ORIGINAL PAPER

Aromatic characterization and enological potential of 21 minorvarieties (Vitis vinifera L.)

Sonia Garcıa-Munoz • Andriani Asproudi •

Felix Cabello • Daniela Borsa

Received: 17 February 2011 / Accepted: 2 July 2011 / Published online: 16 July 2011

� Springer-Verlag 2011

Abstract The homogenization of international wine

market led to a gradual impoverishment of the genetic

pool. In fact, in several Spanish Quality Demarcations, the

most frequent international varieties are replacing the local

ones. As a result, minor varieties, perfectly adapted to the

local environmental conditions, are nowadays at risk of

extinction. The study of minor varieties could provide

useful inputs to satisfy the demand for new and interesting

wine products. This work aims at filling a gap in the

existing literature, focusing on the aromatic potential of

minor varieties. Here, the study of glycosidic volatile

compounds and the evaluation of the influence of several

variables on aroma composition were considered. Fifty-one

glycosidic compounds were identified and quantified. The

results revealed differences for glycosidic compounds

according to cultivars, berry color, clone and sample origin.

The inclusion or not of the cultivars in Quality Demarca-

tion was also discussed. Moreover, the synthesis of some

compounds involved in these differentiations seems to

have a genetic component. The characterization of aro-

matic potential of several minor varieties, achieved for the

first time, revealed that some of these (Argamusa, Quigat,

Pampolat girat, Excursach, Gorgollasssa or Mandon) could

represent an excellent option for winemaking and com-

mercial offer diversification strategies, besides being

important for these cultivars conservation.

Keywords Aroma � Gas chromatography � Glycosides �Grapevine

Introduction

The phenomenon of replacement of local grape varieties

with widely spread international cultivars, such as Caber-

net-Sauvignon or Chardonnay, is coming to a standstill. In

addition, the wine consumers’ taste and preferences have

changed during the last few years, since there are other

values and motivations out of aroma and taste for drinking

wines, e.g., as marketing attributes and new wine style [1].

Several Quality Demarcations are starting to promote

varieties linked to specific locations, which produce ori-

ginal and high-quality wines. Minor varieties, perfectly

adapted to the local environmental conditions, may repre-

sent a good option. Even though they may not be allowed

in any Quality Demarcation, their reintroduction in the

wine industry would not only entail an increase in the wine

offer but also provide an important tool for their conser-

vation. Being the wine aroma one of the most important

attributes for influencing consumer’s preferences, a more

detailed study about the aromatic potential of these tradi-

tional varieties appears to be an important first step.

The qualitative and quantitative volatile compounds in

grapes are important as a source of flavour in wines [2].

The grape and wine aroma, and specifically its intensity,

has long been one of the principal attributes taken into

account for grapevine selection [3]. The wine aroma

compounds depend on the cultivar [4, 5], on environmental

Electronic supplementary material The online version of thisarticle (doi:10.1007/s00217-011-1538-1) contains supplementarymaterial, which is available to authorized users.

S. Garcıa-Munoz (&) � F. Cabello

IMIDRA Finca ‘‘El Encın’’, A2 km 38.200,

28800 Alcala de Henares, Madrid, Spain

e-mail: sonigarmu@gmail.com

A. Asproudi � D. Borsa

CRA-Centro di Ricerca per l’Enologia,

Via Pietro Micca 35, 14100 Asti, Italy

123

Eur Food Res Technol (2011) 233:473–481

DOI 10.1007/s00217-011-1538-1

conditions [6] as well as on enological processes such as

maceration [7], yeast and fermentation [8, 9].

The aromatic compounds have been studied in several

grape varieties [10], as well as the relationship between the

grape variety and the wine obtained [9, 11]. In grapes,

those compounds can be found in their free and bound

form, especially as glycosides [9]. The majority of wines

are made with neutral varieties, characterized by very low,

and sometimes not detectable, values of the free volatile

forms content [10]. Consequently, the contribution of those

compounds to the final wine aroma is often negligible [9].

By contrast, bound compounds, together with fermentation

volatile ones, can contribute to the wine aroma [9], since

they can be hydrolyzed by yeast or exogenous enzymes

during winemaking [9, 12] and aging [2].

Although all volatile compounds contribute to the odor

[13], only a few of them play a role in determining varietal

aroma [14]. This research focuses on the glycosidic vola-

tiles, whose quantification could define the potential wine

aromatic profile, according to the reviewed literature [12].

Previous studies on the use of aroma compounds in wine

characterization have been carried out for Callet, Manto

Negro and Fogoneu, varieties included in several Quality

Demarcations of the Balearic Islands, Spain [11, 15]. For

the remaining ancient cultivars, the contribution of grape

aroma precursor to wine aroma profile has not yet been

studied. In this work, we analyzed the aromatic potential of

21 varieties, mainly native from the Balearic Islands,

Spain. The aims of this study were (1) to characterize the

glycosidic compound in grapevine (Vitis vinifera L.), (2) to

evaluate how different variables such as the berry color, the

accession (or clone) and the origin of the samples can

influence the aroma profile and finally (3) to evaluate the

effect of the cultivar’s inclusion or not in Quality Demar-

cation. Furthermore, improving the knowledge of minor

varieties could represent an interesting option for wine

marketing strategies, aiming at satisfying the evolving

consumer’s demand.

Materials and methods

Plant material and field conditions

We analyzed 32 accessions of Vitis vinifera L. corre-

sponding to 21 different varieties of Vitis vinifera L. since

some accessions or clones of the same variety were studied

[16]. Some of the studied varieties are allowed in several

Spanish Quality Demarcations (Table 1). The varieties are

preserved at the Vitis Germplasm Bank ‘‘Finca El Encın’’

(IMIDRA, Alcala de Henares, Spain) and were collected

mainly in the Balearic Islands (Spain) but also from the

Levante area and Girona (Table 1). The grapes were picked

by hand, in excellent sanitary conditions, at commercial

maturation during 2007 vintage. Approximately 1.5 kg of

grapes were collected from the whole vine, ensuring the

selection of a fully representative sample. Three subsam-

ples of 100 berries were randomly selected from the grape

sample from each accession. They were stored in labeled

glass containers at -30 �C until analyses were performed.

Plants were almost 20 years old and were grown under

the same field conditions. All of them were unirrigated and

grafted onto 41B, trained vessel, and eight buds per vine

were left in winter pruning. The distance between vines

was 2.5 m, and the one within rows was 2.5 m.

Sample preparation

Aroma compounds were extracted following a previously

described method [10] with some modifications. The sam-

ples of 100 berries were frozen until the analyses were

carried out. The seeds were discarded before defreezing the

samples at a temperature of 4 �C. Subsequently, each

sample was homogenized in an Omni-Mixer (Sorvall,

Labequip, Ontario, Canada) where 100 mg Na2S2O5 was

added, centrifuged for 15 min at 4,000 rpm, and the liquid

phase was recovered. The pellet was washed with pH = 3.2

solution and centrifuged again. This operation was repeated

twice. The liquid phases were then assembled up to

300 mL, adding 100 mg of a commercial pectolitic enzyme

(Vinozym, Novo Nordisk Ferment Ltd, Dittingen) without

glycosidase activity. After a period of 6 h, they were filtered

using filtered disks (Whatman 589). About 150 mL of these

extracts was passed through a Sep Pack � Cartridge C18

5 g (Waters, Milford, MA, USA) previously activated with

20 mL of methanol (Sigma–Aldrich Co, St Louis, MO,

USA) and 50 mL of water. After treating each sample, the

cartridge was washed with 50 mL of water and subse-

quently with 30 mL of dichloromethane. The bound

compounds were eluted with 25 mL of methanol. Methanol

was then eliminated under vacuum, and the residue was

solubilized in 5 mL of 0.1 M citrate–phosphate buffer

(pH = 5.0; 51.5% 0.2 M sodium phosphate and 48.5%

0.1 M citric acid). When analyzing red variety samples, 1 g

of polyvinylpolypyrrolidone (PVPP; Sigma, St Louis, MO,

USA) was added to remove the phenolic compounds. The

glycosidically bound fraction was hydrolyzed with 200 lL

of a commercial glycosidase rich enzyme (Cytolase M102,

Ferrari- s.r.l. Italy) at 40 �C for 24 h. A known concentra-

tion of internal standard (1-heptanol at a final concentration

of 100 lg L-1; Fluka, Sigma–Aldrich Co.; St Louis, MO,

USA; purity assay [99.0%) was added to the mixture

containing the aglycons released by enzymatic hydrolysis at

room temperature. The suspension was then centrifuged at

4,000 rpm during 15 min to remove PVPP. The supernatant

was then passed through a Sep Pack � Cartridge C18 1 g

474 Eur Food Res Technol (2011) 233:473–481

123

previously activated with 5 mL of methanol and 10 mL of

water. The free compounds were recovered with 12 mL of

dichloromethane. After drying with anhydrous sodium

sulfate, the obtained volume was evaporated at ambient

temperature and pressure. After concentration, the sample

was ready for GC/GC–MS analysis.

Gas chromatography–mass spectrometry conditions

The separation was achieved using an Innowax 19091-N

(Agilent-J&W Scientific, Santa Clara, CA, USA) capillary

column (30 m 9 0.25 mm ID, 0.25 lm d.f.), using helium

as the carrier gas at 70 kPa. The gas chromatograph was a

Hewlett Packard 5890 series II connected to a selective

detector MSD 5970 (Hewlett Packard, Wilmington, DE,

USA). The injector temperature was 250 �C, and interface

temperature was 230 �C. The oven temperature program

was set at 45 �C for 2 min, then linearly increased, at a rate

of 30 �C min-1 to 60 �C, 2 �C min-1 to 160 �C and 3 �C

min-1 to 230 �C. The final temperature was maintained for

13 min. The injection mode was splitless for 2 min.

Acquisition mass ranged was from 28 to 300 u.m.a., and

ionization energy was 70 eV. Volatile compounds were

identified by comparing the retention time and mass spectra

with those reported in the literature [10, 17]. All mass

spectra were also compared with those of the data system

libraries (Wiley275 and libraries of mass spectra). Semi-

quantitative data were obtained by the ratio of area

of individual compounds versus internal standard area

(1-heptanol).

Table 1 List of the accessions

analyzed; conserved at the VitisGermplasm Bank ‘‘Finca El

Encın’’ (IMIDRA, Alcala de

Henares, Spain)

Acc. accession, Id. code used in

the analysis, Origin PM Palma

de Mallorca, GI Girona, LELevante area, N number of

Spanish Quality Demarcation

where the variety is allowed

– Minor varieties non-included

in a Quality Demarcation

* Allowed out of the Balearic

Islands

Main name Acc. Id. Berry color Origin N

Argamusa E34 ARG.E34 White PM –

Batista E23 BAT.E23 Red PM –

Beba E39 BEB.E39 White GI 1*

Beba O16 BEB.O16 White LE 1*

Beba O17 BEB.O17 White LE 1*

Beba O44 BEB.O44 White LE 1*

Beba E14 BEB.E14 White PM 1*

Beba E32 BEB.E32 White PM 1*

Beba E43 BEB.E43 White PM 1*

Beba roja E19 BER.E19 Rose PM –

Bobal E11 BOB.E11 Red PM 6*

Bobal E21 BOB.E21 Red PM 6*

Callet E26 CAL.E26 Red PM 2

Epero de Gall E37 EPE.E37 Red PM –

Escursach E27 EXC.E27 Red PM –

Fernandella E22 FER.E22 Red PM –

Fogoneu E24 FOG.E24 Red PM 1

Fogoneu E16 FOG.E16 Red PM 1

Giro E31 GIR.E31 Red PM –

Giro E36 GIR.E36 Red PM –

Gorgollassa E25 GOR.E25 Red PM –

Manses de Tibbus E30 MES.E30 Red PM –

Manto Negro E29 MTN.E29 Red PM 2

Manto Negro E20 MTN.E20 Red PM 2

Mandon E15 GAL.E15 Red PM –

Pampolat girat H24 PAM.H24 Red LE –

Pensal Blanca SCL PEN.SCL White PM 2

Quigat E38 QUI.E38 White PM –

Quigat E33 QUI.E33 White PM –

Sabate E35 SAB.E35 Red PM –

Valenci Tinto E18 BEN.E18 Red PM –

Vinate E45 VIN.E45 White PM –

Eur Food Res Technol (2011) 233:473–481 475

123

Statistical analysis

The aromatic compounds and varieties were analyzed

using stepwise forward discriminant analysis (SFDA) and

principal component analysis (PCA). SFDA was used in

order to differentiate white, rose and black varieties, and

the origin of Beba samples. The PCA analysis allowed

establishing a relationship between the different aroma

compounds and the studied grapevines, as well as identi-

fying the most important compounds in white, rose and red

varieties. The correlations between different accessions of

the same variety were evaluated using Pearson correlation

analysis (r). The differences between varieties included and

excluded from Quality Demarcations were checked by

ANOVA followed by Tuckey0s HSD test to enable pairwise

comparisons of means (p \ 0.05). All statistical analyses

were performed using XLSTAT 2009 version.

Results and discussion

In this study, 51 glycosidic compounds from 21 wine

varieties were identified and classified into four categories

(alcohols, benzenes, terpenes and norisoprenoids; Sup-

porting Tables 1, 2; Supporting Figures 1, 2). Total amount

of glycosidic compounds varied from 947 (Fogoneu vari-

ety) to 2,911 lg kg-1 (Sabate variety), being in line with

other grape glycosidic studies [10]. The compounds benzyl

alcohol and 2-phenylethanol were the most abundant

compounds for all the considered varieties, as it has been

appointed by other authors [10]. At the same time, all

varieties presented high values of benzenic compounds,

except Beba (E43 and O44 accessions), Beba roja and Giro

(E31 accession) that instead presented high value of nori-

soprenoid compounds and Giro (E36 accession) that pre-

sented high value of terpenic compounds.

The results found in this study based on glycosidic

compounds were useful for the characterization of the

studied cultivars; some odorant compounds were key for

the differentiation as benzyl alcohol for red cultivar which

has been described as fruity and floral [18] or 3-oxo-a-

ionol for white cultivars, described as spicy [19]. Differ-

ences between the studied clones were found in this study,

corroborating the results obtained by other authors [3, 20,

21]. Terpene and norisoprenoids compounds were the most

important compounds for this differentiation. The Beba

accessions could be grouped taking into account their

geographic origin. The terpene compounds, as a-terpineol,

citronellol or 2-phenylethanol, were the compounds most

important for this dissimilation. All of these compounds

have been proved to be odor-active compounds in musts

[22] and wines [9, 14]. Therefore, it seems possible to

choose cultivars with several aroma profile, since the

compounds found in the grape are related to the final wine

aroma profile [4, 9].

In this study, we have discussed the differences between

cultivars taking into account if the cultivar is included or

not in Quality Demarcation. It is interesting to note that

some considered cultivars non-included in Quality

Demarcation showed significant differences principally

regarding terpene compounds in both red and white culti-

vars. The red cultivars non-included in Quality Demarca-

tion showed a higher content of terpenic compounds

compared to those included, contrary to the obtained for

white ones. These results confirm the results obtained by

other authors, which comment that it is not easy to assess

the differences taking into account the inclusion of a cul-

tivar in a Quality Demarcation [23].

As a consequence, several minor cultivars might be

included in Quality Demarcations, helping to conserve

these minor cultivars and also being an important market-

ing strategy to satisfy the new demand of wine consumers.

Glycosidic compounds’ characterization

Two PCA were done, one for each grape color (one for red

and other one for white and rose varieties). The first PCA

axis for the red varieties (Fig. 1a) explained the 30.2% of

the total variance. The most important compounds in the

first axis were hydroxy geraniol and diol (2,6-dimethyl-3,7-

octadien-2,6-diol), accounting for 6.9 and 5.9% of the total

variance explained by this axis, respectively. These com-

pounds were highly related to Batista, Pampolat girat and

Giro varieties. The second axis explained 19.1% of the

total variance, and it was related to benzyl alcohol (7.3% of

the variance), trans-2-hexenol (7.2%) and benzaldehyde

(6.5%). Trans-2-hexenol has been found to be an odor

activity compound in must, relating with herbaceous and

green aroma [22]. However, benzaldehyde is commonly

associated with the presence of Botritis cinerea [24]. It was

present in Vinate variety with the highest value

(140 lg kg-1); however, no other compound associated

with Botritis cinerea, for instance 1-octen-3-ol [25], has

been found in this study. Benzaldehyde has also been

reported with significant values in other Spanish varieties

[26, 27]; therefore, we could consider a high amount of this

compound to be a characteristic of this variety. This

compound possesses almond, fragrant [7] and roasted

aroma [4]. Sabate was related to benzyl alcohol compound

showing a more than double value with respect to the rest

of the analyzed varieties (Fig. 1a, Supporting Table 1). The

aroma of benzyl alcohol has been described in Chardonnay

wines as fruity and floral [18]. Regarding the mapping of

red cultivars, it highlights that Pampolat girat, Batista,

Gorgollassa and Fernandella varieties were correlated

with norisoprenoid and terpenic compounds. Giro was

476 Eur Food Res Technol (2011) 233:473–481

123

correlated with linalool and cis-8-hydroxy-linalool com-

pounds, reaching the highest values (60 and more than

240 lg kg-1, respectively). Linalool is important aroma

for wine characterization being describing as floral [18],

citrus, sweet and grape-like [7]. Other cultivars as Sabate,

Bobal (E21 accession), Beba negra, Excursach and

Mandon showed a correlation with benzenic compounds.

Finally, the cresol aroma exerted an influence on Bobal

(E21 accession), Callet, Fogoneu and Mandon red cultivars

classification. This compound presents a high odorant

impact acknowledged by neurological studies [13]. This

report is the first study addressing the grapes aromatic

profile characterization of several Spanish minor varieties;

therefore, only Callet and Manto Negro have been studied

previously and the results achieved in this study for these

cultivars were consistent with previous descriptions, even

though different techniques have been used and the varie-

ties were planted in a different location [15]. On the other

hand, methyl syringate, a shikimate derivative, was present

in all red varieties and in only white variety namely Quigat,

although the existence of this compound in other white

cultivar has been previously recorded [28].

As regards the differences among white and rose vari-

eties (Fig. 1b), the first PCA axis explained 40.8% of the

total variation and was associated with diol (2,6-dimethyl-

3,7-octadien-2,6-diol), explaining 4.5% of the variance,

4-vinylguaiacol (4.3%), 1-hexanol (4.2%) and blumenol C

(4.2%). All these compounds have been described in wines

[7, 29, 30]. The compounds 4-vinylguaiacol and 1-hexanol

were important for Vinate and Quigat classification

(Fig. 1b). The 1-hexanol compound is a key factor in the

characterization of grapevines [27], and its content depends

on the cultivar [4]. It has also been found in wines [4] being

its aroma described as herbaceous, grass and woody [22].

Blumenol C played an important role in the characteriza-

tion of white and rose varieties (Fig. 1b), and it is a deri-

vate of blumenin [31], which has been proved to possess

antifungal properties in barley and wheat [32]. As a con-

sequence, this compound could have a function in resis-

tance mechanisms.

The second axis explained 23.5% of the variance being

mainly related to 3-oxo-a-ionol (6.9%), hydroxy citronellol

(6.2%) that separated Argamusa, Pensal Blanca and Quigat

varieties and cis-8-hydroxy-linalool (5.4%). Argamusa was

related to trans-2-hexenol, dihydroconiferyl alcohol and

endiol. All the Beba accessions were linked with nori-

soprenoid and terpenic compounds, especially with 3-oxo-

a-ionol, which attains the highest value ([166 lg kg-1)

and with cis-8-hydroxy-linalool ([60 lg kg-1). Vinate

showed strong relation with benzaldehyde (140 lg kg-1).

Regarding PCA, results of red and white cultivars

revealed that 3-oxo-a-ionol compound was relevant for the

classification of both white and red varieties (Fig. 1). The

red cultivar Pampolat girat and the white one Beba showed

the highest concentration for this compound in red and

white cultivars (234 and [ 166 lg kg-1, respectively).

Thus, 3-oxo-a-ionol was important for the classification of

both cultivars (Fig. 1 and Supporting Tables 1, 2). How-

ever, other authors found the same quantity of 3-oxo-a-

ionol in other 13 different varieties [33]. The presence of

this compound could be related to biological activity [34],

being one of the most important C13-norisoprenoids in

grapevine [35]. This compound has also been found in

Pinot noir wines being its aroma described as spicy [19].

On the other hand, diol (2,6-dimethyl-3,7-octadien-2,6-

diol), a terpene compound, was important for red and white

cultivars classifications being important for the classifica-

tion of the red cultivars Pampolat girat, Batista and Giro

(a)

-15

-10

-5

0

5

10

15

-20 -15 -10 -5 0 5 10 15 20 25

F2

(19.

10 %

)

F1 (30.16 %)

Biplot (axes F1 and F2: 49.26 %)

(b)

-30

-20

-10

0

10

20

30

-50 -40 -30 -20 -10 0 10 20 30

F2

(23.

55 %

)

F1 (40.84 %)

Biplot (axes F1 and F2: 64.39 %)

Fig. 1 Principal component analysis plots for PCA1 and PCA2 of the

aroma volatile compounds used for a red and b rose and whitegrapevines. Symbols: black circles (grapevine varieties) and graycircles (aroma from glycosidic precursors; keys in Supporting

Tables 1–2). The averages of the three replicate samples were used

for the analysis

Eur Food Res Technol (2011) 233:473–481 477

123

(Fig. 1a), and for the classification of all the Beba acces-

sions (Fig. 1b). This compound has also been found in

other Spanish [36, 40], Portuguese [10] and Italian grape-

vines [37], and it was an odorant compound in wine [29].

Therefore, taking into account the results found in this

study, each cultivar is characterized by several compounds,

consequently they might show differences in their aromatic

profiles, since each identified compound is characterized by

an aroma descriptor. As in other studies, the compounds

found in analysed grapes determine the wine aroma profile

[2, 4, 5]; consequently, it is possible to choose cultivars to

provide different aroma profile.

Role of the berry color

The differences between red and white grapevines are

caused mainly by a mutation on two genes [38]. However,

the origin of most of aroma compounds in plants is

unknown, despite the last research done in this field among

the last years [39, 40]. The results of this study are con-

sistent with the reviewed literature, since the varieties were

grouped and differentiated according to the berry color

since the within-class covariance matrices reported differ-

ences between berry color (SFDA analysis, p \ 0.001). All

the studied samples were classified correctly except one

replicate of Beba (O16 accession), which changed from the

white to the rose category. The percentage of correct clas-

sification for the 96 samples analyzed was 98.96% (Fig. 2).

The most suitable variables for this classification were two

alcohols, three benzenic compounds and four terpenes. As

regards the alcohols, hexanol turned out to be higher in red

varieties (p \ 0.001; 65.55 vs. 33.93, 36.76 lg kg-1 per

red, rose and white samples, respectively), which is con-

sistent with previous studies [33]. This compound has a

grape origin [4], and it is scarcely affected by the fermen-

tation process [5, 26]. Tirosol was present exclusively in red

varieties (p = 0.026; 4.42 lg kg-1). As regards the ben-

zenic compounds, 2-phenylethanol was higher in rose

samples (p \ 0.001; 257.77 vs. 186.80, 238.97 lg kg-1 per

rose, red and white samples, respectively), and 4-vinyl-

phenol was higher in red samples (p \ 0.001; 91.36 vs.

10.14 and 14.25 lg kg-1 per red, rose and white samples,

respectively). In this study, 4-vinylphenol was always

accompanied by 4-vinylguaiacol, and it is well known that

these compounds are products of the breakdown of p-cou-

maric and ferulic acids that are present in grapes and musts

[41]. These compounds have also been described in red

wines as almond shells and smokey aroma, respectively

[30]. Finally, methyl vanillate appeared to be higher in red

samples than in white ones (p \ 0.001; 22.20 vs.

3.35 lg kg-1, respectively). As regards the differences

among benzenic compounds between white and red varie-

ties, it could be suggested a link between the biosynthetic

pathway of benzene compounds and the shikimic acid

pathway that leads to the synthesis of polyphenols, having

them a point of contact represented by the phenylalanine

[39, 42]. Finally, the four relevant terpenes were trans-furan

linalool oxide, higher in rose samples (p = 0.033; 16.21 vs.

8.35 and 7.05 lg kg-1 per rose, red and white samples,

respectively), a-terpineol was higher in red followed by rose

and white varieties (p = 0.016; 11.06, 7.21 and

4.76 lg kg-1, respectively), endiol, higher in white sam-

ples than in red varieties (p = 0.002; 2.32 lg kg-1 vs.

0.74 lg kg-1, respectively), and p-menthene-7,8-diol,

higher in red samples (p = 0.026; 28.12 vs. 16.78,

11.64 lg kg-1 per red, rose and white samples, respec-

tively). The synthesis of terpenic compounds has been

appointed to be related to a genetic component [40, 43].

Therefore, the genetic differentiation between red and white

cultivars might be also related to the different grape aroma

compound found in white and red varieties.

The influence of the berry color in the aroma charac-

terization of Spanish varieties has also been studied by

different authors [33, 44]. However, volatile compounds

were different from those found in other studies [33], in

which the grapevines did not easily fall into the identified

groups [33, 44]. This could be explained considering the

larger number of grapevines included in this study with

respect to other studies.

Due to the different aroma composition in grape culti-

vars, the wine made from white and red varieties can be

clearly differentiated, since the aromatic profile depends on

the cultivars [4, 5] and the relationship between grape and

wines aroma compounds has been demonstrated [9, 11].

Red

Rose

White

-6

-4

-2

0

2

4

6

8

F2

(6.8

0 %

)

F1 (93.20 %)

Observations (axes F1 and F2: 100.00%)

Red Rose White Centroids

-8 -6 -4 -2 0 2 4 6 8

Fig. 2 Discriminant analysis (SFDA) of aroma from glycosidic

precursors from grapevines (Vitis vinifera L.) grouping according to

grape color. Symbols: triangles (centroids of each group) and circles(grapevine varieties). Three replicates per sample were used in the

analysis

478 Eur Food Res Technol (2011) 233:473–481

123

Role of the clone and origin of the Beba accessions

Differences between grapevine clones based on aroma

compounds have been appointed by other authors [3, 20,

21]. These results were confirmed in this study, since the

studied cultivars showed differences in aromatic profiles

between accessions of the same variety. The correlation

between accessions of the same variety was higher than

0.90 for all accessions of Bobal, Giro, Manto Negro and

Quigat varieties (r [ 0.97); however, they showed signifi-

cant differences (p \ 0.05) especially regarding terpene

and norisoprenoid compounds (Supporting Tables 1, 2).

Nonetheless, it is highlighted the notable exception of Beba

variety which presented r = 0.81 between the studied

accessions.

The dissimilation between the accessions of Beba variety

could be due to the origin of the accession, since the quality

and value of the glycosidic compounds were statistically

different depending on the origin of the accessions which

could be Peninsular (Girona and Levante area) or island

origin (Balearic Islands). Actually taking into account the

origin of the Beba accessions (Palma de Mallorca, Girona

and Levante; PM, GI and LE, respectively), the result of the

SFDA method indicated that the within-class covariance

matrices were different (p \ 0.001), and all the varieties

were classified correctly (Fig. 3). The clones of Beba

variety differed as far as their glycosidic compounds were

concerned; the variables that showed significant differences

were one benzenic compound, namely homovanillyl alco-

hol, higher in samples from PM (p = 0.002; 26.22 vs.

10.57, 13.82 lg kg-1; PM, GI, LE, respectively) and four

terpenes, namely a-terpineol, trans-pyran linalool oxide,

citronellol and diol (2,6-dimethyl-3,7-octadien-2,6-diol),

thus the terpenic compounds were crucial for the geo-

graphic grouping of Beba accessions. The a-terpineol

compound was higher in samples from PM (p = 0.036;

8.46 vs. 4.53, 5.93 lg kg-1; PM, GI, LE, respectively), this

compound is one of the most odoriferous monoterpene

alcohols not affected by the winemaking procedure [5] and

its aroma has been described in wine as floral, lilac and

sweet [7]. Trans-pyran linalool oxide was higher in PM

samples (p = 0.004; 22.03 vs. 14.68, 15.89 lg kg-1; PM,

LE, GI, respectively). Citronellol, exclusively present in

samples from LE (p = 0.078; 1.92 lg kg-1), has also been

found in other Spanish cultivars as Albarino [45], and

its aroma has been described in wines as citronella

[6]. Finally, diol (2,6-dimethyl-3,7-octadien-2,6-diol) was

lower in samples from PM (p = 0.011; 7.55 vs. 9.72,

10.71 lg kg-1; PM. LE, GI, respectively). It is an aroma

precursor [46] which is an odorant compound in wines [29].

Therefore, it is possible to choose clones with different

aroma profile, confirming the results obtained by other

authors [21].

The influences of the geographic origin over the aroma

compounds in must have been appointed by others authors

[11, 20, 45], highlighting the terpene compounds as a-ter-

pineol and citronellol [20, 45], confirming the results

obtained in this study. The synthesis of some terpenic

compounds has been appointed to have a genetic compo-

nent [3, 43]. Despite the fact that all Beba accessions were

planted in the same location, since they have been con-

served at the Vitis Germplasm Bank ‘‘Finca El Encın’’ from

the beginning of the XX century, they were collected in

several point of Spanish geography (PM, GI and LE). This

ancient variety is recorded in the Balearic Islands since

1730, in Levante area since 1791, or in Girona since 1877

[47]. As a result, the Beba variety could have adapted to

local environmental conditions, improving the synthesis of

the aroma metabolic route with a cumulative effect over

time.

Varieties included and excluded in Quality Demarcation

A comparison between varieties included in a Demarcation

of Quality and those excluded highlighted statistical dif-

ferences for red varieties in norisoprenoids and terpene

compounds (p \ 0.001; 520 vs. 371 and 350 vs.

132 lg kg-1, respectively), being the varieties not

belonging to any Quality Demarcation the richest ones. The

white varieties presented statistically significant differ-

ences (p \ 0.001) in alcohols, benzene and terpene com-

pounds (58 vs. 43; 821 vs. 614 and 324 vs. 190 lg kg-1,

respectively); however, in this case, the varieties included

GI

LE

PM

-5

0

5

10

15

-10 -5 0 5 10 15

F2

(12.

06 %

)

F1 (87.94 %)

Observations (axes F1 and F2: 100.00 %)

PM LE GI Centroids

Fig. 3 Discriminant analysis (SFDA) of aroma from glycosidic

precursors from Beba accessions grouping according to geographical

origin: Palma de Mallorca (PM), Levante area (LE) and Girona (GI)

Symbols: triangles (centroids of each group) and circles (grapevine

varieties). Three replicates per sample were used in the analysis

Eur Food Res Technol (2011) 233:473–481 479

123

in a Quality Demarcation turned out to be the richest ones

in these three groups of compounds. These results con-

firmed that differences between varieties including or not

in a Quality Demarcation are not easy to assess [23]. As a

matter of fact, comparing the aromatic profile between the

varieties included and excluded from Quality Demarca-

tions, we have demonstrated that some excluded varieties

showed greater quantity of glycosidic compounds with

respect to those included in Quality Demarcations (Sup-

porting Table 1, 2). Therefore, some cultivars as the red

ones Pampolat girat, Excursach, Gorgollasssa or Mandon

or the white ones Argamusa and Quigat might be recom-

mended to included in Demarcation of Quality; conse-

quently, they could be an excellent option to satisfy the

new demand of wine consumers. The quality of Gorgo-

llassa variety was probed in the past since their wines

obtained several prizes toward the end of the XIX century

[47]; its cultivation was gave up possible due to its low

production (Garcıa-Munoz et al. in preparation); therefore,

this variety is excluded from all Quality Demarcations

nowadays.

Conclusions

Glycosidic compounds were crucial to differentiate the

aroma potential of 21 cultivars. Differences in the aroma

profile between red and white varieties and clones of the

same variety have been identified as well as differences in

benzenic and terpenic compounds based on the origin of

the Beba variety. Difference between cultivars included

and excluded from Quality Demarcations has been asses-

sed, especially regarding terpene compounds. Based on

these results, it is possible to choose cultivar with different

aroma profile. The results suggest that cultivar, berry color,

and cultivar adaptations to location and environmental

conditions play an important role in the metabolic path-

ways to obtain the aromatic compounds. These metabolic

pathways seem to have a genetic component.

Minor varieties excluded from Quality Demarcations

appear to be more aromatic than the included ones, and as a

consequence, Argamusa, Quigat, Pampolat girat, Excur-

sach, Gorgollasssa or Mandon varieties could be taken into

consideration for the development of new wine market

strategies, which could also play an important role in the

conservation of these cultivars.

However, after this study, which has permitted to indi-

vidualize the more significant compounds that characterize

the potential aroma of those varieties, other criteria might

be analyzed. Agronomic characterization and further

investigations, such as GC–olfactometric studies or quan-

titative data, are needed to verify the impact of the odor-

active compounds identified in this analysis.

Acknowledgments Financial support from INIA project RTA 04

175-C3-3. Sonia Garcıa Munoz was supported by a PhD scholarship

from INIA. We thank Josu G. Alday for statistical advice. Sonia

Garcıa Munoz thanks to CRA-Centro di Ricerca per l’Enologia, Asti

(Italy) for welcoming her during her visiting period.

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