全 文 ：植　　　物　　　研　　　究
BULLETIN OF BOTANICAL RESEARCH
第 18 卷　第 2 期 1998 年　4 月
Vol.18　No.2 April , 　1998
香石竹花可恢复性乙烯反应的研究
姜微波1　Shimon Mayak2　Abraham H.Halevy2
(1.中国农业大学食品科学系 , 北京 100094)
(2.Department of Ho rticulture, Faculty of Ag riculture , The Hebrew University of Jerusalem , P.
O.Box12 , Rehovo t 76-100 , I srael)
摘　要　AOA处理花瓣抑制乙烯合成 ,大大提高了小剂量乙烯(12h x 1μl/ l)诱导的
卷曲花瓣恢复为正常花瓣的比率 ,但对大剂量乙烯(12h x 10μl/ l)诱导的卷曲花瓣
影响很小 ,经小剂量乙烯处理的花瓣中乙烯合成速率很低。卷曲花瓣能否恢复为正
常花瓣取决于花瓣所接受的乙烯剂量及花瓣中内源乙烯的合成能力 。
关键词　香石竹;乙烯;反应;敏感性
THE STUDYOF REVERSIBLE RESPONSES
OF CARNATION FLOWERS(DIANTHUS
CARYOPHYLLUS l.)TO ETHYLENE＊
Wei-Bo Jiang 1　Shimon Mayak2　Abraham H.Halevy2
(1.Corresponding author , Depar tment of Food Science , China Agricultural University , Beijing
100094)
(2.Department of Horticulture , Faculty of Ag riculture , The Hebrew University of Jerusalem , P.
O.Box12 , Rehovo t 76-100 , I srael)
Abstract　When f lowers were pre -t reated w ith AOA to inhibit ethy lene synthesis ,
the recovery of the flowers from inrolling w as markedly enhanced af ter exposure to a
low dosage of ethylene(12h x 1μl/ l),but it was not changed af ter exposure to a higher
dose of ethylene (12h x 10μl/ l).After the f lowers were t reated w ith the low dose of
ethylene , the ethylene production rate in the recovered petals remained at low level sim-
ilar to that in petals not exposed to ethy lene.Ethy lene production rate ,ACC content
and ACC oxidase activity in the non-recovered petals w ere g reatly higher than in re-
covered petals.The recovery abili ty f rom inrolling of petals depends on the dose of
本文是姜微波在 The Heb rew University of Jerusalem 攻读博士研究工作的一部分。 This research was supported in
part by the US-Israel Binational Agriculyural Research and Developmen t Fund(BARD)US-1876-90R.收稿日期:1997-4-29
ethylene and the capability to synthesis endogenous ethy lene.The differences in capaci-
ty to synthesis ethylene among the petals t reated w ith the same dosage of ethy lene may
reflected the dif ferences in sensitivity to ethy lene of the petals.
Key words　Carnation;Ethylene;Response;Sensi tivi ty
Introduction
Carnation f lower w as used as a model system fo r many studies on ethy lene-induced flow er
senescence ,which is indicated by inrolling of the petals(“ sleepiness”)and accompanied by a
marked increase in ethylene production , which was termed climacteric ethy lene production
(Nichols 1968).This increase in ethy lene plays a major role in the regulation of petal senes-
cence.Treatment of f low ers w ith inhibito rs of ethylene synthesis or action prevents typical petal
senescence(Borochov and Woodson 1989).The continued presence of ethylene seem to be a re-
quirement for the development of the senescence symptoms.The later is suppo rted by the obser-
vation that inrolling induced by ethylene in young f lowers may be reversed af ter removal of
ethylene(Mayak and Kofranek 1976).Also ,blocking the sites of ethylene action by norbornadi-
ene has effectively reversed inrolling symptoms(Wang and Woodson 1989).Litt le is known
how this reversal response occur and what is its relationship w ith the climacteric ethy lene pro-
duction.These issues were addressed in the present study.
1.Materials and Methods
1.1　Plant material and treatments.Carnation flowers(Dianthus caryophyl lusL., cvs White
Sim and Sandrosa)were g row n in the g reenhouse of the Department of Ho rticulture as previous-
ly described(Jiang et al 1994a).Ethy lene t reatment and chemical t reatments were carried as de-
scribed before(Jiang et al 1994b).
1.2　Ethylene product ion 、activi ty of ACC oxidase ACC content were measured as described
befo re(Jiang et al 1994a).
1.3　Activity of ACC synthase.ACC synthase activi ty w as determined basically acco rding to
Boller and Kende(1980)w ith some modifications as described below .The petals(1g)were
frozen under liquid N2 and homogenized w ith a mortar and pestle in 3 ml of 50 mM Tris-HCl
buffer containing 1 mM dithiothreitol (DTT) and 1 mM phenylmethylsulfony l fluoride
(PMSF),PH 8.2.The homogenate was centrifuged at 13000g for 20 min , the supernatant w as
collected , and 1 m l of the ex tract w as desalted on a column of Sephadex G-25(1.5×5).Fo r
determination of the activity of ACC synthase ,0.1 ml of the desal ted ex tract w as placed in a
g lass tube together w ith 0.1 mM SAM , 5m py ridoxal-5-phosphate and 10 mM Tris-HCl
buffer containing 1 mM DTT and 1 mM PMSF ,pH 8.2 , final volume 0.5 ml.The mixture w as
kept at 28℃ for 1 h.The ACC content of the samples was determined according to Lizada and
Yang(1979).
2532期 姜微波等:香石竹花可恢复性乙烯反应的研究(英)
2.Results
2.1　Dose related reversible response to ethylene of carnation petals.
When Sandrosa f low er petals w ere treated w ith ethylene for 12h ,petals inrolling occurred
at ethy lene concentration as low as 0.5μl/L (Fig.1).Pre-t reatment wi th AOA had no ef fect
on init ial inrolling of the petals.Recovery from inrolling w as observed after the ethy lene treat-
ment ,which w as inversely related to the applied ethy lene concentration.
F ig.1　Dose related reversible response to ethy lene of cv.Sandrosa car-
nation petals.Petals w ere treated in AOA(2 mM)4h prior to the ethy-
lene treatment for 12h.Extent of inrolling w as evaluated at 0 and 24h
after termination of the treatment.Means of 4 tested samples S.E.
24 h after the ethylene
treatment , more than 90% in
petals treated wi th 0.5μl/L
ethylene had recovered from in-
rolling , but only 30% had re-
covered of petals t reated with
1μl/L ethy lene.When petals
w ere treated w ith 5μl/L o r
higher ethylene , the inrolling of
petals was i rreversible (only
1% in the 5μl/L ethylene
treated petals).In petals pre -
treated w ith AOA , recovery w as
g reatly enhanced.100% of the
petals recovered when treated
w ith 0.5μl/L of ethy lene.More
than 90% or 60% in petals
treated with 1μl/ L or 5μl/L
ethylene respectively.No rever-
sal w as observed , how ever , in
petal t reated w ith 10μl/ L ethy lene.Clearly , the capacity to recover f rom inrolling depends on
the ethy lene dosage applied.The above results and of the previous experiments suggested that
12h of 10μl/L ethy lene treatment is a saturated dosage for induction of irrecoverable inrolling
(senescence)for all the carnat ion f lowers and petals tested.By comparison , in petals exposed to
low ethylene dosage(lower than 5μl/L for 12h)recovery w as apparent in many petals , how ev-
er , some petals did not recover.The reason for the dif ference betw een individual petals is not
clear.The enhancement of petal recovery from inrolling in AOA treated petals suggest that the
suppression of ethylene synthesis may be a factor related to the ability of petals recover from in-
rolling .
2.2　Ethy lene production in recovered and non-recovered petals.
Ethy lene synthesis capacity of the recovered and non-recovered petals w as studied(Figure
2).When the petals were treated w ith 1μl/ L ethylene for 12h , and then transferred to ai r , the
254 植　　物　　研　　究 18卷
Figure 2　Ethylene production in carnation petals of recov-
ered and non - recovered from inrolling induced by ethy-
lene.Inrolling petals were chosen immedia tely af ter 12h
ethylene exposure(1 μl/ L)for the measurement.Petals ex-
posed to air w ere used as contro l.Means of 4 samples ±S.
E.
ethylene production rate in the recovered
petals remained at low level as that in
control petals not exposed to ethy lene.
Twenty four hours af ter the termination of
the ethy lene treatment , the rate of ethy-
lene product ion by inrolling petals show ed
a typical climacteric rise and w as about 40
times higher in non-recovered petals than
in recovered petals.By 48h af ter exposure
to ethylene the production of ethy lene rose
in non-recovered petals to 80 folds that
of recovered petals.
2.3 　ACC content , ACC synthase and
ACC oxidase activity in petals.
Twenty four hours af ter the termina-
tion of the 12h ,1μl/L ethylene t reatment ,
both ACC content and ACC-synthase ac-
tivi ty in petals recovered f rom inrolling re-
mained at low level as those in controls
(Table 1).However , the ACC content and
ACC synthase activi ty in non-recovered
petals were about 15 and 25 folds higher
respectively than in recovered petals.Although ACC oxidase activity in recovered petals w as 30
times higher than that in control , it w as only 18% of that in non-recovered petals.These re-
sults clearly indicate that ethylene synthesis capacity w as an important factor related to the abil-
ity of carnation petals to recover f rom the initial response to ethylene.
Table 1　ACCcontent ,ACCsynthase and ACC oxidase activity in carnation petals recov-
ered and non-recovered from inroll ing induced by ethylene.Inroll ing petals
were chosen immediately after 12h ethylene exposure(1μl/L)and the measure-
ments were done at 24h after the ethylene exposure.Petals exposed to air were
used as control.Means of 3 samples ±S.E.
ACC content
(n mol/ g·h)
ACC synthase activity
(ethylene nl/ g·h)
ACC oxidase act ivi ty
(ethylene nl/ g·h)
cont rol
recovered
non-recovered
0.17±0.05
0.19±0.05
2.88±0.64
0.8±0.1
0.9±0.2
24.5±3.8
1.4±0.6
41.2±6.7
227.4±38.6
3.Discussion
Initial petal inrolling induced by ethylene may be reversed when the ethylene is removed
2552期 姜微波等:香石竹花可恢复性乙烯反应的研究(英)
(Mayak and Kofranek 1976)or by t reating with no rbo rnadiene (Wang and Woodson 1989).
However , severe inrolling of carnation petals is considered unrecoverable(Halevy and M ayak
1979).Under the present experimental condit ions(10μl/L , 24℃, 12h), the severe inrolling of
the control petals w as not reversed by norbo rnadiene(data not show n).
The petal response to ethylene could be divided into two stages , initial phase and stable re-
sponse phase.If the exogenous ethylene t reatment was ceased when the petals are in the initial
response phase , the prog ress in response to ethylene w ould be dominated by their ethylene syn-
thesis capacity(ESC).If the ESC remains at low level as in petals t reated w ith 0.5μl/ L ethy-
lene o r w hen pretreated AOA , relevant processes can be reversed and the petals can recover(Fig-
urel).This view is also suppo rted by the observation that inrolling petals can be reversed by
treatment w ith no rbornadiene , an inhibitor of ethy lene act ion(Wang and Woodson 1989).How-
ever ,when the ESC in petals is high , as in petals t reated w ith higher ethylene concentration ,
they enter the stable phase w hich can not be reversed.Rapid death of the petals could thus not
be prevented in this petals even when treated w ith norbo rnadiene(unpublished results).
In summary of this repo rt ,we show ed that the recovery ability f rom inrolling of petals de-
pends the dose of ethylene and the capability to synthesis endogenous ethylene.The dif ferences
in capacity to synthesis ethylene among the petals t reated w ith the same dosage of ethy lene may
reflected the dif ferences in sensitivity to ethy lene of the petals.
References
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2.Borochov A , Woodson W R.Physiology and biochemist ry of f low er petal senescence.Hort ic.Rev ., 1989 , 11:15-43
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4.Jiang WB , Mayak S , Weiss D , et al., Regulation of a petal-specif ic ethylene-induced 70kD protein f rom carnat ion.
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256 植　　物　　研　　究 18卷