全 文 ：广 西 植 物 Guihaia 26(3)：286～288 2006年 5月
Rapid plant regeneration from Glechoma longituba
HAN Su-iu，LI Yun—xiang ，YANG Zi—song。
JIANG Tian—liang，LI You
(Sichuan Pr。vincial K Y Laboratory of Environmental Scienc and Biological Diversit
Conservation，China West Normal University，Nanchong 637002，China)
Abstract：A successful mlcropropagation technique of Glechoma longituba(Nakai)Kupr was established from
the explants of auxiliary buds and stem tips． The result showed that：(1)MS medium with 0
． 1 mg／L NAA
and 1·5 mg／L BA could promote the growth of vegetative auxiliary buds and regeneration of plentifu1 fatera1
buds．(2)Shoots derived from auxiliary and lateral buds could form roots on MS medium with 1
． 0 mg／L IBA
and 1．0 mg／L KT，the proportion was up to 100 ．
Key words：Glechorna longituba；tissue culture；rapid propagation
CLC number：Q943．1 Document code；A ArticLe ID：1000—3142(2006)03—0286—03
Glechorna longituba(Nakai)Kupr is an impor—
tant medicinal herb (Chinese Pharmacopoqia，
2000)．In the tradi tional system of Chinese medi—
cine，G．1ongituba is a reputed medicinal herb and
contains lots of useful chemical compositions in its
leaves，stems and roots such as flavonoids，ursolic
acid，choline and so on(Wang，et a1．，1995；Jin，
2001)． At the moment，the wild stock has been
markedly depleted because pharmaceutical compa—
nies largely depend upon material procured ftom
naturally occurring stands，which are being deple—
ted rapidly raising concern about possible extinc—
tion of the species and providing j ustification for
the development of in vitro propagation techniques
for plant regeneration from stem—tip and auxiliary
buds of G．1ongituba． In recent years，there has
been an increased interest in in vitro culture tech—
niques which offer a viable tool for mass multipli
cation and germplasm conservation of rare，endan—
gered and threatened medicinal herbs(Li，200 3)．
Since 20th century，the tissue culture and the fast
reproduction technology already widely have ap—
plied in each kind of Labiatae medicinal plant(Du—
an et a1．，2001)Therefore，it is important to deve卜
op an efficient micropropagation technique for G
．
1ongituba to rapidly disseminate superior clones
once they are identified．Tissue culture technique
can play an important role in the propagation of G
．
1ongituba and the elite herb conservation of G．Z0 —
gituba．In addition，compounds from tissue culture
may be more easily purified because of simple ex—
traction procedures to possibly reduce the produc—
tion and processing costs．
1 Material s and methods
1．1 Plant materiaI
Auxiliary buds and shoot—tips were collected
from G．1ong ituba grown in greenhouse．
1．2 Experiment methods
1．2．1 Surface sterilization The materials were
surface sterilized as follows：(1)Flushing for 3 O
收稿 日期：2005—07—26 修回日期：2005—12 25
基金项目：四川省杰出青年学科带头人培养计划项目(O4ZQo26—047)；四川省科技厅基础项 目(03JY029 021—22)[supported by
Outstanding Youth and Leader Cultivated hem(04ZQ026—047)Sichuan Science and Technology Basic Item 0f Sichuan(03JY029
021—22)]
作者简介：韩素菊(1976一)，女，四川资中人，硕士研究生，主要从事药用植物的有关研究。
通讯作者(Author for correspondence，E—mail：yx
_ li@263．net)
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3期 韩素菊等：连钱草组培快繁技术研究 287
min under running tap water followed removal of
1eaves．Shoot tip pieces of about 1_5 cm were ex-
cised from the material；(2)Rinsing with 70 9／6 etha—
nol for 10 s；(3)Sterilizing with 2 0A Javel water for
30 seconds and washing with autoclaved sterilized
distilled water for 5 min(4 times)；(4)Trimming
the explants to about 1．0 cm prior to inoculation in
culture medium．
1_2．2 Culture medium Explants were placed on
solid basal MS medium with sucrOse(3．0 g／L)and
agar(0．8 g／L)supplemented with different concert—
trations and combination of BA(0．5，1．0，2．0，2．5，
3．0 mg／L)and NAA(O．1 mg／L)for the prolifera—
tlon of explants． The pH value of the media was
adjusted to 5．8 using 0．1 N HC1 or 0．1 N NaOH
before autoclaving．M edia were sterilized at 121℃
and 104 kPa for 15 min．After thirty days，the pro一
1iferations of buds were re—divided and transferred
onto another MS medium with 1．0 mg／L IBA and
1．0 mg／L KT to induce rooting and formation of
complete plantlets．
1．2．3 Culture condition AlI the cultures were main-
tained at(25±2)℃ with 12～14 hour’s photoperiod in
CO01 white fluorescent 1ight(1 500~2 000 1x)．
2 Result and discussion
2．1 Effects of sterilization
The sterilizing duration of explants had influ—
ences on the percentage of pollution． The results
of observation and statistics after inoculated for 1O
days can be found in table 1．
It was difficult to sterilize explants because of
its dense epiderma1 hairs．Almost all of the ex—
plants were decontaminated after a short duration
in Javel water．While explants were dipped in the
Javel water for 10 s，the percentage of pollution
was 80 and it was the highest． After sterilizing
for 30 s，it was 10 ．There was no obvious influ—
ence on the percentage while the explants were
dipped for 30～ 50 s． However，the proliferation
buds became thin and weak after explants were
dipped in the Javel water for over 40 s．Consider—
ing the percentage of pollution and the quality of
the proliferation，the best sterilization duration
should be 30 s．
2．2 Proliferation of shoots
Explants were cultured on MS medium with
different cOncentrations and combinations． There
was an obvious sign with the alteration of hor
mones．The results were observed after being cul—
tured for 30 days．
Table 1 Effects of the different sterilization duration
on the percentages of contaminated explants
Table 2 Effects of different c0ncentration 0f
hormones on induction proliferation
Each segment formed 5～ 10 shoots by the
proliferation of apical and axillary meristems．In
Table 2，the different times of proliferation could
be due to the varying concentrations of BA used in
the medium． The times of proliferation increased
with an increase in the concentrati0n of BA and the
rate varied from 2．07 to 13．12，so the higher con—
centration of BA was found to promote the prolif—
eration on plantlets． H owever，the young shoots
became thin，long and weak with increasing in con—
centrations． It makes the surviva1 rate of trans—
plant lower．Considering the rate of proliferation
and the quality of plantlets，the proper eoncentra—
tion of BA should be 2．0 mg／L when inducing the
proliferation of multiple shoots(Figs．1_3：1)，
2．3 Root development in regenerated plants
The proliferation buds were separated and
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288 广 西 植 物 26卷
transferred to the rooting medium tO form COrn—
plete plantlets．Roots were formed on young buds
grown in MS medium containing IBA(1．0 mg／L)
and KT(1．0 mg／L)(Figs．1 3：2)．A high percent—
age of shoots(100 )rooted in the medium．Root
initially formed in 8，～ 1 0 d and developed into a
good root system in 12～ 16 d． Even roots were
formed on MS medium without growth regulators．
This may show that G．1ongituba has inherent hot—
mone tO promote itself to root．
Figs．卜3 1．Buds of clone；2．Roots of cloned plants；3．Survival plants On nutritional soil
2．4 Acelimatization of rooted plant
After being cultured for 1 5～ 20 d in the taking
roots media，when the plantlets were 3～ 4 em
high，with 6～ 9 roots(2～ 3 cm long)，the tubes
were taken tO the outdoor．TWO days later，rooted
plantlets were taken out of the tubes and washed
off the medium．The plantlets were transferred in—
tO pots containing sand：humus in the ratio of 3：1
and the pots were put under the shade． Ten days
later，the plantlets grew well(Figs．1-3：3)．
In conclusion，we have developed the tissue
culture system of an important medicinal herb of
G+longituba．The results of this study will hope—
fully offer US opportunities not only for rapid and
economical propagation Of G．1ongituba，but also
for its conservation of the species．
参考文献：
中华人民共和国卫生部药典委员会．2000．中华人民共和国
药典IS]．北京：化学工业出版社：134—135．
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草药 ，26(10)：557．
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[J]．中医中药，8(11)：5,t．
Duan HJ(段黄金)，Gao JS(高疆生)，Zhao SZ(赵书珍)，et a1．
200 1．A study on propagation of Coleus blumei(彩叶草的组
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(1)：25—27．
连钱草组培快繁技术研究
韩素菊，黎云祥 * ， 杨子松，姜天亮，李 尤
(西华师范大学 四川省环境科学与生物多样性保护重点实验壁，四川 南充 637002)
摘 要：用连钱草无菌茎尖为外植体进行快速繁殖，分别诱导、分化、生根形成再生植株进行快速繁殖，并移
栽成活。结果表明在 MS+6一BA 1．5 mg／L+NAA 0．1 mg／L培养基上诱导丛生芽效果最佳。在 MS+IBA
1．0 mg／1+KT 1．0 mg／L培养基中根的诱导率为 100N。
关键词：连钱草；组织培养；快繁
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