Plant Physiol.

(1972) 50, 585-590

The Level of Phytohormones in Monoecions and Gynoecious

Cucumbers as Affected by Photoperiod and Ethephon1
Received for publication January 28, 1972

J. RUDICH, A. H. HALEVY, AND N. KEDAR

The Hebrew University, Faculty of Agriculture, Rehovot, Israel

ABSTRACT (33). When ethylene evolution from cucumber plants was ex-
amined, more ethylene was evolved from apices of the gynoe-
The endogenous levels of auxin, gibberellin, and inhibitors cious than from those of monoecious type (28).
were followed in monoecious and gynoecious cucumber (Cu- The interactions between auxin and ethylene are complex.
cumis sativus L.) plants, and in plants treated with the ethyl- Many studies have shown that auxins hasten the production of
ene-releasing compound Ethephon (2-chloroethyl phosphonic ethylene (1, 6, 8), that ethylene enhances the decomposition of
acid). Higher auxin inhibitor and lower gibberellin levels were auxin (3), and that it also inhibits the movement of auxin in the
associated with female tendency. The endogenous level of gib- plant (3). A number of effects, in the past attributed to high
berellin and auxin decreased in Ethephon-treated plants. Appli- levels of auxin, are now considered to be a result of ethylene
cation of Ethephon induced a rise in abscisic acid. Root production under the influence of auxin (5, 7).
application of abscisic acid promoted female tendency of gyno- Little is known about the interaction between ethylene and
ecious cucumbers grown under conditions which increase male- gibberellin. Antagonism between them has been found in
ness. High C02 levels, which are known to antagonize ethylene, germination of wheat and the production of both a-amylase
increased maleness of gynoecious cucumbers. The possibility and invertase (31). Their action is also antagonistic in fruit
of interrelationship between gibbereilin, auxin, ethylene, and ripening and in sex expression of flowers (13, 26). We are not
abscisic acid on sex expression are discussed. aware of any publication on the effect of ethylene on endoge-
nous gibberellin levels. Kang et al. (18) found that gibberellin
had no effect on ethylene production in bean seedlings. They
concluded that GA and ethylene effects in the development of
the seedlings were independent.
Since sex expression in cucumbers is influenced by Ethephon
as well as by auxin and gibberellin, we tried to determine in
Studies of hormone systems involved in the regulation of sex the present work the effect of Ethephon on the level of the above
expression in cucurbits have been confined mostly to auxins phytohormones as well as on ABA and other native inhibitors,
and gibberellins. Femaleness of cucumbers has been increased in an attempt to gain additional knowledge on the relation of
by application of auxin (9, 19). When homologous sections of these phytohormones to sex expression.
hermaphrodite and andromonoecious cucumber plants were
compared, higher levels of auxin were found in the hermaphro- MATERIALS AND METHODS
dite type (10). The role of auxin in the development of female Plant Material. Endogenous levels of growth substances were
flowers in cucumbers has been demonstrated (11) by growing, determined in the monoecious and gynoecious 'Bet Alpha' 2
in tissue culture, flower buds of cucumber plants from nodes lines of cucumber (Cucuinis sativus L.) plants. The gynoecious
which produce only male flowers. Addition of IAA to these line was bred from the monoecious one and differs from it in
cultures produced ovaries and stigmas. Short days and low the gene for femaleness (2). Under short day conditions it
temperatures enhanced the femaleness of certain cucumber produces female flowers from the first or second node, under
cultivars (20) and of squash (23). Nitsch et al. (23) assume long day conditions male flowers are produced at the first six
that this was due to high levels of endogenous auxin found nodes, all the rest being female. In one experiment andromo-
under short day conditions. noecious muskmelon plants (cv. Ananas PMR) were used.
Treatments with exogenous gibberellin increased maleness in In most experiments plants were grown in growth chambers
cucumbers (25) or delayed female flower formation (4). More at 8-hr photoperiod under mixed fluorescent and incandescent
direct evidence for the participation of gibberellin influencing lamp light of 3500 ft-c. Long day treatments were given by
differentiation of male flowers was reported (2, 16). They incandescent lamps of 70 to 80 ft-c. Temperature during the
found higher levels of gibberellin in monoecious than in day, 8 hr, was 28 C, at all other times it was 18 C. In some
gynoecious varieties using the diffusion and exudation methods. experiments CO2 level was also controlled in the chambers.
Femaleness has been enhanced in muskmelon by treatment In a few experiments plants were grown in a phytotron
with growth retardants (14) which affect endogenous gibberel- under natural light intensities. Long days were given by ex-
lin levels (27). Cucumber, squash, and muskmelon plants tending the natural day to 16 hr by incandescent lamps of 100
bearing male flowers have been found to produce female ft-c. Temperature during the day was 27 C and at night 22 C.
flowers after treatments with Ethephon (17, 22, 26) which re-
lease ethylene and enhance ethylene production in plant tissue
2
Seeds originating from a line bred at the Department of Plant
Genetics of the Weizmann Institute of Science, Rehovot were
I
This paper represents a part of the Ph.D. thesis of J. Rudich. generously supplied by ZRAIM Gedera Seed Co.
585

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Copyright © 1972 American Society of Plant Biologists. All rights reserved.
586 RUDICH, HALEVY, AND KEDAR Plant Physiol. Vol. 50, 1972
External Application of Growth Regulators. ABA was ap- nolic extract was partitioned with methylene chloride and
plied to plants by two ways: (a) by daily spraying at 1 to 250 chromatographed by thin layer chromatography in the two
mg/I for 2 weeks starting at the first leat stage; (b) by root solvent systems as above. The zone corresponding to authentic
application through nutrient solution. Plants were grown with ABA, detected under an ultraviolet lamp at z54 nm, was
half-strength Hoagland solution which was supplemented five scraped off and eluted with methanol. After methylation with
times at 4-day intervals with solutions containing various con- diazomethane dissolved in hexane, a 1-,ul portion ot the extract
was injected into a Packard 7400 gas chromatograph, using a
centrations of ABA. First application was at the first leaf stage.
Ethephon was sprayed on seedlings at the first leaf stage glass spiral column 1.8 m long X 3.2 mm inner diameter,
(26). For bioassay, plants with two to three leaves and an packed with 1.5% QFI on Gas-chrom Q 60 to 80 mesh. The
elongated first internode were used. The plants were cut just column temperature was 200 C; injection and detector temper-
below the youngest leaf, the tip and youngest leaf were used ature were 225 C and 195 C, respectively. Nitrogen gas flow
for extraction or diffusion into agar. of 42 ml/min served as carrier. An electron capture detector
Diffusion into Agar. Twenty apices were planted in each of was used (29) with radioactive tritium foil. Calibration curve
four Petri dishes containing 1.5% agar. The dishes were kept at relating the amount of cis-ABA methyl ester to the computed
25 C in moist chamber for 24 hr and illuminated with 600 ft-c area of the recorded peak was used for quantitative estimation
of fluorescent light. The agar was deep-frozen at -20 C over- of ABA. Retention time of ABA was 5.14 min. The results of
night and was then thawed and washed with 100% methanol. ABA content are expressed in ,ug ABA per 100 g fresh weight.
Extraction. Approximately 3 g fresh material consisting of Bioassays. The wheat coleoptile test based on Nitsch and
the apex and youngest leaf of three-leaved seedlings were ex- Nitsch (24) was used for testing auxins and auxin inhibitors.
tracted in 150 ml of 80% methanol. An identical number of The wheat variety used was M-852 (supplied by Hazera Co.,
seedlings was used for each assay. The mixture was shaken Haifa). Tests of each RF were replicated three times, each repli-
overnight in a cold room at 3 to 4 C. After decanting, the cate including three coleoptiles, i.e., nine per test.
solids were further extracted by shaking with 150 ml of 100% Barley Endosperm Test for Gibberellins and Inhibitors. Half
methanol for half an hour. The methanol fractions were united seeds of the variety Omer were used. Tests of each RF were rep-
and evaporated. licated three times, each replicate including two half seeds.
Fractionation. Several fractionation methods were tried and Standards were replicated five times. The method used was as
adapted for use with cucumber plants. The method used is indi- described earlier (12, 15). Incubation at 25 C on the rotator (1
cated in the legend to figures. rpm) lasted 32 hr. Reducing sugars in paper chromatographed
Bicarbonate and Ether Fractionation. After ether extraction with the extractants were found to be negligible and no a-
(10), the ether was washed five times with 0.2 M sodium bicar- amylase activity was found in paper tested with the extractants
bonate at pH 8.4. The bicarbonate was acidified to pH 3.0 and solvents separately.
with HCI and was washed five times with peroxide-free ether. Rice Growth Test for Gibberellins. Rice seeds of the variety
The acid ether fraction was used for determination of auxin Tan ginbozu (32) were soaked and germinated in water. On the
activity. Since auxin and inhibitor activity were found in all 5th day seven germinated seeds (with radicles only) were trans-
fractions, other methods in which auxin activity would concen- ferred to each 3- X 10-cm tube containing 5 ml of 1.5% agar
trate in one fraction were sought. In some experiments the on which a chromatogram section equivalent to one RF unit
crude extract was acidified to pH 3.0 and partitioned five times was placed and which was covered with 1 ml of water. The
into ether. seedlings were grown at 25 C at light intensity of 600 ft-c. One
Ethyl Acetate Fractionation for Auxins. Extracting methanol week after transfer the length of the second leaf sheath of the
was evaporated, the aqueous residue was acidified to pH 3 with five most developed seedlings in each tube was measured.
HCl and shaken six times with petroleum ether and then five Cucumber Hypocotyl Test. Thirty to fifty g of fresh material
times with ethyl acetate or ether. Both petroleum ether and were extracted as described above. One and one-half ml of
ethyl acetate (or ether) fractions were tested for activity. Most ether extract were loaded on each strip of paper. Chromatog-
auxin activity was found in the ether or ethyl acetate fraction.raphy was ascending using isopropanol: ammonia: water
Methylene Chloride Fractionation for ABA. The pH of the (8:1:1) as the solvent system. Equal RF values were combined
aqueous residue was raised to 8.3 with NH4OH, centrifuged at and eluted with 80% ethanol. The resulting extract was
10,000g for 15 min, and partitioned four times with methylene poured into a 250-ml beaker containing two layers of filter
chloride. The aqueous phase was acidified to pH 3.0 with HCI paper. The ethanol and water were evaporated to dryness, 3 ml
and again partitioned four times with methylene chloride. The of water were added, and 15 cucumber seeds of the variety Bet
acidic methylene chloride fraction was evaporated to dryness, Alpha were sown in each beaker. After 5 to 7 days of continu-
dissolved in distilled absolute methanol, and used for ABA ous illumination at 600 ft-c, the hypocotyl of the 10 largest
determinations. seedlings was measured. Each test was replicated three times,
Fractionation for Gibberellin Activity. Fractionation method, as were the standards of 50% GA, and 50% GA7 at concentra-
as described by Halevy and Shilo (15), was used, as well as the tions of 10 nM, 0.1 ,tM. and 1 riM.
petroleum ether, ethyl acetate method described for auxins.
Chromatography. Two-cm wide Whatman No. 2 strips were RESULTS
loaded with 1.5-ml aliquots, equivalent to 500 mg fresh weight.
Isopropanol: ammonia: water (8: 1: 1, v/v) was used as solvent. Effect of Daylength and Ethephon on Endogenous Auxin
Thin Layer Chromatography was carried out with plates Level. Under long day conditions endogenous auxin levels in
coated with 250 ,u layer of Silica Gel G (Merck). The plates the gynoecious type were higher than in the monoecious one
were developed in two different solvent systems: (1) n-propa- (Fig. 1). A relatively greater activity of inhibitors was found in
nol: n-butanol: water: ammonium hydroxide 28% (6:2:2:1, the monoecious variety at RF 0.9 to 1.
v/v): (2) benzene:ethvl acetate:acetic acid (50:5:2, v/v). The Greater auxin activity was found in monoecious plants
plates were developed by ascending chromatography to a dis- grown under short day conditions enhancing femaleness than
tance of 15 cm and dried at room temperature in a forced air in those grown under long days (Fig. 2). Auxin levels in the
cabinet. former were approximately 10 times greater than in the latter.
Detection of ABA by Gas Liquid Chromaography. Metha- In all cases auxin activity focused between RF 0.3 and 0.5,

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Copyright © 1972 American Society of Plant Biologists. All rights reserved.
Plant Physiol. Vol. 50, 1972 ETHEPHON AND PHYTOHORMONES IN_SEX EXPRESSION 587

IAA(M) showed gibberellin activity in control plants when tested by

25r the rice assay. GA activity in all three regions disappeared fol-
E
lowing Ethephon treatment (Fig. 5). High levels of inhibitor
E 23 were found in the ether fraction at RF 0.4 to 0.9.
-C

IAA (M)
C 21
24- 710--
a._ E
22K [In I i-
-110-5
IV 19
0 Monoecious Gynoecious -810-

17k-
U1 I I I I -~18
0 0.5 1.0 0 0.5 1.0 Rf
c, 16 Untreated 2 days 7 days
FIG. 1. Chromatograms of auxin-like substances and inhibitors 13 days
in monoecious and gynoecious cultivars of cucumbers. Acidic ethyl K oafter treatment after treatment after treatment
acetate fraction (pH 3.0) of apices of 18-day-old seedlings. Biologi- L
cal activity was determined by the wheat coleoptile bioassay. Each 0 ._ 1.0 0 0.5 1.0 0 05 1.0 0 0.5 1.0 Rf
chromatogram represents 500 mg fresh weight. Values are means of FIG. 3. Chromatograms of auxin-like substances and inhibitors
three replicates. of an andromonoecious cultivar of muskmelon (Ananas PMR), at
various times after a treatment with Ethephon (250 mg/l). Acidic
IAA (M) ethyl acetate fraction (pH 2.5), wheat coleoptile bioassay. Each
25 r 7-10 7
chromatogram represents 500 mg fresh weight.
_ S.D.
E 2 Ethyl acetate fraction (pH 6) Ethyt acetate acidic fraction GA3(M)
10-8 600- (pH 3)
Ec

c 21'1 10-9 400-

a,
200- V]
c-
4,1c 0- C3zciXza1C -W

-200-
3 0.5 1. I i
0 0.5 1.0 0 0.5 1.ORf
FIG. 2. Chromatograms of auxin-like substances and inhibitors Manoecious
in apices of a monoecious cultivar of cucumber seedlings grown - Gynoecious Monoecious Gynoecious
-600-
under long and short day conditions. Acidic ether fraction (pH 3.0),
wheat coleoptile bioassay. Each chromatogram represents 500 mg -800
fresh weight. 0 05 1.0 0 0.5 1.0 0 0.5 1.0 0 0.5 1.0 Rf

while IAA in the same solvent system has an R1 between 0.4 FIG. 4. Chromatograms of gibberellin-like substances and in-
and 0.5. The activity of an inhibitor found at R, 0.9 and 1.0 in hibitors in diffusates of apices from monoecious and gynoecious cu-
cumber seedlings. Activity was tested in the barley endosperm bio-
the monoecious variety was greater under short day than under assary. Each chromatogram represents 40 apices placed on agar for
long day conditions. Auxin activity decreased during the first 24 hr. Values are means of three replicates.
few days after Ethephon treatment of both cucumber and
muskmelon and returned to the previous activity levels approxi- Untreated Ethephon
mately 10 days after treatment (Fig. 3). Concurrent with the 24 GA3 (M)
decrease in auxin level, the activity of the inhibitor at R, 0.9 -10-7
to 1.0 increased and remained above its former level even 13
days after Ethephon treatment (Fig. 3). It should be empha-
sized that all bioassays were performed with apices of seedlings
which had developed three leaves. The apex did not include
leaves or cotyledons which had been treated with Ethephon.
Gibberellin and Gibberellin Inhibitor. Diffusates from the
monoecious variety had greater gibberellin-like activity than
diffusates from the gynoecious one. High gibberellin activity
was especially noted in the acid fraction (Fig. 4). The barley
E 20
-C
a)

-C
o 16
LO)
22

18

14 _
:t w

and a-amylase test also facilitates the determination of endoge-

nous inhibitors. At RF values 0.8 and 1.0 in the neutral fraction
higher levels of such inhibitors were found in the gynoecious
cucumber variety than in the monoecious one. The aqueous
fraction of the diffusate generally showed no gibberellin-like
or inhibitor activity.
Treating the monoecious line with Ethephon resulted in dis-
appearance of gibberellin activity from the acid fraction. These
results, however, were not consistent in the barley endosperm
test.
Three main regions, at RF 0.1 to 0.2, 0.4 to 0.5, and 0.7,
12 .-
0 0.5 1.0 0 0.5 1.0Rf
FIG. 5. Chromatograms showing the effect of Ethephon on
gibberellin-like substances and inhibitor levels in diffusates of
apices from monoecious cucumber seedlings. Activity was tested in
the rice seedling bioassay. Each chromatogram represents 40 apices
placed on agar in three replicates. Acidic ether fraction (pH 3.0).
Ethephon (250 mg/l) was applied to cucumber seedlings at the
first leaf stage, and apices were cut from plants at the third leaf
stage.

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Copyright © 1972 American Society of Plant Biologists. All rights reserved.
588 RUDICH, HALEVY, AND KEDAR Plant Physiol. Vol. 50, 1972

GA3(M) promoting the growth of the ovaries (Table II). The same ABA
--10 3 treatments had no effect on elongation but slightly decreased
36+
node numbers.
Effect of CO, Level on Sex Expression. Higher ethylene
32
_ Untested Ethephon evolution was associated with female tendency of cucumber
E buds (28). In the present study, high CO2 level, which is known
28 to antagonize ethylene, markedly increased maleness by induc-
c ing the formation of male flowers in gynoecious plants (Table
24 III). An average of 2.2 male flowers per plant were formed in
0 the gynoecious plants. The high CO2 level also delayed the
appearance of the pistillated flowers in the gynoecious line.
016 ffi,0
12 Table I. Effect of Ethephoni anid Sex Type onI ABA levels as
determinied by GLC
Leaves (100 g fresh weight) of four to five leaf stage plants grown
0 0.5 1.0 0 0.5 1.0 f
under long day in a phytotron were extracted with methanol and
FIG. 6. Chromatograms showing the effect of Ethephon on partitioned with methylene chloride (pH 3.0). A single spray with
gibberellin-like substances and inhibitors in diffusates from cucum- Ethephon (500 mg/1) was applied at the two leaf stage, 10 days
ber seedling apices. Activity was tested in the cucumber hypocotyl prior to ABA extraction.
bioassay. Treatment diffusion and fractionation procedure was as
in Figure 5. ABA Content
Sex Type
GA (M) Untreated Ethephon
Untreated 0o-7
2E8_ jpg '100 g freslz wt
Monoecious 1.00 10.75
21 6 6.43
Gynoecious 2.23
E
E 21

' 22 Table II. Effect oJ Root-applied ABA oni Sex Expressioni of

-C
LI)
a Gynoecious Linle of Bet Alp/ha Cucutmber
QS 2CO _ Plants were grown in a growth chamber; day temperature was
28 C; (12 hr); night temperature was 18 C. Average was 10 plants
1E per treatment. Values followed by different letters within a
1J column are significant at the 5' level (Student-Newman-Keuls
0 0.5 1.0 0 0.5 10 Pf
FIG. 7. Effect of Ethephon on gibberellin-like substances and
multiple range test).
inhibitors in extracts of monoecious Bet Alpha cucumber leaves, as
No. of Flower Budsi Node of
tested in the rice seedling bioassay. Ethephon (500 mg/i) was -Node Length of _________ First O-r
applied to 10-day-old seedlings, plants were harvested and extracted Treatment NoO. First Five
Nodes
Pistillate
Stain. Pistil late Flw
Ovgar
6 days after treatment. Acidic methylene chloride fraction (pH 3.0). inate Bud
Each chromatogram represents 3.0 g fresh weight. Plant was grown
under long day conditions in the phytotron. CM,
Control 6.9 a 20.4 1.7 a 1.5 b 6.1 a 1.0 b
ABA, I mg 'l 6.3 ab 21.9 1.0 ab 1.9 b 5.0 b 1.4 b
This effect of Ethephon on the level of gibberellins and ABA, 5 mg/i 6.1 b 24.7 0.5 b 4.3 a 3.8 c 3.3 a
inhibitors in diffusates from cucumber was found also with ABA, 10 mg/I 6.0 b 21.5 0.0 b 4.1 a 2.5 d 3.6a
the cucumber test (Fig. 6). In this test most of the gibberellin SE 0.1971 1.232 NS2 0.2653 0.3811 0.2981 0.331'
activity found in control plants appeared at RF 0.5, whereas
treated plants showed no gibberellin activity. In contrast,
'
%0
Significant at 1 level.
2 NS: not significant.
strong inhibition was evident at RF 0.5 and 0.6 and also RF 0.7 3 Significant at 5 c, level.
and 0.8 (Fig. 6).
High inhibitor activity was found in extracts of apices from
monoecious cucumber seedlings sprayed with Ethephon as Table III. Effect of CO2 Level oni Sex Expression of Monioecioius
tested in the rice growth bioassay (Fig.7). antd Gynioeciouts Linies of Bet Alpha Cucumiibers
ABA Level as Affected by Ethephon and Sex Type. ABA Plants were grown in a growth chamber, day temperature (10
content of the gynoecious line was twice that of the monoecious hr) was 26 C; night temperature was 15 C. CO2 was applied for 15
one. Ethephon increased the level of ABA several times in days from the first leaf stage. There were six plants per treatment.
both lines (Table I).
Effect of ABA on Sex Expression. ABA applied as a daily Node of First Pistillate -Node of First Alale
Flower Flower
foliar spray of 1 to 250 mg/l did not affect growth or sex CO2 Level
expression of plants in both monoecious and gynoecious Monoecious Gynoecious Mlonoecious Gynoecious
cucumbers. Root application of ABA (1-10 mg/l) to monoe-
cious line had no effect on type of flowers formed up to the tnl/l
tenth node. However, in a gynoecious line grown under con- 300 -1 3.3 4.3 -1
ditions which promote maleness, root applied ABA markedly 3000 - 5.5 3.0 2.3
enhanced femaleness. This effect was manifested by decreasing
the number of male flowers, increasing the number of pistillate 'No flowers of respective type appeared within the experimental
flowers, advancing their formation at a lower node, and also period.

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Plant Physiol. Vol. 50, 1972 ETHEPHON AND PHYTOHORMONES IN SEX EXPRESSION
Plants grown under high CO2 level were more developed than
those grown at normal Co2 level. This was expressed also in
earlier formation of male flowers in the monoecious plants.
DISCUSSION
We have demonstrated earlier a promotion of femaleness in
cucurbits by external application of the ethylene-releasing
compound Ethephon (26). Higher ethylene evolution was
found in apices of gynoecious plants as compared to those of
monoecious ones and from female buds as compared to male
ones. Plants grown under short day conditions which promote
femaleness evolved more ethylene than those grown under long
day conditions (28). We have suggested (28) that ethylene
participates in the endogenous regulation of sex expression,
promoting femaleness. A further support to this suggestion
is presented here (Table III). High CO2 levels which are
known to antagonize ethylene action (18) strongly enhanced
maleness in gynoecious plants. The gynoecious cucumber
shows greater endogenous auxin activity than the monoecious
one (Fig. 1). This and the relatively higher auxin levels found
in cucumber plants grown under short day conditions which
promote femaleness (Fig. 2) confirm earlier reports (9) and
the hypothesis of Nitsch et al. (23) that higher auxin level
is associated with female sex expression.
Auxin treatments induce higher ethylene evolution of many
plants (1, 5, 6, 8) including cucumbers (30), while ethylene
lowered auxin level (6, and Fig. 3). It thus seems that ethylene
is a more direct regulator of femaleness.
Inhibitors levels seem also to be related to female sex ex-
pression. ABA content of gynoecious plants is higher than
of monoecious ones (Table I). Higher GA-inhibitor levels were
found in the neutral fraction of diffusates from gynoecious
as compared to monoecious plants (Fig. 4). Higher inhibitor
levels in the monoecious plants were also found in the auxin
assays of cucumber extracts from plants growing under short
days (Fig. 2). It has been suggested (9) that the ratio between
auxin and inhibitor levels arising in mature leaves determines
the sex of the flower bud. We should like to point out that
in our experiments apices and young leaves were extracted and
the inhibitor found here may not be identical with that de-
scribed in previous studies (9).
Higher gibberellin content was found in monoecious plants
as compared to gynoecious ones (Fig. 4). This confirms earlier
results (2). Ethephon treatments reduced gibberellin activity
in diffusates as found by the cucumber and rice assays (Figs.
5, 6). Concurrent with the reduction in gibberellin levels, an
increase in native inhibitor levels was found in the gynoecious
line. Bioassay detection of inhibition after Ethephon treatment
may. however, indicate that Ethephon residues themselves
are the inhibiting agent in the growth of the rice and cucumber
hypocotyls. We have checked this possibility. Chromatography
of Ethephon and subsequent bioassay do not confirm this as-
sumption. Furthermore, Ethephon greatly increased ABA con-
tent of cucumber leaves (Table I). The presence of high level
of ABA and the neutral inhibitor in gynoecious plants and the
low inhibitors level in the monoecious plants (Fig. 4 and Table
I) may support the assumption that Ethephon induces also the
formation of ABA as was also found in orange fruits (13) and
rose petals (21) following treatment with Ethephon and
ethylene.
ABA may participate in the regulation of female sex ex-
pression in cucumbers by interacting with gibberellin. This
hypothesis is supported by the increased femaleness of ABA-
treated plants (Table II). The data presented here and in
earlier paper may strengthen the view that four phytohormones
participate in the regulation of sex expression in cucumber:
ethylene, auxin, GA, and ABA. Ethylene may promote fe-
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Copyright © 1972 American Society of Plant Biologists. All rights reserved.
589
maleness directly, by reducing endogenous GA level or by
promoting the production of ABA.
It may of course also be that each hormone stimulates the
production of enzymes affecting stages in flower (and sex)
development, there being no direct correlation between one
growth regulator and the increase or decrease in the level
of other hormones. At this stage we have not proved the cause
and effect relationship between the different hormones and
thus this second hypothesis cannot be ruled out.
Acknouwledgments-The expert technical assistance of Mrs. Hana Jackman is
gratefully acknowledged. Etheplion (Amchem 62-240) was kindly supplied by
Agan Chemicals, Tel Aviv. ABA (RO-08-0095) is a gift of Hoffman La Roche.
Dwarf rice seeds w-er e kindly suppled by Prof. Y. 'Murakami, Tokyo.
LITERATURE CITED
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leaf abscission by auxin. Plant Physiol. 39: 963-969.
2. ATSNo-\, D., A. LA-NG, AN-D E. N. LIGHT. 1968. Contents and recovery of gib-
berellins in monoecious and gynoecious cucumber plants. Plant Physiol. 43:
806-810.
3. BEYER, E. 'M., JR. AN'D P. W. MORGAN. 1969. Time sequence of the effect of
ethylene on transport, uptake and decarboxylation of auxin. Plant Cell
Physiol. 10: 787-799.
4. BUKOV AC, M. J. AN-D S. H. W\ITWER. 1961. Gibberellin modification of flower
sex expression in Cucumis satirus L. Adv. Chem. Series, Gibberellins
28: 80-88.
5. BURG, S. P. AN-D E. A. BURG. 1966. Auxin-induced ethylene formation: its rela-
tion to flowering in pineapple. Science 152: 1269.
6. BURG, S. P. AN-D E. A. BURG. 1966. The interaction between auxin and
ethy-lene and its role in plant growth. Proc. Nat. Acad. Sci. U.S.A. 55:
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