Journal of Ethnopharmacology 108 (2006) 414–422

A new insight into Cannabis sativa (Cannabaceae) utilization from

2500-year-old Yanghai Tombs, Xinjiang, China
Hong-En Jiang a,b,c, Xiao Li c, You-Xing Zhao d, David K. Ferguson e, Francis Hueber f, Subir Bera g,
Yu-Fei Wang a , Liang-Cheng Zhao h , Chang-Jiang Liu a , Cheng-Sen Li a,i,∗
a Laboratory of Systematic and Evolutionary Botany, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China
b Graduate School, Chinese Academy of Sciences, Beijing 100039, China
c Bureau of Cultural Relics of Turpan Prefecture, Turpan 838000, Xinjiang, China
d State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany,

Chinese Academy of Sciences, Kunming 650204, China

e Institute of Palaeontology, University of Vienna, Althanstraβe 14, A-1090 Vienna, Austria
f Department of Paleobiology, Smithsonian Institutions, Wastington DC 20560-0121, USA
g Department of Botany, University of Calcutta, Kolkata 700019, India
h College of Biological Science and Biotechnology, Beijing Forestry University, Beijing 100083, China
i Beijing Museum of Natural History, Beijing 100050, China

Received 9 February 2006; received in revised form 25 May 2006; accepted 30 May 2006
Available online 23 June 2006

Abstract
A cache of shoots, leaves and fruits dated by 14 C at 2500 years B.P. were unearthed in the Yanghai Tombs, Turpan District in Xinjiang, China.
By comparing the morphological and anatomical characteristics of the plant remains found in the tomb and specimens of modern plants, it is
shown that the remains belong to Cannabis. Based on the shamanistic background of the deceased man and ancient customs, it is assumed that the
Cannabis was utilized for ritual/medicinal purposes.
© 2006 Elsevier Ireland Ltd. All rights reserved.

Keywords: Cannabis sativa L.; Palaeoethnobotany; Yanghai Tombs; Xinjiang; China

1. Introduction China. The shamanistic practices of Central Asia were not shared
by a majority of people or openly mentioned in the ancient texts
As one of the oldest domestic plants in the history of mankind, that refer to this region (Touw, 1981). Ritual use of Cannabis
Cannabis has probably been utilized for 10,000 years or more by nomadic tribes in Central Asia has been well described
(Schultes et al., 1974; Merlin, 2003). It was eventually cultivated (Rudenko, 1970), but there is little archaeological evidence of
and selected for multi-purposes including the use of fiber from its such use from northwest China. The discovery of floral and other
stem, edible food/oil from its achene, medicine, and psychoac- remains of Cannabis in the Yanghai Tombs provides physical
tive substances in its resin glands (Schultes et al., 1974). Fiber, evidence of its ethnobotanical significance in ancient Turpan.
food, and medicinal uses of Cannabis by the ancient Chinese Yanghai Tombs (42◦ 48 –42◦ 49 N, 89◦ 39 –89◦ 40 E) are
have been well documented (Keng, 1974; Li, 1974a,b, 1978). located in a stony desert (Fig. 1A–C). The tombs consist of
However, little is known about the ancient magic-religious and three groups: Nos.1–3. The deceased person in Room 90, Tomb
medicinal use of Cannabis in the region of Xinjiang, northwest No. 1 was a Caucasoid male, nearly 40 years in age. The grave
gifts in his tomb were more elaborate than those in others at the
site. Most of the utensils were connected with horsemanship,
∗ Corresponding author at: Department of Palaeobotany, Institute of Botany,
but there were also a bow, arrows, musical instruments and some
Chinese Academy of Sciences, No. 20 Nan xin cun, Xiangshan, Beijing 100093,
wooden cups. A leather basket (specimen number M90:8-1) was
China. Tel.: +86 10 62836436; fax: +86 10 62593385. found near the head of the corpse in this tomb, seemingly within
E-mail addresses: [email protected], [email protected] (C.-S. Li). close reach of the deceased in the afterlife. The lidless basket

0378-8741/$ – see front matter © 2006 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.jep.2006.05.034
 H.-E. Jiang et al. / Journal of Ethnopharmacology 108 (2006) 414–422 415

Fig. 1. The map of the location of the Yanghai Tombs, Xinjiang, northwest China. (A) The location of Xinjiang in China. (B) The location of Turpan (the rectangular
area). (C) The location of the Yanghai Tombs.

is 31 cm in height and 24 cm in diameter (Fig. 2A), was full of Schultes et al., 1974; Hillig, 2004, 2005a,b; Hillig and Mahlberg,
fruits, leaves and shoots (Fig. 2C). A wooden bowl (specimen 2004). However, based on its morphology, anatomy, phytochem-
number M90:8-2), 21 cm in height and 36 cm in diameter, was istry and genetic studies, Cannabis is generally treated as a sin-
also found to contain the same plant remains (Fig. 2B). After a gle, highly polymorphic species, Cannabis sativa L. (Bouquet,
detailed study, the material was determined as Cannabis. 1950; Miller, 1970; Davidyan, 1972; Small, 1975b; Small and
The genus Cannabis produces erect, annual herbs, which Cronquist, 1976; Klimko, 1980; Gilmore et al., 2003; Wu and
are mostly dioecious, rarely monoecious (Miller, 1970; Wu and Raven, 2003). The monotypic species concept is adopted here.
Raven, 2003). The number of Cannabis species is controversial. Cannabis sativa is probably indigenous to Central Asia, but the
Some authors are of the opinion that there are more than one long history of cultivation makes it difficult to pin-point the orig-
species in this genus (Anderson, 1974, 1980; Emboden, 1974; inal distribution of the plant. It is cultivated throughout China

Fig. 2. The leather basket, wooden bowl and Cannabis remains. (A) The lidless leather basket. Scale bar = 10 cm. (B) The wooden bowl showing the holes (Arrows).
Scale bar = 10 cm. (C) Shoots, leaves, and fruits in the basket. Scale bar = 40 mm.
416 H.-E. Jiang et al. / Journal of Ethnopharmacology 108 (2006) 414–422

now, but is considered to be native in Xinjiang (Wu and Raven, 2.3. The measurement of the samples
2003).
Fruits, petioles and shoots, 50 of each, were picked randomly
2. Materials and methods and their size was measured with a vernier caliper. The diameter
of shoots and petioles was measured in their middle portions.
The specimens of Cannabis sativa (No. M90:8), collected The weight of the fruits was obtained by using a Mettler AE160
from a leather basket and a wooden bowl in the Yanghai Tombs, Analytical Balance.
include shoots, fruits and fragmented leaves.
2.4. Dating
2.1. SEM (scanning electron microscope) examination
The samples for dating were collected directly from the
The dried specimens were directly placed on the stubs and plant remains, and dated with an accelerator mass spectrometer
sputter-coated with gold using a SPI-MODULE sputter coater. (AMS) 14 C in Peking University.
The samples were examined and photographed under a Hitachi- For comparative studies, specimens of a modern plant (num-
S 800 SEM at an accelerating voltage of 30 kV. ber BJ200401) were collected in the Botanical Garden, Institute
of Botany, Chinese Academy of Sciences, Beijing, China and
2.2. LM (light microscope) examination stored in the National Museum of Plant History of China.
The botanical terms used in the description of the Cannabis
The shoots selected for examination were boiled for 8 h and specimens came from the following references: (Hayward, 1938;
embedded in polyethylene glycol at a temperature of 60 ◦ C for Mohan Ram and Nath, 1964; Miller, 1970; Vaughan, 1970;
2 days. Then they were sectioned on a sliding microtome at Hammond and Mahlberg, 1973; Anderson, 1974; Ledbetter
15 ␮m intervals, and the sections stained with safranin. Xylem and Krikorian, 1975; Small, 1975a; Dayanandan and Kaufman,
elements were measured with a calibrated ocular micrometer 1976; Small and Cronquist, 1976; Prakash et al., 1979; Klimko,
and photographed under an Olympus 60 light microscope. 1980; Wu and Raven, 2003).

Fig. 3. Scanning electron microscope (SEM) micrographs of fossil and extant shoots, infructescences and leaves of Cannabis. (A)–(C) from the remains and (D)–(F)
from the modern plant. (A) and (D) The shoot, showing the non-glandular trichomes (arrow) and the ribs (arrow). Scale bar = 750 ␮m. (B) and (E) The infructescence,
displaying the receptacle (arrow). Scale bar = 1.5 mm. (C) and (F) The leaves, showing the palmately compound primary veins. The tissue between the major veins
of the blade from (C) has disintegrated leaving only the primary veins. Scale bar of (C) = 1.2 mm; scale bar of (F) = 1 cm. NGT: non-glandular trichomes; RI: rib;
RE: receptacle. Note picture (F) are not SEM photograph.
 H.-E. Jiang et al. / Journal of Ethnopharmacology 108 (2006) 414–422 417

3. Results 49 to 117 ␮m (X̄ = 64 ␮m, N = 50) and the diameter of the base
varies from 68 to 134 ␮m (X̄ = 80 ␮m, N = 50). The trichome
3.1. Description density averages 60 mm−2 .
Two types of trichomes occur on the abaxial epidermis, i.e.
The materials are 789 g in weight, 2475 ± 30 years in age glandular and non-glandular. The non-glandular ones are con-
(specimen number BA04538), and are deposited in the Turpan ical, 85–173 ␮m long (X̄ = 137 ␮m, N = 50) with an enlarged,
Museum, Xinjiang, China. bulbous base. The bases vary from 17.8 to 29.5 ␮m (X̄ =
21.9 ␮m, N = 50) in diameter (Fig. 5B). The trichomes located
3.1.1. Leaves on or near the principal veins display a warty surface, while
Leaves are preserved as brittle fragments of the blades and those between the veins are only slightly warty or smooth. The
some petioles. Leaf stipules could not be identified. The laminae tips of the trichomes are inclined toward the leaf apex at vari-
are yellow-green in color, usually enrolled and wrinkled. The ous angles. Such non-glandular trichomes are more numerous
venation is actinodromous. The principal veins (3–7) diverge (360 mm−2 ) than on the adaxial epidermis. The glandular tri-
from the leaf base at an angle of 10–30◦ (Fig. 3C). The veins chomes are sessile, brown-red in color, rounded in shape with
form prominent ridges on the abaxial surface of the leaves while an average diameter of 37.4 ␮m (N = 50). They are sparsely and
they are represented by a groove on the adaxial surface. uniformly distributed with an average density of 28 mm−2 .
The cuticle is rough with raised epidermal cells. The epider- Petioles are scabrous and furrowed, 5.40–15.40 mm in length
mal cells are irregular in outline with undulate anticlinal walls (X̄ = 11.27 mm, N = 50) and 0.24–0.60 mm in diameter (X̄ =
(Fig. 5A). 0.39 mm, N = 50). Non-glandular and sessile glandular tri-
The adaxial epidermis of the leaves has conical non-glandular chomes occur on the epidermis. The former are conical, curved,
trichomes with a warty surface. Their tips are erect or inclined simple, with a warty surface, and inclined toward the apex of the
towards the leaf apex. Nearly all trichomes have a rounded leaf. The latter are globular, light brown in color, with a smooth
enlarged base (Fig. 5A). The length of the trichomes varies from surface.

Fig. 4. SEM micrographs of fruits. (A)–(C) from the remains and (D)–(F) from the living plant. (A) and (D) The fruit is surrounded by a bract with a beak (arrow).
Scale bar = 1.5 mm. (B) and (E) Lateral view of the fruit displaying the slightly tapered apex and the rounded to truncate base and the finely reticulate venation
(arrow). Scale bar = 1.2 mm. (C) and (F) The oblique view of the fruit base showing the ribs (arrow) and the inserted fruit scar (arrow). Scale bar = 600 ␮m. BK:
beak; RV: reticulate venation; RB: rib; FS: fruit scar.
418 H.-E. Jiang et al. / Journal of Ethnopharmacology 108 (2006) 414–422

Fig. 5. SEM micrographs of the characters of the trichomes from the leaf and bract, respectively. (A)–(C) are from the remains and (D)–(F) are from living
plants. (A) and (D) The adaxial epidermis of the leaf displaying the irregularly shaped epidermal cells and the non-glandular trichomes with enlarged base (arrow).
Scale bar = 120 ␮m. (B) and (E) The abaxial epidermis of the leaf showing the non-glandular trichomes (arrow) and the sessile glandular trichomes (arrow). Scale
bar = 75 ␮m. (C) and (F) The outer surface of the bract with non-glandular trichomes (arrow), sessile-(arrow) and capitate glandular trichomes (arrow). Scale
bar = 120 ␮m. NGT: non-glandular trichomes; SGT: sessile glandular trichomes; CGT: capitate glandular trichomes.

3.1.2. Shoots kinds of trichomes on the epidermis of the shoot, viz. glandu-
The shoots are scabrous and ribbed (Fig. 3A), 7–42 mm lar and non-glandular. The non-glandular trichomes are conical,
long (X̄ = 23.66 mm, N = 50), 0.4–1.5 mm in diameter (X̄ = 49–290 ␮m long (X̄ = 131.43 mm, N = 50), and inclined toward
0.90 mm, N = 50). Ribs are straight and parallel. There are two the stem apex. They are simple and have a conspicuously warty

Fig. 6. SEM micrographs of the characters of the perianth. (A)–(C) are from the remains and (D)–(F) are from the living plant. (A) and (D) The outer epidermis of
the perianth, showing the thread-like trichomes (arrow), the rounded trichome bases (arrow) and the outer surface of the epicarp (arrow). Scale bar = 136 ␮m. (B)
and (E) The aligned perianth cells with straight cell walls and diagonal end walls. Scale bar = 27 ␮m. (C) and (F) The perianth cells with wavy margins. Scale bar,
27 ␮m; PER: pericarp; TB: trichome base; TLT: thread-like trichome.
 H.-E. Jiang et al. / Journal of Ethnopharmacology 108 (2006) 414–422 419

surface and bulbous base (Fig. 7C). Glandular trichomes are rectangular or polygonal, 8–16 ␮m (X̄ = 12.15 mm, N = 50) in
sessile, globular (X̄ = 46 mm, N = 50), brown-red in color. tangential length; 5–18 ␮m (X̄ = 10.94 mm, N = 50) in radial
The cylindrical pith occupies 1/3 of the diameter of the shoot length. Ray parenchyma cells are rectangular, separated by 3–14
(Fig. 7G). Primary xylem is poorly preserved, with few remains layers of xylem fibers.
of vessels. In the region of secondary xylem, vessels are round
(70%), elliptical (20%) or irregular (10%) in shape, 16–57 ␮m in 3.1.3. Fruit
diameter (X̄ = 32.4 mm, N = 200). Sometimes three to five ves- Infructescence is 0.8–1.5 cm long, 0.2–0.4 mm in diameter.
sels are radially contiguous. Xylem fibers are triangular, square, One or two fruits are born in the axil of a leaf. When the fruit

Fig. 7. SEM micrographs of the characters of pericarp, trichomes from the shoot and light microscope (LM) photographs of the transverse section of the shoot.
(A)–(C) and (G) are from the remains and (D)–(F) and (H) are from the living plant. (A) and (D) The inner surface of the endocarp displaying the undulate cell walls.
Scale bar = 25 ␮m. (B) and (E) The longitude section of the pericarp showing its three layers. Scale bar = 50 ␮m. (a) epicarp; (b) mesocarp; (c) endocarp. (C) and
(F) The epidermis of the shoots displaying the non-glandular trichomes with a warty surface. NGT: non-glandular trichomes. Scale bar = 10 ␮m. (G) and (H) The
transverse section of the shoots. Scale bar = 400 ␮m.
420 H.-E. Jiang et al. / Journal of Ethnopharmacology 108 (2006) 414–422

is abscised, the receptacle of the fruit, 0.70 mm in diameter, ference could be found between the subfossils and comparative
remains in the axil (Fig. 3B) of the leaf. Epidermal features of modern plants of Cannabis sativa L.
the infructescence, including the non-glandular and glandular
trichomes, are similar to those of the shoots. 4. Discussion and conclusions
Each fruit is surrounded by a perianth. The fruit together
with its perianth is surrounded loosely or tightly by a bract The remains of hemp (Cannabis sativa L.) have been
(Fig. 4A). Due to preservation, 48% of the fruits have lost the unearthed mainly in Eurasia (Fleming and Clarke, 1998; Merlin,
bract (N = 100). The bract has a pointed, straight beak. Both non- 2003). Most of these remains are carbonized or decayed because
glandular and glandular trichomes occur on the outer epider- of the climate and storage conditions. The remains of Cannabis
mis of the bract. Non-glandular trichomes are erect or inclined from the Yanghai Tombs located in the Turpan Basin, are
toward the apex of the bract. Glandular trichomes are sessile well preserved due to the extremely dry climate with little
(80%) or capitate (20%) (N = 100). Sessile glandular trichomes or no annual rainfall. Many mummies and funerary objects
have similar structures to those on the shoots. Capitate glandular were well preserved in the Yanghai Tombs along with the
trichomes are born on a long stalk, but are otherwise simi- remains of ancient Cannabis. The ancient leaves, shoots and
lar to the sessile ones (Fig. 5C). The non-glandular trichomes fruits, including their cellular structures, are still preserved in
together with the sessile glandular trichomes on the inner epi- three dimensions, without any distortion, including the non-
dermis of the bract have similar characters to those on the outer glandular and glandular trichomes that have remained preserved
epidermis, but mainly occur at the margin and near the apex of for 2500 years. These remains of hemp are the oldest examples of
the bract. Cannabis thus far reported in which macroscopic/microscopic
The perianth is free but adheres closely to the fruit. It is mem- structures can be demonstrated in such great detail. These mate-
branous, usually hyaline, but some parts are mottled brown to rials provide us with significant information on ancient civiliza-
dark brown. The thread-like unicellular trichomes are long, but tion in the Turpan area and about the utilization of hemp.
are not preserved; only their rounded to elliptical bases, averag- Common daily tasks such as food gathering and preparing,
ing 22.5 ␮m in diameter, remain (Fig. 6A). Epidermal cells are cooking, cloth weaving, shelter construction, medicine, ritual
mainly elongate with long axes parallel to that of the fruit. Their and even intoxication have been accomplished primarily through
walls are relatively straight and parallel (Fig. 6B), or undulate the use of plants (Hastorf, 1999). Cannabis has long been espe-
(Fig. 6C). In the former, the epidermal cells are nearly rectan- cially useful because it can be utilized for all these purposes.
gular, with an average length of 92 ␮m and width of 9 ␮m. The fruits, shoots, and leaves of Cannabis in the ancient leather
The fruit is an achene, oval (Fig. 4B), flattened with two basket could be considered as an important gift for the deceased
prominent ribs (Fig. 4C). It has a slightly tapering apex and man laid to rest in the tomb, but why did he need these plants?
a rounded to truncate base, 2.2–3.6 mm long (X̄ = 2.99 mm, If they were meant as a cereal or for oil extraction, the leaves
N = 50), 1.7–2.5 mm wide in the middle (X̄ = 2.19 mm, N = 50). and shoots would have been removed; if fiber production was
The scar of abscission is usually inserted, surrounding the many intended, only the stems needed to be saved. In this connection
vascular bundles (Fig. 4C). it is significant that no hemp textiles have been unearthed in the
The pericarp is crustaceous, 0.08–0.20 mm thick. It consists Yanghai Tombs. Thus, we found no suitable evidence that the
of three layers: epicarp, mesocarp and endocarp (Fig. 7B). The ancient, indigenous people utilized Cannabis for food, oil, or
outer surface of the epicarp is shiny, mostly smooth, but marked fiber.
by some finely reticulate venation (Fig. 4B). The mesocarp con- The glandular trichomes on most aerial surfaces of Cannabis
sists of one to three layers of parenchymatous cells (Fig. 7B). produce terpenophenolic compounds called cannabinoids that
The inner walls of the endocarp cells are heavily thickened, with are unique to Cannabis (Mahlberg and Kim, 2004). THC (delta-
the inner surface of the endocarp appearing undulate under the 9-tetrahydrocannabinol), one of the most abundant cannabi-
SEM. The anticlinal walls of the endocarp cells are sinuous and noids, is psychoactive. The THC content decreases in various
interlocking, and traversed by numerous fine pits (Fig. 7A). plant parts in the following order: bracts, flowers, leaves, smaller
The achene has a single seed. The seeds are wrinkled and stems, larger stems, roots and fruits (Fetterman et al., 1971).
with no detailed structures preserved. The seed coat is white in Most of the samples consist of leaves and small stems, which
color and consists of spongy parenchymatous cells. have a moderate amount of THC. Although Cannabis fruits
are not psychoactive, about 50% of them were still enclosed
3.2. Comparison by a bract, which contains the highest percentage of THC of
any tissue. All the Cannabis remains could be used for psy-
The plant remains from the Yanghai Tombs display ribbed choactive purposes, which would suggest that the deceased man
shoots, palmately compound leaves, flattened fruits, non- knew of the narcotic value of Cannabis. Moreover, based on
glandular and glandular trichomes on all epidermides, finger- the analysis of all the funerary objects in Room 90, Tomb No.1,
print-like ornamentation of the cuticle on the adaxial surface the owner was considered to be a shaman. As mentioned in
of the leaves, a perianth with parallel and undulate epidermal the introduction, the funereal objects in his tomb were richer
cell walls, an epicarp with reticulate venation, an endocarp with than those in the others, which demonstrates his special sta-
an undulate inner surface, etc. These features suggest that the tus. Together with the musical instrument and the Cannabis,
remains belong to Cannabis sativa L. (Cannabaceae), as no dif- which are unique among the Yanghai Tombs, the shamanistic
 H.-E. Jiang et al. / Journal of Ethnopharmacology 108 (2006) 414–422 421

status of the deceased becomes all the more apparent. Due to Acknowledgements
its apparently prolonged use as a pestle, the inner surface of
the wooden bowl containing Cannabis had become smooth, and The authors thank Drs. Karl W. Hillig, Mark D. Merlin, and
one side became perforated (Fig. 2B). The Cannabis was pre- Ernest Small who discussed the characters of Cannabis and sent
sumably pulverized with a mortar before being consumed for copies of their papers and some other useful references; to Drs.
psychoactive purposes. Thus, we assume that the deceased was Loran C. Anderson, Jan J. Wójcicki, Mr. Robert C. Clarke, and
more concerned with the intoxicant and/or medicinal value of Ms. Gong Qiu-Ping who helped with the references. This study
the Cannabis remains. was supported by a grant from the Chinese Academy of Sciences
The ancient medicinal, ritual, and psychoactive uses of (No. 2004CB720200, KZCX2-SW-118).
Cannabis are often difficult to separate (Merlin, 2003). The
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