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Investigating the origin of hazelnut

(Corylus avellana L.) cultivars using

chloroplast microsatellite

 ARTICLE  in  GENETIC RESOURCES AND CROP EVOLUTION · SEPTEMBER 2009

Impact Factor: 1.46 · DOI: 10.1007/s10722-009-9406-6

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Paolo Boccacci

Italian National Research Council

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Roberto Botta

Università degli Studi di Torino

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R E S E A R C H A R T I C L E

Investigating the origin of hazelnut (Corylus avellana  L.)

cultivars using chloroplast microsatellitesPaolo Boccacci

 

Roberto Botta

Received: 14 July 2008 / Accepted: 12 January 2009 / Published online: 31 January 2009

 Springer Science+Business Media B.V. 2009

Abstract   The place and time of European hazel

(Corylus avellana   L.) domestication is not clear,

although it was already cultivated by the Romans. In

this study, 75 accessions from Spain, Italy, Turkey,

and Iran were analysed using 13 chloroplast micro-

satellite to investigate the origin and diffusion of 

hazelnut cultivars. Four loci were polymorphic and

identified a total of four different chlorotypes. Their

distribution was not uniform in each geographical

group. The most frequent chlorotype A was present in

all groups. An increase in chlorotype number anddiversity from Spain eastward to Italy, Turkey, and

Iran was observed. Results suggest that some spread

of cultivars occurred from East to West and that

hazelnut cultivation was not introduced from the

eastern Mediterranean basin into Spain and southern

Italy by Greeks or Arabs. Moreover, the results

suggest considerable exchange of germplasm

between Italy and Spain, probably by the Romans.

Hazelnut appears to have been domesticated inde-

pendently in three areas: the Mediterranean, Turkey,

and Iran.

Keywords   Chlorotype     Corylus avellana  cpSSR    Cultivar diffusion    Cultivation  Domestication   Filbert

Introduction

In Europe, two different  Corylus   species are present:

the European hazel,  C. avellana   L., that has a wide

distribution, and the Turkish hazel,   C. colurna   L.,

restricted to the Balkans, Romania, and northernTurkey (Thompson et al.   1996). The geographical

distribution of the European hazel extends from the

Mediterranean coast of North Africa northward to the

British Islands and the Scandinavian peninsula, and

eastward to the Ural Mountains of Russia, the

Caucasus Mountains, Iran, and Lebanon (Kasapligil

1972). It is the source of important cultivars in Europe

and Turkey, which were selected over many centuries

from local wild populations (Trotter   1921; Tasias

Valls 1975; Thompson et al.  1996). Turkey has long

been the leading producer and exporter of hazelnuts,accounting for about 71% of world production. Italy is

second with over 13%, the United States third with

4.1%, and Spain fourth with 2.8%. Azerbaijan, Iran,

Georgia, China, France, and Greece are other pro-

ducers (FAOstat 2008). About 90% of the world crop

is used as kernels in the food industry, while the

remaining 10% is sold in-shell for fresh consumption.

The present-day distribution of  C. avellana L. was

established about 7,000 years B.P., as result of a

P. Boccacci (&)    R. Botta

Dipartimento di Colture Arboree, Universita degli Studi di

Torino, Via Leonardo da Vinci 44, 10095 Grugliasco,

Torino, Italy

e-mail: paolo.boccacci@unito.it

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postglacial recolonization process starting approxi-

mately 11,000 years earlier (Huntley and Birks 1983).

During the most recent glaciation hazel was restricted

to glacial refugia in southern Europe, but low levels of 

pollen were deposited in central Europe during the full

glacial period (Bennett et al. 1991). Huntley and Birks

(1983) suggested that southern Italy and the areaaround the Bay of Biscay (southwestern France) were

the most important refugia. Between 10,000 and

9,000 years B.P. the amount of  Corylus  pollen found

across Europe sharply increased (http://www.

pierroton.inra.fr/Cytofor/Maps/index.html). The most

plausible explanation for this increase is a very rapid

spread of hazel (1,500 m/year) from these refugia

(Huntley and Birks 1983; Birks 1989). Recent analysis

of chloroplast DNA variation (Palme and Vendramin

2002) indicates a rapid expansion from one large

refugium in the Biscay area or from several scatteredrefugia in western Europe into most of Europe, except

southern-central Italy and the Balkans, and then a local

expansion in the latter two areas.

Nut dispersal during the postglacial recolonization

was caused by animals (small mammals and birds)

and human migration. A prevalent opinion is that

Mesolithic tribes (10,000–6,000 years B.P.) could

have aided, intentionally or more likely accidentally,

the spread of hazel. However, there is no evidence

that they attempted to propagate it  (Tallantire 2002).

In several Mesolithic archaeological findings,abundant nutshell fragments have been recorded,

indicating that hazelnuts were cracked for consump-

tion or some kind of nut-processing (Bakels  1991;

Kubiak-Martens 1999). Most likely the expansion of 

the hazelnut distribution was due to chance spread

during the preparation of ‘‘hazelnut meals’’ by

migratory Mesolithic people (Kuster   2000). During

the spread of agriculture in Europe,  C. avellana   L.

was one of the species domesticated and cultivated

(Forni 1990; Zohary and Hopf  2001). Archaeologists

have repeatedly retrieved nuts, kernels, and shellremains from many Neolithic, Bronze Age, Classical,

and Medieval sites all over Europe (Bakels   1991;

Russel-White 1995; Arobba et al. 2003; Cleary 2003;

Pena-Chocarro et al.   2005; Sostaric   et al.   2006;

Schmidl et al.  2007). Nevertheless, where and when

the domestication of  C. avellana L. was started is not

yet clear; although it was cultivated by the Romans

(Trotter   1921; White   1970; Vaughan and Geissler

1997).

In recent years, the availability of nuclear micro-

satellite or simple sequence repeat (nSSR) markers in

C. avellana  L. (Bassil et al. 2005a, b; Boccacci et al.

2005) provide new possibilities to investigate the

breeding history of hazelnut cultivars. nSSR loci have

been used to fingerprint and to identify mistakes in

hazelnut accessions from several germplasm reposi-tories (Botta et al. 2005; Gokirmak et al. 2008). These

studies investigated genetic relationships among

cultivars grown in important production areas,

identified accessions with identical fingerprints, and

suggested the parentage for several cultivars

(Boccacci et al.   2006,   2008; Ghanbari et al.   2005;

Gokirmak et al.  2008). In particular, Boccacci et al.

(2006) and Gokirmak et al. (2008) concluded that

cultivars from Italy and Spain are genetically related.

On the contrary, Turkish cultivars constitute a

separate germplasm group, indicating little gene flowbetween the eastern and western areas of the

Mediterranean basin.

The chloroplast genome has a lower evolutionary

rate than the nuclear genome. Although its inheri-

tance is paternal in conifers (Vendramin and

Ziegenhagen 1997), inheritance is maternal in angio-

sperms (Dumolin et al.   1995; Arroyo-Garcıa et al.

2002). Thus in angiosperms the chloroplast genome

can only be disseminated by seeds or cuttings.

Chloroplast SSR (cpSSR) markers have been devel-

oped in recent years (Provan et al.   2001) and havebeen used to investigate the origin and diffusion of 

several fruit tree species, such as grape (Arroyo-

Garcıa et al. 2002; Imazio et al. 2005; Arroyo-Garcıa

et al.  2006) and olive (Breton et al.  2006).

In the present work, a total of 75 genotypes from four

different geographical regions were analysed using

cpSSR markers to investigate the history of hazelnut

cultivation and diffusion. Moreover, the usefulness of 

cpSSRs as molecular markers to study genetic rela-

tionships among hazelnut cultivars was evaluated.

Materials and methods

Plant material and DNA extraction

Seventy-five hazelnut accessions were sampled in

field collections assembled by the Dipartimento di

Colture Arboree of Torino (Italy); Cooperativa San

Colombano (Genova, Italy); Istituto di Frutti-

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viticoltura of Piacenza (Italy); Consiglio per la

Ricerca e Sperimentazione in Agricoltura (CRA) of 

Roma and Caserta (Italy); National Clonal Germ-

plasm Repository (NCGR) of Corvallis (Oregon,

USA), Institut de Recerca i Tecnologia Agroali-

mentaries (IRTA) of Constantı   (Tarragona, Spain),

and Seed and Plant Improvement Institute (SPII) of Kamal-Abad (Iran). Trueness-to-type of the acces-

sions was previously verified by Boccacci et al.

(2005, 2008) and Ghanbari et al. (2005) using nSSR

markers. The cultivars were assigned to four groups

on the basis of their country of origin (Table 1).

Cultivars with a direct parental relationship were

excluded, as were synonyms. (i) Group Spain is

composed by 33 cultivars grown in the province of 

Tarragona (Catalonia, northeastern Spain), except

‘Casina’, which is from Asturias (northern coast of 

Spain); (ii) Group Italy includes 22 cultivars grown indifferent Italian regions: Piedmont and Liguria

(northwestern Italy), Latium (central Italy), and

Campania and Sicily (southern Italy); (iii) Group

Turkey is represented by ten cultivars of Turkish

origin, seven grown in the Black Sea coastal prov-

inces of northern Turkey, and three cultivated in

Greece (‘Extra Ghiaghli’, ‘Sivri Ghiaghli’, and

‘Tombul Ghiaghli’); (iv) Group Iran is composed of 

10 accessions from Iran.

Total genomic DNA was extracted from 0.2 g of 

leaves or immature catkins using the modifiedprocedure described by Thomas et al. (1993) i n a

Tris–EDTA–NaCl buffer containing 0.25 M NaCl,

0.2 M Tris pH 7.6, 2.5% PVP 40,000, 0.05 M

Na2EDTA, and 0.1%  b-mercaptoethanol. After puri-

fication, the DNA was suspended in 50 ll Tris–

EDTA buffer.

cpSSR analysis and data elaboration

A total of thirteen cpSSR loci were analysed. Eight

loci (ccmp1, ccmp2, ccmp3, ccmp4, ccmp5, ccmp6,ccmp7, ccmp10) were described by Weising and

Gardner (1999), and five loci (cmcs1, cmcs2, cmcs4,

cmcs11, cmcs13) were described by Sebastiani et al.

(2004). The primer pairs were those designed by

these authors for   Nicotiana tabacum   L. and   Casta-

nea sativa Mill., respectively. PCR amplification was

carried out using a reaction mixture (20  ll) consisting

of 50 ng DNA template, 0.5  lM of each primer,

200  lM dNTPs, 2 mM MgCl2, 2  ll 109 NH4  buffer

[160 mM (NH4)2SO4, 670 mM Tris–HCl (pH 8.8 at

25C), 0.1% Tween-20], and 0.5 U BioTaq DNA

polymerase (Bioline, London, UK). All cpSSR

amplifications were performed using the following

temperature profiles: 3 min of denaturation at 95C,

then 28 cycles of 30 s of denaturation at 95C, 45 s of 

annealing at 55C, and 90 s of extension at 72C; anda final 10 min extension step at 72C. Amplified

fragments were loaded onto a polyacrylamide gel and

run on a semi-automated ABI-PRISM 377 sequencer

(Applied Biosystems, Foster City, Calif., USA).

Results of the run were processed with Genescan

software and allele sizes (in base pairs, bp) were

estimated using a GeneScan-350 ROX size standard

(Applied Biosystems).

Numbers of chlorotypes (N) were directly esti-

mated for each cultivar group. An unbiased estimate

of the chlorotype diversity was calculated as:H ¼   n=   n  1ð Þ   1  Rg2

i

, where gi   is the fre-

quency of the   i-th chlorotype and n is the number

of individuals analysed in each cultivar groups (Nei

1987). A chlorotype median network was constructed

using the program Network v. 4.5 (Bandelt et al.

1999).

Results

cpSSR polymorphism and chlorotype definition

In the 75 hazelnut genotypes, nine of 13 cpSSR loci

were monomorphic. Low levels of variation were

detected with the other four loci (Table 2). Locus

ccmp2 showed three different size variants, whereas

two variants were found at loci ccmp3, ccmp4, and

ccmp10. The allele variations differed by increments

of 1 bp due to variation in the number of A or T

residues within mononucleotide repeats. The same set

of cpSSR loci was also polymorphic in 26 natural

hazel populations (Palme   and Vendramin   2002).Thus, ccmp2, ccmp3, ccmp4, and ccmp10 are useful

loci for studying genetic variability in  C. avellana  L.

Considering the allele variants at the four loci, four

different chlorotypes were detected (Table 3) and

their relationships were analysed under a network 

model (Fig. 1). The chlorotype network indicated the

minimum number of evolutionary events separating

each chlorotype. Chlorotype A was the most frequent

in the 75 cultivars. Chlorotypes B and D were

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Table 1   List of accessions, germplasm collections, and chlorotypes observed in 75 hazelnut cultivars

Cultivar Collectiona

Chlorotype Cultivar Collectiona

Chlorotype

Group Spain  (33 cultivars)

Ametllenca IRTA A Pinyolenc IRTA A

Apegalos IRTA A Planeta IRTA A

Artellet IRTA A Punxenc IRTA A

Casina DCA A Queixal de llop IRTA A

Castanyera (syn. Barcelona) DCA A Queixal de ruc IRTA A

Closca molla DCA A Ratllada IRTA A

Culpla   DCA A Ratolı   IRTA A

Gironell (syn. Grossal) DCA A Ribet IRTA A

Gironenc (syn. Vermellet) NCGR A Ros IRTA A

Grifoll IRTA A Rosset IRTA A

Lluenta IRTA A Sant Jaume IRTA A

Martorella IRTA A Sant Joan IRTA A

Morell DCA A Sant Pere IRTA A

Negret IRTA A Segorbe IRTA A

Negret capellut IRTA A Trenet IRTA A

Negret garrofı   IRTA A Vimbodı   IRTA A

Pauetet DCA A

Group Italy  (22 cultivars)

Camponica DCA A Nocchione DCA A

Catainetto Coop A Noscello Coop A

Del Rosso Coop A Nociara DCA A

Dell’Orto Coop A Riccia di Talanico CRA A

Ghirara DCA A San Giovanni DCA A

Gianchetta IFP A Tonda bianca DCA DIannusa Racinante DCA A Tonda di Giffoni DCA A

Menoia IFP A Tonda Gentile delle Langhe DCA A

Mortarella DCA A Tonda Gentile Romana CRA A

Napoletana DCA A Tonda rossa CRA D

Napoletanedda CRA A Trietta IFP A

Group Turkey  (10 cultivars)

Badem IRTA A Palaz DCA B

Extra Ghiaghli IRTA A Sivri IRTA A

Imperial de Trebizonde (syn. Karidaty) DCA B Sivri Ghiaghli NCGR B

Incekara IRTA B Tombul IRTA A

Kalinkara NCGR B Tombul Ghiaghli NCGR BGroup Iran  (10 cultivars)

Asle Gharebag SPII C Pashmineh SPII C

Dobooseh SPII A Rasmi SPII C

Jorow Gharebag SPII C Shastak-2 SPII C

Mish-pestan SPII C Shirvani SPII C

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associated with chlorotype A, while chlorotype C was

related only to chlorotype B. The intermediate

relationship of chlorotype A to the others suggests

that it is likely the ancestral chlorotype of hazelnut

cultivars.

Chlorotype variation and distribution

in the geographical groups

The chlorotype distribution was not uniform in thefour geographical groups (Table 3). Chlorotype A

was present in all groups with a decreasing frequency

from Spain eastward to Italy, Turkey, and Iran

(Table 3). It was observed in all 33 Spanish and 20

of 22 Italian accessions. ‘Tonda bianca’ and ‘Tonda

rossa’, both of which are cultivated in Avellino

province (Campania, southern Italy), showed the

rarest chlorotype D (Table  1). Chlorotypes B and C

were mainly present in the groups Turkey and Iran,

respectively, and were absent in Spanish and Italian

cultivars (Table 3).

In order to evaluate the chlorotype distribution in

each analysed group, a chlorotype diversity index (H)

was calculated (Table 3). H values were zero in the

Spanish group, and low in the Italian group. On the

contrary, high H values were observed in groups

Turkey (H  =  0.533) and Iran (H  =   0.378), although

a relatively fewer accessions from these groups were

analysed. These data showed that H increases fromWest to East in the Mediterranean basin, unlike what

was observed in grapevine by Imazio et al. (2005).

Discussion

The postglacial migration of   C. avellana   L. was

hypothesized on the basis of pollen records (Huntley

and Birks 1983; Birks 1989) and a recent analysis of 

Table 1   continued

Cultivar Collectiona Chlorotype Cultivar Collectiona Chlorotype

Nakhoni Rood SPII C Tabari Rood SPII B

a  DCA, Dipartimento di Colture Arboree of Torino (Italy);   IRTA, Istitut de Recerca i Tecnologia Agroalimentaries of Reus

(Tarragona, Spain); Coop, Cooperativa San Colombano (Genova, Italy); IFP, Istituto di Frutticoltura of Piacenza (Italy);  CRA, Centro

Ricerche e sperimentazione in Agricoltura of Roma and Caserta (Italy);  NCGR, National Clonal Germplasm Repository of Corvallis(Oregon, USA);  SPII , Seed and Plant Improvement Institute (Karaj, Iran)

Table 2   Chlorotype genotypes at 13 cpSSR loci

Chlorotype ccmp1 ccmp2 ccmp3 ccmp4 ccmp5 ccmp6 ccmp7 ccmp10 cmcs1 cmcs2 cmcs4 cmcs11 cmcs13

A 130 214 119 117 108 98 154 108 103 134 108 224 118

B 130 214 118 117 108 98 154 108 103 134 108 224 118

C 130 215 118 117 108 98 154 108 103 134 108 224 118

D 130 216 119 116 108 98 154 107 103 134 108 224 118

Only four loci (ccmp2, ccmp3, ccmp4, and ccmp10) were polymorphic and identified four chlorotypes

Table 3   Chlorotype

distribution in each

geographical group (with

chlorotype frequency in

parentheses)

n  =  number of samples;

N  =  number of 

chlorotypes;

H  =  chlorotype diversity

Group Spain Group Italy Group Turkey Group Iran Total

n 33 22 10 10 75

Chlorotype A 33 (1.000) 20 (0.909) 4 (0.400) 1 (0.100) 58 (0.773)

Chlorotype B – – 6 (0.600) 1 (0.100) 7 (0.094)

Chlorotype C – – – 8 (0.800) 8 (0.108)

Chlorotype D – 2 (0.091) – – 2 (0.027)

N 1 2 2 3 4

H 0.000 0.173 0.533 0.378 0.391

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chloroplast DNA variation in wild hazelnuts (Palme

and Vendramin 2002). The most likely scenario is an

expansion from southwestern France into most of Europe, except for Italy and the Balkans where

expansions were local. Archaeobotanical remains

indicate that hazel kernels have been an important

constituent in the human diet since prehistoric times.

Kernels are tasty and have a high energy value (600–

680 kcal per 100 g fresh weight) due to their high fat

and protein content. In addition, hazelnuts are easy to

store and transport. Mesolithic tribes could have

aided, intentionally or more likely accidentally, the

spread of hazel. According to Tallantire (2002), they

did not attempt to propagate it.The place and time of hazel domestication is not

clear. The common opinion is based on a statement of 

Pliny the Elder (23–79 A.D.) in the work   Natu-

ralis Historia   that the hazelnut came from Asia

Minor and Pontus (north coast of Turkey). On the

basis of this assertion, the accepted general idea is

that hazelnut cultivation was brought to Italy by the

Greeks and that the specific epithet  avellana  derives

from Abellina in western Asia, allegedly the present

valley of Damascus (Trotter   1921; Rosengarten

1984). Contacts and trade between eastern and

western Mediterranean basin as long ago as the

second half of the XVI century B.C. are documented

through abundant archaeological findings dating back 

to the Mycenaean civilization (1500–1100 B.C.). The

colonization of the Italian peninsula by the Greeks( Magna Graecia) may have contributed to transfer of 

the cultivation techniques and cultivars of grape,

olive, and other species to the local populations and

eventually the Romans (Forni  1990).

The cpSSR results of our analysis show an

increase in chlorotype number and diversity from

West to East. In addition, a decrease in frequency of 

the most common chlorotype A from the groups

Spain and Italy to Turkey and Iran was observed

(Table 3). These results suggest little gene flow from

East to West, and that Spanish and Italian accessionshave a common genetic base. The nSSR analyses of 

Boccacci et al. (2006) and Gokirmak et al. (2008)

support this hypothesis, as both studies indicated a

close genetic relationship between Spanish and

Italian cultivars. On the contrary, Turkish cultivars

were assigned to a separate cluster, indicating little

gene flow between the eastern and western areas of 

the Mediterranean basin. Thus, both cpSSR and nSSR

data indicate that hazelnut cultivation and cultivars

were not introduced from the eastern Mediterranean

basin into Spain and southern Italy by Greeks or byArabs (Trotter 1921; Alberghina 2002). According to

Alfonso (1886) and other Sicilian authors, hazelnut

cultivation was introduced to Sicily by Arabs. Yet

Arabs only dominated the island after the second half 

of the IX century A.D., whereas hazelnut was already

cultivated in Campania (southern Italy) centuries

earlier by the Romans (Trotter  1921).

Evidence of the importance of hazelnut cultivation

during the Roman civilization was reported in several

Latin classics (Trotter   1921). In Cato’s (234–149

A.D.)   De re rustica   the hazelnut was mentioned asnux avellana, and the author recommended cultiva-

tion on farms of ‘‘nuces,   calvas,   avellanas,

 praenestinas et graecas’’. In Columella’s (I century

A.D.)   De re rustica   and Pliny’s (23–79 A.D.)

 Naturalis Historia   this species was reported in the

list of the plants that grown on farms. In the poetic

texts (Eclogues and  Georgics) of Virgil (70–19 B.C.),

hazelnut was mentioned in a mountainous context,

often in association with pastoral activities, as well as

Fig. 1   Chlorotype median network representing all chloro-

types (A, B, C, and D) identified in hazelnut. Circle areas are

proportional to chlorotype frequencies obtained in all 75

samples analysed

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in large-scale agricultural production. In Palladius’s

(IV–V century A.D.)   Opus agriculturae   a multipli-

cation technique of hazelnut was briefly described.

Hazelnut was also mentioned in the alimentary or

medicinal Latin literature. Dioskurides (I century

A.D.) recommended the use of kernels as a cure for

the common cold and persistent coughs (Rosengar-ten,   1984). Apicius (14–37 A.D.) in the manuscript

 De re coquinaria reported a recipe for the preparation

of a dessert similar to a nougat using hazelnuts and

honey (Trotter 1921). The geographical region which

most Latin authors mention in relation to the

cultivation of hazelnut was Campania. The   latifun-

dium, the entrepreneurial management of agricultural

estates using groups of slaves, started after the II

century B.C. in Latium and Campania regions, and

then became widespread under Roman rule (Sirago

1995). On the other hand, the Latin world adopted theterms   abellanae  and   abellinae   to indicate the hazel-

nut tree and its fruit, respectively, from which is

derived the specific epithet   avellana. These expres-

sions derived from  Abella  and  Abellinum, the actual

towns of Avella and Avellino in Campania, respec-

tively, where hazel was widely cultivated during

Roman times. Servius (III–IV century A.D.), in a

comment to the  Georgics   poem by Virgil, reported:

‘‘…sane coryli proprie dicuntur, nam abellanae ad 

 Abellano   ( Abellino   in other documents)   Campaniae

oppido, ubi abundant, nominatea sunt ’’ (Trotter1921). Other evidence of the importance of hazelnut

production in this region during the Roman period are

the archaeological remains and the pollen data. The

presence of carbonized hazelnuts and the representa-

tion of hazel trees and nuts in wall paintings found in

the excavations at Pompeii and Herculaneum (Trotter

1921; Jashemski  1970), indicate that hazelnuts were

consumed and commercialized also in areas rela-

tively distant from the traditional cultivation centers.

Pollen maps indicate that in 2500 B.P. the   Corylus

pollen percentage was low in southern Italy (0–5%),but it increased to 5–10% around 2000 B.P. and to

10–15% around 1500 B.P. (http://www.pierroton.

inra.fr/Cytofor/Maps/index.html), most likely in

relation to an increase in human activity.

The historical documents and archaeological find-

ings indicate that hazelnut cultivation and

consumption were significant during the Roman

period. This suggests that the use of hazelnut was

very likely spread throughout the empire by the

Romans. The presence of chlorotype A in all

analysed groups (Table 3) indicates a common origin

of the accessions; the intermediate relationship of 

chlorotype A to the others suggests that it could be an

ancestral chlorotype (Fig. 1). Thus, it can be hypoth-

esized that Italian hazelnut germplasm was spread by

the Romans to the Iberian peninsula (150–100 B.C.)and to a lesser extent to Asia Minor (133 B.C.) during

their expansion. Moreover, cpSSR and nSSR

(Boccacci et al.   2006)   results suggest that these

exchanges were stronger and frequent between Italy

and Spain. In the province of Tarragona (Catalonia,

northeastern Spain), where 88% of the Spanish

hazelnut area is located, Roman influence was strong.

The Romans improved existing cities, such as

Tarragona (Tarraco), and superimposed the   latifun-

dium  on the local landholding system. This supports

the hypothesis that the Romans may have introducedthe idea of systematically cultivating and using

hazelnut in their dominions. Human migrations and

trade between the central and western Mediterranean

basin were also frequent after the Roman civilization,

because southern Italy was dominated by the Spanish

from the XIV to the XVIII century. The prevalence of 

chlorotype A in Spanish and Italian accessions

suggests that southern Italy, most likely the Campania

region, was an important centre of origin and diffusion

of hazelnut cultivars. The archaeological findings,

historical document, and pollen data already discussedsupport this hypothesis. In addition, the absence

of chlorotypes B and C in the groups Spain and

Italy suggests that hazelnut varieties were domesti-

cated separately in Turkey and Iran, with perhaps

limited germplasm exchange with Italy and Spain.

In conclusion, our results show the usefulness of 

chloroplast markers to study relationships among

cultivars. The chlorotype distribution allowed us to

hypothesize a possible geographical origin and dif-

fusion of hazelnut cultivars. Moreover, the

integration of genetic data with historical, archaeo-logical, and palynological information indicates that

the history of  C. avellana   cultivation in Europe is

similar to that of   Castanea sativa   Mill. In fact, as

observed in chestnut by  Conedera et al. (2004),   our

results indicate a very limited influence of the Greek 

hazelnut cultivation techniques on the Latin and

subsequent Roman civilization, unlike the situation in

grape and olive. Thus, it is probable that systematic

cultivation and use of hazelnut was spread by the

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Romans, probably from the Campania region. We

investigated few cultivars from Turkey and Iran, but

found considerable chlorotype diversity. Our data

point toward separate domestications in these

regions, and limited germplasm exchange with the

western to eastern Mediterranean basin. Further

analyses of wild populations and cultivars from othergeographical areas, using chloroplast and nuclear

markers, would improve our understanding of the

origin and spread of hazelnut cultivars.

Acknowledgements   Authors are grateful to: Dr. Nahla V.

Bassil (USDA-ARS National Clonal Germplasm Repository of 

Corvallis, Ore.), Dr. Roberto De Salvador and Dr. Pasquale

Piccirillo (Consiglio per la Ricerca e sperimentazione in

Agricoltura, Italy), Dr. Silvia Dellepiane (Cooperativa San

Colombano, Genova, Italy), Dr. Alireza Ghanbari (Department

of Horticultural Science, Shahed University, Iran), Dr. Merce

Rovira (Istitut de Recerca i Tecnologia Agroalimentaries,

Tarragona, Spain), and Dr. Virginia Ughini (Istituto di Frutti-

viticoltura, Universita   Cattolica S.C., Piacenza, Italy) for

providing samples of leaves or catkins. Authors thank prof.

Shawn A. Mehlenbacher (Oregon State University, Corvallis,

Ore.) for reviewing the manuscript. The research was funded

by Regione Piemonte Administration (CIPE, 2003) and MIUR

(ex 60%, 2006).

References

Alberghina O (2002) La coltura del nocciolo in Sicilia. Atti II

Convegno Nazionale sul Nocciolo, Giffoni Valle Piana(Salerno), Italy, pp 141–145

Alfonso F (1886) Monografia sul nocciolo. Tipografia dello

Statuto, Palermo, Italy

Arobba D, Caramiello R, Del Lucchese A (2003) Archaeobo-

tanical investigations in Liguria: preliminary data on the

early Iron Age at Monte Trabocchetto (Pietra Ligure,

Italy). Veg Hist Archaeobot 12:253–262. doi:10.1007/ 

s00334-003-0022-7

Arroyo-Garcıa R, Lefort F, de Andres MT, Ibanez J, Borrego J,

Jouve N, Cabello F, Martınez-Zapater JM (2002) Chlo-

roplast microsatellite polymorphism in   Vitis   species.

Genome 45:1142–1149. doi:10.1139/g02-087

Arroyo-Garcıa R, Ruiz-Garcıa L, Bolling L, Ocete R, Lopez

MA, Arnold C, Ergul A, Soylemezoglu G, Uzun HI, Ca-

bello F, Ibanez J, Aradhya MK, Atanassov A, Atanassov I,

Balint S, Cenis JL, Costantini L, Gorislavets S, Grando

MS, Klein BY, McGovern PE, Merdinoglu D, Pejic I,

Pelsy F, Primikirios N, Risovannaya V, Roubelakis-An-

gelakis KA, Snoussi H, Sotiri P, Tamhankar S, This P,

Troshin L, Malpica JM, Lefort F, Martınez-Zapater JM

(2006) Multiple origins of cultivated grapevine (Vitis

vinifera   L. ssp.  sativa) based on chloroplast DNA poly-

morphisms. Mol Ecol 15:3707–3714. doi:10.1111/j.1365-

294X.2006.03049.x

Bakels CC (1991) Western continental Europe. In: van Zeist

W, Wasylikowa K, Behre KE (eds) Progress in old world

palaeoethnobotany. Balkema, Rotterdam, pp 279–298

Bandelt HJ, Foster P, Rohl A (1999) Median-joining networks

for inferring intraspecific phylogenies. Mol Biol Evol

16:37–48

Bassil NV, Botta R, Mehlenbacher SA (2005a) Microsatellite

markers in the hazelnut: isolation, characterization and

cross-species amplification in  Corylus. J Am Soc Hortic

Sci 130:543–549

Bassil NV, Botta R, Mehlenbacher SA (2005b) Additional

microsatellites of the European hazelnut. Acta Hortic

686:105–110

Bennett KD, Tzedakis PC, Willis KJ (1991) Quaternary refugia

of north European trees. J Biogeogr 18:103–115. doi:

10.2307/2845248

Birks HJB (1989) Holocene maps and patterns of tree-

spreading in the British Isles. J Biogeogr 16:503–540. doi:

10.2307/2845208

Boccacci P, Akkak A, Bassil NV, Mehlenbacher SA, Botta R

(2005) Characterization and evaluation of microsatellite

loci in European hazelnut (Corylus avellana L.) and theirtransferability to other  Corylus   species. Mol Ecol Notes

5:934–937. doi:10.1111/j.1471-8286.2005.01121.x

Boccacci P, Akkak A, Botta R (2006) DNA-typing and genetic

relationships among European hazelnut (Corylus avellana

L.) cultivars using microsatellite markers. Genome

49:598–611. doi:10.1139/G06-017

Boccacci P, Botta R, Rovira M (2008) Genetic diversity of 

hazelnut (Corylus avellana L.) germplasm in northeastern

Spain. HortScience 43:667–672

Botta R, Akkak A, Boccacci P (2005) DNA-typing of hazelnut:

a universal methodology for describing cultivars and

evaluating genetic relatedness. Acta Hortic 686:117–124

Breton C, Tersac M, Berville A (2006) Genetic diversity and

gene flow between the wild olive (oleaster,   Olea euro- paea   L.) and the olive: several Plio-Pleistocene refuge

zones in the Mediterranean basin suggested by simple

sequence repeats analysis. J Biogeogr 33:1916–1928. doi:

10.1111/j.1365-2699.2006.01544.x

Cleary RM (2003) Enclosed late Bronze Age habitation site

and boundary wall at Lough Gur, co. Limerick. P R Ir

Acad 103:97–189

Conedera M, Krebs P, Tinner W, Pradella M, Torriani D

(2004) The cultivation of  Castanea sativa  (Mill.) in Eur-

ope, from its origin to its diffusion on a continental scale.

Veg Hist Archaeobot 13:161–179. doi:10.1007/s00334-

004-0038-7

Dumolin S, Demesure B, Petit RJ (1995) Inheritance of chlo-

roplast and mitochondrial genomes in pedunculate oak 

investigated with an efficient PCR method. Theor Appl

Genet 91:1253–1256. doi:10.1007/BF00220937

FAOstat (2008) Agriculture data. Available from:   http:// 

faostat.fao.org/site/408/default.aspx. Accessed 03 April

2008

Forni G (1990) Gli albori dell’agricoltura. REDA, Roma

Ghanbari A, Akkak A, Boccacci P, Talaie A, Vezvaie A, Botta

R (2005) Characterization of hazelnut (Corylus avellana

L.) cultivars using microsatellite markers. Acta Hortic

686:111–115

858 Genet Resour Crop Evol (2009) 56:851–859

 1 3

7/23/2019 Boccacci_09_GRCE_vol56 (1)

http://slidepdf.com/reader/full/boccacci09grcevol56-1 10/10

Gokirmak T, Mehlenbacher SA, Bassil NV (2008) Character-

ization of European hazelnut (Corylus avellana   L.)

cultivars using SSR markers. Genet Resour Crop Evol,

doi: 10.1007/s10722-008-9352-8

Huntley B, Birks HJB (1983) An atlas of past and present

pollen maps for Europe: 0–13,000 years ago. Cambridge

University Press, Cambridge

Imazio S, Labra M, Grassi F, Scienza A, Failla O (2005)

Chloroplast microsatellites to investigate the origin of 

grapevine. Genet Resour Crop Evol 53:1003–1011. doi:

10.1007/s10722-004-6896-0

Jashemski WF (1970) University of Maryland Excavations at

Pompeii, 1968. Am J Archaeol 74:63–70. doi:10.2307/ 

502906

Kasapligil B (1972) A bibliography on  Corylus   (Betulaceae)

with annotations. Annu Rep North Nut Growers Assoc

63:107–162

Kubiak-Martens L (1999) The plant food component of the diet

at the late Mesolithic (Ertebolle) settlement at Tybrind

Vig, Denmark. Veg Hist Archaeobot 8:117–127. doi:

10.1007/BF02042850

Kuster H (2000) The history and culture of food and drink inEurope: northern Europe–Germany and surrounding

regions. In: Kiple KF, Ornelas KC (eds) The Cambridge

world history of food, vol 2. Cambridge University Press,

Cambridge, pp 1226–1232

Nei M (1987) Molecular evolutionary genetics. Columbia

University Press, New York 

Palme AE, Vendramin GG (2002) Chloroplast DNA variation,

postglacial recolonization and hybridization in hazel,

Corylus avellana. Mol Ecol 11:1769–1779. doi:

10.1046/j.1365-294X.2002.01581.x

Pena-Chocarro L, Pena LZ, Gazolaz JG, Morales MG, Sesma JS,

Straus LG (2005) The spread of agriculture in northern

Iberia: new archaeobotanical data from El Miron cave

(Cantabria) and the open-air site of Los Cascajos (Navarra).Veg Hist Archaeobot 14:268–278. doi:10.1007/s00334-

005-0078-7

Provan J, Powell W, Hollingsworth PH (2001) Chloroplast

microsatellites: new tools for studies in plant ecology and

evolution. Trends Ecol Evol 16:142–147. doi:10.1016/ 

S0169-5347(00)02097-8

Rosengarten F (1984) The book of edible nuts. Walker and

Company, New York 

Russel-White CJ (1995) The excavation of a Neolithic and Iron

Age settlement at Wardend of Durris, Aberdeenshire. Proc

Soc Antiq Scot 125:9–27

Schmidl A, Jacomet S, Oeggl K (2007) Distribution patterns of 

cultivated plants in the Eastern Alps (Central Europe)

during Iron Age. J Archaeol Sci 34:243–254. doi:10.1016/ 

 j.jas.2006.05.001

Sebastiani SF, Carnevale S, Vendramin GG (2004) A new set

of mono- and dinucleotide chloroplast microsatellites in

Fagaceae. Mol Ecol Notes 4:259–261. doi:10.1111/ 

 j.1471-8286.2004.00635.x

Sirago VA (1995) Storia agraria romana. I: Fase ascensionale.

Liguori, Napoli, Italy

Sostaric R, Dizdar M, Kusan D, Hrsak V, Marekovic S (2006)

Comparative analysis of plant finds from early Roman

graves in Ilok (Cuccium) and Scitarjevo (Andautonia),

Croatia—a contribution to understanding burial rites in

southern Pannonia. Coll Antropol 30:429–436

Tallantire PA (2002) The early-Holocene spread of hazel

(Corylus avellana   L.) in Europe north and west of the

Alps: an ecological hypothesis. Holocene 12:81–96. doi:

10.1191/0959683602hl523rr

Tasias Valls J (1975) El avellano en la provincia de Tarragona.

EXCMA Diputacion Provincial de Tarragona, Tarragona,

SpainThomas MR, Matsumoto S, Cain P, Scott NS (1993) Repetitive

DNA of grapevine: classes present and sequences suitable

for cultivar identification. Theor Appl Genet 86:173–180

Thompson MM, Lagerstedt HB, Mehlenbacher SA (1996)

Hazelnuts. In: Janick J, Moore JN (eds) Fruit breeding:

nuts, vol 3. Wiley, New York, pp 125–184

Trotter A (1921) Contributo alla storia colturale del nocciuolo

nella Campania. Ristampa di una comunicazione fatta al

Congresso di Arboricoltura Meridionale, Napoli (Italy),

16–20 September, pp 3–19

Vaughan JG, Geissler C (1997) The new Oxford book of food

plants. Oxford University Press, New York 

Vendramin GG, Ziegenhagen B (1997) Characterization and

inheritance of polymorphic plastid microsatellites in Abies. Genome 40:857–864. doi:10.1139/g97-811

Weising K, Gardner R (1999) A set of conserved PCR primers

for the analysis of simple sequence repeat polymorphisms

in chloroplast genomes of dicotyledonous angiosperms.

Genome 42:9–19. doi:10.1139/gen-42-1-9

White KD (1970) Roman farming. Cornell University Press,

Ithaca

Zohary D, Hopf M (2001) Domestication of plants in the Old

World: the origin and spread of cultivated plants in West

Asia, Europe, and the Nile valley. Oxford University

Press, New York 

Genet Resour Crop Evol (2009) 56:851–859 859

 1 3