Table Of Content植物研究雑誌
J. Jpn. Bot.
Originals 81: 63-74 (2006)
Chemical Characterization of Rhizomes of Atractylodes lancea
and A. chinensis Identified by ITS Sequences of nrDNA
Yo ichiro NAKAI，K oya YA NO，M ao SHIBA，K e吋iKONDO，O sami TAKEDA，
Iwao SAKAKIBARA，S usumu TERABAYASHI，S huichi TAKEDA and Minoru OKADA
Pharmacognosy and Medicinal Resources Research Department，Ce ntral Research Laboratories，T sumura & Co.，
3586，Y oshiwara，A rni-machi，In ashikigun，Ibaraki，3 00-1192 JAPAN
E-mail: nakai_youichirou@同mail.tsumura.co.jp
(Received on September 5，2 005)
Ac omparison was made with regard to interspecific and geographical characteristics
of nine pharmacologically active components in dried rhizomes of Atracηlodes lancea，
A. chinensis，a nd the hybrids between them，w hich were identified by sequence data of
intemal transcribed spacer nuclear ribosomal DNA. The chemical constituents of the nine
components were determined. Principal component analysis using the contents data of
these nine constituents revealed that A. lancea differs chemically from A. chinensis. The
hybrids possessed chemical variations within the limits observed in both the species. The
relationships between these chemical variations and geographical localities were
numerically analyzed by the Mahalanobis-Taguchi method，a nd this analysis enabled the
classification of individuals from the localities into four groups.
Key words: Atractylodes chinensis，A tractylodes lancea，h ybrid，l ocal variation，
polyacety lenes.
Atracηlodes lancea (Thunb.) DC. and A. species have been detected.
chinensis Koidz. (Compositae) are perennial The chemical constituents of the dried rhi
国
herbs that are widely distributed in China zomes of A. lancea and A. chinensis have
(Institute of Botany，A cademia Sinica ed. been determined by isolation studies or ana-
1975). Dried rhizomes of both the species， lytical methods，s howing the presence of
which have been used as diuretics and anal- sesquiterpenes (Yo sioka et al. 1959)，p oly-
gesics as well as for treating stomach disor- acetylenes (Nishikawa et al. 1976，L ehner
ders in traditional Kampo medicines，a re et al. 1997)，a nd sugar esters (Yahara et al.
described in the Jap anese Pharmacopeia 1989，K itajima et al. 2003). Among these
(The Society of Jap anese Pharmacopeia components，e lemol (1) (Matsunaga et al.
s-
2001) as the botanical origins of “Sojutsu." 2000)，h inesol (2) (Satoh et al. 2000)，
These plants had been identified based on eudesmol (3) (Tsuneki et al. 2005)，a tracty-
the morphological characters of their aerial lon (4) (Resch et al. 1998)，a tractylodin (5)
parts (Kawanishi et al. 1994)，S hiba et al. (Resch et al. 2001)， atractylodinol (6)
(2006) have reported that A. lancea and A. (Takeda et al. 2001)，a nd acetylatracty
司
chinensis could be more clearly identified by lodinol (7) (Na kai et al. 2003) have been re-
sequences of intemal transcribed spacer ported to possess pharmacological activities.
(ITS) nuclear ribosomal DNA (nrDNA)，in Furthermore，w e have reported that the con-
addition，h ybrid individuals between the two stituents of the dried rhizomes of A.
-63-
64 植物研究雑誌第81巻第2号 平成18年4月
chinensis contain a new polyacetylene Jiangsu，A nhui，H ubei，H ebei，S handong，
atractyloyne (9) and its structural isomer， Henan，a nd Shaanxi provinces of China from
3-isovaleryloxy-tetradeca-4，6，12-triene-8，10- 1990 to 1994 and the contents of the four
diyne-l，14-diol (8)，t hese e'easily deri- sesquiterpenes determined previously
紅
vatized to a phmacologically active (Takeda et al. 1994，1 995a，1 995b，1 996a).
訂
substance，4 ，6，12 -tetradecatriene-8，10 -diyne- The rhizomes were numbered according to
1，3，14-triol (10) (Nakai et al. 2005b). the aerial parts of the plant，w hich were iden-
Although many studies have been carried tified by Ms. Shiba，o n the basis of ITS
out on the above-mentioned pharmacologi- sequence data. The localities，da te of collec-
cally active compounds，o nly few studies tion，a nd the number of test materials used in
have reported on the botanical species con""' the present study are listed in Table 1. The
taining the active principles and their voucher materials are stored in the herbarium
hybrids. Takeda et a .1(1996b) reported that of the Tsumura Central Research Labora-
A. lancea，A . chinensis，a nd A. koreana， tories，T sumura & Co.，J apan.
which grow in China，h ave large intra-
populational and geographical variations Standard compounds
with regard to the essential oils that chiefly Five polyacetylenes，n amely，5 ，6 ，7 ，8 ，
consist of 2a nd 3. However，t he identifica- and 9w hich were used for HPLC as the stan-
tion of these species at the time of reporting dard compounds，w ere isolated from the
was not supported by ITS sequence data but commercial1ya vailable dried rhizomes of A.
by morphological data. In addition，th e phar- lancea or A. chinensis，a s described in the
macologically active polyacetylenes such as previous reports (Nakai et al. 2003，2 005).
6，7 ，a nd 10，t hat were derivatized from 8
and 9，ha ve received little attention. Sample preparations
In the present study，w e used crude drugs The powdered rhizome (100 mg) was ac-
prep edfrom the dried rhizomes of A. curately weighed，e xtracted with methanol
訂
lancea，A . chinensis， and their hybrids， (l.5m L) for 10 min under sonication，a nd
which were identified based on ITS sequence then centrifuged (3000中m，10 min). The
data (Shiba et al. 2006) and their contents of residue was similarly extracted with metha-
pharmacologically active four sesquiterpenes nol (1.5 mL) for 10 min and centrifuged
1-4 described previously by Takeda et al. twice. Three supernatants were combined，
(1994，1 995a，1 995b，1 996a，1 996b)，a ndin- and the solution was made up to av olume of
vestigated five pharmacologically active exactly 5m Lb y the addition of methano .1A
polyacetylenes 5-9. Furthermore，w e rear- 20-JlL aliquot of the filtrate白atwas passed
ranged the results of sesquite中enesbased on through a0 .45-μm filter was then i吋ected
ITS sequence data and summarized the into the HPLC system.
chemical characterization of the species and
the localities in which these plants contain- HPLC analysis
ing these nine compounds grow. HPLC analysis with LC-I0AD pumps，a n
SPD-MI0AVP photodiode-array detector，
島laterialsand Methods and aC TO-I0A column oven (Shimadzu，
Crude drug. materials Kyoto，J apan) was performed using an ODS
The crude drugs were used for performing column (TSK-gel ODS-80Ts，4 .6 mmi. d. X
qualitative assays of five polyacetylenes; 250 mm，T osoh，T okyo，J apan). The f10w
these drugs were prepared from the same rate and the column temperature were 1.0
rhizomes， namely， those collected from ml/min and 40oC，r espectively. The UV
April2006 Joumal of Japanese Botany Vo .l81 No. 2 65
匂
H
1: elemol 2: hinesol
3: s-eudesmol 4: atractylon
。
5: atractylodin (R = H) 'R
6: atractylodinol (R OH)
ニ
7: acetylatractylodinol (R OAc)
ニ
R10
\
\
8:R1ニ isovaleryloxyR2= H OH
1 2
9: R = H R = isovaleryloxy
Fig. 1. Structures of compounds: 1，el emol; 2，hi nesol; 3，s -eudesmol; 4，at ractylon; 5，at ractylodin;
6，a tractylodinol; 7，a cetylatractylodinol; 8，3 -isovaleηloxy-tetradeca-4ム12-triene-8，10-diyne-
1，14-diol; 9，at ractyloyne.
data-recorded from 200 to 400 nm-oft he ef- The solvents employed were 20 mMp hos-
fluent from the column were collected，a nd phoric acid (A) and acetonitrile (B)，a nd the
the peak analysis and assignment were per- analytical conditions were as follows: a
formed using the system analysis software linear gradient of 50 % A and 50 % B that
CLASS-LCI0 (Shimadzu， Kyoto， Japan). changed over ap eriod of 60 min to 0 % A
66 植物研究雑誌第81巻第2号 平成18年4月
Table 1. Localities，d ate of collection，t he number of test materials，a nd identification results based on ITS se-
quences data of Atractylodes lancea，A. chinensis and hybrids
Date of Total number
Localities Abbreviation collection of plants A. lancea A. chinensis Hybrids
Xuebu，Ji ntanxian，J iangsu prov. Xu Oct. 15，1 991 16 11 5
Liyan，Ji angsu prov. Li Oct. 27，1 990 12 7 5
Lianyungang，J iangsu prov. Yu Oct. 18，1 990 6 6
Huangshan，T aiping，A nhui prov. Hu Oct. 13，1 991 23 20 3
Yingshan，H ubei prov. Ta Oct. 4，1 991 4 4
Suizou，H ubei prov. Ca Oct. 1，1 991 6 6
Danjiangkou，H ubei prov. Wu Sep.29，1 991 15 4 11
Ludiang，H ubei prov. Lu Sep. 28，1 991 27 3 2 22
Zanhuang，H ebei prov. Za Sep. 1，1 992 12 12
Chengde，H ebei prov. Da Sep. 11，1 992 37 35 2
Chongli，H ebei prov. Ch Sep. 8，1 992 14 14
Taian，S handong prov. Tt Sep. 18，1 992 17 16
Qingdao，S handong prov. La Sep. 16，1 992 25 20 5
Songshan，H enan prov. So Sep. 23，1 994 15 14
Lushixian，H enan prov. Ba Sep. 13，1 994 42 21 21
Linshi，H enan prov. Sh Sep. 20，1 994 38 38
Huayin，S haanxi prov Hs Sep. 17，1 994 41 41
Changan，S haanxi prov. An Sep. 15，1 994 7 6
357 75 194 88
，一not estimated.
and 100 % B. The amount of each compound Results
in the methano1 extract was calcu1ated based Interspecies comparison of sesquiterpenes
on the peak area at 314 nm for 8a nd 9，at contents
336 nmf or 5a nd 6，a nd at 370 nmf or 7. Tab1e 2s hows the mean contents and de-
tectab1e frequencies of the four sesquiter-
Statistical analysis penes in the dried rhizomes of A. lancea，A.
The principa1 component ana1ysis (PCA) chinensis，a nd their hybrids，w hich were
and the test of significance were carried out identified by ITS sequence data. The data
using Microsoft Exce1 with an add-in pro- presented in Tab1e 2r epresent the outcome
gram for statistica1 ana1ysis (Excel statistics of rerangingthe data that had been re-
紅
2002) provided by the Socia1 Survey ported previous1y，b ased on ITS sequence
Research Information Co. Ltd.， Tokyo， data. First，A. lancea was compared with A.
Japan. In addition，n umerica1 ana1ysis by the chinensis. The mean contents of 1 and 4
Maha1anobis-Taguchi (MT) method was per- were significant1y higher in A. chinensis than
formed using Microsoft Exce1 with an add-in in A. lancea， whereas that of 2 was
program (MTS for Windows) provided by significant1y 10wer in A. chinensis (each P<
Ohken Co.，T okyo，Ja pan. The methodo1ogy 0.01). However，t he detectab1e frequencies
for numerica1 ana1ysis using the MTm ethod of 1-4 were not accepted as ad istinct differ-
has been described in our previous reports ence in the interspecies comparison. The hy-
(Nakai et al. 2005a). brids were then compared with A. lancea and
A. chinensis. The mean contents and the fre-
quencies of the hybrids showed on1y minor
differences from both species. On ana1yzing
Apri12006 Journal of Japanese Botany Vol. 81 No. 2 67
Table 2. Mean contents (%) and detectable frequencies (%) of sesquite中enesobtained from the dried rhizomes of
Atracかlodesspecies
Components 1 2 3 4
A. lanceα 0.077 ::t 0.110 1.574 ::t 1.970 1.396 ::t 1.544 0.343 ::t 0.431
n = 75 (37) (63) (72) (49)
A. chinensis 0.172::t 0.170* 0.437 ::t 0.695* 1.148 ::t 0.898 0.126::t 0.245*
n = 194 (64) (62) (96) (32)
hybrid 0.138::t 0.119 1.582 ::t 1.397 1.682 ::t 1.337 0.181 ::t 0.335*
n = 89 (65) (79) (88) (29)
Values are represented as mean ::t S.D. of 357 rhizomes and the detectable frequencies of the 357 rhizomes are pro-
vided in parentheses. *p 0.01compared with A. lancea (Dunnet test).
く
the four sesquiterpenes，a lthough it was ob- 95 %， respectively). In A. lanca，w here 8a nd
served that significant difference was accept- 9c ould be detected，t he mean contents of
able with regard to the components 1，2 ，a nd these compounds were 0.021 % and 0.037
4i n A. lancea and A. chinensis，t he hybrids %， respectively，a nd these values were simi-
showed only slight differences from both lar to those of A. chinensis. Since the
specles. detectable frequencies of 8a nd 9w ere low，
the mean contents of these constituents were
Interspecies comparison of polyacetylene also considered to be low. Typical HPLC
contents profiles of A. lancea and A. chinensis are
Table 3s hows the mean contents and de- shown in Fig. 2. Next，th e hybrids were com-
tectable frequencies of the five pared with A. lancea and A. chinensis.
polyacetylenes in methanol extracts of the Although the mean contents of the constitu-
dried rhizomes of A. 1αncea，A . chinensis， ents 5-7 in the hybrids differed slightly from
and their hybrids，w hich were identified by those of A. lancea and A. chinensis，t he de-
ITS sequence data. First，A. lancea was com- tectable frequencies of 8 and 9 were not
pared with A. chinensis. With regard to the similar to those of A. lancea and A. chinensis
mean contents of 5-7 in A. lancea and A. and showed av alue of approximate1y 70 %.
chinensis， the range of variation in the Along with compound 8，c ompound 9c ould
interspecies comparison was small，w hereas also be detected in all the hybrids. In the hy-
the contents of 8 and 9w ere significantly brids，th e mean contents of these constituents
higher in A. chinensis than in A. lancea (P < were 0.026 % and 0.044 %， respectively，an d
0.01). The detectable frequencies of 5-7 these values were similar to those of the two
were greater than 80 % in both the species， species. The hybrids were specifically sepa-
whereas the detectable仕equenciesof both 8 rated into two groups with regard to the de-
and 9s howed aw ide range of variation in tectable frequencies of 8a nd 9. The analysis
the interspecies comparison. Besides，t he of the five polyacetylenes showed that the
constant correlation between 8a nd 9w as ob- detectable frequencies of 8a nd 9d iffered in
served as follows: 9c ould be detected with- the two species and their hybrids.
out reference to the species whenever 8
could be detected. The detectable frequen- Chemical characterization of the two spe-
cies of compounds 8a nd 9i n A. lancea were cies and their hybrids
approximately 9 % and were considerably To comprehensively characterize the two
lower than those in A. chinensis (88 % and species and their hybrids with regard to their
68 植物研究雑誌第81巻第2号 平成18年4月
Table 3. Mean contents (%) and the detectable frequencies (%) of polyacetylenes in the dried rhizomes of
Atrac砂lodesspecies
Components 5 6 7 8 9
A. lancea 0.137 :t 0.108 0.046 :t 0.056 0.034 :t 0.045 0.002 :t 0.010 0.006 :t 0.029
n = 75 (100) (100) (83) (9) (9)
A. chinensis 0.126 :t 0.092 0.062 :t 0.062 0.034 :t 0.046 0.024 :t 0.024* 0.120:t 0.111*
n = 194 (100) (100) (93) (88) (95)
hybrid 0.096 :t 0.102 0.081 :t 0.065* 0.032土0.035 0.018 :t 0.022* 0.078 :t 0.084*
n = 89 (100) (100) (92) (70)
Values are represented as mean :t S.D. of 357 rhizomes and the detectable frequencies of the 357 rhizomes are pro-
vided in parentheses. *p <0 .01 compared with A. 1αncea (Dunnet test).
constituents，P CA was applied to the con- A. lancea and A. chinensis hybrids have been
tents of the four sesquiterpenes (1-4) and the plotted as small symbols. The hybrids did
five polyacetylenes (5-9) determined by GC not possess the intermediate chemical char-
and HPLC， respectively. PCA was per- acteristics between A. lancea and A.
formed using the data of 357 rhizomes ob- chinensis; the variation was observed within
tained from the two species and their the limits of the characteristics that were ob-
hybrids，a nd it yielded two new principal served in both the species.
components. The individual scores of A.
lanceα，A. chinensis，a nd their hybrids based Local variations in the contents of poly-
on the two new variables (PC 1 and PC2) acetylenes and sesquiterpenes
were plotted as shown in Fig. 3. Although Since the results of PCA showed ac hemi-
these scores suggested the division of A. cal variation in A. lancea，A. chinensis，a nd
lanceαindividuals into two groups，P CA en- their hybrids，t he relationship between the
abled the chemical differentiation between A. chemical variations and localities was nu-
lancea and A. chinensis. The eigenvectors of merically analyzed. Although various pattern
each chemical component that affects PC1 information processing techniques have been
and PC2 eshown in Fig. 4. It was sug- used for the numerical analysis，t he MT
紅
gested that the chemical component 4w as method，w hich represents the coincidence of
mainly responsible for the characterization the pattem from the standard group (unit
of PCl in the negative direction，w hereas the space) as the Mahalanobis distance，w as
chemical components 2( positive vector) and used. A unit space (locality in which So is
both 8a nd 9( negative vectors) were mainly found) was constructed as the standard group
responsible for the characterization of PC2. based on the regions described in the a
These results suggest that constituents 2a nd ailable Jap anese literature. The distances be-
4a re typically involved in the determination tween each rhizome from 17 localities were
of the chemical characteristics of A. lancea determined by the inverse matrix method.
and enable the classification of A. lancea The chemical components were selected
into two groups，wh ile the incidence of com- from each locality and classified based on
pound 8a nd 9e nable the chemical differen- the differences in the contribution ratio of
tiation of A. chinensis. Additionally，t hese the signal-to-noise ratio and sensitivity of the
speculations were well supported by the re- nine components. As a result， it was
sults presented in Tables 2a nd 3. Further，as observed that the localities could be roughly
shown in Fig. 3，th e individual scores of the classified into four groups based on the con-
〉
宮
・
口
。。。M
旬。
zgmw-
。
?
『
岳
山
口
2
0
回。
S
E
U『
〈
。
戸
∞
H
。・Z
M
(min)
Fig. 2. HPLC profiles of Atractylodes lancea (top) and chinensis (bottom). 5，at ractylodin; 6，at ractylodinol; 7，ac etylatractylodinol; 8，3 -isovaleryloxy-
tetradeca-4，6，12 -triene-8，lO -diyne-l，14 -diol; 9，at ractyloyne.
、。
。、
70 植物研究雑誌 第81巻 第2号 平成18年4月
6
ロ
町田 《
M
@ g
4
@ち ∞ ω
-1. ち Z
自t • (
(ま2o ".E さ0)g渓 AIと。a=
m hr. {z
.@村)nt @85~F 。 SEEOo言。 .コ.J '‘-OJ)
色NU SZE ω.c
o '"" 8s
・ミ 。 円- =
0 。 υ- 3:
。
4 a
-6
ω
4 -2 0 2 4 6 8
PC1. (44.1. %) N12
a
Fig. 3. Chemical characterization of the Atractyloses ca
0
lancea，A. chinensis，a nd their hybrids using PCA -
results of the content data of the nine compo-
トー
nents: (・)A. lancea; (0) A. chinensis; and (・)A.
lancea and A. chinensis hybrids.
伺
4
長1=
箇。6 a
-2 コ ‘02-
〉 M
0.4
コ
NU 0.2 ・-3
nhO』 。 7 -6 ト圃偲
L -
F a
o-3ωm』 Zコ =。』
-4 -1 国
Zω〉 0.2 旬。
四
ω四一 -5 凶 o It) C」
ー0.4 ・ .4n "rI 0
9• 8 (%) SlU司UO:J
。
ー0.6
-0.4 -0.2 0.2 0.4 0.6
Fig. 5. The contents of 4( atractylon) and the total
eigenvectors of PC1 contents of 8 and 9 (3-isovaleryloxy-tetradeca-
4，6，12 -triene-8，1O - diyne-1，14 -diol and atractyl-
oyne，r espectively) in the dried rhizomes of
Fig. 4. Eigenvectors for PC1 a nd PC2 using the PCA
Atracηlodes plants collected from 18 areas. Each
results of the content data of the nine compo-
column represents mean::f: S.D.
nents. 1，e lemol; 2，h inesol; 3，s -eudesmol; 4，
atractylon; 5，a tractylodin; 6，a tractylodinol; 7，
acetylatracty lodinol; 8，3- isovalery loxy -tetradeca-
4，6，12 -triene-8，1O -diyne-1，14 -diol; and 9，a trac-
tyloyne.
Apri12006 Journal of Japanese Botany Vo .l81 No. 2 71
tents of 4a nd the total contents of 8a nd 9. that the component contents of the two spe-
The contents of 4a nd the total contents of 8 cies and their hybrids possessed the chemical
and 9i n each locality in the four groups are characteristics in every area. Moreover，it
shown in Fig. 5. The localities in the four was possible to roughly classify the localities
groups，n amely，g roup 1( 口:Ca，H u，a nd Ta)， into four groups on the basis of the contents
group II (0: Li，Y u，X u，a nd La)，g roup III of 4a nd the total contents of 8a nd 9a s an
(・:Tt，D a，Z a，C h，a nd So)，a nd group IV index. Further，th e hybrid was detected in all
(・:Wu，S h，L u，B a，H s，a nd An) are shown the four groups，a nd it could be concluded
in the map (Fig. 6). Both the closed symbols that the chemical characteristics of the
(・ and・)indicate the localities in which hybrids depend on the local variation.
the component. 8o r 9w as detected，w hile
both the open symbols (口 and0) indicate Discussion
the localities in which the components 8a nd In the present study，w e used crude drugs
9 were not detected. Similarly，b oth the prepared from the dried rhizomes of A.
square symbols (・ and口)indicate the 10- lancea，A . chinensis， and their hybrids，
calities in which component 4w as detected， which were identified using ITS sequence
while the circular symbols (・ and0) indi- data and their contents of four pharmacologi-
cate the localities in which component 4w as cally active sesquite中enes1-4 described
not detected. Thus，it could be concluded previously by Takeda et al. (1994，1 995a，
つ
1I11Jf
/rf|/\'\//ノ
汁へ~T fjj) rI .:. ";，._.(.可 j.J"--，
レ叫'i.ゐr ヂT\ k ヘ、;> Da 、、
__---\ト 1~~ ) :イ目\
w r・
Hebei '---'~~:
，:/ ! t___\..... :~，' \\ ~ ~ \'1 Za，j
.......、. ".-、.~句..，::-一..~-"
\\\jfγ一-1.{ 〈「ノ() .
\ ・，/ノShandong
L\)一「，、j'，--(-)'Hsj (，/--'---ノ J.，-----イ，<.，，':，----'-i.，IーY/u
」・\-.
くSh制nx 50 J<~~\ \'--"ーすiangsu
.Sh
、:';fム 咽.Ba P Aヘ、\圃n "r-{' r，戸\一r一一:._.一'-.~-f一---(cid:631) ，叫(cid:631).. 、W.圃....，.‘.. ".固 、..u1L'"、u\、口\C百"、-一a(cid:631)へ、-- ，、\、"，(cid:631)ペ，、寸〆一-、 u山I \\Lし戸ρT--..'.L....i.'.ノ、、点/
，_---/) H山 i 一百\日u口f
--ノ L，'ーγ
\、.'/..-......._5
-......_..."\....，'..~..""\/.....，ノ\
1 1 f
‘1
，，副旬 ，
500 km
Fig. 6. Local variation in the components of Atracηlodes plants examined.
Symbols and abbreviations indicate the localities and their names: (口)
Group 1; (0) Group II; (・)Group III; and (園)Group IV.
72 植物研究雑誌第81巻第2号 平成18年4月
1995b，1 996a，1 996b)，a nd investigated five gest that compounds 2a nd 4 etypically in-
紅
pharmacologically active polyacetylenes volved in the determination of the chemical
5-9. Furthermore，w e rearranged the results characteristics of A. lancea and enable the
of sesquite中enesbased on ITS sequence classification of A. lancea into two groups，
data and summarized the chemical charac- while compounds 8a nd 9e nable the chemi-
terization of the species and the localities in cal differentiation of A. chinensis. Additio-
which these plants containing these nine nally，th ese speculations were well supported
components grow. by the results of the mean contents and de-
A comparison of the sesquiterpenes con- tectable frequencies of each component.
tents in A. lαncea and A. chinensis showed Although ac omparative study of A. lancea
that significant differences were present in and A. chinensis has been performed with a
their contents of 1，2 ，a nd 4. Although some specific focus on essential oils，th e results of
comparative studies of A. lancea and A. sesquiterpenes along with those of poly-
chinensis with af ocus on essential oils have acetylenes enabled the recognition of the two
been performed (Nishikawa et al. 1976b， species in this study.
Yasuda et al. 1996，T akeda et al. 1996b，Ji Moreover. these chemical characteristics
et al. 2004)，th e results of the analysis of the of the A. lancea and A. chinensis hybrids，
two species identified using ITS sequence which was identified by ITS sequence data，
data revealed as ignificant difference in these were investigated for the first time. Although
three compounds. Although several studies the components 1-7 detected in the hybrids
have reported on the detection of 4-a charac- were common to A. lancea and A. chinensis，
teristic component of A. japonica and A. the detectable frequencies of 8a nd 9i n the
ovatα-in A. lancea and A. chinensis hybrids differed from those observed in the
，
(Nishikawa et al. 1976b， Takeda et al. other species. The result of the PCA，i n
1996b，Ji et al. 2004)，w e observed that the which the contents of the nine components
compound was detected in the two species were used as variables，w as not helpful in
and their hybrids as distinguished by ITS identifying the chemical characteristics of
sequence data. The analysis of poly- the hybrids. It was recognized that the
acety lenes showed that compounds 8a nd 9 hybrids did not possess the intermediate
were considered to be specific and were not chemical characteristics between A. lancea
detected in al arge population of A. lancea， and A. chinensis， and a variation was
but were detected in A. chinensis although accepted within the limits observed in both
polyacetylenes 5-7 were detected as com- the species.
mon costituents in the two species. In order to investigate the chemical varia-
Therefore，it is reasonable to presume that tion in the two species and their hybrids，th e
compounds 8a nd 9m ay serve as one of the relationship between the chemical variations
indices for the differentiation of A. chinensis and localities was numerically analyzed. The
from A. lancea. results of the MTm ethod suggested the clas-
To chemically characterize the two sification of the examined localities in China
species，P CA was applied to the relative con- into four groups based on the total contents
tents of above-mentioned nine compounds. of 8a nd 9a s well as the content of 4. These
Although these scores suggested the division results also supported the local grouping
of A. lancea individuals into two groups， based on 4，w hich was reported by Takeda
PCA enabled chemical differentiation et al. (1995b)，a nd facilitated am ore detailed
between A. lancea and A. chinensis. classification;
Furthermore，t he resultant eigenvectors sug- Generally， qualitative and quantitative
