全 文 ：Factors affecting induction of shoot cluster of Artemisia annua
GENG Sa
1 , JI Sheng-dong
1 , YU AN Jin-yun
1 , LU Long-dou
1 , Ye He-chun
2 , L I Guo-feng
2
( 1. Co lleg e of Life Science, Henan No rmal Univ er sity , Xinx iang 453002, China; 2. Resea rch Cente r of Plant Molecular
and Developmenta l Bio lo gy , Institute of Botany , Chinese Academy o f Sciences, Beijing 100093, China )
Abstract: Object　 To study the facto rs af fecting the induction of shoo t cluster of Artemisia annua.
Methods　 Conventional plant tissue culture tech nolog y w as applied to regula te the biosynthetic
metabolism of seconda ry metabolism product, ar temisinin of A. annua. Results　 Effects o f genotype, ho r-
mones, basal medium on the induction of A . annua shoo t cluster were no table, w hile tha t of light intensi ty
ranging betw een 1 000— 6 000 lx and tempera ture ranging betw een 20℃— 30℃ were no t. Among the fiv e
A. annua st rains used, the induction ra tio of st rain 025 w as the best. The optimal concentration fo r induc-
tion of shoot cluster w as 6-BA 2. 0 mg /L and N AA 0. 15 mg /L. Ions w ere also found to be very impo rtant
in the shoot cluster induction and the biosynthesis of artemisnin. Conclusion　 Effectiv e regula tion of in-
duction of A. annua shoo t cluster and artemisinin biosynthesis can be carried out via the regulation of
physical and chemical facto rs.
Key words: Artemisia annua L. ; shoo t cluster; induction ratio; a rtemisnin
青蒿丛生芽诱导影响因素的研究
耿　飒 1* , 姬生栋 1 , 袁金云 1 , 卢龙斗 1 , 叶和春2 , 李国凤 2
( 1. 河南师范大学生命科学学院 ,河南 新乡　 453002; 2. 中国科学院植物研究所植物分子发育研究中心 ,北京　 100093)
摘　要: 目的　对影响青蒿丛生芽诱导因素进行基础性研究。 方法　把常规的植物组织培养技术应用于调控青蒿
中次生代谢产物青蒿素的生物合成代谢。结果　青蒿的基因型 ,激素和基本培养基对丛生芽的发生有显著影响 ,而
光强在 1 000～ 6 000 lx和温度在 20℃～ 30℃对丛生芽的发生影响不大 ;在 5种基因型的青蒿中 , 025丛生芽的诱
导率最高 ;诱导丛生芽的激素组合是 6-BA 2. 0 mg /L和 NAA 0. 15 mg /L;另外 ,离子在青蒿丛生芽的诱导和青蒿
素的生物合成过程中起着非常重要的作用。 结论　组织培养条件下 ,青蒿丛生芽的诱导及青蒿素的生物合成可以
通过理化因子有效地进行调控。
关键词: 青蒿 ;丛生芽 ;诱导率 ;青蒿素
中图分类号: R286. 02　　　文献标识码: B　　　文章编号: 0253 2670( 2004) 05 0566 07
1　 Introduction
In recent y ears, applied studies have been
more and mo re ex tensiv ely used in plant bio tech-
nolog y , so as to modi fy th e plant characteristics,
especially medicinal plants
[1, 2 ] . Artemisinin is a
colo rless, needle fo rm o f crystal, i t s melting point
is 156 ℃— 157 ℃ , it s molecular fo rmula is
C15 H22O5 , and i t i s a sesquiterpene lactone wi th a
peroxy g roup
[3 ]
, which is easy to decompose by
heat. Artemisinin has characteristics of fast cura-
tiv e effects and low side-ef fect in malaria
[ 4] . Cur-
rent ly , there are tw o billion people suf fering f rom
the infection of malarial parasi tes every year in the
w orld. In t ropical regions, especially in the South-
easter n Asia and Western Africa, chlo ro-quine-re-
sistant malaria had become the great enemy of pub-
lic h ealth and has also become the main cause of
death
[5 ] . Artemisinin has been a recommended
medicine by the World Heal th Orga- nization.
Artemisinin also is immunode-pressiv e, cell im-
muno logical enhancing , and an effector of anti-in-
fluenza v irus[4 ] .
·566· 中草药　 Chinese T raditional and Herbal Drug s　第 35卷第 5期 2004年 5月
收稿日期: 2003-09-15基金项目:国家“九五”攻关项目 ( 96-C02-03-02)作者简介:耿　飒 ( 1971— ) ,男 ,副教授 ,博士 ,主要从事药用植物组培及分子生物学研究。
* 通讯作者　 E-mai l: gengs a@ yahoo. ca　 Fax: ( 0373) 3326524
Recently , medicinal artemisinin w as direct ly
ex t racted f rom the leaves and buds of Artemisia an-
nua L. The ex t ractiv e w as done w ith complex
links, taking much time and g rea t ef fo rt: the qual-
ity of A. annua in di fferent regions also made a
g rea t di fference. Although artemisinin could be
synthesized a rtificia lly, i t could not be put into
production due to high cost and st rong side-effect.
The methods of callus and cell cul ture have been
tested, but the synthesis of artemisinin, because of
the nondi fferentia tion status of i ts cells, could only
be achieved af ter tissue dif ferentiation. The experi-
mental resul ts w ere disappointing [6 ] . Woerdenbag
used buds fo r cultiva tion. The content of
a rtemisinin could be up to 0. 08% o f dry w eight of
the cul ture, but the g row th cycle w ould last
w eeks
[7 ] . In this paper, by using the explants of
A. annua , the shoo t cluster cul ture system w as es-
tablished and also the existence o f artemisinin in
the cul tures w as detected, w hich probably w ill be
a new method for the indust riali zed production of
artemisinin.
2　Materials
Plant materials. High a rtemisinin producing
strains 001, 014, 021, 025, 032 w ere collected
f rom Sichuan Province of China. The seeds w ere
germinated in aga r aseptically and sterile seedling s
of A. annua were cultured on the Murashige and
Skoog
[8 ]
basal medium with 0. 7% Sigma agar con-
taining 3% sucrose in g row th chambers at 26℃
fo r 16 h day leng th. Sterile seedlings w ere used fo r
inducing shoot clusters in M S basa l medium sup-
plemented w ith ho rmones in di fferent concen- tra-
tions ( Table 1) .
Table 1　 Effect of plant hormones in diff erent concentration induced ratio of A. annua on multiple shoots
Hormones / ( mg· L- 1 )
6-BA KT ZT 2, 4-D N AA IBA
Ex plantsa and mul tiple sh oots formation explan ts w ith shoots /% b
LDc L LS SF
　　 0. 0 　　 0 　　 0 　　 0 　　 0 　　 0 　　　　 0 　　　　 0 　　　　 0 　　　　 0
0. 4 0 0 0 0. 8 0 75. 2 68. 6 76. 0 48. 2
0. 8 0 0 0 0. 4 0 89. 1 80. 3 91. 1 56. 5
1. 0 0 0 0 0. 2 0 93. 5 85. 8 94. 4 63. 3
2. 0 0 0 0 0. 15 0 96. 8 89. 4 95. 8 68. 7
2. 0 0 0 0 0. 1 0 97. 4 92. 5 97. 8 74. 2
2. 0 0 0 0 0. 05 0 98. 7 93. 9 99. 8 76. 1
2. 0 0 0 0 0 0 79. 2 72. 5 83. 4 50. 0
1. 0 0 0 0 0 0 48. 2 44. 7 55. 7 38. 1
0 1. 0 0 0 0. 05 0 72. 1 65. 2 76. 0 55. 0
0 1. 0 0 0 0. 5 0 68. 4 61. 3 71. 8 46. 3
0 2. 0 0 0 0. 1 0 62. 2 54. 3 66. 6 42. 2
0 4. 0 0 0 0. 2 0 53. 2 48. 9 55. 4 38. 9
0. 8 0 0 0. 1 0 0 86. 3 81. 5 89. 5 75. 8
1. 0 0 0 0. 5 0 0 78. 5 72. 3 81. 7 67. 4
2. 0 0 0 1 0 0 66. 2 62. 8 69. 9 57. 8
0 0 1. 0 0 0. 1 0 80. 2 76. 4 82. 5 64. 1
0 0 2. 0 0 0. 2 0 78. 5 73. 1 80. 6 69. 7
0 0 4. 0 0 0. 5 0 74. 1 70. 4 78. 0 65. 8
1. 0 0 0 0 0 0. 5 56. 3 54. 0 59. 7 50. 9
1. 0 0 0 0 0 1. 0 45. 2 40. 2 49. 8 36. 3
2. 0 0 0 0 0 0. 5 60. 9 57. 4 63. 3 55. 4
2. 0 0 0 0 0 1. 0 58. 4 57. 6 64. 1 56. 5
　　　　 a All result s w ere analyzed f rom 200— 300 explants　 b Data w ere tak en af ter 4 w eeks of culture in ev ery case
　　　　 c LD, L, LS, and SF represented leaf di sc, leaves, leaf w i th leaf s talk and stem fragmen t
3　Methods
3. 1　 Measurement of shoo t cluster g row th: Four
shoot clusters, about 0. 5 g ( f resh w eight , FW )
induced in the same condi tion w ere inoculated into
each triangle flask w eigh ted in advance (W 1 ) . The
t riang le flask af ter inocula ting was w eighed (W 2 ) ,
the dif ference of the tw o w eigh t is that of ino-cu-
lation material (ΔW = W 2 - W 1 ) . FW o f shoo t
cluster wa s obtained in the same g row th condi tion,
the fresh shoot cluster was dried at 50℃ and oven
·567·中草药　 Chinese T raditional and Herbal Drug s　第 35卷第 5期 2004年 5月
dried to constant w eight , and the dry w eight ( dry-
ing weight , DW) was got so the DW of the inocu-
lation could come up in the ra tio o f FW to DW
( FW /DW) . The shoot clusters w ere sub cul- tured
fo r 4 w eeks later and ha rv ested after cultured in
the subculture medium fo r 6 w eeks.
　 　 Accounting method fo r the shoot clusters
g row th:
　　 Grow th increm ent ( GI, g /flask ) = Harvest w eigh t
( HW , g /flask, DW or FW) - Inocula ting w eight ( IW , g /
fla sk, DW or FW)
3. 2　 Measurement of light and tempera ture effect
on induction o f shoo t clusters: Solid induction
medium 30— 40 mL was contained and 28 leaf discs
w ere inoculated in each 90 mm cultiv ation disc.
1. To leave the disc in g row th chamber a t ( 25± 1)
℃ fo r 16 h day leng th wi th 1 000, 2 000, 3 000,
4 000, 5 000, and 6 000 lx light intensity and dark-
ness sepa rately , then analy ze the ef fect of lig ht on
shoot cluster induction ra tio; 2. To leave the disc
in g row th chamber at 3 000 lx light intensi ty fo r 16
h day leng th at 15℃ , 20℃ , 25℃ , 30℃ , 35℃ ,
40℃ separately, then analy ze the ef fect of temper-
ature on shoo t cluster induction ratio. Best grow th
o f 4 w eeks o ld shoot clusters w ere used fo r subcul-
ture test.
3. 3　 Measurement o f di fferent facto rs in the effect
on g row th and artemisinin biosynthesis of shoot
clust res: Clusters 3— 4 of induced shoot cluster
w ere inoculated into so lid subculture medium with
di fferent combina tion. Cul tiv ated in grow th cham-
ber a t ( 25± 1)℃ fo r 16 h day leng th wi th 1 000 lx
ligh t intensi ty. Ef fect on the g row th o f shoo t clus-
ters w as concluded acco rding to the GI dif ference.
In the same time, content o f artemi- sinin wa s
measured. So di ffer- ent factors in dif ferent con-
centration af fecting the g row th and artemisinin
biosynthesis could come out.
3. 4　 Measurement of a rtemisinin content: UV
a rtemisinin determina tion w as perfo rmed as des-
cribed by Shen et al [ 9] .
4　 Results
4. 1　 Effects of dif ferent ho rmone concentra tion on
induction ratio of shoo t clusters: Put leaf discs,
leav es, leaves wi th stems and stem fragments f rom
seedlings of 2— 3 w eeks old into the M S medium
supplemented w ith di fferent hormones a t dif ferent
concentration ( Table 1) . Resul ts indicated that
shoo t cluster induction ratio w as best using 6-BA
and N AA combination to di fferent explants. Re-
sults also told us that shoo t clusters induction ratio
could reach to 98. 7% , 97% , 96. 8% when 6-BA
2. 0 mg /L w as combined wi th N AA 0. 05 mg /L,
N AA 0. 1 mg /L, and N AA 0. 15 mg /L, respec-
tiv ely. Further study show ed that g row th of shoot
cluster was good, but v erification a t di ffer-ent de-
grees was found when concentration varied ( Fig.
1) . Veri fication ratio of shoo t cluster w as about
15. 4% , 10. 5% , and 5. 4% when N AA concentra-
tion w as 0. 05, 0. 1, and 0. 15 mg /L res-pectiv ely.
So combination of 6-BA 2. 0 mg /L and N AA 0. 15
mg /L w as preferred w hen induc- tion and verifica-
tion ra tio w ere taken into account. In this paper,
M S medium supplemented w ith 6-BA 2. 0 mg /L
and N AA 0. 15 mg /L was called M-I1.
1-ex plants wi th sh oots /%　 2-vit rifi ed sh oots /%
Fig. 1　 Ef fects of dif ferent NAA concentration
in combination with 6-BA at 2. 0 mg /L
on shoot cluster induction ratio and
vitr if ied shoot ratio of A. annua
4. 2　 Effects of A. annua with dif ferent g enotype
on induction ratio of shoot clusters: Put leaf disc
explants of di fferent geno type at same age into
M -I1 medium , the results w ere included in Fig. 2,
Fig. 2 rev ealed tha t shoo t cluster induction ra tio of
025 st rain could be 98% , that o f 032 line about
80% , and that of the three o ther lines betw een
60%— 70% . Resul ts indicated that th e same genus
plants wi th di fferent g eno type possessed dif ferent
perception ability to the same exogenous ho rmone.
·568· 中草药　 Chinese T raditional and Herbal Drug s　第 35卷第 5期 2004年 5月
So in o rder to g et ef ficient induction ra tio of shoot
cluster, di fferent ho rmone combina tion should be
used to induced shoo t cluster acco rding to different
g eno type plant o f the same genus.
Fig. 2　 Shoot cluster induction rat io of A. annua
with dif ferent genotype
4. 3　 Effects of dif ferent explant ag e on induction
ra tio of shoo t clusters: Put leaf disc explants f rom
di fferent part of 025 line o f 1— 8 w eeks o ld, re-
spectiv ely into M-I1 medium, the resul ts dis-
played in Fig. 3. Fig. 3 demonst ra ted tha t induc-
tion ratio o f shoo t cluster in M-I1 medium could
reach up to mo re than 95% using explants betw een
1 and 8 w eeks old, which indicated tha t the age of
seedling s had less effect on the induction o f shoot
clusters. Fig. 4 a and b demonst rated tha t induc-
tion ratio w as the same, almost 100% , using
young and o ld seedling s as explants.
Fig. 3　 Effects of culture time of explant plant on
shoot cluster induction ratio of A. annua
4. 4　 Effects o f light and temperature on induction
ra tio o f shoo t clusters: Laid leaf disc explants on
M-I1 medium in dif ferent culture condi tion f rom
darkness to 6 000 lx light intensi ty , at 25℃ , ef-
fect o f ligh t intensi ty on induction ratio of shoot
cluster w as reco rded in Fig. 5. Results indicated
that shoo t cluster induction ratio of M-I1 medium
was almost dependent on light intensity. Just
w hen in darkness, induction ratio o f shoot cluster
a. shoot clus ter ind uct ion f rom leaf dis c explan ts aged 3 w eeks;
b. s hoot cluster induction f rom leaf di sc ex plants aged 8 w eeks;
c. s hoot clus ter induced in light; d. sh oot clus ter ind uced in
dark nes s; e. sh oot cluster induced f rom leaf di sc of s eed lings
w i th no rmal cul tu re condi tion; f. shoot cluster induced f rom
leaf dis c of seedlings w i th cont inuous ligh t cu lture condi tion; g.
s hoot clus ter induced f rom leaf di sc of seedlings t reated at nor-
mal ligh t leng th anew af ter con tin uous ligh t cul ture
Fig. 4　 Shoot cluster induction
was a li tt le low er than tha t induced in light , also
could be more than 80% . Shoot cluster induced in
da rkness g rew faster than that in light , but shoo t
clusters in da rkness w as slimmer and each shoo t
cluster contained few er li t tle shoo t compared w ith
tha t of in ligh t ( Fig. 4 c and d) . Also, laid leaf
disc explants on M-I1 medium in di fferent cul ture
·569·中草药　 Chinese T raditional and Herbal Drug s　第 35卷第 5期 2004年 5月
condition f rom 15℃ to 40℃ , in 6 000 lx light in-
tensi ty , ef fect of tempera ture on induction ratio of
shoot cluster w as recorded ( Fig. 6 ) . Results
show ed that too high or low temperature had
st rong effect on induction ra tio of A. annua shoot
cluster, while the induction ratio was the highest ,
up to 98% , at 25℃ . Further research indicated
that v aria tion ratio of± 3℃ had li t tle effect on the
induction o f shoo t cluster.
Fig. 5　 Effects of light intensity on shoot
cluster induction ratio of A . annua
Fig. 6　 Ef fects of temperature on shoot cluster
induction ratio of A. annua
4. 5　 Effect of explants from seedling s t reated in
di fferent light leng th on the shoo t cluster induction
ra tio: Befo re inoculation, put sterile seedlings in
g row th chamber wi th dif ferent ligh t leng th 8, 12,
16, 20 fo r 24 h at 25℃ for 4 w eeks. Result of
shoot cluster induction was included in Table 2.
Table 2 revealed that continuous light on seedling s
could badly rest rain the occurrence of shoot clus-
ters, ev en the 6-BA concentra tion in the induction
medium was increased up to 8 mg /L, but result
w as unal terable. How ever, t reating the seedling s
exposed by long light leng th in the g row th cham-
ber for 2 w eeks at 16 h light leng th, shoo t clusters
could be induced from leaf disc explants ( Fig. 4 e,
f , and g ) . Probable reason o f the result is that
continuous ligh t inhibi ted the expression of cy-
tokinin receptor or recepto r-related gene in A. an-
nua seedling s, so the leaf disc exhibi ted defects in
exogenous cy tokinin-mediated shoo t induction.
Yet w hen no rmal light leng th resumed, the re-
strainabi li ty of cy tokinin recepto r or receptor-relat-
ed gene expression w as released f rom the continu-
ous light and th e shoot cluster could also be in-
duced successfully. In Arabidopsis study , tw o mu-
tants crel and w ol also w ere found to exhibit de-
fects in cy tokinin-mediated shoo t induction f rom
callus and roo t v ascula r mo rpho- genesis
[10, 11 ] ,
w hich demonst rated that ex t racellu- lar cytokinin
could activ ated AHK4, a cy tokinin recepto r. In ad-
dition, explants f rom seedlings in relativ e sho rt
light leng th cul tivation could lead to low induction
ratio of shoot cluster. Maybe genetic, g enomic,
and biochemical experiments wi ll elucidate the de-
tails in cytokinin perception and target g ene ex-
pression essentia l in forma tion of shoo t cluster.
　　　 Table 2　 Ef fect of light cycle on shoot cluster
induction of A. annua
Leaf disc
L L* * * / h
Seedlings LL* * * / h
8 12 16 20 24
8 62* /126* * 79 /130 94 /139 76 /132 0 /132
12 68 /125 86 /132 121 /136 108 /134 0 /135
16 76 /128 92 /129 139 /142 124 /129 5 /178
20 75 /126 93 /130 121 /128 111 /123 2 /154
24 70 /129 89 /125 112 /118 109 /119 1 /128
　　* Inducing m ultiple shoot number; * * Explan t number;
　　* * * LL-light length
4. 6　 Effect of ag ar concentra tion on the induction
of shoot cluster: Agar w as used as not only solidi-
fying material in medium in common use but also
regulato r to regulate the humidi ty in culture flask.
Because, v eri fied shoots w ere detained by aga r in
ti ssue cul ture efficiently th rough regulated humidi-
ty in medium[ 12] . So , in present study , ag ar w as
used as a regulator to regulate the induction,
g row th of shoo t cluster, and ar temisinin biosyn-
thesis. Fig. 7 and Fig. 8 elucidated that 0. 8% a-
gar concentration fav o ri ted the induction and
g row th o f shoo t cluster, no rmal shoot cluster ratio
w as about 95% . When the concentra tion of aga r
w as too high , shoo t cluster w as short of w ater and
veri fication ra tio w as low , moreover, shoo t cluster
w ould sco rch rapidly. When the concentration ra-
tio w as to o low , ra tio of FW /DW would be over
·570· 中草药　 Chinese T raditional and Herbal Drug s　第 35卷第 5期 2004年 5月
20, shoo t cluster show ed w ater- log ged station and
verifi- cation w as serious. Determination of
a rtemisinin suggested that a rtemisinin content be
the highest in 0. 8% agar concentration when shoot
cluster ra tio o f FW /DW was about 10, which indi-
cated tha t bio synth esis o f artemisinin was closely
rela ted to the g row th o f shoo t cluster. It seemed
complica ted w hy veri fica tion w as decrea sed when
the concentration of aga r w as increased. Debergh
et al . [ 12] fo und that the increase o f aga r could lead
to rema rkable increase of Ca
2+
, K
+
, Na
+
, M g
2+
,
Mn
2+
, Cu
2+
.
1-vit rifi ed sh oots / to tal shoots /%
2-normal shoots /total sh oots /%
Fig. 7　 Effect of agar on growth
of A. annua shoot cluster
1-FW /DS　 2-artemisinin conten t
Fig. 8　 Effect of agar concentration on growth
and artemisinin biosynthesis of A. annua
shoot clusters
4. 7　 Effect of pH value on the shoo t cluster induc-
tion and ar temisinin bio synthesis: Inoculated leaf
discs in induction medium with dif ferent pH value,
resul t w as summarized in Fig. 9. Fig. 9 elucidated
that the induction of shoo t cluster w as improved
when pH value w as between 5. 5 and 5. 8. How ev-
er induction of shoo t w as rest rained when pHvalue
w as higher than 5. 8, o r low er than 5. 5. Bio syn-
thesis of artemisinin could be promo ted when pH
va lue o f medium was 5. 8, other pH values inhibi t-
ed artemisinin bio synthesis in differ- ent deg ree,
especially low er than 5. 3.
1-induced ratio of m ultiple shoots /% 2-ar temisinin / ( mg· g- 1)
Fig. 9　 Effect of initial pH value on shoot
cluster induction ratio of A. annua
and artemisinin biosythesis
5　 Discussion
Optimal phy sical and chemical facto rs of af-
fecting the induction o f A. annua shoo t clusters
w ere found in the present study , M S basal medium
supplemented 6-BA 2. 0 mg /L, N AA 0. 15 mg /L,
0. 8% aga r, and pH 5. 8 w as found to be ef ficient
in g row th chamber in 16 h light leng th , 3 000 lx
light intensiy , and at 25℃ . Seedlings used as ex-
plants should be t rea ted in about 16 h light leng th
2 w eeks before induction o f shoot clusters. Only in
this condi tion, could shoot cluster g row w ell and
production of artemisinin also be im- proved. In
previous study, the rule that the biosynthesis of
artemisinin could only be achiev ed after a certain
deg ree of dif ferentiation fo r the A . annua tissue
w as revealed[1, 13 ] . The synthesis of ar temisinin has
been studied in suspension cell
[14 ] , ca llus
[6, 15 ] ,
shoo t[16 ] , and hai ry ro ot cul ture[ 17] . Al l related
studies indica ted that the artemisinin pro- duction
obtained by dedif ferentiated cul tures of A. annua
i s v ery poo r, a certain deg ree of develop-ment is a
prerequisite fo r artemisinin bio synthesis. In the
present study , shoo t clusters in optimal condition
could also become dif feren- tiated and synthesis
artemisinin. And shoot clusters in dif ferent g row th
stage and culture condition also produced dif ferent
content of artemsinin. Further study of A. annua
shoo t cluster wi ll be to discover proper medium fo r
the g row th of shoo t cluster and biosynthesis of
artemisinin respectively , w hich maybe the founda-
·571·中草药　 Chinese T raditional and Herbal Drug s　第 35卷第 5期 2004年 5月
tion of the industrialized pro- duction of
a rtemisinin th rough tw o-step method.
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特异性真菌作用于龙血树材质形成血竭的研究
杨　靖 1 ,江东福 2 ,马　萍 2
( 1. 湛江海洋大学 现代生化实验中心 ,广东 湛江　 524088; 2. 云南大学微生物研究所 ,云南 昆明　 650091)
摘　要: 目的　探讨剑叶龙血树血竭的形成与微生物活动之间的关系。 方法　用分离自剑叶龙血树根部的内生真
菌 9568D镰孢霉接种于剑叶龙血树材质 (经灭活处理 )。结果　保湿培养 4～ 5个月后 ,在接种部位有红色血脂颗粒
形成 ,经 UV、 IR光谱分析及抗菌活性实验 ,初步证实该血脂与来自剑叶龙血树的天然血竭无本质差异。 结论　特
异性真菌作用于龙血树材质可促成血竭的形成。
关键词: 9568D镰孢霉 ;龙血树材质 ;血竭
中图分类号: R286. 02　　　文献标识码: B　　　文章编号: 0253 2670( 2004) 05 0572 03
Study on formation of Dragon s blood inDracaena cochinchinensis
inoculated withFusarium 9568D
YANG Jing
1
, JIANG Dong-fu
2
, M A Ping
2
( 1. Experim enta l Center o f Biochemistr y , Zhanjiang Ocean Univ ersity, Zhanjiang 524088, China;
2. M icrobiolog y Institute of Yunnan Univ ersity, Kunming 650091, China )
Key words: Fusarium 9568D; Dracaena cochinchinensis ( Lour. ) S. C. Chen; Dragon s blood
　　血竭 ( Dragon s blood)是中药传统品种之一 , 其性味甘、温、咸、平 ,兼有活血、化瘀、止血、补血的
·572· 中草药　 Chinese T raditional and Herbal Drug s　第 35卷第 5期 2004年 5月
收稿日期: 2003-07-04作者简介:杨　靖 ( 1967— ) ,男 ,苗族 ,湖南永顺人 ,助理研究员 ,硕士 ,主要从事微生物生态及应用微生物研究。 Tel: ( 0759) 2383477
* 通讯作者　 E-mai l: yh sn e@ hotmai l. com