全 文 ：广 西 植 物 Guihaia 27(6)：932— 936 2007年 11月
induction and its survival rate
WANG Ya～LiI，2，W ANG Xiao—DongI，ZHAO Bing ，W ANG Yu—Chun
(1．State Key Laboratory of Biochemical Engineering，Institute of Proces Engineering，Chinese Academy of Sciences，
Bering 100080，China；2．Graduate School of the Chinese Academy of Sciences，Bering 100049，China)
Abstract：Maca(Lepidium meyenii)，belongs to the family of Brassicaceae，has high value of nutrition and medicine．
In the process of micropropagation，light had remarkable influences on maca callus growth，shoot induction and surviv—
a1．Green light and blue light not only made against maca calus growth but also inhibited shoot induction and surviv—
a1．White light，red light and yellow light improved callus growth remarkably．Under these light conditions，the shoot
induction rates were from 6O％ to 80％，and the shoot survival rates were from 29 to 36 ．Appropriate prolonging
of the time under light increased the shoot induction rate，and the appropriate photoperiod was 16h per day．But higher
intensitv of light decreased the st~vival rate of the shoots，and the appropriate light intensity was 24-41／~mol／m2。s．
Key words：spectral quality；photoperiod；light intensity；maca callus shoot
CLC number：Q813．1 Document code：A Article ID：1000-3142(2007)06—0932—05
1 Introduction
Maca(Lepidium meyenii)，a species belonging to
the family of Brassicaceae，is mainly domesticated in the
Andes Mountains in Peru where the climate is harsh and
the altitude is 3 000— 5 000 n．1． Maca was cultivated
more than 2 000 years ago and used by An dean Indians
as food and folk medicine to enhance fertility and sexual
performance of men and women(Rea，1992)．Recently，
the consumption of rl~ca is increasing as its value has
been reappraised．The research on nlaca was mainly a—
bout its compositions and pharmacological function．The
tests of pharmacology showed that rl~ca could enh ance
fertility(Kuo et a1．，2003)and sexual performance，im—
prove sexual dysfunction(Zheng et a1．，2ooo)，regulate
incretion and nourish offspring(Canales et a1．，2000)．
Analyzing the compositions of D-／~ca hypocotyls showed
that it was abundant in protein．essential amino acids，
free fattv acids and ma ny kinds of vitamin and minerals
(DinJ et a1．，1994)，some secondary metabolite such as
ma caene and ma camide(Zheng et a1．，2000)，alkaloids
(Cui et a1．，2003)and glucosinolate(Li et口Z。，2001)
were also identified in rl~ca．
Studies on the tissue and cell culture of maca have
been rarely reported．Our present work investigated
the effects of light intensity，spectral quality and photo—
period on the maca callus growth，shoot induction and
jts s1】rvival rate．
2 Material and method
2．1 Plant material
Maca callus was induced in our lab as folow：
Maca seeds were immersed in 7O (V／V)ethanol for
2 H1inutes，and rinsed three times with sterile distilled
water．Subsequently，they were put into 2 (V／V)
SOdiurfl hypochlorite solution for 2O minutes and rinsed
three times with sterile distiled water．
For germination，disinfected seeds were placed in
150 mL conical flasks containing 50ml hormone-free
MS(Murashige and Skoog’s medium，1962)medium
Received date：2006—02—24 Accepted date：2006—10～12
作者简介：2E~gg(1971一)，女，山西垣曲人，在读博士，主要从事植物细胞工程和植物快繁的研究，(E-mail)zkywyl@yahoo·com·an．
*通讯作者(Author for correspondence，E-mail：ycwang45@yahoo．corn)
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6期 王亚丽等：光对马卡愈伤组织生长、丛生芽诱导和存活的影响 933
(pH= 5．85- 5．90)and the medium was solidified by bout 0．3 cm long and cultured m l00 n comcal ilasks
a础 tion of 6 g／L agar．The seeds were cultured about for 30 days under 16 h ligh (1igh im n Y was 24
45 davs under 16h light per day at 25_+2℃．The light ~mol／m2)at 25±2℃·Each flask c。m imd 50 mL
intensity 0f 24 m。l／m was provided by white fluo— MS medium which supplemented with l mg／L 6-
rescent lamps． benzylaminopurine(6-BA)0·5 mg／L naph h l n
The mots of disinfected plantlets were cut into a— acetic acid(NAA)and 0-1 mg／L 24-dichloro’phenoxy
Table 1 Main parameters of different lamps
All lamp powers were 30 W
(2，4一D)and solidified by addition of 6 g／L agar．The
calli were subcultured every 30 days on the same cut—
ture conditions as the former．
2．2 Effects of spectral quality and photoperiod on maca
callus growth
In the tests of effects of spectral quality on 1TlaCa
callus growth。the main parameters of the lights provided
bv five kinds of lamps are listed in Table 1．The time of
irradiation was 16 h per da y．In the tests of photoperi—
od。the time of irradiation was 4 h，8 h，12 h and 16 h re—
spectively and the light intensity was 24／xmol／m2．s
which was provided by fluorescent lamps．0．6 g fresh
weight calli were cultured on 50 n MS medium (pH-
5．85—5．90)in 100 mL conical flask for 30 days at 25±
2℃ under diferent light conditions．The medium was
supplemented with 0．5mg／L 6-BA，0．25mg／L NAA and
0．1 mg／L 2，4-D，and solidfied by addition of 6 g／L
agar．Each cuhure condition had three repetitions．The
callus fresh weights(FW)in each flask were weighted
and then dried in an oven at 50℃ until their dry weights
(DW )were constant．
2．3 Effects of spectral quality。photoperiod and light in。
tensity on shoot induction
The conditions of spectral quality and photoperiod
were the same as in the former experiments． In the
tests of light intensity，the light intensities were 58
#tool／m2·s，24 b~mol／mz·s，17 gmol／mz·s and 14
#mol／m ·s respectively．Al induction temperatures
were 25±2℃．The differentiation medium was MS
supplemented with 2 mg／L 6-BA，0．25 mg／L NAA．
Each conical flask contained five pieces of calli，and
each calus piece was about 0．7 cma．Alter 25 days，
the shoot induction rates were calculated．
2．4 Effects of spectral quality，photoperiod and light in。
tensity on shots survival rate
The shoots which were induced on differentiation
media under white light at 25± 2℃ for 25 days were
cuItured 0n subculture media under different light con-
ditions at 1 5± 1℃ ．The subculture medium was the
same as differentiation medium． After 25 days，the
well—growing shoots were counted and the shoot liva—
bilities were calculated． The parameters of spectral
quality and photoperiod were the same as the form er．
The light intensities were 96／~mol／mz．s，81#mol／mz．
s。53#mol／m2．s，41 b~mol／m2．s and 24#mol／mz．s re—
spectively．
3 Result
3．1 Effects of spectral quality and photoperiod on maca
callus growt h
In the tests of light influence on callus growt h，
different photoperiod had 1ittle distinct effect on callus
growth (Table 2)，but spectral quality could affect
maca callus growth remarkably (Table 3)． The bio—
mass of maca calli decreased remarkably under blue
light and green light than under white light，but they
had no discrepancy under yellow light，red light and
white light．
3．2 Effects of spectral quality，photoperiod and light in。
tensity on shot induction
In the experiments of light influence on shoot in—
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934 广 西 植 物 27卷
duction，the shoot induction rates slightly increased as
fight intensities decreased (Fig．1)and photoperiods
prolonged(Fig 2)．Shoot induction rates were different
when the calli cultured under different spectral qualities
(Fig．3)．Under red light and yellow light，the shoot
induction rates were 60 which resembled that under
white light(8O )．Green light and blue light made a—
gainst shoot induction．Under these two lights respec-
Table 3 Effects of spectral
tively，the shoots almost died or grew abnormally，and
the shoot induction rates were nearly zero
．
Table 2 Effects of photoperiod on maea callus growth
FW 5．252：t：0．113 5．359=1：0．050 4．466=t：0．195 3．8515=0
． 387
DW 0．406士0．034 0．357士O．035 0．346=t=0．012 0．344~0．089
Note；Indicates mean standard deviation
quality on maca calus growth
Note：Compared with white fight，significant differences(t=0．01)are indicated with*·；+indicates mo
_~tl srandard deviati0n
§80
70
60
C
．
o
． 50
4J
40
_c 30
4
o
J 20
兰 l0
∞
0
14 1 7
kight intens
24 58
ty (umoI／m ·s)
Fig．1 Effects of light intensity on maca
shoot induction rate
4 8 1 2 1 6
Photoper i od(h)
Fig．2 Effects of photoperiod on maca shoot induction rate
3．3 Effects of spectral quality，photoperiod and light in-
tensity on HIaca shoot survival rate
Spectral quality，light intensity and photoperiod all
had remarkable effects on shoot surviva1 rate．There
was no discrepancy in shoot survival rate when the
shoots were cultured under red light(29 0A)and white
light(33 )．Under yellow light，the shoot survival
wk RL YL GL BL
SpecttaI quaI itY
Fig．3 Effects of spectral quality on maca
shoot induction rate
rate(36 )was higher than those under other lights．
Contrarily，the shoot surviva1 rate was zero when they
were cultured under green light and blue light(Fig．
4)．In the tests of light intensity on shoot Surviva1．the
shoot surviva1 rate decreased as light intensity in—
creased．The highest survival rate of shoots was 33
when the light intensity was 24／~mol／m。·s and 41
gmol／m2·s． The phenomenon showed that tOO
strong light was harmful to shoots(Fig．5)．The re—
suhs of photoperiod tests showed that shoot surviva1
rates increased from 10 to 29 when the photopefi—
od was prolonged from 4 h to I6 h(Fig．6)．The opti—
mized photoperiod to shoot survival was 16h per day．
4 DiSCUSS
Light can influence plant growth through three
aspects：light intensity，photoperiod and spectral quali一
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936 广 西 植 物 27卷
shoot livability．Under red light and yellow light，the
shoot induction rates resembled that under white light．
The survival rate was higher under yellow light than
those under other lights． But under green light and
blue light，the shoot induction rate and its survival rate
were zero respectively．This phenomenon supports our
estimation：there are some photoreceptors in maca cal—
lus which can utilize the long wavelength lights more
effectively．
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光对马卡愈伤组织生长、丛生芽
诱导和存活的影响
王亚丽1，2，王晓东1，赵 兵1，王玉春1
(1．中国科学院 过程工程研究所 生化工程国家重点实验室，北京 100080；2．中国科学院 研究生院，北京 100049)
摘 要：马卡属于十字花科独行菜属，具有极高的营养价值和药用价值。在快繁过程中，光对马卡愈伤组织生
长，丛生芽的诱导和存活有显著的影响。绿光和蓝光既不利于愈伤组织的生长也不利于丛生芽的诱导和存活。
白光、红光和黄光能明显促进愈伤组织的生长，在这些光照条件下丛生芽的诱导率为 6o ～8o ，丛生芽存活率
为29 ～36 。适当延长光照时间可提高丛生芽的存活率，合适的光照时间为 16 h／d。但是过强的光照可使丛
生芽的存活率降低，合适的光照强度为 24~41／zmol／m s。
关键词：光质；光周期；光强；马卡；愈伤组织；丛生芽
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