Albilla’ Cultivar from Galicia and Its Relationships with Other Spanish ‘Albillos’

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CHARACTERIZATION OF THE NEARLY EXTINCT ‘ALBILLA’ CULTIVAR FROM GALICIA AND ITS RELATIONSHIPS WITH OTHER SPANISH ‘ALBILLOS’

Emilia DÍAZ-LOSADA 1*, Sandra CORTÉS-DIÉGUEZ 1, Inmaculada RODRÍGUEZ-TORRES 2, José Manuel MIRÁS-AVALOS 1, Ignacio ORRIOLS-FERNÁNDEZ 1 and Santiago PEREIRA-LORENZO 3 1:INGACAL-Estación de Viticultura y Enología de Galicia, Ponte San Clodio s/n, 32420 Ourense, Spain 2: Instituto Canario de Investigaciones Agrarias (ICIA), Apdo. 60, La Laguna 38200 Santa Cruz de Tenerife, Spain 3: Escola Politécnica Superior. Departamento de Producción Vexetal, Universidad de Santiago de Compostela, Campus de Lugo, 27002 Lugo, Spain.

Abstract Résumé Aims : This work contributes to the knowledge of the Objectifs : Ce travail contribue à la connaissance de la genetic diversity of Vitis vinifera L. with the characterization diversité génétique de Vitis vinifera L. à travers l’évaluation of the non-referenced cultivar ‘Albilla’ by ampelographic de la variété inconnue ‘Albilla’ en utilisant une description description and molecular markers. This will be useful for ampélographique et des marqueurs moléculaires. Cette étude the preservation, identification and propagation of this pourra être utile pour la préservation, identification et cultivar in the future. propagation de cette variété dans le futur. Methods and results : Six microsatellites (SSRs), 55 OIV Méthodes et résultats : Six microsatellites (SSRs), (International Organisation of Vine and Wine) 55 descripteurs morphologiques de l’OIV (Organisation morphological descriptors and 45 phylometric traits were Internationale de la Vigne et du Vin) et 45 caractères evaluated in two accessions of ‘Albilla’ preserved in the phyllométriques ont été évalués sur deux accessions Germplasm Bank of EVEGA (Estación de Viticultura e d’‘Albilla’ préservées dans la banque de matériel Enoloxía de Galicia), Xunta de Galicia. Additionally, we set phytogénétique de l’EVEGA ( Estación de Viticultura e out to determine whether this cultivar was distinct from the Enoloxía de Galicia ), Xunta de Galicia. Nous avons de plus yet uncharacterized ‘Albillo’ cultivars found in the Canary déterminé si cette variété était distincte de la variété encore Islands. These methods allowed us to describe the cultivar non caractérisée ‘Albillo’ présente aux Îles Canaries. Ces known as ‘Albilla’ and to compare it with other homonym méthodes nous ont permis de décrire la variété dénommée cultivars. comme ‘Albilla’ et de la comparer avec d’autres variétés homonymes. Conclusions : The ‘Albilla’ cultivar is different from other genotypes denominated ‘Albillo’ and any other genotype Conclusions : La variété ‘Albilla’ est différente des described until now. This may indicate that it could be an génotypes nommés “Albillo” et distincte des génotypes autochthonous cultivar from Galicia. décrits jusqu’à aujourd’hui. Ceci indique qu’elle pourrait être une variété autochtone de Galicia. Significance and impact of the study : The characterization of cultivars is still necessary in order to identify any Signification et impact de l’étude : La caractérisation de homonyms or synonyms, develop measures for their variétés reste nécessaire pour trouver des homonymes ou conservation (germplasm collection) and evaluate their synonymes, agir pour leur sauvegarde et évaluer leur potential for producing quality wines. potentiel pour produire des vins de qualité. Key words : Vitis vinifera , ampelography, microsatellites Mots clés : Vitis vinifera , ampélographie , microsatellites

manuscript received 6th September 2012 - revised manuscript received 11th June 2013

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INTRODUCTION by ampelography and molecular markers (Cervera et al. , 2001), are preserved in the grapevine collection of Geographical diversity, climate conditions and history El Encín (Spain) : i) ‘Albillo’ (official prime name have contributed to the large number of cultivars that ‘Albillo Real’), cultivated in Ávila, Madrid, León and were conserved and spread throughout Europe before Zamora, with its synonyms ‘Nieves Temprano’ in the nineteenth century (This et al. , 2006), that is, when Zaragoza, ‘Temprano de Campo Real’ or ‘Temprano phylloxera, downy mildew and powdery mildew de Mora’ in Madrid, ‘Albillo de Toro’ in Zamora and caused severe genetic erosion. ‘Albillo de Cebreros’ in Ávila and its homonym ‘Albillo de Granada’ (prime name) ; ii) ‘Albillo Galician grapevines originate from western Europe Mayor’ from Palencia, with its synonyms ‘Turruntés’ and are composed of a mixture of autochthonous and in La Rioja (Rodríguez-Torres et al. , 2000), ‘Albillo’ allochthonous cultivars, mainly of French origin, in Burgos and Valladolid, and ‘Blanco del País’ in introduced into the region with the establishment of Burgos ; and iii) ‘Albillo de El Bierzo’ (official prime religious institutions (Huetz de Lemps, 1967) . Due to name ‘Chasselas’ in the Registro de Variedades the problems caused by phylloxera and downy and Comerciales), with its synonyms ‘Albillo’ in Asturias powdery mildew in the nineteenth century, only a few and Salamanca, ‘Malvasía’ in León, and ‘Temprano’ of the most productive cultivars were promoted for or ‘Temprano Blanco’ in Valladolid. wine production, and this process continues. However, autochthonous cultivars are also able to produce both Cabello et al. (2003) have reported that the ‘Albillo’ quality and distinct wines. Thus, the identification and cultivated in Galicia is a synonym of ‘Chasselas characterization of such cultivars is still necessary in Doré’. Rodríguez-Torres et al. (2000) considered the order to discover any homonyms or synonyms and ‘Albillo’ cultivated in the D.O. Ribera del Duero to be evaluate their potential for producing quality wines. synonymous with the ‘Turruntés’ from the D.O. La Rioja. Martínez and Pérez (2000) have described Some traditional cultivars from north-western Spain ‘Albillo’ in Asturias, which has been recently have recently been characterized, but this analysis did identified as ‘Chasselas’(Moreno-Sanz et al. , 2008). not include ‘Albilla’ (Santiago et al. , 2005). ‘Albilla’ is a minor cultivar found in north-western Spain. It is The basic methodology for cultivar characterization is in danger of extinction since its potential to produce ampelography complemented with the analysis of quality wine is unknown. ‘Albilla’ has been cultivated molecular markers, mainly microsatellites (SSRs) since ancient times in the Ribeiro region (Galicia) and (Thomas et al. , 1994) . The International Organisation has been mentioned by the oldest growers, though the of Vine and Wine (OIV) provides a list of earliest written evidence was found in ‘Catastro morphological descriptors and recommends the Vitícola’ (1983). ‘Albilla’ is authorized in the analysis of six microsatellite loci for cultivar Designation of Origin (D.O.) Ribeiro, but it is not yet identification (OIV, 2009). included in the National Register of Commercial Cultivars (Cabello et al. , 2011). Therefore, it cannot In this context, the aim of this study was to be commercially propagated. ‘Albillo’ (not ‘Albilla’) characterize the ‘Albilla’ cultivar conserved in the has been reported by García de los Salmones (1914) in Germplasm Bank of EVEGA, Xunta de Galicia. eastern Galicia, whereas ‘Albilla’ has been reported in Ampelography and microsatellite analysis were used the ‘Catastro Vitícola y Vinícola (D.O. Ribeiro)’ in order to establish the distinctiveness and (1983) specifically in the localities of Beade and ampelographical characteristics of the cultivar so that Cenlle, where a few scarce accessions can still be it could be registered in the future and to contribute found. towards maintaining the genetic diversity of the area. Additionally, we determined whether this cultivar has The complexity of the ‘Albillo’ denomination in Spain been present in the Canary Islands, considered as a goes back over two centuries due to the general refuge for Spanish cultivars, where phylloxera would meaning of the word (i.e., white). To our knowledge, not have affected it ( Zerolo et al. , 2006). in Galicia, ‘Albillo’ was first cited in 1914 by García de los Salmones in eastern Galicia, and more MATERIALS AND METHODS specifically in Fonsagrada (province of Lugo) and 1. Plant material Puebla de Trives (province of Ourense). Recently, many authors have studied the complexity of The plant material used in this study comprises two ‘Albillo’, including synonymies and homonymies. accessions of ‘Albilla’ from EVEGA, located in Leiro- The different genotypes named ‘Albillo’ cultivated in Ourense (Galicia, Spain) and three accessions named the Iberian Peninsula, which have been characterized ‘Albillo Criollo’, ‘Albillo Forastero’ and ‘Albillo

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Monte Lentiscal’ from the collection of ICIA 9 ) 7

e G 3 3 9 5 5 3 9 7 (Tenerife, Canary Islands, Spain). The ‘Albilla’ f i 4 4 4 5 4 4 4 5 A r 2 2 2 2 2 2 2 2 Z accessions at EVEGA were obtained by clonal e r n V propagation of one-hundred-year-old grapevines from e T

( 9

the D.O. Ribeiro (north-western Spain). Two French 7

A G I 7 7 9 9 5 7 3 9 3 3 4 4 4 3 4 4

cultivars from the EVEGA collection were included as A C 2 2 2 2 2 2 2 2 I Z

references for the SSR analyses : ‘Pinot noir’ and r V m o

‘Syrah’. r 2 f

s G 5 5 5 3 7 3 3 3 r 9 9 9 9 8 9 9 0 a 2. Morphological analysis A 1 1 1 1 1 1 1 2 v Z i r t l V u

Different traits, corresponding to young shoots, mature c

2 ’ 6 o

leaves, clusters and berries, were studied in 2006, l G l 7 7 7 7 5 7 7 3 i 8 8 8 8 8 8 8 9 A b 1 1 1 1 1 1 1 1

2007 and 2009 on the two accessions of ‘Albilla’, l Z r A V ‘

according to the descriptors defined by the OIV

e 7 (2009). e r 4

h G 9 9 5 1 9 5 7 5 t

6 6 6 7 5 6 6 6 , A ) 1 1 1 1 1 1 1 1

Ten leaves from the middle third of the shoot were Z r d l V collected from each accession and evaluated for o

7 . n 45 phylometric traits, as proposed by Cid-Alvarez et 4 i

A , G 5 5 1 5 7 1 5 1 a 6 6 6 6 5 6 6 6 G A

al. (1994) using NIS-Elements BR 2.30 software. i 1 1 1 1 1 1 1 1 c Z E i r l V V a E

For the quantitative analysis of leaf parameters, we G 7 ( n

estimated the average, the coefficient of variation and D A 1 1 7 5 5 1 7 5 d 4 4 3 5 4 4 3 4 M n G the significant differences by ANOVA between the 2 2 2 2 2 2 2 2 V u E o V f V

accessions studied, using SPSS 15.1 software (SPSS

s E 7

Inc for Windows, Chicago, IL, USA). c 7 7 7 7 7 7 7 7 m n 3 3 3 3 3 3 3 3 M o e 2 2 2 2 2 2 2 2 r r V f

e ’

3. Molecular analysis V f a e

l l r 5

h D b l The six SSRs recommended by the OIV (2009) were 6 6 6 4 6 4 8 2 c 3 3 3 2 3 3 2 3 M n A 2 2 2 2 2 2 2 2 ‘ e V

used in this study. Polymerase chain reaction (PCR) r f V o F

was conducted according to the methodology defined s e 5 n e

D r by Martín et al. (2003). The PCR products were o 4 4 8 8 2 4 2 4 i h 3 3 1 1 3 2 2 2 s M t s 2 2 2 2 2 2 2 2

separated and analysed on a 3730 DNA Analyser V e d c V n (Applied Biosystems, USA). The size of the amplified c

a a

0 0 0 2 0 0 0 0 o products was determined using the Peak Software S 5 5 3 4 5 5 3 4 V w 1 1 1 1 1 1 1 1 t

version 1.0 ( http://www.appliedbiosystems.com ). V n

s 0 0 0 0 2 4 0 0 S e 3 3 3 3 4 3 3 3 t

RESULTS AND DISCUSSION V i 1 1 1 1 1 1 1 1 l V l e

n n a

Both ‘Albilla’ accessions from EVEGA showed the n n n o o s i i o o o i i i o

t t t t t r c c same genotype based on six SSRs and were different n n n c c c e e c o o o e e e l l i i i i l l l l l t t t l l l o o

from ‘Albillo Criollo’, ‘Albillo Forastero’, and c c c m o o o

e e e C C l l l C C C

x l l l

‘Albillo Monte Lentiscal’ from ICIA (Table 1). They i o o o s A A

A A A

C C C n f G G

i were also different from ‘Chasselas’ from EVEGA, G G G o g

E E E E E A A A i ) I I I r V V V V V which corresponded to the genotype from the p C C C O E E I I I E E E b

(

‘Colección de El Encín’ reported by Martín et al. ’ s l e a z c

(2003). Therefore, ‘Albilla’ from EVEGA is different i s s i

e n

from the other cultivars previously reported ’ l e

o e

r l L ’

(http://www.sivvem.monbyte.com/ e e o t e l t A s l

m a n . o

sivvem.asp ; http://www.vivc.de ; Rodríguez-Torres et r i a o ’ ’ 1 r

o r s n i

e a C F M ’ ’ l o

r al. , 2000 ; Borrego et al. , 2002 ; Cabello et al. , 2003 ; l

e a a n ’ a o o o b

l l l l l s v t l l h l l l a s i i i i i i

Ibañez et al. , 2003 ; Martín et al. , 2003 ; Jiménez- a o t a T r b b b b b l n l l l l l h i y u P Cantizano et al. , 2006 ; Santana et al. , 2008 ; Díaz- A A A A A S C Losada et al. , 2012). C ‘ ‘ ‘ ‘ ‘ ‘ ‘ ‘

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The ampelographic characteristics of shoots, young OIV 602, N3-OIV 603) ; the number of teeth between and mature leaves, bunches and berries of ‘Albilla’ are the tooth tip of N2 and the tooth tip of the first presented in Table 2 and Figure 1. Ampelography secondary vein (ND-OIV 616) ; the distance between confirmed the singularity of ‘Albilla’ observed by the petiole point and upper sinus (U-OIV 605) ; the microsatellites with respect to other ‘Albillo’ types length of the tooth N1 to the half point at its base reported in Spain . Cabello et al. (2003) identified (L1) ; the distance between the tips of N2 and N3 ‘Albillo’ as a synonym of ‘Chasselas’; however, (ENM) ; the projection of the distance between N3 ‘Chasselas’ presents bronze young leaves with very and N1 over the leaf axis (L) ; the sum of the lengths low density of hairs and mature leaves with very deep N2 on the large side and N1 on the small side of the lateral sinuses, as opposed to ‘Albillo’ from EVEGA. leaf (N2S) ; the angle between the extremes of N1 and ‘Albillo Mayor’, cultivated in the D.O. Ribera del N2 (AP) ; and the distance between the bases of the Duero and Cigales (Rubio et al. , 2003), can be upper sinuses (DS1). These variations can be distinguished from the ‘Albillo’ maintained at considered as clonal variations. EVEGA by its large mature leaves with 7 lobes, very overlapped petiole sinus and lateral sinuses, Microsatellite genotyping and ampelographic data cylindrical bunches, and berries with Muscat flavor. showed that ‘Albilla’ from EVEGA, propagated from Finally, ‘Albillo’ from EVEGA is different from Galician grapevines that are more than 100 years old, ‘Albillo Real’ (Rubio et al. , 2003 ; Cabello et al. , is different from the grapevine preserved in the 2003) found in Madrid, Zamora and Ávila since the ‘Colección de Variedades de Vid de El Encín’ and is latter presents overlapped lateral sinuses, opened also different from ‘Albillo Criollo’, ‘Albillo petiole sinus, and spherical berries. Forastero’ and ‘Albillo Monte Lentiscal’ from the Canary Islands. Therefore, ‘Albilla’ from EVEGA is a Table 3 gives the phylometric measurements of minor cultivar. ‘Albillo’ is the name cited in previous ‘Albilla’ leaves. Both of the ‘Albilla’ accessions from references and corresponds to the genotypes included EVEGA presented 77.7% of the ampelographic traits in the European Vitis Database, but we have used the with no significant differences, which indicates the name ‘Albilla’ because this is the name given by the stability of these traits. The main significant variations growers to the genotype from EVEGA, the true-to- were found in : the lengths of the central vein and the type variety authorized in the D.O. Ribeiro, which is upper and lower lateral main veins (N1-OIV 601, N2- different from other genotypes named ‘Albillo’.

Figure 1 -Young shoot (a), upper side of young leaf (b), upper side of mature leaf (c), lower side of mature leaf (d), cluster (e), berries (f), cross-section of the grape berry (g) and seeds (h) of ‘Albilla’ from EVEGA, Galicia.

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Table 2 - Ampelographic traits analysed in a young shoot, mature leaf, bunch and berry of the ‘Albilla’ cultivar (OIV, 2009) (Expression indicates mode with minimum and maximum values in parenthesis).

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Table 3 - Leaf traits of the ‘Albilla’ cultivar from EVEGA (mean and F values, standard deviation [S.D.], coefficient of variation [C.V.] in %). The references from OIV are shown in parentheses.

n.s., *, **, *** non-significant and significant at P ≤ 0.05, 0.01 or 0.001, respectively.

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García de los Salmones (1914) cited ‘Albillo’ in grapevine accessions known as ‘Albillo’. Am. J. Enol. eastern Galicia, while the ‘Albilla’ from EVEGA was Vitic. , 52 , 2, 127-135. localised in the Ribeiro region. There is no description Cid-Alvarez N., Boursiquot J.M., Saa-Otero M.P. and for the ‘Albillo’ cited by García de los Salmones Romani-Martinez L., 1994. Différenciation des (1914) to compare with the ‘Albilla’ from EVEGA. cépages autochtones du Nord-Ouest de l’Espagne The ‘Albillo’ genotypes from previous studies are (Galice) et élaboration d’une clé de détermination different from the ‘Albilla’ genotype included in the basée sur l’ampélométrie. J. Int. Sci. Vigne Vin , 28 , 1, EVEGA and used in the Ribeiro region, where it is 1-17. still cultivated and cited by the oldest growers. Díaz-Losada E., Tato Salgado A., Ramos-Cabrer A.M., Moreover, in the ‘Catastro Vitícola’ (1983), it is Díaz-Hernández B. and Pereira-Lorenzo S., 2012. included with the denomination ‘Albilla’ from Ribeiro. Genetic and geographical structure in grapevines from For all these reasons, we recommend maintaining the northwestern Spain. Ann. Appl. Biol. , 161 , 1, 24-35. name ‘Albilla of Ribeiro’ (the name of the accessions García de los Salmones N., 1914. Memoria General de las in the EVEGA collection) to avoid any confusion with Sesiones del Congreso y Ponencias Presentadas . other genotypes named ‘Albillo’. Congreso Nacional de Viticultura, Pamplona, Spain. Huetz de Lemps A., 1967. Vignobles et Vins du Nord-Ouest CONCLUSIONS de l’Espagne, tome premier . Ed. Impressions Bellenef, Bordeaux, France. This study provides descriptive data of the ‘Albilla’ cultivar, which is currently in danger of extinction due Ibáñez J., de Andrés M.T., Molino A. and Borrego J., 2003. Genetic study of key Spanish grapevine varieties to the impossibility of commercializing unregistered using microsatellite analysis. Am. J. Enol. Vitic. , 54 , plants for new plantations, even though it has been 1, 22-30. considered a preferred variety in the D.O. Ribeiro. Jiménez-Cantizano A., Martínez-Zapater J.M., García de This cultivar is different from other genotypes Luján A. and Arroyo-García R., 2006. Caracterización denominated ‘Albillo’ and any other genotype molecular de accesiones de vid del banco de described until now, as proved by the six SSR analysis. germoplasma del Rancho de la Merced. In Proc del This may indicate that it could be an autochthonous XXIX Congreso Mundial de la Viña y el Vino , cultivar from Galicia. Logroño, Spain.

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