全 文 ：现代生物医学进展 www.biomed.net.cn Progress in Modern Biomedicine 2009 Vol.9 No.8
·实验研究·
Research on the Callus Induction and the Cell Growth and Metabolism
Characteristics of Capparis spinosa L. *
WANG Yan-ting, GAN Lu, LIU Wei, YU Long-jiang, LI Mao-teng*
（College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074）
ABSTRACT: Different kinds of explants and plant hormones were applied to study their effects on the callus induction of Capparis
spinosa L., and different proportions of plant hormones were also employed to explore the effects on callus propagation as well as the
growth and metabolism characteristics of suspension cells. These results revealed that, the fresh leaf was the best explant for inducing the
callus formation, and MS medium with 0.5 mg/L 2, 4-D +1.0 mg/L 6-BA and 1.0mg/L 2, 4-D +1.5 mg/L6-BA was optimum for callus in-
duction and subculture. The growth cycle was about 30 days and the growth showed curve of S type, the proliferation rate was about
2.8 times. On the other hand, during the growth cycle of the suspension cell, concentrations of both sucrose and total soluble sugar were
decreased steadily, concentration of reducing sugars was first increased and then decreased, and POD activity was accordingly posi-
tive-correlated with the concentration of sucrose.
Key Words: Capparis spinosa L.; Callus; Suspension cell; Growth and metabolism characteristics
Chinese Library Classification（CLC）: 282.71 Document code: A
Article ID: 1673-6273（2009）08-1453-06
*Foundation items: Natural Science Foundation of China（No. 90202016）
Author introduction: WANG Yan-ting,（1983-）, femal, master, Mainly engaged in plant biotechnology research
**Correspoonding author: LI Mao-teng, E-mail: limaoteng426@163.com
（Received: 2009-02-10 Accepted:2009-03-08）
Capparis spinosa L., Capparidaceae Shan Gan herb, mainly
distributes in Spanish, the Mediterranean Sea bay area, as well as
broadly distributes in such place as Xinjiang, Gansu and Tibet in
China [1]. Capparis spinosa L. is believed as one of typical eremo-
phytes for its developed root system and the xylem, by which it
can use the groundwater resources highly effectively [1,2]. The
chemical analysis indicates that the effective constituents of Cap-
paris spinosa L. include volatile oil, alkaloids, flavonoids, terpene,
mustard oil glycoside and so on, all these constitutes have certain
effects at antibiotics, anti-inflammation, anti-oxidation, anti-hyper-
tension, lower the blood sugar and blood fat, diuresis, as well as
uarthritis and rheumatism treatment [3-8]. The on-spot investigation
shows that, the eremophytes community whose constructive
species are Capparis spinosa L. almost distributes by small frag-
mentary area. Meanwhile, the hierarchical structure of the commu-
nity is extremely simply and the species composition is also very
deficient [1]. The vegetable cell suspension culture is the microbio-
logical form of the vegetable cell growth, it has several advantages
such as good dispersivity, the shape and size of the cell group are
approximately same, moreover, it grows rapidly and is easy to re-
produce and control. So far, the suspended cell culture has become
one of the most useful methods in the plant biotechnology [9].
Therefore, the callus induction of Capparis spinosa L., the con-
struction of suspension cell system and the analysis of its growth
and metabolism characteristics, will pave the foundation for pro-
ducing correlative medicinal stuffs through the large scale cell cul-
ture, the callus induction of Capparis spinosa L. and the establish-
ment of cell line have not been reported yet until now. So the cal-
lus induction and propagation of Capparis spinosa L. were studied
in different explants, different surface disinfection methods and
plant hormones, and accordingly the cell growth curve, the sugar
metabolism and peroxide enzyme activity were also analyzed.
1 Materials and methods
1.1 Plant materials
The materials for callus induction include the seed of Cap-
paris spinosa L., seedlings and the plants grown in the experimen-
tal field in Huazhong University of Science and Technology.
The materials for cell growth curve determination and metabolism
process research are the cell line grown stably in the liquid media
transferred from the exuberant and pultaceous callus grown on the
solid media for more than two generations.
1.2 Experimental methods
1.2.1 The induction and formation of callus
1.2.1.1 Selection and disinfection of explants Firstly, made
the seeds, the young stems and the leaves of Capparis spinosa L
soaked in the biotex solution for 30 min, scrubbed the surface of
the explants and flushed them 5-6 h with the running water; Then
soaked in 70% ethyl alcohol for 15-20 s and flushed 3 times with
aseptic distilled water in the ultra-clean bench; After surface steril-
ization in 0.1% mercuric chloride, flushed these explants 3-4 times
with the sterilized water.
1.2.1.2 Induction of callus The leaves and the stems were cut
to 0.5 cm X 0.5 cm scraps, 1.0 cm long slices, while the seeds
were cut into equal halves. Three kinds of these explants were sub-
cultured on MS medium supplemented with 1.5 mg/L 2, 4-D + 3.0
1453· ·
DOI:10.13241/j.cnki.pmb.2009.08.020
现代生物医学进展 www.biomed.net.cn Progress in Modern Biomedicine 2009 Vol.9 No.8
mg/L 6-BA +3 % sucrose (pH 6.0), incubated in the dark for one
week and then continued to culture in the light, and meanwhile
compared the callus growth of different explants in induction
medium. To study the callus induction of Capparis spinosa L. un-
der the plant hormones with different levels and combinations, the
leaves were focused to apply for the singlet factorial experiment
and orthogonal experiments with 4 factors and 3 levels. The ran-
dom design for the concentration levels was taken, 30 explants in
each treatment were cultured for 30 d, and the induction percent-
age of the callus was counted for statistical analysis.
1.2.1.3 Multiplication of callus Based on the optimum induc-
tion combination experiment, different concentrations of 2, 4-D
and the 6-BA were designed to find the optimum culture condition
for callus propagation. The callus were subcultured in 4 bottles ac-
cording to each process and advanced for two generations, count-
ed the multiplicity rate of propagation.
1.2.1.4 Parameters analysis of the culture results After the
explants materials been cultured and subcultured, the contamina-
tion rate, browning rate and induction rate were assessed in 30d,
and the statistics indexes were as follows: contamination rate %=
(contaminated explants /total explants) ×100%; browning rate %
= (brown explants / total explants)×100%; Survival rate %= (sur-
vival explants / total explants) ×100% ; induction rate %= (in-
duced callus / total explants) ×100% ; proliferation rate = (the
fresh weight of callus grown for 30d / (the fresh weight of callus
before subcultured)×100%
1.2.2 Analysis on cell growth and metabolism characteristic
of Capparis spinosa L. suspension cells
1.2.2.1 Culture condition MS liquid media supplemented with
1.0 mg/L 2, 4-D + 1.5 mg/L 6-BA +30 g/L sucrose (pH5.8), inocu-
lation quantity was 10%, illumination intensity was 2100 Lx, 14
h/d, temperature was 25 ℃±1℃, rotational speed of the shaking
bed was 100 -120 rpm.
1.2.2.2 Determination of biomass 3.0g callus were subcul-
tured in liquid media for each bottle and cultured continuously for
32d, then measured its fresh weight on 1/1000 precision electronic
balance after suction filtration at the interval of 4d.
1.2.2.3 Determination of the sugar concentration in nutrient
solution of cell suspension culture Concentration of sucrose,
reducing sugar, total soluble sugar and POD activity were deter-
mined as described by Li Hesheng[10].
2 Results and analysis
2.1 The effect of different time of HgCl2 to different kinds
of explants
The processing time of explants was not completely the same
because of certain differences in the physiological condition. As
shown in Table 1, as the disinfection time of 0.1% mercuric chlo-
ride extended, the contamination rate of various kinds of explants
degraded but browning rate increased, and disinfection for 8 min
was the best for leaf explants that survival rate reached 75.1%. In
the case of stem explants, the disinfection for 8-15 min could not
achieve a good effects, contaminate situation was under control
but the browning rate was greatly enhanced by prolonging the dis-
infection process. When the seed explants was thoroughly disin-
fected, they had a high survival rate without yellowing, however it
still had some disadvantages such as: the 15-25min disinfection
time of mercuric chloride for seeds explants was the twice that of
leaf explants, the poison of disinfectant was relatively serious and
the disinfection procedure was tedious. Owing to all above de-
scribed, the fresh leaf was the best explants for the callus induction
of Capparis spinosa L., and the optimum disinfection time of mer-
curic chloride was 8 min.
Table 1 Influence of treatment time of HgCl2 on the stem, leaf and seed explants of Capparis spinosa L.
Sterilization
time (min)
Leaf Stem Seed
Contamin
ation rate (%)
Browning
rate (%)
Survival rate
(%)
Contamin
ation rate (%)
Browning
rate(%)
Survival rate
(%)
Contamin
ation rate(%)
Survival rate
(%)
5 31.6 19.8 48.6 41.3 17.6 41.1 34.2 65.8
8 8.6 16.3 75.1 36.6 9.5 53.9 21.5 78.5
15 9.7 25.0 65.3 31.2 12.4 56.4 10.6 89.4
25 5.4 46.2 48.4 11.3 37.2 51.5 11.8 88.2
2.2 The effects of different explants of Capparis spinosa L.
to the rate of callus induction
Table 2 demonstrated the influence of different explants on
callus induction on MS media with 1.5 mg/L 2, 4-D + 3.0 mg/L
6-BA +3 % sucrose. It showed that, the callus induction of Cap-
paris spinosa L. resulted from the leaf, stem and seed explants, and
the leaf was the best explants with the highest induction rate simi-
lar to the stem, but the callus induced by the seed was loose and
soft which was not suitable for subculture. The induction time was
almost the same for different explants. The incision position start-
ed to expand at 4-5d after they were subcultured on medium, and
resulted in callus hereafter they continued to culture for 4-5d a-
Explant
types
Induction rate
(%)
Callus formation
time (d)
Callus state
Leaf 37.6 9
flavovirens，
rarefaction
Stem 28.8 8
Light flavovirens，
rarefaction
Seed 33.3 11 white，soft
Table 2 The effects of different explants of Capparis spinosa L. to callus
induction rate
1454· ·
现代生物医学进展 www.biomed.net.cn Progress in Modern Biomedicine 2009 Vol.9 No.8
gain, but there was some had slight differences in the color and
state of callus for different explants.
2.3 The influence of different plant hormone combination
on callus induction of Capparis spinosa L.
To study the influence of different hormone combinations on
callus induction of Capparis spinosa L., the leaf explants was used
and treated by 6-BA, 2, 4-D and NAA with L9(34) orthogonal de-
sign method, 30 explants were subcultured for each treatment
(Table 3).
Based on different concentration combination treatment of 2,
4-D, 6-BA as well as NAA, we found that, different combinations
had great effects on callus induction of Capparis spinosa L. In the
Table 3, the influence degree of three kinds of hormones on callus
induction of Capparis spinosa L. were rated in the order of 6-BA,
2, 4-D and NAA.
The variance analysis results indicated that, Apart from the
other insignificant factors, 6-BA had the most remarkable influ-
ence on callus induction of Capparis spinosa L. (Table 4). The op-
timum plant hormone combination on callus induction of Capparis
spinosa L. was: A2B2C1, namely, MS +1.0mg/L 6-BA + 0.5 mg/L
2, 4-D. Further experiment revealed that the callus induced by this
design presented three kinds of different morphology and color
shades (Fig. 1).
Note: ** denotes difference at an obviously significant level (P<0.01)
Treatment
A B C D
Induction rate (%)
6-BA(mg/L) 2, 4-D(mg/L) NAA(mg/L) Black
1 1(3.0) 1(1.0) 1(0) 1 53.4
2 1(3.0) 2(0.5) 2(1.0) 2 44.5
3 1(3.0) 3(0.1) 3(3.0) 3 14.2
4 2(1.0) 1(1.0) 2(1.0) 3 67.5
5 2(1.0) 2(0.5) 3(3.0) 1 69.5
6 2(1.0) 3(0.1) 1(0) 2 65.6
7 3(0.5) 1(1.0) 3(3.0) 2 41.2
8 3(0.5) 2(0.5) 1(0) 3 53.4
9 3(0.5) 3(0.1) 2(1.0) 1 23.1
M1 M11=42.4 M12=54.0 M13=57.5 M14=48.7
M2 M21=62.5 M22=55.8 M23=45.0 M24=50.4
M3 M31=39.2 M23=34.3 M33=41.6 M34=45.0
R R1=23.3 R2=21.5 R3=15.9 R4=5.4
Table 3 The results of orthogonal experiment
Variance origin df SS S2 F F 0.05 Significant
A 2 957.1 478.6 20.9 19.0 **
B 2 853.6 426.8 18.6
C 2 420.6 210.3 9.2
D 2 45.7 22.9
Error 2 45.7
Table 4 Variance analysis of orthogonal experiment results
Fig. 1 The different states of calli
a White and pulpy calli; b yellow and nubby calli; c: Kelly and loose
granule calli
2.4 Multiplication of callus
In this experiment, six kinds of different hormone combina-
tions were applied to multiply the callus with similar growth state.
It indicated that, multiplication coefficient of callus from explants
was between 2-3 times for each combination of 2, 4-D and 6-BA,
but the callus multiplied faster in T4 and T2 treatments than other
treatments; Apart from T5 treatment, the growth state of the callus
on subculture medium with several other kinds of hormone combi-
nations were approximately the same. According to the callus
state, the optimum medium for callus multiplication considered as
MS medium with 1.5 mg/L 6-BA and 1.0 mg/L 2, 4-D. And mean-
while, as 2, 4-D concentration increased in the range of 1.0-2.0
mg/L, the induction rate of callus was enhanced correspondingly.
2.5 The growth curve of cell suspension culture of Capparis
spinosa L.
1455· ·
现代生物医学进展 www.biomed.net.cn Progress in Modern Biomedicine 2009 Vol.9 No.8
Generally, the growth curve of cell suspension culture of
Capparis spinosa L. was S type (Fig. 2), and its growth process
includes three stages: lag phase, log phase and decline phase.
Once subcultured, the first 4 days were called as cell lag period for
the lowest cell multiplication quantity, possibly because the cell
had not adapted to the new environment completely and could not
absorb nutrients sufficiently. From the 4th day to the 24th day was
the log phase, the multiplication speed of the cell was relatively
fast during this phage. After the 24 day, the multiplication speed of
the cell slowed down gradually and the medium began to change
muddy, the cell also turned dark-brown from xanthos. By this
time, if the medium had not been replaced, the cell would easily
turn brown and die. Therefore, 24 days of cell culture was the
most appropriate for subculture from the viewpoint of both high
growth quantity and the optimum growth state. For the suspen-
sion-cultured cell, the cell fresh weight proliferation rate of Cap-
paris spinosa L. was about 2.8 times from initial period to the de-
cline phase.
2.6 Sugar metabolism in the suspension cell of Capparis
spinosa L.
The carbon source was one of the most important nutrients in
the medium. The most commonly used carbon source in the cell
culture was sucrose [11]. It was known from Fig. 3 that, at the initial
time, the sucrose showed little decomposition with a concentration
of 28.71 g/L. Since the cell lag period, the sucrose in medium was
hydrolyzed to glucose and fructose by invertase, and in about 28
days, there was extremely little sucrose surplus in the nutrient flu-
id. In the entire process of cell culture, the average daily consump-
tion rate of sucrose was 0.89 g/L. In lag period, sucrose consump-
tion was quick with the average daily consumption rate of 2.73
g/L. About 60 % sucrose was consumed in the log phase, and so
there was only extremely little sucrose surplus in decline phase.
Judging from the sucrose consumption situations of the three
stages, the lag period, the consuming rate was highest, and simul-
taneously, the production rate of reducing sugar was also highest,
and the average daily growth rate achieved 1.57 g/L. the concen-
tration of reducing sugar began to decrease in 16th day later so
that it left only little in the 32nd day later. During the entire cell
cycle, the concentration of total sugar showed the trend to decline.
In the initial period, the consumption of total sugar was small with
5.50 g/L during the initial 4 days. In log phase, the consumption
rate of total sugar accelerated to 56% in 24 days. During the 32
day of cell culture process, the quantity of total sugar consumption
was 24.03 g/L. (Fig. 3) the total sugar consumption curve and the
cell growth curve were found closely related. The consumption of
total sugar was low in log period, and correspondingly, cell grew
slowly. In the log phase, large quantity of the total sugar was con-
sumed up, the cell growth was also biggest in quantity. It was thus
clear that the carbon source was the primary energy for accumula-
tion of cell biomass.
2.7 Enzyme activities of POD in the suspension cell of Cap－
paris spinosa L.
Our results indicated that, as culture time extended, POD ac-
tivities of suspension cell fluctuated (Fig.4), under condition with
3% sucrose, POD activities of suspension cell appeared the peak
Table 5 The influence of various 6-BA and 2,4-D levels on callus multiplication
Treatment Concentration (mg/L) Multiplication coefficient Callus state
T1 6-BA 1.0+2, 4-D 0.5 2.33± 0.24 xanthos、granulo
T2 6-BA 1.0+2, 4-D 1.5 2.82± 0.36 stramineous、white、loose and soft
T3 6-BA 1.5+2, 4-D 0.7 2.32± 0.32 flavovirens、granulo
T4 6-BA 1.5+2, 4-D 1.0 2.89± 0.26 flavovirens、granulo
T5 6-BA 2.0+2, 4-D 0.5 1.78± 0.34 xanthos、massive
T6 6-BA 2.0+2, 4-D 1.0 2.05± 0.42 dark xanthos、massive
Fig. 2 The growth curve of cell suspension culture of Capparis spinosa L.
Fig. 3 The time course change in the sugar consumption of Capparis
spinosa L.
1456· ·
现代生物医学进展 www.biomed.net.cn Progress in Modern Biomedicine 2009 Vol.9 No.8
value in 12th day, 20th day and the 28th day, respectively. But it
happened in 12th day and the 28th day under the condition with
5% sucrose. Fig.4 also showed that under the concentrations with
both different sucrose conditions, POD activities were almost
same at 20th and 28th days. Analyzed From the overall level, POD
activities under condition of 5 % sucrose was slightly higher than
that in condition of 3% sucrose, this was possibly because the high
concentration sucrose condition brought about a stress environ-
ment for suspension cell of Capparis spinosa L., POD often acted
as the one of key enzymes in plant defense system under adverse
condition [12], and therefore, the increase in the peroxide enzyme
level in the suspension cell could be regarded as the adaptation re-
sponse to high concentration sucrose condition.
3 Discussions
The induction time of callus and the large different rate of
callus to subculture differ along with organs with different sources,
all these determinant factors are very important for explants selec-
tion and cell line establishment. In this experiment, leaf, stem and
seed explants were chosen to assess the influence of disinfection
methods and efficiencies. Our results indicated that, 70% ethyl al-
cohol followed by the suitable time process of the mercuric chlo-
ride could enhance the disinfection rate effectively, but different
explants required the disinfection time. The leaf was the best ex-
plant for induction of callus of Capparis spinosa L., and its opti-
mum disinfection time of mercuric chloride was 8 min. Leaf, stem
and seed explants were also employed to analyze the influence of
different explants of Capparis spinosa L. on callus induction, the
young leaf approved to be the best explant for induction of callus.
During the process of cell culture, the in vitro cells often lack the
ability to biosynthesize plant hormone auxin and cytokinin, so dif-
ferent exogenous hormones should be applied into the culture
medium to induce cell proliferation, differentiation and morpho-
genesis. The influence of different kinds and concentrations of
plant hormones on embryo callus multiplication was quite differ-
ent. In many plant explants, 2, 4-D was indispensable for the in-
duction of callus dedifferentiation. A dicotyledonous medicinal
plant, Medlar only needs 0.2 mg/L 2, 4-D, but in the ginseng cell
culture, the highest concentration of 2, 4-D reaches 12 mg/L [13].
In this experiment, the effect of hormone induction on callus in-
duction of Capparis spinosa L. was revealed too. The results indi-
cated that, plant hormone had the obvious effects during the callus
induction of Capparis spinosa L. In the experiments with the hor-
mone concentration proportion, the optimum culture condition for
callus induction and subculture was MS medium with 0.5 mg/L 2,
4-D + 1.0 mg/L 6-BA and 1.0mg/L 2, 4-D +1.5 mg/L6-BA, su-
crose concentration 30 g/L, pH 6.0.
The research of suspension cell culture was the essential
foundation for the medicinal plant cell industrialized production,
compared with the solid medium, the cell suspension culture had
several advantages such as high speed of cell reproduction, large
culture scale, and the cell cultures with synchronized state. It was
the liquid culture technology, which could cause callus cultures
multiplied efficiently and easy to regulate the metabolite product.
Our results indicated that, the growth curve of suspension cell of
Capparis spinosa L. was S type and the growth cycle was about
30 days [14,15]. Compared with the common liquid suspension cell,
the vegetative cycle was slightly longer, we conjectured the fol-
lowing two reasons: at first, growth cycle of the cell was affected
by vegetation type, density of primitive cells, nutrients ingredient
type as well as recipe; Secondly, subculture times of suspension
cell was limited, the cell was possibly in the process of the adapta-
tion and adjustment regarding the liquid condition. Through the in-
crease in the subculture times, the growth cycle of suspension cell
could be reduced gradually. In the plant cell culture, the sugar was
the best carbohydrate origin in the cellular metabolism pathway,
had a tremendous influence on the cell growth. Our results indicat-
ed that, sucrose completely transformed into reducing sugars such
as glucose, fructose in the cell in log phase. Peroxide enzyme is
generally present in higher plants, participates in plant respiration,
biosynthesis of plant lignin and ethylene, and is closely related to
plant growth and reproduction [16]. Under different sucrose concen-
tration in this experiment, certain concentration of sucrose could
enhance POD activities. Our research provided some basic foun-
dations for large-scale cell culture of Capparis spinosa L.
References
[1] ZHANG LY, YANG C. A aeolian protection plants- Capparis spinosa
L [J]. Plants, 2004, 31(1): 3-4
[2] LEVIZON E, DRILLAS P, KYPARISSIS A. Exceptional photosyn-
thetic performance of Capparis spinosa L under adverse conditions of
Mediterranean summer[J]. Photosynthetica, 2004, 42(2): 229-235
[3] KHANAFAR M A, SABRI S S, ZARGA M H. The chemical con-
stituents of Capparis spinosa of Jordanian origin [J]. Nat Prod Res,
2003, 17(1): 9-14
[4] AL-SAID M S, ABDELSATTAR E A, KHALIFA S I, et a1. Isolation
and identification of an anti-inflammatory principle from Capparis
splnosa[J]. Pharmazie, 1988, 43:640-641
Fig. 4 The time course change in enzyme activities of POD of Capparis
spinosa L.
1457· ·
现代生物医学进展 www.biomed.net.cn Progress in Modern Biomedicine 2009 Vol.9 No.8
[5] BERTRAND M, MUSA O. Glucosinolate Composition of young
shoots and flower buds of capers (Capparis species) growing wild in
Turkey[J]. J Agrie Food Chem, 2002, 50: 7323-7325
[6] JOUAD H, HALOUI M, RHIOUANI H, et a1. Ethnopharmacological
survey of medicinal plants used or the treatment of diabetes,cardiac
and renal diseases in the north centre region of Morocco (Fez-Boule-
mane)[J]. Journal of Ethnopharmacology, 2001, 77: 175-182
[7] ALI M, RAMACHANDRAM R, RAFIULLAH M R M, et a1. Preven-
tion of carbon tetrachloride-induced hepatotoxicity by the ethanol ext-
ract of Capparis moonii fruits in rats [J]. Pharmaceutical Biology,
2004, 42: 286-288
[8] EDDOUKS M, LEMHADRI A, MICHEL J B. Hypolipidemic activity
of aqueous extract of Capparis spinosa L in normal and diabetic rats[J].
Journal of Ethnopharmacology, 2005, 98: 345-350
[9] LI T T, SHI J S, CHEN J H. Developmental synchronization of somatic
embryogenesis under suspension culture condition. Molecular Plant
Breeding, 2007, 5(3): 436-442
[10] LI H S. The experimental principal and technology of plant physiology
and biochemistry. Beijing: High Education Press, 2001
[11] COURNAC L, DIMON B, CARRIER D et al. Growth and photosyn-
thetic characteristics of Solanum tuberosum plantlets cultivated in vit-
ro in different conditions of aeration, sucrose supply and CO2 enrich-
ment [J]. Plant Physiol, 1991, 112: 111-117
[12] LIANG Y R, HU X H, ZHANG Y L. Progress on physiological func-
tion research of plant peroxidase. Journal of Inner Mongolia Agricul-
tural University, 2003, 24(2): 110-113
[13] CHUI K R, DAI R L. The molecular of plant somatic embryogene-
sis[M]. Beijing: Scientific Press, 2000, 49-53
[14] HOU X W, GUO Y. The effects of carbon sources on the growth and
respiration of suspension Roselle cell. Guihaia, 1999, 19(2):150-153
[15] GUO Z G, DU J, LIU R Z. Kinetic investigation of Taxus cell growth
and nutrient consumption. Journal of Tsinghua University (Science
and Technology), 2002, 42(5): 599-602
[16] MONICA QUIROGA, CONSUELO GUERRERO, MIGUEL A et al.
A tomato peroxidase involved in the synthesis of lignin and suberin [J].
Plant Physio1, 2000, 112: 1119-1127
药用植物刺山柑愈伤组织诱导及细胞生长代谢特征研究 *
王艳婷 甘 露 刘 薇 余龙江 栗茂腾 **
（华中科技大学生命科学与技术学院 武汉 430074）
摘要：本文研究了不同外植体及激素对刺山柑愈伤组织诱导的影响，不同激素配比对愈伤组织增殖培养以及悬浮细胞的生长与
代谢特征。结果表明：以刺山柑叶片作为诱导愈伤组织的材料，效果较佳；愈伤组织诱导和继代的适宜培养条件是分别是 MS+
0.5 mg/L 2, 4-D +1.0 mg/L 6-BA和MS +1.0mg/L 2, 4-D +1.5 mg/L6-BA。刺山柑悬浮培养细胞的生长周期为 30天左右，细胞生长
曲线呈 S形，生物量增长 2.8倍左右；细胞生长周期内，碳源消耗规律表现为蔗糖和可溶性总糖的浓度持续降低，而还原糖则表
现为先升高后降低；过氧化物酶活测定显示酶活水平与蔗糖浓度的高低呈一定程度的正相关。
关键词：刺山柑；愈伤组织；悬浮细胞；生长代谢特征
中图分类号：282.71 文献标识码：A 文章编号:1673-6273（2009）08-1453-06
*基金项目：国家自然科学基金项目资助（No.90202016）
作者简介：王艳婷，（1983-），女，硕士，主要从事植物生物技术方面的研究
**通讯作者：栗茂腾，E-mail: limaoteng426@163.com
（收稿日期：2009-02-10 接受日期：2009-03-08）
1458· ·