Plant Growth Regulation 9 : 19-26, 1990.

© 1990 Kluwer Academic Publishers . Printed in the Netherlands.

Regulation of plant productivity I : Improved seedling vigor

and floral performance of Phalaenopsis by 2-(3,4-
dichlorophenoxy)triethylamine IDCPTAI

J.H. KEITHLY* & H . YOKOYAMA

United States Department of Agriculture,t Agricultural Research Service, Fruit and
Vegetable Chemistry Laboratory, 263 S. Chester Avenue, Pasadena, CA 91106, USA

Received 20 May 1989 ; accepted 23 October 1989

Key words . bioregulator, orchid, seedling survival, plant maturity

Abstract . Application of the tertiary amine bioregulator, 2-(3,4-dichlorophenoxy)-

triethylamine [DCPTA] to seedling Phalaenopsis plants increased seeding survival, long-term
vegetative plant growth, and greatly accelerated flowering when compared to controls .
Application of 30µM (10 ppm) DCPTA during the transfer of orchid seedlings from sterile
agar culture to greenhouse community pot culture caused a 2 to 3-fold increase in root and
leaf growth of seedling plants over the growth of controls . After 2 years greenhouse culture,
the total dry weight of 30 pM DCPTA-treated plants was doubled when compared with
controls . Flowering of 30 pM DCPTA-treated plants was increased significantly (p < 0 .05)
over that of controls after 11 and 18 months plant growth . During a 2 year growth period,
the total number of racemes produced by 30pM DCPTA-treated plants was doubled when
compared with the floral performance of controls . The effects of DCPTA on plant flowering
appeared to function independently of DCPTA-mediated effects on plant growth .

1. Introduction

Advances in clonal orchid propagation [11, 13] and hybridization programs

[3] have promoted the use of Phalaenopsis orchids as flowering plants for
pot-sales in the commercial florist industry . Unfortunately, hybrid
Phalaenopsis production requires 2 to 3 years growing time to produce
mature, flowering plants . The use of synthetic plant growth regulators to
reduce the growing time to flowering of Phalaenopsis has been generally
unexplored .
The tertiary amine bioregulator, 2-(3,4-dichlorophenoxy)triethylamine
[DCPTA] has been shown to increase the growth and yield of a wide variety

* Author to whom correspondence should be sent .

t Reference to a company or product name does not imply approval or recommendation of
the product by the U .S . Department of Agriculture to the exclusion of others that may be
suitable .
20
of crop plants [2, 5, 6, 7, 8] . Application of DCPTA to ungerminated seeds
or to seedling plants appears to regulate chloroplast development during
exponential plant growth [7] . In DCPTA-treated plants, chloroplast com-
partment size [7, 8], ribulose-1,5-bisphosphate carboxylase/oxygenase
(Rubisco) activity [7, 8], and carbon assimilation per unit leaf area [2, 8] are
increased in mature leaves when compared to controls . In addition, the
growing time to harvest of DCPTA-treated crop plants is often reduced
significantly when compared with controls [5, 6] . In this report, the first in
a series which examine the effects of DCPTA on the vegetative growth and
floral productivity of ornamental plants, the effects of DCPTA on the
vegetative growth and floral maturity of Phalaenopsis are investigated .

2. Materials and methods

2 .1 Chemicals

DCPTA was synthesized and purified according to the methods Echols et al .

[1] and Poling et al . [10] .

2 .2 Phalaenopsis hybrids

Phal . X . cv. # 1609, (Herbert Hager x Best Rose) x Snow Leopard ; Phal.
X. cv . # 12, (Capitola `Moonlight' FCC/AOS x Joseph Hampton `Diana'
AM/AOS) ; Phal. X . cv . # 27, (Barbara Moler 'Gertie' AM/AOS x
Zauberrose)

2 .3 Plant material and experimental protocol

Seedlings of Phal . cultivars # 12 and # 27 were purchased as Elnmeyer

flask cultures (60 to 75 plants per 14 month-old flask culture) . Plants of Phal.
cultivar # 1609 were purchased as community pots (25 plants per pot, plant
age out of flask : 3 months) . Experimental plants were selected for uniformity
of plant size prior to DCPTA-treatment . Through random selection, ex-
perimental plants were divided into 20 seedlings per treatment block
replicated four times . Orchid seedlings were totally immersed for 2 h in
solutions ranging from 3 to 300 µM DCPTA containing Tween 80 (0 .1 %,
w/v) as a surfactant . Control seedlings were soaked for 2 h in Tween 80
(0 .1 %, w/v) . Seedlings were not rinsed prior to potting in a commercial
orchid seedling growth medium containing fir bark and perlite (Stewart's
Seedling Mix, Steward Orchids, San Gabriel, CA) . For seedling growth
 21
studies, each DCPTA-treatment block was planted into a 15 x 38 cm rec-
tangular plastic pot (20 plants per pot) . Replicate community pots were
arranged as a randomized complete block and each plant was considered to
be an experimental unit . Plants were greenhouse grown under 40% saran
shade cover and received a photosynthetic photon flux density of 600 to
800 µcool m-2 s- ' photosynthetically active radiation . Temperature control
was maintained at 28 ± 4'C day and 20 ± 2'C night . Plants were fer-
tilized every 10 days with 185 mg 1 - ' N, 53 mg 1 - ' P, and 53 mg 1 - ' K . For
long term Phalaenopsis culture, DCPTA-treated seedlings of cultivar # 1609
were potted individually into 5 cm square plastic pots . Plants were repotted
after 8 months growth into 13 cm clay pots that were filled with a mixture
of redwood bark, coarse perlite, and New Zealand moss (2 :1 :1, v/v) .

2 .4 Plant growth determination

Seedling survival and vegetative growth were determined 4 months after

DCPTA-treatment . Dry weights of root and shoot samples were obtained
after the plant samples were oven dried at 70 °C for 48 h. Leaf areas were
determined using a Delta-T video leaf area (Decagon Devices Inc ., Pullman,
WA) . For long-term plant growth determinations using Phal . cultivar
# 1609, six randomly selected plants were harvested from each DCPTA-
treatment group. In flowering plants, the size (width) of mature flowers, the
number of flowers per raceme, and the number of racemes per plant were
determined . Analysis of variance was performed for all growth and floral
performance data . Percentile data were transformed to arcsine values prior
to statistical analysis [14] . Duncan's multiple range test (p < 0 .05) was used
to compare treatment means [14] . Data represent typical values from three
independent experiments .

3. Results

The application of DCPTA to Phalaenopsis seedlings during routine trans-

planting resulted in statistically significant (p < 0 .05) increases in seedling
survival and seedling growth when compared to controls (Table 1) . For the
three cultivars tested, the largest numerical increases in seedling survival
were observed in the 30µM DCPTA treatment groups . However, general
improvements in seedling survival were observed in all DCPTA treatment
groups when compared to controls . Four months after bioregulator treat-
ment, 30 µM DCPTA-treated seedlings showed a 2 to 3-fold increase in root
and leaf growth when compared with controls . Of the concentrations tested,
22

Table 1 . Growth and survival of seeding Phalaenopsis determined 4 months after DCPTA
treatment

Cultivar DCPTA Root dry wt. Leaf dry wt . Leaf area Seedling
µM g g dm' survival

# 1609 0 0 .18 (b)a 0 .19 (b) 0 .21 (b) 48 .6 (c)

3 0 .21 (b) 0 .20 (b) 0 .22 (b) 66 .3 (b)
30 0 .52 (a) 0 .46 (a) 0.44 (a) 79 .5 (a)
300 0 .19 (b) 0.22 (b) 0 .24 (b) 65 .6 (b)
# 12 0 0 .15 (b) 0.17 (b) 0.15 (c) 58 .3 (bc)
3 0.26 (b) 0.24 (b) 0 .22 (b) 69 .2 (b)
30 0 .47 (a) 0 .34 (a) 0 .36 (a) 73 .7 (a)
300 0.18 (b) 0.14 (b) 0.12 (c) 58 .9 (bc)
#27 0 0.21 (b) 0.20 (b) 0 .21 (b) 47 .6 (c)
3 0.20 (b) 0.22 (b) 0 .20 (b) 66 .6 (b)
30 0 .41 (a) 0 .35 (a) 0 .33 (a) 83 .3 (a)
300 0 .13 (b) 0.18 (b) 0 .14 (c) 61 .9 (b)

a Letters in parentheses indicate significant differences among means according to Duncan's

multiple range test (p S 0 .05) .

/e

H
3
04
H
O
O

4 8 12 16 20 24
MONTHS AFTER TREATMENT
Fig . 1 . Effect of DCPTA on the long-term vegetative growth of Phalaenopsis X . cv. # 1609 .
Data represents the mean ± SE of six replicate plants . Control, 0 ; 3 µM DCPTA, ∎ ; 30 µM
DCPTA, 4 ; 300µM DCPTA, A .
 23

Table 2. Effect of DCPTA on the flowering of Phalaenopsis X cv . # 1609

DCPTA Flowering Racemes Flowers Flower sizes

Months after
treatment µM plants % per plant per raceme cm

11 0 8 .3 (c)b 0 .1 (c) 7 .0 (a) 8 .2 (a)

3 41 .7 (b) 0 .4 (b) 6 .2 (a) 7 .9 (a)
30 83 .3 (a) 0 .8 (a) 7 .2 (a) 8 .6 (a)
300 33 .3 (b) 0 .3 (b) 4 .3 (b) 7 .5 (a)
18 0 16 .7 (c) 0 .2 (d) 9 .5 (b) 7 .9 (a)
3 100 .0 (a) 1 .2 (b) 12 .7 (ab) 8 .1 (a)
30 100 .0 (a) 1 .8 (a) 14.5 (a) 8 .4 (a)
300 58 .3 (b) 0.6 (c) 10.1 (b) 8 .0 (a)
24 0 100.0 (a) 1 .3 (a) 10.8 (b) 8 .2 (a)
3 ND` ND ND ND
30 100 .0 (a) 1 .5 (a) 22 .8 (a) 8 .7 (a)
300 ND ND ND ND

I Total width of flower petals measured horizontally across the center of mature flowers .
b Means (n = 12) followed by the same letter are not significantly different according to
Duncan's multiple range test (p < 0 .05) . Data are collected 11, 18, 24 months after DCPTA
treatment were analyzed separately .
Not determined.

Fig . 2 . Effect of DCPTA on the floral maturity of Phalaenopsis X . cv . # 1609 . Plants were
greenhouse grown and were photographed 11 months after DCPTA treatment . Left, control;
Right, 30µM DCPTA.
24

application of 300 µM DCPTA to seedling plants appeared to inhibit seed-

ling plant growth .
Phalaenopsis X. cultivar # 1609 was used to examine the long-term effects
of DCPTA upon vegetative plant growth (Figure 1) . Compared to controls,
root and leaf development of 30µM DCPTA-treated plants were increased
significantly and the increased plant growth rate was sustained 2 years after
DCPTA treatment . Twenty four months after DCPTA treatment, the mean
total dry weight of 30 pM DCPTA-treated plants was doubled when com-
pared to controls . The long-term root and leaf growth of 3 and 300µM
DCPTA-treated plants was statistically similar to control values (Figure 1) .
The growing time to flowering of DCPTA-treated plants was reduced
significantly when compared to the flowering of controls (Table 2, Figure 2) .
Application of 30 pM DCPTA to seedling plants caused a 10-fold increase
in raceme production over that of controls after 11 months plant growth
(Table 2) . Eleven months after bioregulator treatment, raceme production
by 3 and 300µM DCPTA-treated plants showed a 5 and 4-fold increase,
respectively, over the flowering of control plants . During a 24 month grow-
ing period, the total raceme production by 30 pM DCPTA-treated plants
was doubled when compared with the floral performance of controls . The
number of flowers per raceme produced by mature, 30 pM DCPTA-treated
plants was increased significantly over controls during the second and third
flowering periods . Flower size was statistically similar among all DCPTA
treatment groups at all plant flowering periods (Table 2) .
The floral performance of Phalaenopsis plants used in this study was not
absolutely dependent upon either initial seedling size or upon the rate of
plant growth . Prior to DCPTA treatment, many of the original Phal.
cultivar # 1609 seedling plants were too large for use as experimental
material . After 11 months growth, these large, non-experimental plants were
often greater in total plant dry weight than many of the 30 pM DCPTA-
treated plants . However, these large, non-treated plants exhibited only an
8.3% raceme production after 11 months plant growth . In addition, the
flowering of 3 and 3001M DCPTA-treated plants was increased signifi-
cantly when compared with controls after 11 and 18 months plant growth
(Table 2) . However, the long-term vegetative growth parameters of the 3
and 300 pM DCPTA-treated plants were statistically similar to controls
(Figure 1) .

4. Discussion

Routine orchid culture involves the repetitive transfer of seedling orchid

plants from sterile agar cultures to greenhouse community pot culture, and
 25
then from community pot to individual pot culture as the seedlings increase
in size . Seedling mortality and reduced seedling growth after transplanting
can greatly reduce the efficiency of orchid production and may increase the
greenhouse growing time to produce mature, flowering plants . The results
of this study demonstrate that seedling survival and vegetative plant growth
of Phalaenopsis are increased significantly after the application of DCPTA
to seedling plants during routine transplanting (Table 1, Figure 1) . Applica-
tion of 30µM DCPTA improved the overall performance of seedling plants
that were transferred from sterile agar cultures to community pots, and of
community pot seedlings that were repotted to individual pot culture . This
study and the results of previous studies [2, 5, 6, 7, 8] demonstrate that plant
growth induction by DCPTA results in an overall increase in plant dry-
matter, and that DCPTA-enhanced plant growth rates are maintained
across the entire vegetative development of the plant . The optimal dosage of
DCPTA used in this study was 30 pM, which has been widely demonstrated
to accelerate the root and shoot development of crop plants [5, 6, 7, 8] .
The regulation of floral crop maturity by synthetic plant growth
regulators generally involves the coordinate regulation of vegetative plant
growth and flower bud initiation [4, 9, 12] . Time of application and growth
regulator dosage are critical in order to adequately control vegetative plant
growth, to reduce the days to harvest, and to maintain flower quality [4, 9] .
In this study we found that 30 mM DCPTA, when applied to seedling
Phalaenopsis, increased vegetative plant growth and greatly reduced the
greenhouse growing time to produce flowering plants (Figures 1 and 2,
Table 2) . However, application of 3 and 300 pM DCPTA to seedling
Phalaenopsis resulted in significant improvements in plant flowering without
coordinate improvements in plant growth (Table 2, Figure 1) . These results
suggest that the regulation of plant maturity and floral productivity by
DCPTA may be independent of the effects of DCPTA on vegetative plant
growth . Regulation of vegetative plant growth by DCPTA appears to
involve the regulation of chloroplast biogenesis and Rubisco activity [7, 8] .
However, the mode of action of DCPTA on the accelerated reproductive
maturity of ornamental plants is unknown .
The effects of DCPTA upon plant productivity have important applica-
tions to the commercial orchid industry . The improved seedling vigor of
DCPTA-treated plants would improve the efficiency and economic return of
seedling production operations . The significantly reduced growing time to
flowering of DCPTA-treated plants would potentially improve orchid pro-
duction for both the cut-flower and pot-sales industries .
26

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