wine production,
first and second fermentation strategies are also being explored, based on novel and/or
indigenous yeast starters [
2
]. In this review, we summarize relevant studies of sparkling
wine production, innovating both in grape varieties and in yeast starters, and using the
traditional method.
Fermentation 2021,7, 321. https://doi.org/10.3390/fermentation7040321 https://www.mdpi.com/journal/fermentation

Fermentation 2021,7, 321 2 of 16
Fermentation 2021, 7, x FOR PEER REVIEW 2 of 17
studies of sparkling wine production, innovating both in grape varieties and in yeast
starters, and using the traditional method.
Figure 1. Schematic representation of sparkling wine production using the traditional method. During the first fermen-
tation, the grape juice is converted into a base wine, which is then subjected to a second fermentation following the ad-
dition of wine, sucrose, ethanol-adapted yeast cells, nutrients, and a clarifying agent, in characteristic bottles. After the
second fermentation and aging, riddling is performed to allow accumulation of the lees at the bottle neck, facilitating
their mechanical elimination. Disgorging (i.e., lees removal) is followed by the addition of a liqueur de expedition, bottle
corking, and labelling.
Table 1. Conventional grape varieties used for sparkling wine production by the traditional
method.
Country
Sparkling Wine
Color(s)
Grape Varieties
France
Champagne
White, rosé
Chardonnay, Pinot noir, and Pinot meunier
France
Crémant
White, rosé
Chardonnay, Pinot noir, Chenin blanc,
Cabernet franc, Mauzac, and Pinot blanc
Spain
Cava
White, rosé
Macabeo, Xarel.lo, Parellada, Pinot noir, and
Chardonnay
Italy
Talento
White, rosé
Chardonnay, Pinot nero, and Pinot bianco
Italy
Lambrusco
Red
Lambrusco Grasparossa, Lambrusco Maestri,
Lambrusco Marani, Lambrusco Montericco,
Lambrusco Salamino, and Lambrusco Sorbara
Australia
Shiraz
Red
Shiraz
(*)
New sparkling
White, rosé
Chardonnay and Pinot noir
(*) Argentina, Australia, Brazil, Chile, New Zealand, South Africa, Uruguay, USA.
Table 2. Non-conventional grape varieties used for sparkling wine production by the traditional
method.
Country
Grape Variety
Reference
Argentina
Torrontés Riojano
[14]
Brazil
Goethe
[15,34]
Manzoni
[15,34]
Moscatel
[10,34]
Niagara
[15,34]
Villenave
[15,34]
Moscato Giallo
[11]
Syrah
[32]
Chenin Blanc
[32]
Chile
País
[21]
Germany
Sauvignon blanc
[23]
Figure 1.
Schematic representation of sparkling wine production using the traditional method. During the first fermentation,
the grape juice is converted into a base wine, which is then subjected to a second fermentation following the addition of wine,
sucrose, ethanol-adapted yeast cells, nutrients, and a clarifying agent, in characteristic bottles. After the second fermentation
and aging, riddling is performed to allow accumulation of the lees at the bottle neck, facilitating their mechanical elimination.
Disgorging (i.e., lees removal) is followed by the addition of a liqueur de expedition, bottle corking, and labelling.
Table 1.
Conventional grape varieties used for sparkling wine production by the traditional method.
Country Sparkling Wine Color(s) Grape Varieties
France Champagne White, roséChardonnay, Pinot noir, and Pinot meunier
France Crémant White, roséChardonnay, Pinot noir, Chenin blanc,
Cabernet franc, Mauzac, and Pinot blanc
Spain Cava White, rosé
Macabeo, Xarel.lo, Parellada, Pinot noir, and
Chardonnay
Italy Talento White, roséChardonnay, Pinot nero, and Pinot bianco
Italy Lambrusco Red
Lambrusco Grasparossa, Lambrusco
Maestri, Lambrusco Marani, Lambrusco
Montericco, Lambrusco Salamino, and
Lambrusco Sorbara
Australia Shiraz Red Shiraz
(*) New sparkling White, roséChardonnay and Pinot noir
(*) Argentina, Australia, Brazil, Chile, New Zealand, South Africa, Uruguay, USA.
Table 2.
Non-conventional grape varieties used for sparkling wine production by the traditional
method.
Country Grape Variety Reference
Argentina Torrontés Riojano [6]
Brazil
Goethe [7,8]
Manzoni [7,8]
Moscatel [8,9]
Niagara [7,8]
Villenave [7,8]
Moscato Giallo [10]
Syrah [11]
Chenin Blanc [11]
Chile País [12]
Germany Sauvignon blanc [13]

Fermentation 2021,7, 321 3 of 16
Table 2. Cont.
Country Grape Variety Reference
Italy
Bombino bianco [14]
Grillo [15]
Malvasia di Candia [16]
Nero di Troia [14]
Maresco [17]
Verdicchio [18]
Pinot gris [19]
Pignoletto [19]
Portugal Baga [20,21]
Fernão-Pires [20,21]
Romania Muscat Ottonel [22]
Feteasca Neagra [23]
Spain
Airén [24]
Albarín [25–28]
Bobal [29]
Garnacha [25–28,30]
Godello [25–28]
Malvasía [25–28]
Monastrell [30–32]
Pedro Ximénez [33]
Pietro Picudo [25–28]
Tempranillo [24,28,34–36]
Verdejo [25–28]
Viura [25–28]
Turkey Dimrit [37]
Emir [38]
2. Non-Conventional Grape Varieties for Sparkling Wine Production Using the
Traditional Method
Sugar content (i.e., 16–21
◦
Brix), titratable acidity (i.e., >8 g/L tartaric acid) and pH
(i.e., 2.9–3.2) of grape musts are significant chemical parameters when harvesting grapes
destined for sparkling wine production [
39
,
40
]. This “ripened” stage is dramatically in-
fluenced by the grape variety, and by the soil and climate (i.e., terroir) [
39
]. Organoleptic
analyses of grape musts and base wines produced with various non-conventional grape
varieties (Table 2) have shown highly valuable chemical and sensory profiles for the pro-
duction of white, rosé, and red sparkling wines (Table 3), comparable to those obtained by
using the gold standard grape varieties (Table 1) [
3
,
4
,
41
]. In addition to market innovation,
the use of alternative grape cultivars would enable, for example, development of this
industry in geographic areas where climatic conditions do not allow more conventional
grape cultivars to reach optimal grape maturity [15].

Fermentation 2021,7, 321 4 of 16
Table 3.
Chemical profiles of base wines obtained using non-conventional grape varieties in sparkling wine production by
the traditional method.
Grape Variety Wine pH Titratable
Acidity (g/L)
Volatile
Acidity (g/L)
SO2Total
(mg/L) Ethanol (% v/v)Sugars
(g/L) Ref.
Grillo
White
3.07 ±0.04 17.03 ±0.07 0.39 ±0.05 42.00 ±2.00 10.05 ±0.08
1.75
±
0.04 [
15
]
Albarin
White
2.79 9.5 0.28 56 11.1 ND
[
25
]
Viura
White
2.95 8.2 0.16 53 10.7 ND
[
25
]
Godello
White
2.84 7.6 0.31 64 11.7 ND
[
25
]
Malvasía
White
2.99 8.0 0.27 65 10.6 ND
[
25
]
Verdejo
White
2.93 8.2 0.21 56 10.3 ND
[
25
]
Torrontés
Riojano
White
3.3 5.1 ND ND 10.7 1.83 [6]
Villenave
White
2.69 ±0.1 9.13 ±0.23 0.18 ±0.01 24.72 ±0.08 10.03 ±0.05 ND [7]
Niagara
White
3.11 ±0.1 5.24 ±0.09 0.19 ±0.01 11.28 ±0.08 10.07 ±0.02 ND [7]
Manzoni
White
3.07 ±0.1 7.59 ±0.09 0.09 ±0.02 61.92 ±0.08 12.07 ±0.06 ND [7]
Goethe
White
3.32 ±0.1 6.03 ±0.08 0.17 ±0.02 48.08 ±0.08 10.47 ±0.04 ND [7]
Pedro Ximénez
White
3.24 5.67 0.39 ND 12.4 2.4
[
33
]
Airén
White
3.36 5.40 0.31 ND 9.49 0.36
[
24
]
Muscat Ottonel
White
2.90 ±0.02 6.3 ±0.07 0.30 ±0.02 72 ±0.47 12.5 ±0.03 3.4 ±0.13
[
22
]
Garnacha Rosé3.03 7.5 0.18 63 11.8 ND
[
25
]
Pietro Picudo Rosé3.08 8.5 0.17 30 11.5 ND
[
25
]
Tempranillo Red 3.7 ±0.1 4.6 ±0.2 0.39 ±0.05 ND 13.0 ±0.2 ND
[
34
]
Tempranillo Red 3.45 ±0.1 5.5 ±0.2 0.55 ±0.06 ND 12.3 ±0.2 ND
[
36
]
Tempranillo 1Red 3.47 5.10 0.32 ND 11.1 ND
[
35
]
Tempranillo 2Red 3.71 4.80 0.20 ND 13.0 ND
[
35
]
Tempranillo Red 3.36 5.20 0.48 ND 9.74 2.21
[
24
]
Chardonnay
White
2.80 ±0.00 7.5 ±0.0 ND ND 11.0 ±0.1 6.1 ±0.5
[
30
]
Pinot meunier
White
2.85 ±0.01 7.6 ±0.0 ND ND 10.9 ±0.1 ND
[
30
]
Riesling
White
3.3 6.9 0.26 ND 10.5 ND
[
42
]
Conventional grape cultivars for sparkling wine production are shown in gray;
1
Premature and
2
mature Tempranillo grapes; ND: not
determined.
2.1. White Sparkling Wines
White and/or red grape varieties can be used in the production of white sparkling
wines [
3
,
39
]. Golden yellow sparkling wines are typically manufactured using white grapes
(i.e., blanc de blanc) [
3
,
39
,
40
]. On the other hand, when red grape varieties are used, they are
vinified as base white wines by eliminating or greatly reducing must maceration, to produce
sparkling wines known as blanc de noirs [
3
,
39
,
40
]. In addition to traditional grape cultivars
(Table 1), several non-conventional varieties are used for the production of white sparkling
wine (Tables 2and 4). Interestingly, Muscat-related sparkling wines have a worldwide
distribution [
9
,
10
,
22
], and the volatile profile of these wines has revealed a homogeneous
signature for this grape family [
9
,
10
,
22
]. Major classes of volatile compounds in these
sparkling wines are terpenes, a predominant class of compounds associated with their floral
profiles, and higher alcohols and esters, which contribute to their fruity aromas [
9
,
10
,
22
]. In
particular, Moscato Giallo sparkling wines from Brazil have shown high concentrations of 2-
phenylethanol, ethyl octanoate, linalool, and
α
-terpineol [
10
] (Table 5). Similarly, Romanian
Muscat Ottonel sparkling wines show high levels of ethyl octanoate and ethyl decanoate,
as well as high levels of linalool, regardless of the yeast used for second fermentation [
22
]
(Table 5). In Argentina, Torrontés Riojano (Listan Prieto
×
Moscatel de Alejandría), an
aromatic cultivar with rich and variegated profiles comparable to that of Muscat in terms
of intensity and bouquet [
43
], is also used for sparkling wine production [
6
]. The volatile
fraction of Torrontés Riojano sparkling wines revealed a high concentration of volatile
compounds associated with fruit aromas, as well as terpene compounds associated with
the floral Muscat-like aromas [
6
] (Table 5). In fact, the final sensory properties of Muscat,
Moscato Giallo, Muscat Ottonel, and Torrontés Riojano sparkling wines appears to depend
on their unique terpenic profiles [
5
,
7
,
22
,
44
,
45
]. As shown in Table 5, even if differences in
total terpene concentrations are evident between the various Muscat-type varietals, their

Fermentation 2021,7, 321 5 of 16
total contribution is higher (above 0.5 mg/L) than in other varietals used for sparkling
wine production, such as Chardonnay and Riesling [
46
,
47
]. In the last sparkling wines, the
composition of volatile compounds show a relevant contribution of esters, alcohols, and
acetates [46,47].
Other cultivars, such as the white grapes Albarín, Fernão-Pires, Godello, Malvasía,
Verdejo, and Viura, as well as the red grapes Bobal, País, and Baga, which are traditional
varieties used to produce still wines, have also been employed for the production of high-
quality white sparkling wines [
12
,
13
,
19
,
25
,
29
] (Tables 3and 4). Studies on the organoleptic
profiles of these grape varieties, their various ripening stages, the influence of the alternative
soils, as well as other factors, have been conducted for the production of Spanish (i.e.,
Albarín, Viura, Malvasía, Verdejo, and Godello), German (i.e., Sauvignon blanc), Chilean
(i.e., País), and Portuguese (i.e., Fernão-Pires and Baga) sparkling wines [
12
,
20
,
21
,
25
–
27
,
51
].
The grape variety and/or soil have been shown to significantly influence the relative
contribution of various volatile and phenolic compounds in these wines [
12
,
21
,
25
,
51
]. These
parameters influenced their organoleptic profiles more than shorter or longer aging times
on lees [
12
,
27
]. Although sparkling wines made with Verdejo, Viura, Malvasía, Albarín,
and Godello presented excellent physicochemical parameters (Table 4), Albarín and Verdejo
wines showed more color and olfactory intensity than those elaborated with the other three
grape varieties [
25
]. Albarín also had the highest contents of catechins, proanthocyanidins,
and, together with Viura, hydroxycinnamates, all of which contribute to their phenolic
profile [
25
]. Differences in the abundance and glycosylation profiles of grape glycoproteins
could influence foam production and foam quality in non-conventional Sauvignon blanc
Sekt versus conventional Riesling Sekt [
13
]. Portuguese Fernão-Pires sparkling wines show
higher contents of volatile compounds that contribute to their varietal character, including
linalool, hotrienol,
α
-terpineol, geraniol, and nerol, than Baga sparkling wines [
21
]. Baga
wines, however, showed a higher maximum foam height than Fernão-Pires [
20
]. A detailed
sensory characterization of Chilean País sparkling wines highlighted strong floral nuances,
dependent on ethyl isobutyrate, isoamyl acetate, ethyl hexanoate, and
β
-phenylethanol,
which are high-impact aromatic compounds [
12
]. Taken together, these studies reveal how
the character of non-conventional grape cultivars may influence the innovative arena of
sparkling wine production.
Distinctive characteristics of white grape cultivars can also be enhanced, for example,
by applying skin pre-fermentative maceration, a technique that is usually performed in
red and rosésparkling wine production [
39
]. This strategy has been used to manufacture
Spanish Pedro Ximenez sparkling wines, increasing the extraction of grape skin compounds
that influence their ester profiles [
33
]. In the study by Ruiz Moreno et al. (2017), wines
elaborated with pre-fermentative maceration displayed higher contents of ethyl esters of
branched acids and cinnamates, while the absence of maceration rendered higher levels
of ethyl esters of fatty acids and higher alcohol acetates [
33
]. Thus, classic methods used
for red wine production may constitute alternative strategies for the elaboration of white
sparkling wines with improved organoleptic characteristics.

Fermentation 2021,7, 321 6 of 16
Table 4. Chemical profiles of sparkling wines.
Grape Variety Sparkling Wine Aging Time
(Months) pH Titratable
Acidity (g/L)
Volatile Acidity
(g/L)
SO2Total
(mg/L) Ethanol (% v/v) Sugars (g/L) Ref.
Albarin White 9 2.76 7.2 0.39 35 12.2 ND [25]
Viura White 9 2.89 7.4 0.24 43 11.6 ND [25]
Godello White 9 2.82 7.2 0.32 43 12.2 ND [25]
Malvasía White 9 3.10 7.4 0.28 31 11.6 ND [25]
Verdejo White 9 2.94 7.4 0.30 33 11.6 ND [25]
Torrontés
Riojano White 10 3.25 ±0.04 4.85 ±0.09 ND ND 11.97 ±0.09 2.07 ±1.31 [6]
Pedro Ximénez White 9 3.22 5.59 0.35 ND 12.6 2.4 [33]
Airén White 4 3.39 ±0.01 5.17 ±0.06 0.36 ±0.00 ND 10.75 ±0.50 0.08 ±0.01 [24]
Manzoni White 18 ND 8.49 ±0.13 0.36 ±0.11 ND 10.8 ±0.2 8.2 ±0.2 [8]
Villenave White 18 ND 4.36 ±0.15 0.55 ±0.10 ND 10.8 ±0.3 1.0 ±0.1 [8]
Moscato White 18 ND 5.26 ±0.25 0.63 ±0.12 ND 12.3 ±0.4 8.7 ±0.2 [8]
Niágara White 18 ND 4.88 ±0.4 0.57 ±0.12 ND 9.1 ±0.2 1.0 ±0.1 [8]
Goethe White 18 ND 5.60 ±0.17 0.75 ±0.08 ND 11.4 ±0.4 1.0 ±0.1 [8]
Muscat Ottonel White 15 3.1 ±0.01 6.70 ±0.02 0.30 ±0.01 56 ±0.47 11.6 ±0.07 0.7 ±0.02 [22]
Moscato Giallo White 10 ND 6.69 ±0.20 0.62 ±0.05 ND 10.9 ±0.1 1.10 ±0.09 [10]
Chenin blanc White 6 3.42 ±0.02 9.68 ±0.17 0.46 ±0.03 ND 12.35 ±0.37 2.60 ±0.08 [11]
Syrah White 6 3.58 ±0.02 8.18 ±0.07 0.48 ±0.04 ND 13.20 ±0.30 2.67 ±0.15 [11]
Pinot gris and
Pignoletto White 12 3.11 ±0.01 5.51 ±0.01 0.28 ±0.01 ND 11.30 ±0.03 ND [19]
Garnacha Rosé9 2.85 6.9 0.25 44 12.3 ND [25]
Pietro Picudo Rosé9 3.02 7.1 0.29 21 12.5 ND [25]
Tempranillo Red 9 3.5 ±0.1 4.7 ±0.2 0.32 ±0.04 ND 13.7 ±0.2 ND [34]
Tempranillo Red 9 3.42 ±0.1 5.6 ±0.2 0.52 ±0.05 ND 13.3 ±0.2 ND [36]
Tempranillo 1Red 9 3.49 5.2 0.32 ND 12.3 1.50 [35]
Tempranillo 2Red 9 3.70 4.9 0.30 ND 14.0 1.70 [35]
Tempranillo Red 4 3.38 ±0.01 5.03 ±0.06 0.45 ±0.01 ND 10.76 ±0.27 0.12 ±0.10 [24]
S. blanc White 18 ND 6.31 ±0.12 0.65 ±0.08 ND 12.9 ±0.2 10.6 ±0.3 [8]
Riesling Renano White 18 ND 8.64 ±0.25 0.40 ±0.09 ND 11.2 ±0.3 4.5 ±0.3 [8]
Pinot grigio White 18 ND 6.60 ±0.13 0.71 ±0.09 ND 10.5 ±0.2 1.0 ±0.1 [8]
Pinot noir White 18 ND 5.33 ±0.15 0.75 ±0.10 ND 11.4 ±0.2 1.0 ±0.1 [8]
Chardonnay White 18 ND 6.39 ±0.12 0.77 ±0.11 ND 11.8 ±0.1 1.0 ±0.1 [8]
Conventional grape cultivars for sparkling wine production are shown in gray; 1Pre-mature and 2mature Tempranillo grapes; ND: not determined.

Fermentation 2021,7, 321 7 of 16
Table 5. Aromatic compounds in Muscat-related and Chardonnay, Pinot noir, and Riesling sparkling wines.
Compound 1Muscat Ottonel Moscato Giallo Moscato
Embrapa
Torrontés
Riojano Chardonnay Riesling Pinot Noir Descriptor OPT 2
Ethyl hexanoate ND 748.198 ±10.000 841.6 ±29.3 424.7 ±146.0 744.0 ±8.0 750.0 ±40.0 154.1 ±170.0 Apple, fruit 14 [48]
Ethyl octanoate 7998.72 ±0.15 1229.184 ±35.021 954.4 ±32.1 322.0 ±160.0 712.0 ±7.0 670.0 ±30.0 59.8 ±6.9 Pineapple, pear,
soapy 5 [49]
Sum esters 310,634.41 2614.719 2569.9 10,515 ±195 75,921.6 28,771 267.4
2-Phenylethanol ND 8226.558 ±12.664 11824.2 ±162.7 29.9 ±6.7 11.6 ±<0.1 9.1 ±0.1 2990.5 ±163.8 Rose, honey,
woody 14,000 [8]
Nerol oxide ND 26.303 ±2.516 84.9 ±5.3 88.6 ±48.0 ND ND 15.5 ±2.5 Flower 400 [8]
Linalool 138.86 ±0.06 1732.887 ±7.311 169.0 ±28.0 7.7 ±4.0 <5 11.2 ±0.1 42.2 ±2.7 Flower, lavender 0.8 [50]
α-Terpineol 42.79 ±0.40 1211.424 ±11.521 97.1 ±8.4 166.8 ±23.0 3.4 ±<0.1 25.8 ±0.3 9.5 ±1.5 Citrus 250 [8]
Sum terpenes 3181.65 3659.781 588.1 2890 ±187 28 89.3 98.1
1
Ester and terpenic compounds (
µ
g/L) identified in Muscat Ottonel (Ref. [
22
]), Muscat Giallo (Ref. [
10
]), Moscato Embrapa (Ref. [
8
]), Torrontes Riojano (Ref. [
6
]), Chardonnay (Ref. [
42
]), Riesling (Ref. [
42
]), and
Pinot noir (Ref. [
8
]).
2
OPT (Odor Perception Threshold;
µ
g/L) and references are indicated.
3
Total esters and terpens as reported in the references. Conventional grape cultivars for sparkling wine production
are shown in gray. ND: not determined.

Fermentation 2021,7, 321 8 of 16
In warm and dry climates, like traditional Mediterranean winemaking regions, harvest-
ing at optimal grape maturity may be associated with insufficient acidity for sparkling wine
production [
39
,
40
]. Innovative base wines manufactured with Maresco and Grillo grape
musts, however, show high total acidity even when fully ripened grapes are used [
15
,
17
].
Maresco grapes, a minor variety grown in the Apulia region of Italy, besides high acidity
values, also showed floral, fruity, and fatty notes, as a result of the presence of linalool and
phenyl acetate (floral descriptors), isoamyl acetate, ethyl hexanoate, ethyl octanoate, and
ethyl decanoate (fruity descriptors), and octanoic acid (fatty descriptors) [
17
]. Sicilian Grillo
grape musts, also from a dry Mediterranean region, have shown optimal concentrations of
tartaric and malic acids, with the experimentally produced wines presenting remarkable
values of total acidity and low pH values (Table 3) [
15
]. Similarly, in tropical semi-arid
winemaking regions in Northeastern Brazil (i.e., São Francisco Valley), Syrah and Chenin
Blanc are adapted varieties for the production of high quality sparkling wines (Table 4) [
11
].
These Syrah (blanc de noir) and Chenin Blanc (blanc de blanc) wines showed different varietal-
related volatile profiles: Chenin Blanc was characterized by the presence of 2,3-butanediol,
3-ethoxypropan-1-ol, diethyl succinate, and ethyl decanoate, while Syrah by benzaldehyde,
butyric acid, and some acetates [
11
]. Interestingly, due to high temperatures, solar radiation
rates, and irrigation throughout the year, one vine can produce two harvests per year in
the São Francisco Valley region [11].
In geographic regions where the agricultural conditions do not favor the proper devel-
opment of most V. vinifera grapes, V. non-vinifera species and their hybrids are commonly
cultivated [
52
]. In Brazil, non-vinifera grape varieties (e.g., Villenave -V. labrusca
×
Riesling
renano-, Niagara, Goethe) and innovative V. vinifera varieties, such as Manzoni Bianco,
mostly employed in the elaboration of grape juices and wines, express interesting char-
acteristics of aroma and a good balance between acidity and sugar content, resulting in
appealing alternatives for sparkling wines [
7
–
9
]. In this context, studies of changes in
phenolic composition, browning index, and glutathione content during 18 months of bio-
logical aging sur lies, for sparkling wines produced with Villenave, Niagara, Manzoni, and
Goethe cultivars, revealed highly suitable changes in the phenolic profiles during the aging
period [
7
]. For example, (+)-catechin and (
−
)-epicatechin, flavanols with an important
influence on the astringency and color of wines, show concentrations of
1.31–14.05 mg/L
and 2.64–5.70 mg/L, respectively, in these non-conventional sparkling wines [
7
]. Catechin
and epicatechin levels of 3.52–5.80 mg/L and 1.56–2.15 mg/L, respectively, were measured
in sparkling wines made with Sauvignon blanc, Riesling Renano, Pinot Grigio, Pinot noir,
and Chardonnay [
7
]. These studies show that non-vinifera grape varieties could enable
sparkling wine production in latitudes where V. vinifera cultivars are more sensitive to
different pests, environmental stresses and/or high annual rainfall [15].
2.2. RoséSparkling Wines
For the production of rosésparkling wines, red grape pressing conditions and/or
maceration strategies render base wines with varying colors, depending on the concentra-
tion of color-determining compounds extracted from the grape skins [
3
]. The impact of
various non-conventional red grape varieties on the quality of the resulting rosésparkling
wines has been evaluated. Pioneer studies using the red varieties Trepat and Monastrell,
for the manufacture of rosésparkling wine in Spain, largely contributed to the current use
of these grapes for Cava production [
31
] (Tables 3and 4). These original studies showed
that roséTrepat was more similar to the white Cava wines (using a conventional blend of
Xarel.lo, Macabeo, and Parellada) than to the roséwines manufactured with Monastrell [
31
].
Organoleptic characterization of these sparkling wines showed that Trepat and Monastrell
had very good sensory attributes and even slightly better foam profiles than the white
Cava [
31
]. Interestingly, it was also reported that the foamability and color intensity of the
sparkling wines significantly increased when Trepat base wines were blended with white
varieties (Macabeo, Xarel.lo, and Parellada) to manufacture Cava [
32
]. Thus, these qualities
of Trepat grapes validated their use for elaborating either blanc de noirs sparkling wines or

Fermentation 2021,7, 321 9 of 16
novel versions of roséCava, and highlighted the value of autochthonous grape varieties
for sparkling wine production.
Other Spanish cultivars such as Prieto Picudo, Tempranillo, and Garnacha, tradi-
tionally used to produce still wines, have also been characterized for their potential for
manufacturing high quality rosésparkling wines using the traditional method [
25
,
27
,
28
,
53
]
(Tables 3and 4). These wines are rich in typical ethyl esters and alcohol acetates that
contribute to their fruity aroma [
51
] and preserve their varietal characteristics, even after
a long aging time (i.e., thirty months) [
27
]. Furthermore, the sensory and analytical char-
acteristics of five Garnacha Tinta rosésparkling wines, where second fermentation was
performed using five alternative yeast strains, showed that their organoleptic properties
mostly depend on the grape variety itself rather than on the strain used for second fermen-
tation [
53
]. Garnacha roséhas also shown high phenolic potential and hydroxycinnamic
acid concentrations, while Prieto Picudo rosésparkling wines had high color intensity
and anthocyanin concentrations, with high olfactory intensity and freshness in sensory
analyses [25]. Also, these sparkling wines, besides optimal chemical parameters (Table 4),
presented good foaming characteristics, similar to high quality sparkling wines like Cham-
pagne and Cava, with low levels of biogenic amines, which are desirable attributes in
sparkling wine production [25,27,51,53].
2.3. Red Sparkling Wines
Red sparkling or semi-sparkling wines represent a small fraction of the sparkling
wine production worldwide, and are mostly characterized by a slight red color and a
relatively poor complexity [
34
,
35
], because red grapes harvested at the proper time for
sparkling wine production (i.e., <21
◦
Brix), have not yet achieved adequate phenolic
maturity and, due to the relatively low final content of alcohol (i.e., 10.0–11.5% v/v;
Table 3
),
the extraction of phenolic compounds and aroma precursors from the grape berries into
the base wine is relatively low [
35
]. To face many of the challenges associated with
red sparkling wine production, various agricultural and enological practices have been
proposed, including alternative harvest dates, mixing of different grape varieties, pre-
fermentative cold maceration, rack and return, carbonic maceration, and treatment of
pre-mature grapes by applying pectolytic enzymes [23,34–36].
The impact of Tempranillo grape maturity on the alcohol concentrations, acidity, pH,
and color intensity of red sparkling wine production and aging definitely depends on the
ripening stage of grapes (Tables 3and 4). Levels of soluble polysaccharides and oligosaccha-
rides in Tempranillo base wines also increased with grape maturity, suggesting that these
compounds are more easily extracted during maceration-fermentation [
35
]. Following
these results, Pérez-Magariño et al. (2019) applied pre-fermentative cold maceration of
pre-mature Tempranillo grapes, using dry ice to favor the extraction of skin compounds
into the must [
34
]. The resulting wines showed a volatile composition similar to those pro-
duced from mature grapes, and also had good values in foam and sensory descriptors [
34
].
The study, however, also showed that the differences in overall volatile composition of the
wines were impacted more by grape maturity than by the use of enological techniques [
34
].
In fact, sparkling wines made with mature Tempranillo grapes, despite their high alcohol
content, showed better volatile composition and foam characteristics than those obtained
from pre-mature Tempranillo grapes [
34
]. The use of pectinolytic enzymes and/or carbonic
maceration on premature Tempranillo grapes, intended to contribute to the extraction
of polyphenols and varietal aromas, did not improve the chemical and sensory qualities
of the sparkling wines compared with those manufactured using mature Tempranillo
grapes [
36
]. Moreover, Tempranillo red sparkling wines made with mature grapes have
shown high contents of polyphenols, ethyl esters, alcohol acetates, and total volatile acids,
as well as foam stability [
34
,
36
], while wines made using unripe grapes have vegetal aroma
notes [
34
,
36
]. These studies suggest that mature grapes and traditional winemaking prac-
tices are options to be preferred for elaborating red sparkling wines using the traditional
method (Tables 3and 4).

Fermentation 2021,7, 321 10 of 16
3. Indigenous Yeast Starters for First and Second Fermentation of Sparkling Wines
Using Non-Conventional Grape Varieties and the Traditional Method
Yeast starters used in both grape must fermentations and secondary fermentations
of base wines are exposed to highly hostile physicochemical environments. Grape juices
and musts destined for base wine production have high osmotic pressure (i.e., 16–21
◦
Brix),
high titratable acidity (i.e., >8 g/L tartaric acid), low pH (i.e., 2.9–3.2), growing ethanol and
glycerol amounts, presence of sulfites, and progressive consumption of nutrients [
2
]. On the
other hand, yeast starters used for second fermentation must meet additional technological
requisites to those needed for first fermentation [
2
]. Second fermentation of base wines in
bottles requires yeast strains capable of growing under high ethanol contents (10–12% v/v),
low pH (2.9–3.2), low temperature (10–15
◦
C), high CO
2
pressure (up to 6 atmospheres),
relatively high levels of SO
2
(50–80 mg/L), and high total acidity (7–10 g/L; measured as
tartaric acid) (Table 3) [
2
]. In addition, these second-fermentation yeast strains should be
able to properly flocculate and autolysate during aging [
1
,
3
,
54
,
55
], allowing the wines to
remain in contact with the lees (i.e., mostly yeast cell debris and clarifying agents), shaping
their sensory complexity [
4
,
56
,
57
]. The aging process also contributes to other sparkling
wine properties such as foam (i.e., maximum height and stability) and bubble (i.e., size and
persistence) profiles [
25
,
27
,
39
,
58
]. Thus, because sparkling wine organoleptic properties
largely correlate with the physiological and metabolic characteristics of the yeasts used for
fermentation, a proper selection of starters is essential [14,54,59].
Although a wide variety of Saccharomyces cerevisiae strains are commercialized for first
and second fermentation of grape musts and base wines, respectively, there is increased
interest in indigenous S. cerevisiae and non-Saccharomyces yeasts to improve the sensory
attributes of the resulting wines [
15
,
54
,
59
]. Producing both base and sparkling wines with
native yeast starters can give typicality to the wines and may offer valuable technological
alternatives to the application of commercial starters that can lead to wine flavor stan-
dardization [
54
,
59
]. Interesting technological screening methods have been proposed to
identify, characterize, and select novel and indigenous yeast strains with valuable potential
in the sparkling wine industry [
2
,
6
,
14
,
54
,
57
,
59
–
61
]. Enological and technological traits
(i.e., fermenting power and vigor, SO
2
tolerance, alcohol tolerance, flocculence, production
of acetic acid, glycerol, H
2
S, and volatile compounds, autolytic capacity, and foaming
properties), sensory analyses, and genotypic screenings are currently being used as se-
lection strategies for these yeast strains [
6
,
14
,
54
,
57
,
59
,
60
] (Figure 2). As a result, various
non-commercial S. cerevisiae strains for first and second fermentation have been proposed
to provide regional character to sparkling wines as a driver of innovation/segmentation in
this market [14,54,57,59,60].
Producing sparkling wine with the use of alternative indigenous yeast strains renders
wines with varying organoleptic profiles [
14
,
15
,
60
]. What is more, native S. cerevisiae strains
have revealed similar [
53
,
59
] and even improved [
54
,
62
] physicochemical parameters
when compared with commercial starters for second fermentation [
53
,
54
,
59
,
62
] (
Figure 2
).
Studies by Di Gianvito et al. (2018) compared the second fermentation performance of
the commercial strains FI (i.e., a mixture of strains IOC-18-2007 and FRC) and EC1118
with six indigenous Italian S. cerevisiae wine strains (i.e., F6789, F6030, F10599, F10477,
F10471, and F7101), selected on the basis of their degree of flocculation and autolytic
capacity [
54
]. Their results showed that the indigenous yeast strains had an enological
performance comparable, in terms of fermentation rate and the maximum pressure reached,
to the commercial strains [
54
]. This study also highlighted the phenotypic differences of
some yeasts, in terms of their autolysis profiles, as well as the different levels of aromatic
compounds released after six months of aging [
54
]. Sparkling wines obtained with strains
F10471 and F10477 presented the highest amount of esters (e.g., 3-methylbut-1-yl ethanoate,
ethyl ethanoate, ethyl octanoate, ethyl decanoate and ethyl dodecanoate), which can be
attributed to the strong autolytic ability of these strains [
54
]. In similar studies, Martí-Raga
et al. (2016) showed that Cava wines fermented with the indigenous yeast strain P29,
isolated from the Penedes grape-growing region in Spain, reached a remarkable foam

Fermentation 2021,7, 321 11 of 16
height and sensory profiles compared to sparkling wines elaborated using the commercial
S. cerevisiae strains PDM and ARM [62].
Fermentation 2021, 7, x FOR PEER REVIEW 12 of 17
Figure 2. Influence of yeast starters, used in secondary fermentation of base wines, on the sensory qualities of white, rosé,
and red sparkling wines. (A) Mean ratings of the attributes of Torrontés Riojano white sparkling wines after 10 months of
aging on lees, obtained using commercial yeasts EC1118 (gray dashes), bayanus C12 (black dots), and mutant S. cerevisiae
strain IFI473I (gray line). Figure A was prepared based on data from reference [14]. (*) Attributes with significant differ-
ences between treatments (p < 0.05). (B) Mean ratings of the attributes of Verdicchio white sparkling wines obtained using
the following single or combined starters: S. cerevisiae DiSVA 527 (black line), T. delbrueckii DiSVA130 (black dashes),
DiSVA 313 (black dots), S. cerevisiae DiSVA 527 plus T. delbrueckii DiSVA 130 (gray dashes), and S. cerevisiae DiSVA 527
plus T. delbrueckii DiSVA 313 (gray dots). The figure was licensed from reference [58]. (*) Attributes with significant dif-
ferences between treatments (p < 0.05). (C) Mean ratings of the attributes of Garnacha tinta rosé sparkling wines obtained
with the following yeast starters: S. cerevisiae IMIA-2010 (black line), IMIA-2011 (black dashes), IMIA-2012 (black dots),
IMIA-2013 (gray dashes), and EC1118 (gray line). The figure was prepared based on data from reference [38]. (D) Mean
ratings of the attributes of Tempranillo red sparkling wines obtained using the following yeast starters: S. cerevisiae 7VA
(black line), Saccharomycodes ludwigii 979 (dark gray line), and Schizosaccharomyces pombe 938 (light gray line). Values in the
same line with the same letter are not significantly different (p < 0.05). The figure was licensed from reference [59].
4. Conclusions
Innovation and diversification in the manufacturing of high-quality sparkling wines
is largely associated with the use of non-conventional grape varieties and yeast starters.
A worldwide repertoire of prestigious grape cultivars, traditionally used in the produc-
tion of high quality still wines, has demonstrated suitable enological properties for elab-
orating white, rosé, and red sparkling wines. These non-conventional grape varieties can
positively contribute to the sensory diversification of sparkling wines, while showing
comparable physicochemical profiles to Champagne and Cava. The diversity of these
Figure 2.
Influence of yeast starters, used in secondary fermentation of base wines, on the sensory qualities of white, rosé,
and red sparkling wines. (
A
) Mean ratings of the attributes of Torrontés Riojano white sparkling wines after 10 months of
aging on lees, obtained using commercial yeasts EC1118 (gray dashes), bayanus C12 (black dots), and mutant S. cerevisiae
strain IFI473I (gray line). (
A
) was prepared based on data from reference [
6
]. (*) Attributes with significant differences
between treatments (p< 0.05). (
B
) Mean ratings of the attributes of Verdicchio white sparkling wines obtained using the
following single or combined starters: S. cerevisiae DiSVA 527 (black line), T. delbrueckii DiSVA130 (black dashes), DiSVA
313 (black dots), S. cerevisiae DiSVA 527 plus T. delbrueckii DiSVA 130 (gray dashes), and S. cerevisiae DiSVA 527 plus T.
delbrueckii DiSVA 313 (gray dots). The figure was licensed from reference [
18
]. (*) Attributes with significant differences
between treatments (p< 0.05). (
C
) Mean ratings of the attributes of Garnacha tinta rosésparkling wines obtained with the
following yeast starters: S. cerevisiae IMIA-2010 (black line), IMIA-2011 (black dashes), IMIA-2012 (black dots), IMIA-2013
(gray dashes), and EC1118 (gray line). The figure was prepared based on data from reference [
53
]. (
D
) Mean ratings of
the attributes of Tempranillo red sparkling wines obtained using the following yeast starters: S. cerevisiae 7VA (black line),
Saccharomycodes ludwigii 979 (dark gray line), and Schizosaccharomyces pombe 938 (light gray line). Values in the same line
with the same letter are not significantly different (p< 0.05). The figure was licensed from reference [24].
In a detailed study of first and second fermentation of Apulian grape varieties (i.e.,
Nero di Troia and Bombino bianco) using native yeast strains for the production of white
and rosésparkling wines, Garofalo et al. (2018) reported a yeast strain-dependent profile
of the release of volatile compounds [
14
]. Interestingly, they also showed that alternative
native yeast strains influenced the final CO
2
pressure values of white Bambino bianco

Fermentation 2021,7, 321 12 of 16
sparkling wines, confirming the suitability of native S. cerevisiae strains for improving the
quality of these wines [
14
]. Indigenous Italian yeast strains have also been tested for the
production of Grillo base wines in dry Italian Mediterranean climates [
15
]. These native
strains (i.e., CS182, GR1, MSE13 and MSE41) showed their ability to start fermentation
at low pH and in the presence of high amounts of organic acids, contributing to some
herbaceous and vegetal, floral, and exotic fruit descriptors that clearly distinguished each
of the Grillo base wines produced [
15
]. Taken together, these results support the idea that
indigenous S. cerevisiae yeasts could be exploited as starter cultures, differentiating between
sparkling wines, and linking them with their region of production.
Besides the selection of native S. cerevisiae strains, Saccharomyces non-cerevisiae and
non-Saccharomyces yeasts have also been proposed as alternatives for improving the eno-
logical features and flavor complexity of second fermentations of base wines [
37
,
38
,
63
,
64
].
Saccharomyces bayanus and Saccharomyces oviformis have been used both in free forms and
immobilized within coated alginate beads as inoculums for the second fermentation of
Turkish Emir and Drimit sparkling wines [
37
,
38
]. During the second fermentation of Emir
and Drimit base wines, no significant differences were found in free amino acids and
amino acids in peptides between the use of immobilized and free yeasts [
37
,
38
]. However,
significant differences in these compounds were found due to the aging time and the yeast
strains used [
37
,
38
]. Free amino acids were higher in the Emir sparkling wine made with S.
oviformis than with S. bayanus [
38
], and in the Drimit sparkling wine fermented with in S.
cerevisiae than with S. bayanus [37].
Among non-Saccharomyces species, Metschnikowia pulcherrima (Flavia
®
strain) and
Torulaspora delbrueckii (Biodiva
™
strain) have been tested, in sequential inoculations with
S. cerevisiae, for their impact on the composition and quality of Macabeo base wine for
sparkling wine production [
64
]. This study showed that base wines fermented with the
addition of M. pulcherrima resulted in an increase in foam persistence and changes in the
aromatic profile, characterized by smoky and flowery notes [
64
]. Sequential addition of T.
delbrueckii increased glycerol concentration, reduced volatile acidity, and exerted a positive
effect on foaming properties when compared with control base wines fermented with S.
cerevisiae [
64
]. Further studies showed that the foaming properties of the sparkling wines
obtained by sequential inoculation with these T. delbrueckii and S. cerevisiae strains resulted
in sparkling wines with significantly higher maximum foam heights than conventional
S. cerevisiae inoculation, probably due to the autolysis of T. delbrueckii cells in the base
wine [
65
]. Another report used T. delbrueckii strains (i.e., Td130/313) to conduct second
fermentations of Verdicchio base wine, both in mixed fermentations with S. cerevisiae and
as pure inocula [
18
] (Figure 2B). The T. delbrueckii strains tested were able to complete the
secondary fermentation and confirmed the previously reported behavior of T. delbrueckii in
still wines, leading to a reduction in acetaldehyde and some higher alcohols, and increasing
the production of ester compounds [
18
]. Thus, T. delbrueckii sparkling wines showed overall
different aromatic compositions and sensory profiles to those of pure S. cerevisiae starters,
with higher scores for positive aromatic descriptors [18] (Figure 2B).
Finally, physicochemical properties and sensory evaluations of sparkling wines made
by second fermentation in bottle with Saccharomycodes ludwigii and Schizosaccharomyces
pombe have been studied by Ivit et al. (2018) [
24
] (Figure 2D). These non-Saccharomyces
yeasts properly completed the second fermentation of Airén white base wine and of
Tempranillo red base wine, showing significant differences in acidity parameters, non-
volatile compound levels, and sensory evaluations compared to control sparkling wines
produced with S. cerevisiae [
24
] (Figure 2D). These studies demonstrated the potential of
non-Saccharomyces yeasts to develop flavor complexity in sparkling wine production. The
wide diversity of non-Saccharomyces species and strains constitute a great resource in the
arena of sparkling wine innovation. Further studies are needed to define the potential of
these non-Saccharomyces yeast starters [2].

Fermentation 2021,7, 321 13 of 16
4. Conclusions
Innovation and diversification in the manufacturing of high-quality sparkling wines
is largely associated with the use of non-conventional grape varieties and yeast starters. A
worldwide repertoire of prestigious grape cultivars, traditionally used in the production of
high quality still wines, has demonstrated suitable enological properties for elaborating
white, rosé, and red sparkling wines. These non-conventional grape varieties can positively
contribute to the sensory diversification of sparkling wines, while showing comparable
physicochemical profiles to Champagne and Cava. The diversity of these non-conventional
grape varieties opens a window of opportunities for winemakers to innovate and satisfy
consumer expectations. Novel and/or indigenous yeast starters for first and/or second
fermentation can make interesting contributions. Native S. cerevisiae strains and/or S. non-
cerevisiae and/or non-Saccharomyces yeasts offer new tools for innovation. The creative use
of alternative technological operations in winemaking would also contribute to diversifying
the sensory profiles of the growing number of non-conventional sparkling wines.
Author Contributions:
M.L.R.E. and A.L.R. equally contributed to the conception, drafting, revising
of the manuscript. All authors have read and agreed to the published version of the manuscript.
Funding:
This research was funded by Universidad Católica de Córdoba grant SIV-2015. M.L.R.E. is
supported by a postdoctoral fellowship from the Argentine National Research Council (CONICET)
and A.L.R. is a Principal Investigator of CONICET (Argentina).
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Acknowledgments:
We thank J. Seballe for a critical reading of the manuscript and J. Heywood for
English editing.
Conflicts of Interest: The authors declare no conflict of interest.
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