全 文 ：△ 广西壮族自治区科技厅科技攻关项目(桂科攻 0322023-3)
作者简介:刘园(1982-),女 , 广西柳江人 , 在读硕士研究生。 ＊通讯作者 、硕士生导师
Studies on tissue culture and plant regeneration of Emilia sonchifolia(L.)DC.
LIU Y uan 1 , WU Qing-hua 2 , QIN Wen-liu1 , LING Zheng-zh u2＊.
1.Guang xi U niversity (Nann ing 530005), China;
2.Guangxi Botanical Garden of Medicinal Plan t 、Inst itute of Guang xi Med icina l P lant 、
The Guangxi Branch , Inst i tute of medicinal Plant , Chinese Academy of Medical Sciences(Nanning 530023)
　　[ Abstract] 　Objective　To explore if the quick propagation of Emilia sonchi folia(L.)DC.in vitro culture could be used to provide
tube-cultured seedlings in short time.Methods　Axial buds w ere used as explants and cultivated on Murashige and Skoog basal medium
by adding different por tions of hormones.Results　After 30-day cultured., axial buds w hich were cultivated in MS+BA1.5mg·L+
NAA0.1 mg·L-1 could induce fascicular buds , and each node derived 4 to 5 fascicular buds.The induced fascicular buds w ere trans-
ferred to the medium of 1/2 MS+NAA1 mg·L-1 to induce roots.Af ter the test-tube plantlet transplanted , there w as no yellowing phe-
nomenon(caused by yellow virus).Conclusion　This research on tissue culture of Emilia sonchi folia(L.)DC.could supply tube-cul-
tured seedlings for large-scale planting.
　　[ Key Words] 　Emilia sonchi folia(L.)DC.;Axial bud;T issue culture;Regenerated plant
一 点 红 组 织 培 养获 得 再 生 植 株 的 研 究 △
刘　园1　吴庆华2　覃文流1　凌征柱2＊
1.广西大学　(南宁　530005)　2.广西药用植物园 、广西药用植物研究所中国医学科学院药用植物研究所广西分所　(南宁　530023)
　　[ 摘要] 　目的　对一点红进行组织培养研究 ,探讨在短时间内快速繁殖一点红种苗的方法。方法　以一点红带有腋芽的茎
段为外植体 ,用 MS+BA1.5 mg/ L+NAA0.1mg/ L 培养基。结果　经 30 d诱导培养 , 可得丛生芽 4 ～ 5个;小芽苗转入 1/ 2MS+
NAA1mg/ L培养基上诱导生根 , 20 d后可长成具有 3～ 4 条根系的再生植株。结论　此途径为人工栽培一点红提供优质种苗。
　　[ 关键词] 　一点红;腋芽;组织培养;再生植株
　　[ 中图分类号] 　R282.2
　　Emilia sonchifolia(L.)DC., named lilac tasselflower in
English , belongs to the feverfew family[ 1] .Each part of lilac tas-
selflow er can be used as medicine.Lilac tasselflow er has such
medicinal effects as clearing heat and expelling M iasma , reducing
inflammation , discharging of urine , relieving pain and reducing
fever.The plant can be used fo r the treatments o f dysentery and
many kinds of inflammations , such as the inflammation of respi-
ratory disease , pneumonia , tonsillitis and mastitis[ 2 , 3] .Fo r ex-
ample, Huahongpian , making from lilac tasselflower , is a good
medicine for the treatment of gynecological disease.Now adays ,
the lilac tasselflow er are in great demand in medical industry.The
natural resource of lilac tasselflower lessens because the environ-
ments that are suitable fo r the grow th of lilac tasselflow er are be-
ing destroyed for the reclamation and cultivation.In fact , the
quality of lilac tasselflow er is hard to guarantee on account of the
er roneous collection of wild lilac tasselflower.Therefo re, the cul-
tiv ation of lilac tasselflower is imperative.In o rder to provide the
young plants of lilac tasselflow er for large-scale planting and the
materials of lilac tasselflower for the medicine industry , the re-
search on tissue culture of lilac tasselflow er was done to supply
tube-cultured seedlings for large-scale planting.
1　Materials
　　Explants of Emilia sonchifolia(L.)DC.were taken from
one-year-old healthy and strong plants g rown in Guangxi Botani-
cal Garden of Medicinal P lants.
2　Culture medium and conditions
　　The culture medium was Murashig e and Skoog(1962)basal
medium supplied with different concentrations of phy tohormone.
The induction medium:(1)MS+BA 1.0mg·L-1+NAA 0.1mg
·L-1;(2)MS+BA 1.5mg·L-1+NAA 0.1mg·L-1;(3)MS+
BA 2.0mg·L -1+NAA 0.1mg·L-1;(4)MS+BA 2.5 mg·L -1
+NAA 0.1 mg·L-1 。 The subculture medium:(5)MS +BA
798 Guangxi Medical Journal , Jun.2006 , Vol.28 , No.6
1.0mg·L-1+NAA 0.1mg·L-1;(6)MS+BA 1.5mg·L -1+
NAA 0.1mg·L-1.Rooting medium:1/ 2 MS +NAA 1.0mg·
L-1.The medium was supplemented with 2.0% sucrose and so-
lidified with 0.5%(W/ V)agar , and the PH was adjusted to 5.8
to 6.0 before autoclaving at 121℃ and 108 kPa for 20min.All
culture were incuba ted at(25±2)℃ under 10-hour daily illumi-
nation w ith white fluorescent light(1 500 to 2 000 Lx.).
3　Methods and Results
3.1　The Treatment of Material　The stem segments with axi-
ally bud o f lilac tasselflow er were so aked and brushed in scour ,
and then washed tho roughly for 30 min under running tap w ater.
They were surface sterilized in 0.1% Mercuric Chloride solution
containing a few drops of Tween-20 for 8 to 10min , follow ed by
4 to 5 rinses in sterile distilled w ater.This part o f stem with axi-
ally buds were cut into 0.3 cm long single node pieces , and then
placed respectively in No.1 to No.4 medium.
3.2　Clusterbuds induction　Explants were cultured fo r 7 day s
in induction culture medium , the bottom of stem expanded and
new buds expanded , then sprouted bourgeon after 6 to 8 days cul-
ture.After 25-day cultured , explants in No.1 medium could be
induced 2 to 3 shoo ts per node;and in No.2 culture medium , 4
to 5 buds could be obtained.The buds induced in No.3 and No.4
culture medium w ere more than that in No.1 and No.2 culture
medium , but the shoo ts vitrification occurred , especially in No.4
culture medium.Thus , lilac tasselflower was sensitized with the
using concentration o f cy tokinin.Higher concentra tion of cy-
tokinin could be helpful to induce cluster buds , but could bring
about shoots verification in later stage.Moderate concentration of
cy tokinin is necessary to get cluster buds and to avoid verification.
3.3　Multiplication of shoot cultures　When the average height
of cluster buds were 2 to 3 cm after 35-day induction culture , the
cluster buds were cut into single bud and cultivated in No.5 and
No.6 culture media for further multiplication.After 14-day mul-
tiplication cultured , the sing le bud cultivated in No.5 culture
medium grew new cluster buds.The grow th coefficient w as 6 to
7 times in a multiplication period of 30 days.The rate of multipli-
cation of single bud cultivated in No.6 culture medium was higher
than that in No.5 culture medium.As cluster buds g rew , they
got vitrified because of high concentration of BA.In the early
stag e of induction culture , the buds were cultivated in the medi-
um w ith BA , so the buds could accumulate BA.As a result , the
concentration of BA in subculture should be lower than that in in-
duction culture.
3.4　Rooting Culture and Transplanting　When cluster buds
g rew in 5 to 6 cm high , cluster buds w ere separated and cultured
individually in No.7 medium.In the effect of NAA , the bases of
sing le bud appeared initial roo ts af ter 7 to 10 days , and then be-
came small roots after 20 day s cultured.These roots w ere 2 to 3
cm in long.Each sing le bud had 3 to 4 roots.The rooting rate
reached 90%.At this point , Plantlets w ere trained by opened the
bottle caps at normal atmosphere temperature indoo r , in the natu-
ral illumination conditions for 2 or 3 day s.Plantlets with well-de-
veloped roo ts were removed from culture medium and then
washed the roots under running tap w ater to remove agar.
Plantlets were transferred to sand seedbed to ensure higher sur-
vival rate.I n this way , the survival rate could be 95%.The tis-
sue-cultured seedlings w ere transplanted in April 20.The plant
bloomed 50 day s and the seeds ripened 65 day s after transplant.
The cluster buds of Emilia sonchifolia(L.)DC.
4　Brief Summaries
　　The research on the quick-propagation in vitro culture o f E-
milia sonchi folia(L.)DC.showed that the ra te of multiplica tion
could be 6 times.The quick-propagation in vitro culture could be
used to provide lilac tasselflow er in short time fo r the cultiva tion
in large scale.The main stem of test-tube plantlet budded in 50
days af ter transplantation and the seeds ripened in 70 days.I n the
grow th period of test-tube plantlet , there w as no yellowing
(caused by yellow virus)w hich happened commonly in the culti-
vation by seed.Without infected of yellow virus , the quick-prop-
agation in vitro culture of lilac tasselflow er could provide purified
provenance, high-quality seeds for standardized and cultiva tion in
large area.
References
1　Insti tute of Botany , Academia Sinica.Iconographia Cormophytorum
Sinicorum , Volume Ⅳ[M ] .S cience Press , Beijing , 1996.551.
2　Compilers of《The Compi lation of National Chinese Herbal Medicine》 .
The Compi lation of National Chinese Herbal Medicine , Volume One
[ M] .People s Medical Publishing House , Beijing , 1996.1.
3　Edited by Health Service of the Revolut ion Committee of Guangxi
Zhuang Autonomous Region.The Selected Works of Chinese Tradi-
tional Herbs in Guangxi , Volume One[ M] .People s Medical Publish-
ing House of Guangxi , Nanning , 1974.94.
(收稿日期:2006-4-20　修回日期:2006-5-28)
799广西医学　2006 年 6月　第 28 卷　第 6 期