Journal of Essential Oil Bearing Plants

ISSN: 0972-060X (Print) 0976-5026 (Online) Journal homepage: https://www.tandfonline.com/loi/teop20

Effects of Ethephon on Terpenoids in Cannabis

sativa L. in Vegetative Stage

Hakimeh Mansouri, Fatemeh Salari, Zahra Asrar & Fatemeh Nasibi

To cite this article: Hakimeh Mansouri, Fatemeh Salari, Zahra Asrar & Fatemeh Nasibi (2016)
Effects of Ethephon on Terpenoids in Cannabis sativa L. in Vegetative Stage, Journal of Essential
Oil Bearing Plants, 19:1, 94-102, DOI: 10.1080/0972060X.2015.1004122

To link to this article: https://doi.org/10.1080/0972060X.2015.1004122

Published online: 07 Mar 2016.

Submit your article to this journal

Article views: 203

View related articles

View Crossmark data

Citing articles: 3 View citing articles

Full Terms & Conditions of access and use can be found at

https://www.tandfonline.com/action/journalInformation?journalCode=teop20
 TEOP 19 (1) 2016 pp 94 - 102 94
ISSN Print: 0972-060X
ISSN Online: 0976-5026

Effects of Ethephon on Terpenoids in Cannabis sativa L. in Vegetative Stage

Hakimeh Mansouri*, Fatemeh Salari, Zahra Asrar, Fatemeh Nasibi

Department of Biology, Faculty of Science, Shahid Bahonar University of Kerman, Kerman, Iran
Received 01 September 2014; accepted in revised form 31 December 2014

Abstract: In the present study we investigated the effect of ethephon on changes in the amount of
many terpenoid compounds including, tetrahydrocannabinol (THC), cannabidiol (CBD), chlorophyll,
carotenoids, α-tocopherol and pyruvate as one of substrate in plastidic terpenoids biosynthesis in Cannabis
sativa at vegetative stage. Treatment with 1 and 100 μM ethephon increased THC content up to 9 fold in
vegetative stage. Plants treated with 10 μM ethephon had more CBD content in comparison with control
plants. The amount of CBD decreased in plants subjected to 5 and 100 μM ethephon. Leave area in plant
treated with ethephon did not show significant changes in comparison to control plants. Ethephon treatment
caused a declined in the amount of α-tocopherol. The levels of chlorophylls and carotenoids raised in treated
plants with 1 μM ethephon. With regard to the fact that ethephon had not negative effect on the surface of
leaves and chlorophyll content thus its application can be a suitable way for increasing of THC and α-
tocopherol in Cannabis sativa L.

Key words: Ethephon, terpenoid, cannabis, tetrahydrocannabinol (THC), cannabidiol (CBD),

α-tocopherol.

Abbreviations serve in communication and defense, for example,

THC Δ9-Tetrahydrocannabinol
CBD Cannabidiol
MEP 2-Methyl-D-erythritol-4-phosphate
MVA Mevalonate
IPP isopenthenylpyrophosphate
Introduction
The terpenoids constitute the largest family of
natural products and play diverse functional roles
in plants as hormones (gibberellins, abscisic acid),
photosynthetic pigments (phytol, carotenoids),
electron carriers (ubiquinone, plastoquinone),
mediators of polysaccharide assembly (polyprenyl
phosphates), and structural components of
membranes (phytosterols). In addition to these
universal physiological, metabolic, and structural
functions, many specific terpenoid compounds
(commonly in the C10, C15, and C20 families)
as attractants for pollinators and seed dispersers,
competitive phytoxins, antibiotics, and herbivore
repellents and toxins. Members of the terpenoid
group also include industrially useful polymers
(rubber, chicle) and a number of pharmaceuticals
(artemisinin, taxol) and agrochemicals (pyrethrins,
azadirachtin) 16 . Isoprenoids are derived by
consecutive condensations of five carbon pre-
cursors, isopentenyl diphosphate (IPP) and its
isomer dimethylallyl diphosphate (DMAPP). In
plants two distinct pathways synthesize IPP. The
acetate/mevalonate (MVA) pathway which is
shared with animals and fungi occurs in the
cytoplasm where sesquiterpenes (C15) and tri-
terpenes (C30) such as sterols are produced. The
more recently identified MEP pathway (2C-
methyl-D-erythritol 4-phosphate) occurs in
plastids and is also found in protozoa, most

*Corresponding authors (Hakimeh Mansouri)

E-mail: < h_mansuori@yahoo.com > © 2016, Har Krishan Bhalla & Sons
 Hakimeh Mansouri et al., / TEOP 19 (1) 2016 94 - 102
bacteria, and algae 8.
Cannabis plants are interesting to human society
in more ways. As a fiber plant, cannabis produces
some of the best and most durable fiber of natural
origin. The oil of hemp seed was found to be well
balanced in regards to the ratio of omega-3 and 6
fatty acids for human nutrition 10 and it is ideal as
an ingredient for body oils and lipid-enriched
creams 19. The medicinal use of cannabis has a
very long history 10.
Cannabis seems a virtual factory for the pro-
duction of secondary metabolic compounds. A
variety of alkanes have been identified as well as
nitrogenous compounds, flavonoids and other
miscellaneous compounds. Terpenes appear in
abundance and contribute to the characteristic odor
of the plant. The compounds which comprise the
active drug ingredients are apparently unique to
this genus and are termed cannabinoids. Canna-
binoids were originally thought to exist as the
phenolic compounds 18. Cannabis classified as
fiber-type and drug-type. The main difference
between these two is found in the content of the
psychotropically active component, Δ9-tetrahydro-
cannabinol (THC) and its acidic precursor
tetrahydrocannabinolic acid (THCA): a high
content of THC+THCA classifies as drug-type
cannabis. Fiber-type cannabis contains component
that has been found to be biologically active, most
notably the cannabinoid cannabidiol (CBD) 9.
Ethylene is an unsaturated hydrocarbon that
controls many of physiological processes in plant
28
. Ethylene controls some processes such as
germination 13, growth 28, photosynthesis 30 and
falling 6. Also, ethylene involves in the plant
response to biotic and abiotic stress such as
mechanical wounding, chemicals and metals,
drought, extreme temperatures, and pathogen
infection 11,12. On the other hand, terpenoids play
important roles in plant-plant, plant-environment
and plant-animal interactions. Thus in this study
we investigate the effect of ethephon as an ethylene
donor on cannabinoids and terpenoids in cannabis
plants at vegetative stage.
Materials and methods
Plant material
Cannabis (Cannabis sativa L.) plants were
grown from seed in pots (15 cm i.d, filled with
95
perlite under controlled conditions (a photoperiod
of 12 h, Day and night temperatures were 22oC
and 18oC, respectively). The plants were fertilized
regularly with a Hoagland’s nutrient solution every
three days.
Ethephon treatment of plants
Cannabis plants were treated when they had
seven pairs of leaves. Ethephon treatment done
by spraying the whole plants with 1, 5, 10 and
100 μM ethephon solutions. Control plants
sprayed with tap water. The treatment took place
with eight times spraying at three days intervals.
Sample preparation for cannabinoids measure-
ment
The third leaves pair were collected and used
for cannabinoid measurement. Samples were dried
at room temperature in darkness. Sample material
(100 mg) was placed in a test tube with 1 ml
chloroform. Sonication was applied for 15 min.
After filtration, the solvent was evaporated to
dryness and the residue was dissolved in 0.5 ml
methanol.
Chromatographic separation of cannabinoids
was performed as descript by Rustichelli et. al.,
22
. Cannabinoid peaks were identified by cannabi-
noids standards (THC and CBD) that were a
generous gift from Pr. Jun Szopa Wroclaw
University, Wroclaw, Poland.
Chlorophyll and carotenoid determination
Chlorophyll and carotenoids were extracted from
leaves with 80 % acetone and quantified by
measuring the absorbance at 663.2, 646.8 and 470
nm as described by Lichtenthaler, 15.
Tocopherol extraction and chromatographic
conditions
Tocopherols were extracted by grinding and
homogenizing 25 mg of freeze-dried leaves
material in 100 μl 100 % methanol. After 20 min
of incubation at 30oC, the samples were centri-
fuged at 14,000 rpm for 15 min, the supernatant
was transferred to new tubes, and the pellet was
re-extracted twice with 250 μl 100 % methanol at
30oC for 30 min, pooling all supernatants.
HPLC analysis was performed with liquid
chromatography equipped with a Fluorescence
 Hakimeh Mansouri et al., / TEOP 19 (1) 2016 94 - 102 96
detector. C18 was used for separation. Methanol Fiber study
(100 %) was used as mobile phase; the flow rate The cross section pieces from third internod were
was 1 ml min -1. Fluorescence detector was cut with microtome. They placed on the distilled
accomplished at 295-335 nm for α-tocopherol 23. water until microscopic survey. The cuts were
Injection volume was 10 μl. The identities of α- analyzed under light microscope (company
tocopherol in plant samples were determined by Olympus, Provis AX 70). In overview (40-fold
comparing with (±)-α-tocopherol standard was enlargement) we qualitatively studied the changes
purchased from Sigma (Germany). in primary fibers in the cross section of the stems.

Determination of pyruvate of chloroplast Data analysis

Extraction of chloroplast
In order to measurement the amount of pyruvate
in chloroplasts, we have to extract chloroplasts
from leaves tissue. First, the large vein was
isolated. 0.5 g of leaves with 3 ml of Tris-sucrose
buffer (0.3 M sucrose, 0.2 M tris HCL, 5 mM
MgSo4) was pulverized. The resulting solution was
filtered and for 5 min was centrifuged (1000 rpm).
Supernatant was removed and residues were
dissolved in 2 ml of Tris-sucrose buffer. The diluted
suspension of chloroplasts was maintained for
measurement of pyruvate by Anthon and Barrett
method 3.
Measurement of leaves area
For measuring leaves area, we made paper
copies of the third leaves. Then the weight of
square 1*1 of paper was measured. Based on the
proportional relationship between leaves weight,
its surface was calculated.
Cannabinoid content (mg g-1 dw)
Data were subjected to statistical analysis using
statistical software package SPSS version 14. One
way analysis of variance (ANOVA) followed by
Duncan multiple range test were employed and
the differences between individual means were
deemed to be significant at p<0.05. The values
are ±SD for three samples in each group.
Result
Effect of ethephon on cannabinoid content
Our results showed that THC and CBD contents
were equal at leaves (Fig. 1). Treatment with
ethephon increased THC content. In this regard
the lowest and highest concentration of ethephon
had stronger effect. The leaves THC content in
these treatments was 8.9 fold of untreated plants.
However, treatment with 5 and 10 μM ethephon
increased THC content in leaves but it was not
considerably. The amount of CBD in plants treated
with 1, 5 and 100 μM ethephon decreased in com-

μM)
Ethephon (μ
Fig. 1. The effects of ethephon on THC and CBD content of leaves in cannabis plants.
Values are means of three replications ± SD. Asterisks indicate the significance
of difference at P< 0.05 level according to Duncan’s multiple range test
 Hakimeh Mansouri et al., / TEOP 19 (1) 2016 94 - 102
parison to control but treatment with 10 μM ethe-
phon increased CBD content up to 4 fold (Fig. 1).
Effect of ethephon on chlorophyll and caroten-
oids content
Since ethylene may function as major switch
between growth and defense and the associated
changes in resource allocations; we investigated
this probably relationship between plastidic
primary and secondary terpenoids with chloro-
phyll, carotenoid and α-tocopherol measurement.
Chlorophyll content (mg g-1 dw)
97
The amount of chlorophyll a, b and total
increased in 1 μM ethephon treatment. In other
ethephon concentrations, the changes in chloro-
phyll content were not significant in comparison
to control plants (Fig. 2).
The changes in carotenoids content in plant
treated with ethephon were similar to those of
chlorophyll content (Fig 3). Treatment with 1 μM
ethephon stimulated carotenoid accumulation on
leaves, but 5, 10 and 100 μM ethephon treatments
were not affective in this regard.

Ethephon (μ μM)
Fig. 2. The effects of ethephon on chlorophyll content of leaves in cannabis plants.
Values are means of three replications ± SD. Asterisks indicate the significance
of difference at P< 0.05 level according to Duncan’s multiple range test
Carotenoids content (mg g-1 dw)

Ethephon (μμM)
Fig. 3. The effects of ethephon on carotenoids content of leaves in cannabis plants.
Values are means of three replications ± SD. Asterisks indicate the significance
of difference at P< 0.05 level according to Duncan’s multiple range test
 Hakimeh Mansouri et al., / TEOP 19 (1) 2016 94 - 102 98
Effect of ethephon on á-tocopherol content Effect of ethephon on leaves area
The amount of α-tocopherol in treated plants Ethephon treatment with this method and
with 5, 10 and 100 μM ethephon was lower than concentration did not effect on decreasing of
control plants. Treatment with 1 μM ethephon did leaves area of plants in comparison to control
not effect on α-tocopherol (Fig. 4). plants (Fig. 6).

Effect of ethephon on pyruvate Effect of ethephon on primary fibers in stem

Control plants had the highest pyruvate content.
Pyruvate content of plants treated with ethephon
was lower than the control plants (Fig. 5).
μg g-1 dw)
α-Tocopherol content (μ
The amount of the individual layers in the cross
section of the stem and the number of fiber cells
per layer increased in 1 μM ethephon treatment

μM)
Ethephon (μ
Fig. 4. The effects of ethephon on α-tocopherol content of leaves in cannabis plants.
Values are means of three replications ± SD. Asterisks indicate the significance
of difference at P< 0.05 level according to Duncan’s multiple range test
Pyruvate content (mg g-1 dw)

Ethephon (μμM)
Fig. 5. The effects of ethephon on pyruvate content of leaves in cannabis plants.
Values are means of three replications ± SD. Asterisks indicate the significance
of difference at P< 0.05 level according to Duncan’s multiple range test
 Hakimeh Mansouri et al., / TEOP 19 (1) 2016 94 - 102 99

leaf area (cm2)

Ethephon (μ μM)
Fig. 6. The effects of ethephon on leaves area in cannabis plants. Values are means
of three replications ± SD. Asterisks indicate the significance of difference
at P< 0.05 level according to Duncan’s multiple range test
also cell wall diameter had been increased in 1 were amorphous; the wall between the fiber cells
μM ethephon treatment. But in other treatments was separated in this treatment (Fig. 7).
(5, 10 and 100 μM) area of primary fiber cells
and their cell lumen was increased. In 100 μM Discussion
ethephon treatment, the fiber and cambium cells In this study, we were evaluated the effect of

Fig. 7. The effect of ethephon on fiber cells on the cross section of stem third internod
in cannabis plants. (40 fold enlargement). a) control, b) 1 μM ethephon, c)
5 μM ethephon, d) 10 μM ethephon, e) 100 μM ethephon.
 Hakimeh Mansouri et al., / TEOP 19 (1) 2016 94 - 102
ethylene on some primary and secondary terpen-
oids compounds in Cannabis sativa L. The mount
of THC increased in all ethephon treatments but
the mount of CBD increased only by 10 μM ethe-
phon treatment and other treatments decreased the
mount of CBD. Concurrent with our results Bate
et. al., 4 reported that ethylene stimulates synthesis
of ginsenoside in ginseng. Ethylene is known to
enhance the production of terpenoid phytoalexins
in tobacco cultures 31. Also considerable increases
in the levels of notkaton (sesquiterpene) were
observed in the picked and unpicked grapefruits
treated with ethylene. Tomas et. al., 29 suggested
that ethylene regulates nootkatone biosynthesis by
accelerating the maturation-senescence processes
in grapefruit rind. El-Keltawi and Croteau 7
reported the influence of ethephon on the consti-
tuents of monoterpenes of Mentha piperita. Most
of the growth hormone studies on M. piperita or
M. arvensis impact on the enzymes of biosynthetic
pathways 26.
The phytyl (C20) conjugates such as chloro-
phylls and tocopherols, and carotenoids (C40) are
produced by MEP pathway in chloroplast. In order
to investigation the ethephon effects on the MEP
pathway regulation, we measured the main end
products of this pathway in ethephon-treated
plants. Ethephon treatment resulted in increase in
chlorophylls and carotenoids content. An increase
in chlorophyll content has also detected in
cucumber cotyledons 2, Sugarcane 14, carnation
flowers 5. Also many reports showed that an
exogenous application of ethylene to green plant
materials induced a decrease in chlorophyll con-
tent 21,9. However, we think in these works high
concentration of ethylene or longer time of
treatment was used in comparison with our
experiment. Because ethephon did not effect on
decreasing of leaves area and photosynthetic
pigments, it can be supposed that these ethephon
concentrations and treatment method could not
inhibit plant growth. Thus low concentrations of
References
1. Alexieva, V.S., Sergiv, I.G., Todorova, D.A., Karanov, E.N., Smith, A.R. and Hall, M.A.
(2004). Effect of ethylene and its antagonist 1-MCP on the senescence of detached leaves of
Arabidopsis thaliana. Biol. Planta. 48(4): 293-295.
2. Alscher, R.G. and Castelfranco, P.A. (1972). Stimulation by ethylene of chlorophyll biosynthesis
100
ethephon can suitable treatment to increase
cannabinoids in cannabis plants.
Carotenoids content enhanced in the lowest
ethephon concentration (1 μM) similar to chloro-
phyll responds. In coincidence with our results,
ethylene induced carotenoid content in Kinnow
mandarin peels 17 and fig fruits 20 but Alexieva et.
al., 1 reported a decrease in carotenoid content on
detached leaves of Arabidopsis thaliana.
The amount of α-tocopherol decreased by
ethephon treatment. Tocopherols are involved in
protecting lipids and the photosynthetic apparatus
against created oxidative damage by variety of
biotic and abiotic stresses. In cannabis tetrahydro-
cannabinolic acid (THCA) is biosynthesized from
cannabigerolic acid (CBGA). This reaction
depends on molecular oxygen and produces
hydrogen peroxide and THCA at a molar ratio of
1:1 (CBGA + O2JTHCA + H2O2) 27. Probably,
THCA biosynthesis reaction can act as a defense
mechanism by using molecular oxygen instead of
α-tocopherol in this condition.
Pyruvate and glyceraldehydes-3-phosphate are
precursors of IIP biosynthesis in MEP pathway.
In this work, ethephon treatment resulted in
decrease in pyruvate content that this may be due
to the role of ethephon on stimulation of bio-
synthesis pathway of plastidic terpenoids.
Our result showed that the lowest concentration
of ethephon (1 μM) probably has a beneficial effect
on primary fiber cells. This treatment increased
the cell wall diameter. Of course to confirm this
subject we need to quantitative analysis of fibers.
Also Shi et. al., 25 reported a major role for ethylene
in cotton fiber cell elongation. They suggested that
ethylene may promote cell elon-gation by
increasing the expression of sucrose synthase,
tubulin, and expansin genes.
We concluded that probably low concentrations
of ethephon can use either for increasing canna-
binoids or fiber quality in Cannabis sativa without
to have adverse effect on plant growth.
 Hakimeh Mansouri et al., / TEOP 19 (1) 2016 94 - 102 101
in dark-grown Cucumber cotyledons. Plant Physiol. 50: 400-403.
3. Anthon, G. and Barrett, D.M. (2003). Modiûed method for the determination of pyruvic acid
with dinitrophenylhydrazine in the assessment of onion pungency. J. Sci. Food Agr. 83: 1210-
1213.
4. Bae, K.H., Choi, Y.E., Shin, C.G., Kim, Y.Y. and Kim, Y.S. (2006). Enhanced ginsenoside pro-
ductivity by combination of ethephon and methyl jasmoante in ginseng (Panax ginseng C.A. Meyer)
adventitious root cultures. Biotechnol. Lett. 28: 1163-1166.
5. Cook, E.L. and Vanstaden, J. (1983). Senescence of cut carnation flowers: Ovary development
and CO2 fixation. Plant Growth Regul. 1: 221-232.
6. Corbineau, F., Rudnicki, R.M., Goszczynska, D.M. and Come, D. (1995). The effect of light
quality on ethylene production in leaves of seedlings (Avena sativa L.). Environ. Exp. Bot. 35:
227-233.
7. EI-Keltawi, N.E. and Croteau, R. (1986). Influence of ethephon and daminozide on growth and
essential oil content of peppermint and stage. Phytochem. 25: 1285-1288.
8. Estevez, J.M., Cantero, A., Reindi, A., Reichler, S. and Leon, P. (2001). 1-Deoxy-D-xylulose-
5-phosphate synthase, a limiting enzyme for plastidic isoperenoid biosynthesis in plants. Biol.
Chem. 276: 22901-22909.
9. Gestein, S. and Thimann, K.V. (1981). The role of ethylene in the senescence of oat leaves. Plant
Physiol. 68: 349-354.
10. Hazekamp, A. (2009). Cannabis review. Department of Plant metabolomics Leiden University.
Leiden the Netherlands ISBN 1442-8247.
11. Johnson, P.R. and Ecker, J.R. (1998). The ethylene gas signal transduction pathway: a molecular
perspective. Annu. Rev. Genet. 32: 227-254.
12. Kende, H. (1993). Ethylene Biosynthesis. Annu. Rev. Plant Physiol. Plant Mol. Biol. 44: 283-
307.
13. Lacher, W. (2001). Physiological plant ecology. Springer-Veray Berlin Heidelberg New york.
Germany. 103: 1-7.
14. Li, Y.R. (2004). Beneficial effects of ethephon application on sugarcane under sub-tropical climate
of China. Sugarcane Agr. 6(4): 235-240.
15. Lichtenthaler, H.K. (1987). Chlorophyls and Caretenoids: pigments of photosynthetic biomemb-
ranes. Methods Enzymol. 148: 350-382.
16. McGarvey, D.J. and Croteau, R. (1995). Terpenoid metabolism. Plant cell. 7: 1015-1026.
17. Nagar, P.K. (1993). Effect of plant growth regulators on the natural and ethylene induced pigmen-
tation in Kinnow mandarin peels. Biol. Planta. 35(4): 633-636.
18. Pate, D.W. (1994). Chemical ecology of cannabis. Int. Hemp Assoc. 2(29): 32-37.
19. Pinarkara, E., Kayis, A.S., Hakki, E. and Sag, A. (2009). RAPD analysis of seized Marijuana
(Cannabis sativa L.) in Turkey. Electron. J. Biotechno. 12: 1-13.
20. Puech, A.A., Rebeiz, C.A. and Crane, J.C. (1974). Pigment changes associated with appliccation
of ethephon ((2-chloroethyl) phosphonic acid) to fig (Ficus carica L.) fruits. Plant Physiol. 57:
504-509.
21. Purvis, A.C. and Barmore, C.R. (1981). Involvement of ethylene in chlorophyll degradation in
peel of citrus fruits. Plant Physiol. 68: 854-856
22. Rustichelli, C., Ferioli, V., Baraldi, M., Zanoli, P. and Gamberini, G. (1998). Analysis of
cannabinoids in ûber hemp plant varieties (Cannabis sativa L.) by high performance liquid chro-
matography. Chromatographia 47, 215-222.
23. Sattler, S.E., Cahoon, E.B., Coughlan, S.J. and Dellapenna, D. (2003). Characterization of
tocopherol cyclase from higher plants and cyanobacteria. Evolutionary implications for tocopherol
synthesis and function. Am. Soc. Plant Biol. 132: 2184-2195.
 Hakimeh Mansouri et al., / TEOP 19 (1) 2016 94 - 102 102
24. Schaller, G. and Kieber, J. (2002). Ethylene. Am. Soc. Plant Biol. 10: 1-17.
25. Shi, Y.H., Zhu, S.W., Mao, X.Z., Feng, J.X., Qin, Y.M., Zhang, L., Cheng, J., Wei, L.P.,
Wang, Z.Y. and Zhua, Y.X. (2006). Transcriptome proûling, molecular biological and physiological
studies reveal a major role for ethylene in Cotton fiber cell elongation. The Plant Cell. 18: 651-
664.
26. Singh, P., Srivastava, N.K., Mishra, A. and Sharma, S. (1999). Influence of ethereal and gib-
berellic acid on carbon metabolism, growth and essential oil accumulation in spearmint (Mentha
spicata ). Photosynthetica. 36(4): 509-517.
27. Sirikantaramas, S., Morimoto, S., Shoyama, Y., Ishikawa, Y., Yoshiko, Y. and Shoyama, Y.
(2004). The gene controlling marijuana psychoactivity molecular cloning and heterologous express-
ion of tetrahydrocannabinolic acid synthase frome Cannabis sativa L. Biol. Chem. 279: 39767-
39774.
28. Tholen, D., Voesenek, L. and Poorter, H. (2004). Ethylene insensitivity does not increase leaf
area or relative growth rate in Arabidopsis, Nicotiana tabacim and Petunia hybrida. Plant Physiol.
134: 1803-1812.
29. Tomas, A.O., Garcia-Puig, D., Sabater, F., Porras, I., Garcia-Puig, A. and Rio, J.A. (1993).
Influence of ethylene and ethephon on the sesquiterpene Nootkatone production in Citrus paradise.
Am. Chem. Soc. 41: 1566-1569.
30. Trebitsh, T., Goldschidt E.E. and Riov, J. (1993). Ethylene induces de novo synthesis of chloro-
phyllase, a chlorophyll degrading enzyme, in citrus fruit peel. Proc. Natl. Acad. Sci. USA. 90:
9441-9445.
31. Yahia, A., Kevers, C., Gaspar, T., Chenieux, J.C., Rideau, M. and Creche, J. (1998). Cytokinins
and ethylene stimulate indole alkaloid accumulation in cell suspension cultures of Catharanthus
roseus by two distinct mechanisms. Plant Sci. 133: 9-15.