全 文 ：广 西 植 物 Guihaia 24(5)：466—470 2004年 9月
通风对好好吧无性累芽的增殖能力
和抗旱能力的影响作用
周延清1，2 ’ 贾敬芬2
(1．河南师范大学生命科学学院，河南新乡 453002；2．西jE大学生命科学学院，陕西西安 710069)
摘 要：该文旨在研究通风在不同好好吧无性系芽增殖阶段对它们增殖体的叶面积、地上部分总面积和失水
能力的作用，筛选抗旱性好的好好吧无性系。研究结果表明，在 13个好好吧无性系中，通风使他们的叶面积、
总面积和抗失水力均有增加。其叶面积分别增加了 72．2 到 470．5 ，其总面积分别增加了 39．0 到
145．8 ，其抗失水力分别增加了(失水分别减少了)22．2 到 138．0 。从中选择了一个叶面积和总面积都增
加最多(分别为470．5 与 145．8 )和失水减少次多(86．7 )的耐旱好好吧无性系CQ63，可供沙漠和旱地种植。
关键词：好好吧；增殖；通风；失水
中图分类号：Q944．6 文献标识码：A 文章编号：1000—3142(2004)05—0466—05
Effect of ventilation on the proli feration
and drought·。resistance of j oj oba
／ 一 ● l ● ■ ● ● ＼ l
【、 lm m ona 口 C，z nensis Cl0neS
ZHOU Yan—qing 一，JIA Jing-fen
(1．College 0，L fe Sciences，Henan Normal University，Xinxiang 453002，China；
2．College of Life Sciences，Northwest University，Xi an 710069，China)
Abstract：The purpose of this research is tO study effect of ventilation on leaf area，shoot area and water lOSS in
different jojoba clones，and select some drought—resistant jojoba clones from those at their proliferation stage．
The research results showed that ventilation helped increase their leaf areas from the percentage of 72．2 tO
470．5 ，total area of jojoba propagules from 39 tO 145．8 and their resistance tO(decrease in)water loss
from 22．2 tO 138．0 in 13 jojoba clones．One good drought—resistant jojoba clone，CQ63，was selected with
the most increase of 470．5 in Iear area and 145．8 in total area as well as a greater decrease of 86．7 (the
second-most one among 1 3 clones)in water lOSS tO be planted in arid desert and land．
Key words：jojoba(Simmondsia chinensis)；proliferation；ventilation；water lOSS
Jojoba(Simrmndsia chinensis L．)is fl native
oilseed shrub of the Sonoran desert of the south—
western USA and northern Mexico(Vermaut Sa—
bien et a1．，1995)，an evergreen crop of high eco—
nomic interest which is grown for the exceptional
quality of its seed oil(Amarger and Mercier，1995)．
Native jojoba populations have growing tempera—
ture ranging from 9℃ tO 5O℃ (Gentry HS，1958)，
being drought—resistant and salt—resistant(Mills
and Benzioni，1977；Yermanos，1977)． The dry
Received date：2003-10-22 Accepted date：2004—-02—-16
First author：ZH0U Yan-qing(1963一)，Male，born in Dengzhou city，Henan Province，Doctoral student，associate Professor，grad—
uate advisor，working in plant cel engineering and crop genetic breeding． Author for correspondence E-mail：WangXL@edu．
nw u．cn
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5期 周延清等：通风对好好吧无性系芽的增殖能力和抗旱能力的影响作用 467
seeds of jojoba contain about 5O oA lipids in the
form of simple wax esters that have properties sim—
ilar to those of sperm whale oil，which are used in
cosmetics，pharmaceuticals and polish industries
(Jaeoboni and Standardi，1987；Benzioni et a1．，
1992)．Therefore，jojoba was introduced to many
countries during the late 1970s and early 1980s．
Currently，the main jojoba oil producers are Is—
rael，USA，Argentina，Australia and Mexico．Now—
adays，increasing attention has been paid to Jojoba
plantation and breeding． New jojoba plantations
are grown from vegetatively propagated material
and seeds，but in vitro propagation is fl good alter—
native method offering some important advantages
such as mass propagation without the limitation by
the number of explants and the production of
pathogen—-free plants and new good·。quality breeds
or cultivars(Mills et a1．，1997)． Because tissue
culture technique is being widely used for crop im—
provement and clonal multiplication，the selected
jojoba clones are multiplied by micropropagation
method(Benzioni et a1．，1992；Sardana and Batra，
1998；Farook，1998；Roussos eta1．，1999)，andjojo—
ba micropropagation has been described(Mills et
口Z．，1997；Roussos et a1．，1999)．However，micro—
propagated jojoba plantlets often lose their ability
to control water loss caused by high relative hu—
midity and limited gas exchange(Mills et a1．，2001；
Ziv，1 9 9 1)．The improvement of the ability through
ventilation(by using specific membranes)at the elonga—
tion stage had been reported(M．1ls et a1．，2001)．But
up to now，the efect of ventilation on jojoba clones at
the proliferation stage has not been shown．Therefore，
in the present study，effect of ventilation on different
jojoba clones is studied and drought-resistant j 0joba
clones are selected from them．
1 Materials and Methods
1．1 Plant materiaI and culture conditions
Jojoba clones used in the present stIJdy are mi—
co-propagated and provided by DAVID Mills and
ALIZA Benzioni’s plant tissue culture laboratory
in Institutes for Applied Research，Ben—Gurion U—
niversity of the Negev，Israe1． They include
Clone64，Clone154，Clone Q63，Clone78，Clone85，
Clone F，Clonel41，Clone96，Clone OMER，Clone
Male，Clone K120，Clone32，Clone Kalial1．They
were euItured in 5 mE of modified MS medium
(Mills et a1．，1997)for proliferation in fl growth
room．After fl culture of 7 weeks，they were cut in-
to single nodal segments，which had opposite axil—
lary buds and leaves and contained two parts：one
above buds and leaves，about 0．5 mm long~another
under them，about 10 mm．Eight nodal segments
excised from each clone were cultured in 70 mL of
the same medium as above for proliferation in fl
growth vessel，magenta box(375 mL in volume)．
Every growth vessel was sealed with one lid which
had one 1 6 mm diameter membrane at the center，
corresponding to fl membrane area of 207 mm ，to
create ventilation treatments．In the control treat—
ment，every growth vessel was sealed with one lid
without any membrane，and every vessel also con—
tained eight nodal segments from the same clone．
AIl the vessels were sealed with adhesive tape at
the lid edge．Nodal segments inside them were cul—
t ured for twenty eight days in fl growth room at a
temperature of 25+ 1℃ under 45～55 pmol m。 s。
photon flux supplied by cool white fluorescent
bulbs for fl photoperiod of 16 h light．
1．2 Growth measurements
After fl culture of twenty eight days，the fol—
lowing parameters were determined：shoot area，
leaf area and water lOSS．For area determination，
shoots and leaves were photocopied，the image was
scanned，and the area was measured with the image
analysis software NIH Image．For water loss de—
termination，propagules were taken out of the yes—
sel s separated from callus at the base and weighed
every ten minute(6O minutes／seven times)in an
electric balance under room conditions(usually 23
±2℃ and 61±3 RH)．Water lOSS was ealeulat—
ed by dividing the loss of weight after designated
periods by the propagule surface area．
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468 广 西 植 物 24卷
1．3 Statistics
In each experiment，one vesse1 containing eight
propagules was examined for every treatment．Six—
teen propagules(control and treatment)in every
clone were sampled for area and water loss．Exper—
iments were repeated 2～3 times．Data were evalu—
ated by analysis of variance(Fisher s Protected
LSD；Probability of 5 )using super ANOVA．
Figures were prepared with Exce1．
2 Results and Discussion
All segments at the multiplication stage were
cultured in magenta vessels with 16 mm diameter
membrane or closed vessels(no membrane)．After
twenty eight days，the above thirteen jojoba clones
(Fig．1)were examined for leaf area，shoot area
and warer lOSS．
Fig．1 Jojob clone Kaliall’。propagules cultured for
28 days in growth vessels and in growth room
Upjevery vessel sealed with oQc-lid which had ole
1 6 mm dlBreeder br at the center
Downt every wesse]sealed svith one dosed l试
2．1 Elfect of ventilation on leaf area and total area
of propagules in jojoba
As shown in Fig．2 and Fig．3，ventilation had
a posilive effect on the growth of teaf area and total
shoot area of propagules in different jojoba clones．
According to the equation：area increase rate leaf
area or total shoot area of tested propagule／leaf ar—
ea or tota1 shoot area of control propagule．From
clone C64 to clone Kalial 1：(1)Leaf areas of jojoba
propagules from the vessels with 16 mm diameter
membrane in turn increased 247．3 ，227．5 ，
470．5 ．347．4 ，148．8 ，192．9 。73．9 ，
72．2 ，92．1 ，95．8 ，250 ，265．7 and
155．4 over than that of those from the closed
ones without membrane．Among them，leaf area of
done C63 increased most，by 470．5 ．The next
one was clone C78．whose leaf area increased by
347．4 ．Leaf area of clone C96 increased least，by
72．2 i(2)Total areas of jojoba propagules from
the vessels with 16ram diameter membrane in turn
increased l 67 ．50 ．145．8 ，81．5 ，69．2 ，
80％，57 ，39 ，4O ，50 ．76．8 ，47．6 and
122 over that of those from the closed ones with—
out membrane． 0ut of them，tota1 area of clone
CQ64 increased most，by 167 ．The next was
clone CQ63 and clone Kalial 1．whose total area in—
creased by 145．8 and 122％，respectively．That
in cloneC96 increased least。only by 39 ．The in—
crease of Leaf area and shoot area caused by ventila—
tion might be due to two aspects：one was that yen—
tilation could decrease the inhibition of such vola—
tiles as ethylene，ammonia，methyljasmonate and so
forth on propagules，which could not be accumula—
ted because of ventilation；the other was that venti—
lotion could improve the carbon dioxide supply in
growth vessels so that phot0synthe sis was en—
hanced and propagules could grow wel【． Mean—
while，leaf color was light green in closed vessels
and darker under ventilation in the above clones
like Rose and hosta plantlets(Shallanon and
Maziere，1992；Murphy et a1．，1998)． Relatively
high water content is a characteristic of hyper—
hydric plants(Z v．1991)．Reduction of water con-
tent by venti[ation was also reported for carnation
(Majada et a1．，1997)．In papaya and potato，it
XVRS also showed that ventilation made leaf area in-
creased(Lai eg a1．，1 998)．However，because of re—
duced RH。reduction of leaf area in potato was also
re口orted(K0sai et at．．1993)．The di[ferenee in re—
sponse to ventilation among different species might
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5期 周延清等：通风对好好吧无性系芽的增殖能力和抗旱能力的影响作用 469
be due to the differences in ventilation rate and
media or plant genotypes(Mills et a1．，2001)．
2．2 Effect of ventilation on water loss of jojoba
propagules in thirteen different clones
As shown in Fig．4，all the propagules in dif—
ferent jojoba clones from the vessels with 16mm di一
蔓
30 0
250
200
l 50
l 00
5 0
0
ameter membrane lost water at slower rate．This
demonstrated that ventilation had a negative effect
on water loss of the propagules in different jojoba
clones．According to the equation：water loss rate
— water loss weight of control pr。pagule／water
loss weight of tested propagule．From clone C64 to
_
． ． ． ． ． ． I．广} ．广J| ； ．j．『E ．r} ．广
c64 c1 54 cO63 c78 c 85 cloncF C 14 I C96 COMER CMale CK 1 2O C 32 Ckalia
Clone
Fig．2 Effect of ventilation on leaf area in different jojoba clones grown for
twenty eight days at the proliferation stage
Values mean+SE of eight shoots
C64 C1 54 CQ63 C78 C85 CloneF C l4l C96 C0MER CM ale CKl2 0 C 32 Ckalial
CIo13e
Fig．3 Effect of ventilation on total area of shoots(stem and leaves)in different jojoba
clones grown for twenty eight days at the proliferation stage
Values mean~SE of eight shoots
clone Kalial 1，during sixty minutes，their water
loss in turn decreased 138 9／5，41 9／5，86．7％，52 9／5，
22．2 9／5，37．6 9／5，58．3 ，47．2 ，7O．3 ，23．5 ，
76．9 ，24．3 ，8O and 70 over than that of
those from the closed ones without membrane．A—
mong them，the clone C64 decreased most，by
138 9／6．The next one was clone CQ63，whose water
loss decreased by 86．7 ．That in cloneF decreased
least，only by 22．2 ．Therefore，the response of
different clones to ventilation was rather different．
The higher its water loss rate was，the less a clone
lost water．Such resistance can result from closure
of stomata，which was caused by the decrease of
their density when humidity declined in the culture
tubes(Ghashyghaie et a1．，1992)or a decrease in
cuticular transpiration due to increased deposition
of epicuticular wax on the leaf surface or both(Ziv，
1991 Ziv，1986；Majada et a1．，1998)． The im—
provement of stomatal function in response to yen—
tilation is due to a decrease in K+c0ncentrati0n in
guard cels(Majada et a1．，1998)．
In conclusion，al the thirteen jojoba clones ex—
amined had increase in leaf area。total area(1eaf and
stem area)and water lOSS under ventilation，so yen—
tilation wil help jojoba clones grow and resist wa—
ter loss．It is possible that drought—resistant jojoba
clones can be selected through ventilation． Here
one good drought—resistant jojoba clone，CQ63，was
∞ 如 鲫蜘 如 ∞ 印们 0
uI鼋 —耍
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470 广 西 植 物 24卷
。
g 40
岛
g 30
、 -一
罾20
一
墨lo
砖
≥ o
C64 CI54 cQ63 c7$ cg5 CloneF CI4I C96 COMER CMale CKI20 C32 CKalial I
Clone
Fig．4 W ater loss during 60 minutes
Effect of ventilation at the proliferation stage on subsequent water loss of shoots in clones C64一Ckalial 1．Shoots were removed from
vessels，kept under loom conditions and weighed periodicaly．Values mean~SE of eight shoots per treatment and per clone．
selected with a larger increase of both 470．5 in
leaf area and 145．8 in total area as well as a
greater decrease of 86．7 in water lOSS．
Acknowledgements We thank Dr．DA D Mills and
Prolessor ALIZA Benzioni of Institutes ior Applied
Research，Ben-Gurion University of the Negev，Israel，
for their jojoba plant materials，instruction and help
during this study in their laboratory．
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