Vitis 56, 173–180 (2017)
© The author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Share-Alike License
Correspondence to: Dr. M. Crespan, CREA - Centro di ricerca per la viticoltura e l’enologia, Viale 28 Aprile, 26, 31015 Conegliano
(TV), Italy. Fax: +39 0438 738489. E-mail: email@example.com
Identication and characterization of Romanian grapevine genetic resources
C. F. PopesCu1), E. Maul2), L. C. Dejeu3), D. Dinu4), R. N. GheorGe1), V. lauCou5),
T. laCoMbe5), D. MiGliaro6) and M. Crespan6)
1) National Research and Development Institute for Biotechnology in Horticulture Ştefāneşti-Argeş, România
2) JKI - Julius Kühn-Institut, Institut für Rebenzüchtung Geilweilerhof, Siebeldingen, Germany
3) Faculty of Horticulture, University of Agronomical Science and Veterinary Medicine, Bucharest, România
4) Research and Development Station for Viticulture and Oenology Drāgāşani-Vâlcea, România
5) INRA, UMR 1334 AGAP, Equipe Diversité, Adaptation et Amélioration de la Vigne, Montpellier, France
6) Council for Agricultural Research and Economics - Centre of Viticulture and Enology Research, Conegliano (TV), Italy
The research focused on old Romanian varieties of
local distribution, with very limited growing area and
also on those nowadays neglected, to identify, charac-
terize and compare these cultivars with one another and
on a larger geographic scale. Ampelograc characteriza-
tion with 48 descriptors, photographs of shoot tip, leaf
and bunch and genotyping using 13 SSR markers were
applied together with literature references to conrm
trueness to type. The sixty one accessions belonged to fty
one varieties. New synonymies were detected (e.g. 'Ne-
gru românesc' = 'Bātutā neagrā'; 'Cârcioasā' = 'Balint
weiss'; 'Galbenā māruntā' = 'Kakotrygis'), assignments
were modied (e.g. 'Galbenā uriaşā' = 'Mirkovaca' and
≠ 'Galbenā de Odobeşti' as previously assumed), mis-
nomers identied and unique genotypes were detected
which had never been described before. Molecular data
supported previous pedigree relationships and allowed
new parent-offspring hypotheses to be formulated. The
results were shown to be useful for updating the infor-
mation on old Romanian grapevine germplasm.
K e y w o r d s : SSR; genotyping; autochthonous cultivars;
There is a long history of grapevine cultivation and
wine production on Romanian territories, documented from
the 7th century BC onwards (ConstantinesCu et al. 1970). It
is today one of the most important fruit crops with a vine-
yard area of about 192,000 ha, 95 % for wine and 5 % for
table grapes (Romanian National Ofce of Vine and Wine
Products - NOVWP, 2016, http://www.onvpv.ro/). The rst
research papers with scientic descriptions of grapevine
varieties were published in Romania during the 19th century
(IonesCu 1868, niColeanu 1900) and the rst ampelographic
collections were established at Pietroasa (1895), Bucharest
(1925), Valea Cālugāreascā (1925), Huşi (1925), Drāgāşani
and Miniş (1939). Other small collections were established
in six research stations and four university centres aiming
to maintain the highest number of grapevine varieties as
possible under secure conditions. Today these collections
hold around 1,381 wine and table grape accessions (17.8 %
considered as Romanian varieties and clones), 42 rootstock
varieties and 85 interspecic varieties for fruit production.
Over time, these collections have been the source of refer-
ence plant material for thorough comparative studies and
also for most research activities involving ampelographic
characterization, phenology and evaluation of yield and oe-
nological potential of grapevine varieties and clones. Proof
of these intensive studies are the seven volumes of "Ampe-
lograa Republicii Populare Romne" published between
1959 and 1970 (ConstantinesCu et al. 1959, 1960, 1961,
1962, 1965, 1966, 1970), which are still an international
reference in the eld of ampelography. Description and
evaluation of varieties considered of economic importance
for wine production were of primary interest (InDreas and
Visan 2001, Rotaru 2009). Less effort was made for old
Romanian varieties of local distribution and for those now-
adays neglected or with very limited growing areas.
In the last years, the ampelographic descriptions have
been enriched with molecular characterisations, especially
for cultivars of scientic and economic value, aiming to
accurately identify the autochthonous varieties (GheorGhe
et al. 2008, boDea et al. 2009, butiuC-Keul et al. 2010,
Coste et al. 2010), or to facilitate the registration of Ro-
manian cultivars in the European Vitis Database (Gheţea
et al. 2010 and 2012).
The aim of this study was to obtain a molecular and
ampelographic characterization of 61 accessions, most of
them presumed autochthonous Romanian varieties, selected
on the basis of the following criteria: a) varieties grown since
ancient times according to old documents and turned into
international cultivars due to their biological competence; b)
major local cultivars, of local importance, extensively grown
in Romania; c) minor cultivars, of local importance, grown
especially in private vineyards; d) neglected local cultivars,
at risk of extinction and retained in germplasm collections.
They encompassed mainly varieties recommended for wine
174 C. F. popesCu et al.
production and some table and wine/table cultivars. The re-
search was developed within the framework of COST Action
FA1003 "East-West collaboration for grapevine diversity
exploration and mobilization of adaptive traits for breeding"
(Failla 2015), with the aim of identifying, characterizing
and comparing the selected Romanian varieties with one
another and on a larger geographic scale. Knowledge of
grapevine varieties provenance is useful to better understand
their movement through different countries over time, even
when the memory of those transfers had been lost, together
with the original name of the variety. As a consequence,
the information available on specic varieties may become
broader and can be retained or rejected, when aware of the
true identity of the cultivars under study.
Genotyping was applied to identify possible synonyms,
homonyms, misnomers and unique genotypes, using a set
of 13 SSR markers encompassing the nine SSR markers
recommended for common use by the European project
GrapeGen06 (Maul et al. 2012). The SSR proles also
allowed comparison of the Romanian grapevine germplasm
with literature data and molecular databases, searching for
synonyms with respect to neighbouring countries. Prelim-
inary indications obtained from molecular data were then
compared with available morphological information.
Material and Methods
P l a n t m a t e r i a l : Sixty-one accessions (Tab. 1)
were characterized, belonging to three germplasm collec-
tions: Faculty of Horticulture - University of Agronomical
Science and Veterinary Medicine, Bucharest (UASVM);
Research and Development Station for Viticulture and
Oenology Drāgāşani-Vâlcea (RDSVO); National Research
and Development Institute for Biotechnology in Horticulture
Ampelographic description: The ampelo-
graphic description was carried out for two or three consec-
utive years, in accordance with the 2nd edition of the "OIV
Descriptor list for grapevine varieties and Vitis species" (OIV
2009) with 48 descriptors and following the standardized
methodology reported in Rustioni et al. (2014). The mor-
phological characteristics recorded in the three germplasm
collections referred to the following aspects: 7 for young
shoot (OIV 001, 003, 004, 006, 007, 008 and 016), 17 for
young and mature leaf (OIV 051, 053, 067, 068, 070, 072,
074, 075, 076, 079, 080, 081-1, 081-2, 083-2, 084, 087 and
094), 15 for type of ower, bunch and berry aspects (OIV
151, 155, 202, 204, 206, 208, 209, 220, 221, 223, 225, 231,
235, 236 and 241) and 9 for phenology, growth, quality and
quantity of grape yield (OIV 301, 303, 351, 502, 503, 504,
505, 506 and 508).
Genotyping and identification: Genomic
DNA was isolated from 100 mg of young leaf using Qia-
gen DNeasy Plant mini-kit (Qiagen, Hilden, Germany),
following the manufacturer's protocol. DNA concentration
and quality were checked by spectrophotometric analysis
and electrophoresis in 1 % agarose gel. Thirteen SSR
markers were used for genotyping: the nine proposed as
common grape markers for international use within the
framework of the Grapegen06 European project (VVS2,
VVMD5, VVMD7, VVMD25, VVMD27, VVMD28,
VVMD32, VrZAG62, VrZAG79) (Maul et al. 2012), plus
ISV2 (VMC6e1), ISV3 (VMC6f1), ISV4 (VMC6g1) and
VMCNG4b9 (MiGliaro et al. 2013). The SSR analyses
were performed following the protocol detailed in MiGliaro
et al. (2013), using uorescent primers and an ABI3130xl
genetic analyzer (Applied Biosystems, Foster City, CA),
with some minor modications for 11 SSR markers. The
remaining two (VVMD25 and VVMD32) were analyzed
separately, as single markers. SSR allele calling was per-
formed using ABI Prism GeneMapper software version 3.0,
Accession list, general information and SSR matches
Accession no. DNA no. Accession name
Matches by SSR
ROM045-003 180.12.R Alb românesc (misnomer) H green W1961 360 2, 3 Sarba 10738 Sarba
ROM051-237 178.12.R Ardeleancā H green W1959 360 2, 3 Bakator belyi 904 Bakator belyi
ROM06-0011 163.12.R Bacator FFr rose W1959 360 1, 3 Bakator roz 905 Bakator roz
ROM051-238 161.12.R Bābeascā neagrā H blue black W1959 320 1, 2, 3 Babeasca neagra 843 Babeasca negra
ROM051-239 203.12.R Bāșicatā H green W1961 360 2, 3 Basicata 1022 BASICATA
ROM045-025 206.12.R Bātutā neagrā Hblue black W/T 1959 360 2 Batuta neagra 1042 Batuta neagra
ROM045-172 208.12.R Negru românesc black W1966 360 2, 3
ROM051-240 212.12.R Berbecel H green W1959 360 2, 3 Berbecel 1148 Berbecel
ROM045-037 181.12.R Braghinā albā* FFr green W- 360 2, 3 1645 Braghina alba
ROM045-036 182.12.R Braghinā roz FFr rose W1959 360 2 Braghina rosie 1644 Braghina rosie
ROM06-0024 157.12.R Busuioacā de Bohotin H red W1960 320 1, 3
Muscat à petits
8248 Muscat à petits
ROM06-0026 160.12.R Cadarcā H black W1959 320 1, 3 Kadarka kek 5898 Kadarka kek
ROM045-048 184.12.R Cârcioasā FFr green W1965 360 2, 3 Balint weiss 935 Balint weiss
Identication and characterization of Romanian grapevine genetic resources 175
Tab. 1, continued
ROM051-241 185.12.R Crâmpoşie FFe green W/T 1959 330 2, 3 Crimposie 3237 Crimposie
ROM045-051 201.12.R Ceauş alb FFr green T 1959 360 2, 3 Chaouch blanc 10196 Chaouch blanc
ROM051-242 202.12.R Ceauş roz FFe rose T 1959 360 2, 3 Chaouch rozovyi 2507 Chaouch rozovyi
ROM051-243 183.12.R Cioinic FFe green W/T 1961 360 2, 3 Cioinic 2674 Cionic
ROM051-246 177.12.R Coarnā albā FFr green W/T 1959 360 1, 3 Coarna alba 2724 Coarna alba
ROM051-248 204.12.R Coarnā roșie FFr red T 1961 360 2, 3 Coarna rosie 2728 Coarna rosie
ROM051-247 176.12.R Coarnā neagrā FFr black-red T 1959 320 1, 2, 3 Coarna neagra 2726 Coarna neagra
ROM051-244 213.12.R Coada oilor/Ovis H green W1962 360 2, 3 Juhfark 5852 Juhfark
ROM06-0046 169.12.R Creațā H green W/T 1959 360 1, 3 Kreaca 6501 Kreaca
ROM051-249 200.12.R Creațā de Banat 1959 360 2, 3
ROM051-250 187.12.R Cruciuliţā H green W1961 360 2, 3 Cruciulita 3267 Cruciulita
ROM051-251 162.12.R Feteascā albā H green W1959 310 1, 2, 3 Feteasca alba 4119 Feteasca alba
ROM051-252 158.12.R Feteascā neagrā H black W1959 310 1, 2, 3 Feteasca neagra 4120 Feteasca neagra
ROM051-253 165.12.R Feteascā regalā H green W1959 310 1, 2, 3 Feteasca regala 4121 Feteasca regala
ROM051-25458 172.12.R Frâncușā H green W1959 320 1, 2, 3 Francuse 4221 Francuse
175.12.R Galbenā de Odobești
H green W
1959 320 1, 2, 3
Odobesti 12727 Galbena de
758.16 Galbenā de Odobești
756.16 Galbenā de Odobești
174.12.R Zghiharā de Huși
1960 320 1, 2, 3759.16 Zghiharā de Huși
ROM051-274 757.16 Zghiharā de Huși
ROM045-100 186.12.R Galbenā māruntā H green W- 360 2 Kakotrygis 5920 Kakotrygis
ROM051-256 188.12.R Galbenā uriaşā H green W1961 360 2, 3 Galbena uriasa 4322 Galbena uriasa
ROM045-106 189.12.R Gordan
1961 360 2, 3
Iordan 5544 IordanROM06-0069 168.12.R Iordanā green 1960 320 1, 3
ROM045-253 198.12.R Zemoasā green 1962 360 2, 3
ROM051-258 190.12.R Gordin H green W1959 360 2, 3 Gordin 4901 Gordin
ROM06-0065 166.12.R Grasā de Cotnari H green W1959 310 1, 3 Grasa de Cotnari 4948 Grasa de Cotnari
191.12.R Lampāu (misnomer) H green W- 360 2 Tompa Mihaly 12564 Tompa Mihali
ROM051-259 170.12.R Majarcā albā H green-rose W1960 360 1, 3 Slankamenka
bela 11866 Slankamenka
ROM045-146 207.12.R Moroştinā* H green W- 360 2, 3 Morostina 8007 Morostina
ROM051-260 167.12.R Mustoasā de Māderat
(misnomer) H green W1960 360 2, 3 42198
ROM051-261 192.12.R Negru mare
(questionable)* FFe black W/T 1962 360 2, 3 42199 Negru mare
ROM051-262 193.12.R Negru moale H black W1960 360 2, 3 Negru moale 8464 Negru moale
ROM051-263 194.12.R Negru vârtos H black W1960 360 2, 3 Mavrud
Varnenskii 7540 Mavrud
159.12.R Negru vârtos
(questionable) H black W- 360 1 42197 Negru vartos
ROM045-179 195.12.R Om rāu* H green W1962 360 2, 3 8765 Om rau
ROM051-265 215.12.R Pârciu H green W1962 360 2, 3 Pirciu 9300 Pirciu
ROM06-0103 173.12.R Plāvaie Hgreen W1960 330 1, 3 Plavay 9553 Plavay
179.12.R Alb rotund green W- 360 2
ROM045-206 196.12.R Românie* H green W- 360 2, 3 Romanie§ 10177 Romanie
ROM06-0134 164.12.R Tāmâioasā româneascā H green W1960 310 1, 2, 3 Tamaiosa
rominesca (faux) 25546 Tamaioasa
ROM045-234 214.12.R Tâţa caprei albā H green T 1962 360 2 Tsitsa kaprei 16449 Tsitsa kaprei
ROM051-268 210.12.R Tâţa caprei neagrā H black T - 360 2, 3 Hora 5423 Hora
ROM051-269 211.12.R Tâţa vacii albā FFe green T 1962 360 2, 3 Halholyag 6419 Halholyag
ROM045-237 209.12.R Tâţa vacii neagrā* H black T - 360 2 Kozi Cici
cherveni 25547 Kozi Cici
ROM045-238 199.12.R Teişor (misnomer) H green W1959 360 2, 3 Ezerjo 4027 Ezerjo
ROM051-272 197.12.R Vulpea H black W/T 1962 360 2, 3 Vulpea 13186 Vulpea
(1) H = hermaphrodite; FFr = female functionally with reexed stamens; FFe = female funcionally with erect stamens.
(2) W = wine, T = table.
(3) 310 = local cultivar, spread all over, international cultivar; 320 = major local cultivar, of local importance, but extensively
grown; 330 = minor local cultivar, of local importance, fairly utilized; 360 = local neglected cultivar, at risk of extinction.
(4) 1 = UASVM, 2 = RDSVO, 3 = NRDIBH; *genotype not present in Vassal.
§ in agreement with Zulj et al. 2013.
176 C. F. popesCu et al.
with a home-made bin set produced with reference varieties.
Preliminary indications about cultivar identity were obtained
by comparison of the genetic proles with literature data,
VIVC (Vitis International Variety Catalogue, http://www.
vivc.de/), CREA-Viticulture and Enology molecular data-
base, partially published in the Italian Grapevine Catalogue,
http://catalogoviti.politicheagricole.it/ and in the Italian Vitis
Database, http://www.vitisdb.it/, and with the INRA molec-
ular database of the Vassal collection (LauCou et al. 2011).
S t a t i s t i c s : Cervus software (KalinowsKi et al.
2007) version 3.0 (http://cervus.software.informer.com/3.0/)
was used for preliminary indications on possible par-
ents-progeny trios and rst degree relationships.
Results and Discussion
The 61 accessions analysed in the study belonged to
51 different genotypes. This was conrmed by both micro-
satellite markers and ampelography. Precise morphological
descriptions of the Romanian grape germplasm, literature
references, ancient records in grapevine collections and
the availability of SSR-marker databases were of great
Ampelographic description: All recorded
data were compared with the detailed ampelographic de-
scriptions from old documents to obtain a preliminary con-
rmation of the authenticity of the studied accessions, with
the exception of nine varieties: 'Galbenā māruntā', 'Lampāu',
'Moroştinā', 'Negru vârtos' (questionable), 'Alb rotund', 'Ţâţa
caprei neagrā' and 'Ţâţa vacii neagrā', for which no informa-
tion was found, and 'Braghinā albā' and 'Românie', having
only partial information (brezeanu 1912, Gorjan and botu
2013). The ampelographic descriptions, available in the
European Vitis database (http://www.eu-vitis.de), revealed
great differences among the studied varieties, except for
a few traits, such as number of consecutive tendrils (OIV
016), specic for V. vinifera, intensity of esh anthocyanin
coloration in the berry (OIV 231) and formation of seeds
(OIV 241). The morphological and agronomic characters of
the studied accessions mostly conrmed previous ampelo-
graphic documents. Few differences, at only one level of the
notes corresponding to a certain characteristic were found,
such as intensity of anthocyanin coloration on prostrate
hairs of the shoot tip (OIV 003), density of prostrate hairs
on the shoot tip (OIV 004), colour of upper side of blade
(4th leaf) (OIV 051), shape of blade of the mature leaf (OIV
067), degree of opening/overlapping of petiole sinus (OIV
079), shape of base of petiole sinus (OIV 080), density of
prostrate hairs between main veins on lower side of blade
(OIV 084), bunch density (OIV 204), bunch shape (OIV
208), berry shape (OIV 223), sugar content of must (OIV
505) and total acidity of must (OIV 506). These differenc-
es could be ascribed to the range of expected variability
of morphological and agronomic characteristics linked to
environment, cultural conditions, health status of vines, and
might also depend on interpretation by ampelographers.
Table and table/wine accessions, in comparison with
varieties commonly used for wine production, were char-
acterized by larger and heavier bunches, bigger berries and
juice with lower acidity and sugar content.
Among the 51 genotypes found in this study, thirteen
were characterized by functionally female owers. The
percentage (25.5 %) observed in this study was a relatively
high proportion taking into account the average number
observed world-wide (i.e. about 8 %, Boursiquot et al.
1995) and highlighted the originality of the Romanian gene
pool. This trait is considered as ancestral and related to
Vitis vinifera domestication, however it could be inherited
through segregation by sexual reproduction, given that
cultivated grapevines frequently carry a female allele (Hf),
while homozygous hermaphroditic vines (HH) are rare
(FeChter et al. 2012). In addition, this peculiar character
in grapevine has aroused the interest of breeders, because
it simplies cross hybridizations in breeding programmes
(Chaϊb et al. 2010). Morphologically, only eight of them
produce owers with well-formed ovaries, stigma, style and
anthers with shorter laments, reexed outwards from the
ovary: 'Bacator', 'Braghinā roz', 'Braghinā albā', 'Cârcioasā',
'Ceauş alb', 'Coarnā albā', 'Coarnā roşie', 'Coarnā neagrā'.
The other ve develop hermaphrodite owers with anthers
inclined outwards: 'Crâmpoşie', 'Ceauş roz', 'Cioinic', 'Ne-
gru mare' and 'Ţâţa vacii albā' (Tab. 1). These owers, with
stamens sloping outwards from the ovary, are considered
apparently normal, but functionally female (ConstantinesCu
1958). All these varieties have common characteristics: a)
the ratio between length of stamens and length of pistil is
≤ 1; b) the pollen is sterile, abundant and sometimes with
acorn shape ('Braghinā roz', 'Ceauş roz', 'Ceauş alb', 'Coarnā
albā'); c) given pollen sterility, yield is variable, depending
on weather conditions during anthesis; moreover, they re-
quire male or hermaphroditic vines nearby, having the same
owering period to enable pollination. The inconstancy of
grape yield was the main reason for the declining interest in
these varieties. Nowadays only 'Coarnā neagrā' is grown on
large areas (over 100 ha) being remarkable for the special
appearance and taste of grapes, and also 'Crâmpoşie' for its
very good qualities as both table and wine grapes.
Many of these autochthonous varieties, expressing
high sugar content and equilibrate acidity of the must, have
aroused breeders' interest. Elite clones with special quali-
ties were selected and approved for wine production, like
in 'Crâmpoşie', 'Tāmâioasā româneascā', 'Feteascā albā',
'Feteascā neagrā', 'Fetească regală', 'Braghină roz', 'Bragh-
ină albă', 'Gordan', 'Berbecel', 'Băşicată', 'Negru moale' and
'Negru vârtos', or for table grapes with pleasant appearance
and avour, like in 'Coarnă albă', 'Coarnă neagră', 'Coarnă
roşie' and 'Ţâţa caprei albă'.
Genotyping and identification: Fifty-one
distinct SSR proles were found (see Table, suppl. data).
Interestingly, 'Iordanā' accession showed a slightly differ-
ent SSR prole from the other two synonyms 'Gordan' and
'Zemoasā' at VVS2 locus, being heterozygous (133-143),
instead of homozygous (133-133); 'Plāvaie' was triallelic at
VVMD32 locus (253, 265 and 273). A preliminary indication
on genotype identity was rst obtained by comparison of
each SSR prole with literature data, VIVC molecular data-
base, CREA-Viticulture and Enology and INRA molecular
Identication and characterization of Romanian grapevine genetic resources 177
databases. On the basis of genotyping results, ampelographic
comparisons were made with available descriptions and
historical documents for appropriate cultivar identication.
Most accessions were identied by conrming already
known information. In the following we only comment on
the new identications obtained.
Updating of previously uncorrect
i d e n t i f i c a t i o n s : 'Busuioacā de Bohotin' is a major
local cultivar, grown on 216 ha. Different opinions exist
about its origin: a) it is the result of empirical selection
from wild grapevines present along the Prut River; b) it
was brought from ancient Greece together with 'Tāmâio-
asā româneascā'; c) its Romanian origin is proved by the
connected name – the village of Bohotin. While the SSR
prole of 'Busuioacā de Bohotin' matched that of 'Muscat à
petits grains blancs', the red berry colour and ampelograph-
ic description showed it as being very similar to 'Muscat à
petits grains rouges' (VIVC 8248). For these reasons we
concluded that 'Busuioacā de Bohotin' is the red somatic
variant for berry colour of 'Muscat à petits grains blancs'
and that it does not correspond to 'Muscat rouge de Ma-
dere', as previously stated by ConstantinesCu et al. (1960).
The same authors and the "International List of Vine Varie-
ties and their Synonyms" (OIV 2013) considered 'Tāmâio-
asā româneascā', one of the most important and appre-
ciated grapevine varieties in Romania, to be identical to
'Muscat à petits grains blancs'. However, in this study it
turned out that the analyzed accession (ROM06-0134) did
not share the SSR prole of 'Muscat à petits grains blancs',
but matched the 'Tamaiosa rominesca' accession FRA139-
0Mtp1091, grown in Vassal and introduced from the Bu-
charest collection in 1961. After genotyping and owing to
the information given by ConstantinesCu et al. (1960), the
French accession was considered a misnomer and renamed
as 'Tamaiosa rominesca faux #3426' (faux means wrong in
French). Ampelographic distinction of this genotype and
'Muscat à petits grains blancs' is evident, despite their close
genetic relationship because present molecular data and
laCoMbe et al. (2013) showed that this genotype is most
likely a 'Muscat à petits grains blancs' offspring (Tab. 2).
Two further 'Tāmâiosā' accessions entered Vassal, one
from Bulgaria (1950) and another from Romania (1954)
and were identied as 'Muscat à petits grains blancs'. In
addition, 'Tāmâioasā româneascā' from Craiova Univer-
sity grapevine collection, genotyped by Žulj Mihalević
et al. (2013), matched the 'Muscat à petits grains blancs'
genetic prole, leading to the assumption that 'Tāmâioasā
româneascā' perfectly described by ConstantinesCu et al.
in 1960 is really 'Muscat à petits grains blancs'. Moreover
a handwritten record by the ampelographer Paul Truel of
Vassal (Figure) tells that when the 'Tamaiosa rominesca'
accession entered the collection three plants were 'Muscat
à petits grains blancs' with strong muscat avour and two
were different with slight muscat avour and of unknown
identity, pointing to a mixture of the two varieties. The lat-
ter being kept under the accession ID FRA139-0Mtp1091.
As a result of these ndings a new name for 'Tamaiosa
rominesca faux #3426' was proposed: 'Tamaioasa Bucureş-
ti'. For the future a suggestion is to study the mix of varie-
ties in old 'Muscat à petits grains blancs' vineyards all over
Possible rst degree relationships inside Romanian varieties and
with 'Heunisch weiss' comparing 13 SSRs without mismatching
loci, computed with Cervus software and ordered by descending
pair LOD scores
Putative rst degree related varieties Pair LOD
Coarna neagra Hora 1,14E+15 yes
Coarna rosie Tsitsa kaprei 9,90E+14 yes
Muscat à petits
bucuresti 9,47E+14 yes
Balint weiss Galbena uriasa 7,61E+14 yes
Iordan Crimposie 7,36E+14 yes
Coarna alba Kreaca 7,36E+14 yes
Gordin Braghina rosie 7,24E+14
Chaouch blanc Chaouch rozovyi 7,14E+14 yes
Iordan Heunisch weiss 7,11E+14 yes
Iordan Plavay 6,99E+14 yes
Coarna alba Gordin 6,84E+14 yes
Berbecel Pirciu 6,64E+14 possible
Negru moale Heunisch weiss 6,51E+14 yes
Bakator belyi Bakator roz 6,31E+14 yes
Braghina alba Braghina rosie 5,77E+14
Morostina Heunisch weiss 5,56E+14 yes
Balint weiss Slankamenka
bela 5,35E+14 yes
Juhfark Romanie 5,18E+14
Bakator belyi Negru moale 4,93E+14
Francuse Basicata 3,85E+14
Francuse Heunisch weiss 3,07E+14 yes
Grasa de Cotnari Heunisch weiss 2,88E+14 yes
the country to clarify the situation and thus be of benet for
'Galbenā uriaşā' was considered an autochthonous vari-
ety and clonal variant of 'Galbenā de Odobeşti' (NiColeanu
1900). It has been grown here and there in Moldavia and
Transilvania regions and today is maintained in two grape-
vine collections. 'Galbenā uriaşā' SSR prole matched that
of 'Mirkovaca', an endangered Croatian variety described by
Maletić et al. (1999 and 2015), but was different from that
of 'Galbenā de Odobeşti', therefore being another variety. A
rst degree relationship between them can also be excluded.
The 'Om rāu' accession analysed here is certainly the one
described by ConstantinesCu et al. (1962) and is different
from the one genotyped by Žulj Mihalević et al. (2013).
Varieties absent in the Romanian am-
pelographic literature: 'Galbenā māruntā' is one
of the varieties in danger of extinction and is today main-
178 C. F. popesCu et al.
tained in only two grapevine collections. No documents were
available about its origin or descriptions for comparison. Its
SSR prole matched that of 'Kakotrygis', a Greek variety.
Preliminary ampelographic observations and comparisons
performed on the mature leaf and bunch showed that 'Gal-
benā māruntā' is very similar to the 'Kakotrygis' described
by Kotinis (1984). 'Moroştinā' is another very rare variety
in Romanian vineyards, and no reference documents were
available for description. 'Ţâţa caprei neagrā' SSR prole
matched that of a better known Bulgarian variety named
'Hora' and 'Tâţa vacii neagră' matched that of 'Kozi Cici
cherveni', a variety grown at the Bulgarian Institut de Vit-
iculture et d'Oenologie in Pleven. Regarding 'Braghinā',
ConstantinesCu et al. (1959) mentioned the long standing
presence of this variety with rose grapes in Romania, where
there were many populations with high variability, for ex-
ample showing leaves with entire or with more lobes, with
star-shaped owers, and also populations with different
berry colour and size. The two accessions 'Braghinā albā'
and 'Braghinā roz' showed two different molecular proles
so are therefore different varieties; interestingly, they could
be rst degree related (Tab. 2). 'Braghinā albā' analyzed by
Žulj Mihalević et al. (2013) is a distinct variety.
N e w s y n o n y m s : 'Bātutā neagrā' and 'Negru
românesc' were considered different varieties and were thus
described separately by ConstantinesCu et al. in 1959 and
1966, respectively. The analyzed accessions shared the same
molecular prole. The ampelographic descriptions of the
two cultivars from the old documents, in comparison with
those performed in our collections, conrmed the molecular
results and supported their synonymy.
'Cârcioasā' was described by ConstantinesCu et al.
(1965) as an old autochthonous variety, rarely grown and
without any synonym. However, our results showed that
'Cârcioasā' shared the same genotype with 'Balint weiss',
therefore being an additional, new synonym. 'Coada oilor'
was described as 'Ovis' by ConstantinesCu et al. (1962).
Considered as autochthonous and grown since long before
the phylloxera invasion, the cultivar is today maintained
only in collections. Its SSR prole matched that of 'Juhfark'
(GalbaCs et al. 2009, jahnKe et al. 2009). Comparison of
ampelographic features of 'Juhfark' and 'Ovis' as given in the
literature (neMeth 1970, ConstantinesCu et al. 1962) and
'Juhfark' and 'Ovis' accessions described by Geilweilerhof
and Ştefāneşti respectively likewise attested to their identity.
'Gordan', 'Iordanā' and 'Zemoasā' accessions showed
identical microsatellite proles, proving to be synonyms.
All these varieties are considered autochthonous and have
been grown since before the phylloxera invasion. Constan-
tinesCu et al. in 1960 and 1961 mentioned the synonymy
between 'Gordan' and 'Iordan' (in Romanian literature
present as 'Iordanā'), their names having a common origin
from Iordan, becoming Giordan and after that Gordan. Our
results are in accordance with ndings in the French Vassal
and in the European Vitis Database (http://www.eu-vitis.
de). Today, 'Iordanā' is a major cultivated variety grown
mostly in western and central Romania, 'Gordan' is grown
in a small area especially in the southeastern wine region
and 'Zemoasā' is only rarely present in private vineyards or
in repositories. The SSR proles of 'Gordin' and 'Gordan'
proved to be different, excluding the possibility of being
synonyms, unlike the Žulj Mihalević et al. (2013) ndings
and supporting the old descriptions (ConstantinesCu et al.
1959 and 1961).
Interesting results were obtained with two major cul-
tivars of local importance, extensively grown in Romania,
'Zghiharā de Huşi' and 'Galbenā de Odobeşti', nowadays
considered to be different varieties. ConstantinesCu (1958)
wrote that "'Galbenā de Odobeşti' is one of the old Romanian
varieties with a large growing area in vineyards of Odobeşti
and Panciu; it resembles 'Bātutā neagrā', but has a different
grape colour and is a synonym of 'Zghiharā de Huşi'". Even
though ConstantinesCu et al. described 'Galbenā de Odo-
beşti' and 'Zghiharā de Huşi' separately in 1959 and 1960
respectively, our SSR data conrmed this synonymy, but
exclude a parent-offspring relationship between 'Galbenā
de Odobeşti' and 'Bātutā neagrā'.
Questionable genotypes: Some accessions,
being true-to-type on the basis of ampelographic descrip-
tors, turned out to be critical after comparison of their SSR
proles with literature data. 'Negru mare' was classied as
questionable, because it differs from the one grown in Vassal.
Further observations are necessary to check the authenticity
of this variety with respect to the available ampelographic
description. 'Negru vrtos' (at present 'Negru vârtos') has
been grown in Romania since long before the phylloxera
invasion. ConstantinesCu (1958) mentioned two biotypes
of 'Negru vrtos', one with functionally female owers
and another with hermaphrodite owers, the latter being
morphologically very similar to 'Mavrud Varnenski'. The
genetic prole of our 'Negru vârtos' accession (ROM051-
263) matched that of 'Mavrud Varnenski', conrming the old
information. Two years later ConstantinesCu et al. (1960)
wrote that there are many 'Negru vrtos' biotypes. So we
Figure: Handwritten record by paul truel (1924-2014), former
ampelographer at the INRA Vassal collection, concerning the ac-
cession 'Tamaiosa romanesca #0Mtp1091', which means: "In plot
4.27S12, plants 1 and 2 are different from plants 3 to 5. Plants 3 to
5 are 'Muscat de Frontignan' [i.e. 'Muscat à petits grains blancs'].
Plants 1 and 2 are much more vigorous, without muscat avour
on 4 August 1981 (véraison stage) whereas 'Muscat de Frontignan'
has muscat avour”.
Identication and characterization of Romanian grapevine genetic resources 179
hypothesized that 'Negru vârtos' could represent a larger
group of homonyms, encompassing 'Mavrud varnenski',
the female genotype grown in Vassal (turned out to be true-
to-type) and this third variety (159.12.R) genotyped in the
M i s n o m e r s : 'Alb românesc' (ROM045-003) was
shown to be a misnomer as the SSR prole and ampelo-
graphic traits both match 'Sarba', a new bred Romanian
cultivar. 'Teişor' (ROM045-238) was regarded as a misnomer
expected to be a synonym of 'Harslevelu' (ConstantinesCu et
al. 1959), and not 'Ezerjo'. In fact, 'Teişor' is the diminutive
of 'Tei', meaning lime, and many synonyms of 'Harslevelu'
have names referring to the shape of its leaf, similar to that
of the lime tree, like 'Lindenblättriger', 'Feuille de Tilleul',
'Frunzā de Tei'. 'Mustoasā de Māderat' did not match the
true-to-type grown in Vassal; our genotype is original and
no matches were found with other already genotyped va-
rieties. So it remained anonymous. 'Lampāu' was another
misnomer matching 'Tompa Mihaly' SSR prole and the
ampelographic description of this accession also did not
correspond to literature data.
Possible trios for parents and off-
springs and first degree relationships:
Possible parents-offspring trios and rst degree relationships
were found. Cervus software indicated two possible trios
without mismatchings for candidate parents and progeny.
The rst one was that 'Feteasca regala' could be the prog-
eny of 'Feteasca alba' and 'Francuse', with a LOD score of
1.49 E+15, supporting with 6 additional SSRs what laCoMbe
et al. (2013) already found using 20 SSRs. This information
was partially different from the genitors supposed by Con-
stantinesCu et al. (1959), i.e. 'Feteasca alba' and 'Grasa de
Cotnari', this latter variety being excluded with certainty by
molecular data. The second trio indicated 'Braghina alba' as
the progeny of 'Coarna alba' x 'Galbena de Odobesti', 'Coarna
alba' being the putative mother given the functionally female
sex of owers and 'Galbena de Odobesti' the possible father
with hermaphrodite owers. In this case the LOD score was
a little bit higher: 1.71 E+15. Cervus software indicated a
list of possible rst degree relationships (PO), reported in
Tab. 2. Some of them were partial parentages of already
completed trios, such as 'Coarna rosie' and 'Tsitsa kaprei',
full parentage being 'Coarna rosie' = 'Tsitsa kaprei' and 'Par-
mak crven'; 'Crimposie' and 'Iordan', full parentage being
'Crimposie' = 'Iordan' and 'Beala Debela'; 'Balint weiss' and
'Slankamenka bela', full parentage being 'Slankamenka bela'
= 'Balint weiss' and 'Razachie rosie' (LaCoMbe et al. 2013).
The other PO relationships were evaluated by comparison
with the larger INRA database, apart from two, because
the varieties involved, 'Braghina alba' and 'Romanie', were
not present in Vassal. The preliminary PO indications were
mostly conrmed, but three cases discarded by addition-
al markers. Five cultivars, namely 'Francuse', 'Grasa de
Cotnari', 'Iordan', 'Morostina' and 'Negru moale' showed a
PO relationship with 'Heunisch weiss'/'Gouais blanc'. So,
'Heunisch weiss'/'Gouais blanc', one of the most prolic
founders of the present grapevine assortment (Maul et al.
2015) seems also to have played an important role in the
birth of some Romanian cultivars.
Both morphological descriptors and SSR markers
proved to be efcient at conrming or detecting synonyms,
homonyms, questionables, misnomers and unique genotypes
in Romanian grapevine germplasm collections, therefore
helping to update the information about the grapevine ger-
mplasm preserved there. Together, these two methods also
claried some previous suppositions about the origin of
local/autochthonous varieties and brought out new aspects
to be analyzed.
Ampelographic methods used to characterize varieties
through standardized description, based on phenotypic
traits, were applied to obtain an up-to-date description of
the studied accessions. These ampelographic descriptions
together with genetic proles and photos are available via
the European Vitis Database. They represent a reference for
the authenticity of the studied accessions and their respective
varieties. A wide range of variability was determined among
the studied cultivars regarding certain morphological and
agronomic characters, which are especially valuable for
the autochthonous grape varieties in danger of extinction.
The molecular data complemented conventional de-
scriptions, succeeding in identifying almost all the analyzed
accessions, improving the knowledge on Romanian grape-
vine varieties. The accurate identication obtained allowed
the provenance of some varieties previously considered as
autochthonous to be known and, viceversa, to detect autoch-
thonous varieties previously considered as imported. The
pedigrees of some of them were conrmed and additional
information was produced about possible PO relationships,
delegated to future research.
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laboration for Grapevine Diversity Exploration and Mobilization
of Adaptive Traits for Breeding" for a short-term scientic mission
(STSM). The research was supported by the Service for grapevine
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Received May 3, 2017
Accepted August 31, 2017