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Investigating the origin of hazelnut (Corylus avellana L.) cultivars using

chloroplast microsatellites

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DOI: 10.1007/s10722-009-9406-6

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Genet Resour Crop Evol (2009) 56:851–859
DOI 10.1007/s10722-009-9406-6
RESEARCH ARTICLE
Investigating the origin of hazelnut (Corylus avellana L.)
cultivars using chloroplast microsatellites
Paolo Boccacci Æ 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 and
diversity 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.
P. Boccacci (&)
R. Botta
Dipartimento di Colture Arboree, Università degli Studi di
Torino, Via Leonardo da Vinci 44, 10095 Grugliasco,
Torino, Italy
e-mail: paolo.boccacci@unito.it
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 northern
Turkey (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
123
852
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 area
around 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 (Palmé and Vendramin
2002) indicates a rapid expansion from one large
refugium in the Biscay area or from several scattered
refugia 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 shell
remains from many Neolithic, Bronze Age, Classical,
and Medieval sites all over Europe (Bakels 1991;
Russel-White 1995; Arobba et al. 2003; Cleary 2003;
Peña-Chocarro et al. 2005; Šoštarić 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).
123
Genet Resour Crop Evol (2009) 56:851–859
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; Gökirmak 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;
Gökirmak et al. 2008). In particular, Boccacci et al.
(2006) and Gökirmak 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 flow
between 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-Garcı́a 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 have
been used to investigate the origin and diffusion of
several fruit tree species, such as grape (Arroyo-
Garcı́a et al. 2002; Imazio et al. 2005; Arroyo-Garcı́a
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-
Genet Resour Crop Evol (2009) 56:851–859
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-
mentàries (IRTA) of Constantı́ (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 in
different 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 modified
procedure described by Thomas et al. (1993) in 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
853
[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; and
a 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=ðn  1Þ 1  Rg2i , 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 (Palmé 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
123
854 Genet Resour Crop Evol (2009) 56:851–859

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
Culplà DCA A Ratolı̀ 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 garrofı̀ IRTA A Vimbodı̀ 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 D
Iannusa 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 B
Group 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

123
Genet Resour Crop Evol (2009) 56:851–859 855

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 Agroalimentàries 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
Group Spain Group Italy Group Turkey Group Iran Total
distribution in each
geographical group (with n 33 22 10 10 75
chlorotype frequency in
parentheses) 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)
n = number of samples; Chlorotype D – 2 (0.091) – – 2 (0.027)
N = number of N 1 2 2 3 4
chlorotypes; H 0.000 0.173 0.533 0.378 0.391
H = chlorotype diversity

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 the
four 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 from
West 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

123
856
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
chloroplast DNA variation in wild hazelnuts (Palmé
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
123
Genet Resour Crop Evol (2009) 56:851–859
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 accessions
have a common genetic base. The nSSR analyses of
Boccacci et al. (2006) and Gökirmak 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 by
Arabs (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 as
nux 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
Genet Resour Crop Evol (2009) 56:851–859
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 the
terms 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’’ (Trotter
1921). 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
857
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 introduced
the 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 discussed
support 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
123
858
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 other
geographical 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. Mercé
Rovira (Istitut de Recerca i Tecnologia Agroalimentàries,
Tarragona, Spain), and Dr. Virginia Ughini (Istituto di Frutti-
viticoltura, Università 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).
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