Extractable Amounts of trans-Resveratrol in Seed and Berry
Skin in Vitis Evaluated at the Germplasm Level
XIAODONG LI,
†,‡
BENHONG WU,
†
LIJUN WANG,
†
AND SHAOHUA LI*
,†
Institute of Botany, The Chinese Academy of Sciences, 100093 Beijing, People’s Republic of China,
and College of Agronomy and Biotechnology, China Agricultural University, 100094 Beijing, People’s
Republic of China
Extractable amounts of resveratrol in berry skins and seeds were studied in 120 grape (Vitis)
germplasm cultivars during two consecutive years to determine the distribution of resveratrol among
the main grape genotypes. Interspecific rootstock cultivars had much higher extractable amounts of
resveratrol in skin and seed than all other grape genotypes studied in both years. Extremely high
extractable amounts of resveratrol in berry skins [>100 µgg
-1
of skin fresh weight (FW)] and seeds
(>20 µgg
-1
of seed FW) were observed on two rootstock cultivars obtained from hybrids of V.
monticula × V. riparia. Extractable amounts of resveratrol in berries of rootstock cultivars that are
the descendants of V. riparia were also very high. The cultivated European type (V. vinifera) cultivars
and their hybrids with V. labrusca had relatively low levels of extractable resveratrol in berry skin and
seed, and the extractable amounts of resveratrol in berry skin and seeds were, with a few exceptions,
<2 µgg
-1
of skin or seed FW. Extractable amounts of resveratrol in berry skin and seeds were
closely related with fruit traits or purpose of uses and climate. Significantly higher extractable amounts
of resveratrol in berry skin were found in seeded cultivars than in seedless ones, in both berry skin
and seeds in winemaking grapes than in table grapes, and in red grapes than in green ones. Moreover,
rainfall during fruit development resulted in higher extractable amounts of resveratrol in berry skin,
whereas resveratrol synthesis and accumulation in grape seeds were not related to climate change.
KEYWORDS: Resveratrol; grape germplasm; fruit traits
INTRODUCTION
trans-Resveratrol (3,5,4′-trihydroxystilbene) is one of the
major stilbene phytoalexins, originally identified as an active
ingredient of oriental folk medicines used for treatment of a
wide variety of diseases (1, 2). This compound has been found
in at least 72 plant species distributed among 31 genera and 12
families (3), and a number of these are components of the human
diet, for example, grapes, wine, grape juice, cranberries (4),
peanuts (5), and chocolate and cocoa (3). Grapes and grape
products, however, are considered to be the most important
human dietary sources of resveratrol (6-8).
The interest in resveratrol in grape was originally sparked
by epidemiological studies indicating an inverse relationship
between moderate consumption of red wine over a long period
of time and risk of coronary heart disease, the so-called “French
paradox” (9). This biological attribute has been ascribed to
resveratrol (10), and several kinds of evidence have accumulated
in support of resveratrol’s pharmacological activities. Resvera-
trol’s reported biological attributes include anti-inflammatory
(11), cardioprotection (12), cancer chemopreventive (13), and
antioxidant properties (14) and inhibition of platelet aggregation
(10).
Resveratrol’s synthesis can be accomplished in the laboratory
via the Heck reaction (15), but the use of synthetic food additives
in the food industry is often severely restricted. Consequently,
there is more and more interest in natural resveratrol extracted
from plants, especially from grape and grape products.
The grape is the world’s second largest fruit crop, with
>66.41 million metric tons produced in 2005 (FAO STAT
Database at www.fao.org). About 13.4% of the total grapes
harvested are for fresh consumption (table grapes), whereas
86.6% of the crop is processed, especially for winemaking (16).
Grapes cultivated throughout the world today mainly belong to
three types, the European type (Vitis Vinifera L.), the American
bunch type (Vitis labrusca L. and its derivatives, especially the
hybrids obtained from V. labrusca and V. Vinifera), or the
Muscadine type (Vitis rotundifolia Michx). All of these grape
germplasm resources can provide abundant natural resveratrol
products. Therefore, it is important to evaluate resveratrol
content in grape germplasm in order to utilize those germplasm
resources to produce resveratrol or in breeding programs to
obtain new grape cultivars with high levels of resveratrol in
their berries.
* Author to whom correspondence should be addressed (telephone +86
10 62836026; fax +86 10 62836026; e-mail shhli@ibcas.ac.cn).
†
Institute of Botany.
‡
China Agricultural University.
8804 J. Agric. Food Chem. 2006, 54, 8804−8811
10.1021/jf061722y CCC: $33.50 2006 American Chemical Society
Published on Web 10/25/2006

In grape berries resveratrol is primarily synthesized and
located in the skin and seed cells with little or none in the fruit
flesh (7, 8). Although there have been several studies of
resveratrol in grape berry skins and seeds (6-8), none has yet
investigated resveratrol in berry skins and seeds at the grape
germplasm level. Germplasm resources are very rich, and there
are at least 70 species in the genus Vitis (16). Moreover, cultivars
are numerous, with >10000 in the world, due to a long history
of cultivation and an extensive geographical distribution (16).
Grape germplasm resources should play an important role in
resveratrol synthesis, which may be genetically controlled. The
purpose of the present study is to explore the variations in
extractable amounts of resveratrol in skins and seeds in grape
germplasm. This should provide basic data (1) for further study
of the genetic basis of grape resveratrol metabolism, (2) to find
new medicinal resources, and (3) to exploit the grape germplasm
with high levels of resveratrol in breeding programs to obtain
new cultivars rich in resveratrol.
MATERIALS AND METHODS
Plant Material. There were 120 grape cultivars used in this study
in 2003 and 2004, and 70 were studied in two successive years. The
total included 52 table grapes, 1 juice grape, and 1 wine grape hybrid
between V. labrusca and V. Vinifera, 21 wine grapes and 31 table grapes
of V. Vinifera, 3 wine grape hybrids of V. Vinifera × V. amurensis,2
juice grape hybrids of V. thunbergii × V. Vinifera, 1 juice grape hybrid
of V. amurensis × V. lubrusca, 7 rootstock hybrids from V. amurensis
× V. riparia (1), V. berlandier × V. riparia (1), V. riparia × V. lubrusca
(1), V. monticula × V. riparia (3), and V. riparia × V. rupestris (1),
and a Chinese wild grape species V. amurensis var. dissecta (Table
1). All samples were collected from the Germplasm Repository for
grapes in the Institute of Botany of the Chinese Academy of Sciences
located in Beijing. All of the cultivars were planted in the spring of
1993. The vines, trained to bilateral cordons, were spaced 1.5 m apart
within the row and 2.5 m apart between rows with a north-south row
orientation. All were subjected to the same management practices, such
as irrigation, fertilization, soil management, pruning, and disease control.
Grapes were hand-harvested from mid-July to late September at the
ripening stage of each cultivar. The berry ripening stage was evaluated
on the former year’s ripening date and as judged from seed color change
to dark brown without senescence of berry tissue. Berries were sampled
from three clusters, randomly chosen in thee vines of each cultivar as
three replications. The samples were taken to the laboratory, and the
skins and seeds were separated by hand immediately. The skins and
seeds were then frozen in liquid nitrogen, ground to a powder, and
then stored at -40 °C.
Extraction of Resveratrol from Skins and Seeds. trans-Resveratrol
was extracted according to the method of Li et al. (17). Three gram
samples were ground using a porcelain mortar and pestle in 15 mL of
extraction solvent (ethyl acetate for berry skins and methanol for seeds;
both reagents were of analytical grade and purchased from Beijing
Chemical Plant, Beijing, China). The samples were extracted in the
dark at 25 °C for 48 h and then centrifuged at 10000g for 10 min. The
supernatants were evaporated to dryness by rotary vacuum evaporation
at 40 °C. Dried samples were then dissolved in 1 mL of methanol and
stored at -40 °C. The samples were filtered through a 0.45 µm PTFE
membrane filter before resveratrol analysis.
Determination of Resveratrol Concentration. Extractable amounts
of resveratrol were analyzed using a Dionex Summit HPLC system
including a Dionex P680 pump, a Dionex TCC-100 thermostated
Table 1. Grape Cultivars Used in This Study
genotype group
a
no. of
cultivars cultivar
b
table grape of LV 52 Beniyamabiko (1), Benizawa (2), Benizuiho (3), Bennifuji (4), Black Olimpia (5), Canadice (6), Catawba (7),
Delaware (4×) (8), Guixiangyi (9), Himrod (10), Jifeng (11), Jingchao (12), Kangtai (13), Kunitachi
Seedless (14), Mars Seedless (15), Pondicherry (16), Queenora Seedless (17), Ryuho (18), Swenson
Red (19), Takasuma (20), Tano Red (21), Triumph (22), Vehava 180 (23), Venus (24), Yigawa 1014
(25), Yigawa 1015 (26), Yigawa 1025 (27), Yigawa 1050 (28), Yigawa 1055 (29), Yigawa 1060 (30),
Beni Sajku (31)
†
, Beniizu (32)
†
, Fujiminori (33)
†
, Honey Red (34)
†
, Hongboduo (35)
†
, Hyuga (36)
†
,
Izunishiki (37)
†
, Kyoho (38)
†
, Rhodo Berry (39)
†
, Roudingxiang (40)
†
, Ruby Niagara (41)
†
,
Urbana (42)
†
, Vehava540 (43)
†
, Violet Vehara (44)
†
, Wase Takasumi (45)
†
, Fenghou (46)
§
, Harata
314 (47)
§
, Jingya (48)
§
, Jingyou (49)
§
, Premium (50)
§
, Red Queen (51)
§
, White Olympia (52)
§
juice grape of LV 1 Honey Juice (53)
§
wine grape of LV 1 Super Hamburg (54)
§
table grape of V 31 Centennial Seedless (55), Early Muscat (56), Gloria Hungariae (57), Gros Colman (58), Jingdajing (59),
Jingkejing (60), Jingxiu (61), Jingzaojing (62), Kuratsufuri (63), Misket Dounvaski (64), Muscat
Hamburg (65), Fenghuang 51 (66), Red Globe (67), Ruby Seedless (68), Su161 (69), Suffolk (70),
Thompson seedless (71), Zaoyu (72), Fenniu (73)
†
, Guibao (74)
†
, Hiro Hamburg (75)
†
, Jingyu (76)
†
,
Jingzijing (77)
†
, Otilia (78)
†
, Wajisilibaiyu (79)
†
, Zaomanao (80)
†
, Xiangfei (81)
§
, Huaze
Lizamate (82)
§
, Queen of Vineyards (83)
§
, Shiyaira (84)
§
, Superior Seedless (85)
§
wine grape of V 21 Bujiesuli (86), Carigane (87), French Blue (88), Meichun (89), Merlot (90), Semillon (91), Su162 (92), Ugni
Blanc (93), White Suntory (94), Wuyuezi (95), Zexiang (96), Baiyu (97)
†
, Cabernet Franc (98)
†
,
Chardonnay (99)
†
, Guoho2 (100)
†
, Italian Riesling (101)
†
, Pannoniavinesa (102)
†
, Cabernet
Gernischet (103)
§
, Lion Riesling (104)
§
, Suntory (105)
§
, Yan73 (106)
§
wine grape of VA 3 Beichun (107), Beihong (108), Beiquan (109)
juice grape of TV 2 Beifeng (110)
†
, Beizi (111)
†
juice grape of LA 1 Russia Concord (112)
rootstock grape of RR 1 101-14 (113)
rootstock grape of BR 1 5A (114)
rootstock grape of AR 1 ARH2 (115)
rootstock grape of MR 3 188-8 (116), Zhi 166 (117), Zhi 168 (118)
rootstock grape of LR 1 Beta (119)
§
Chinese wild grape
species Ad
1 Yanshan (120)
a
LV, hybrids between V. labrusca and V. vinifera;V,V. vinifera; VA, V. vinifera × V. amurensis; TV, V. thunbergii × V. vinifera; LA, V. lubrusca × V. amurensis; RR,
V. riparia × V. rupestris; BR, V. berlandier × V. riparia; AR, V. amurensis × V. riparia; MR, V. Monticola × V. riparia; LR, V. labrusca × V. riparia; Ad, V. amurensis var.
dissecta.
b
Number in parentheses following the cultivar indicates the sample order;
†
indicates used in 2003 only;
§
indicates use in 2004 only.
trans-Resveratrol in Vitis at Germplasm Level J. Agric. Food Chem., Vol. 54, No. 23, 2006 8805