Scientia Horticulturae 238 (2018) 7–13

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Scientia Horticulturae
journal homepage: www.elsevier.com/locate/scihorti

Effects of GA3 on promotion of flowering in Kalanchoe spp. T

⁎
Ming-Zong Chang, Chang-Hai Huang
Department of Plant Industry, College of Agriculture, National Pingtung University of Science and Technology, 1, Shuefu Road, Neipu, Pingtung, 912, Taiwan, ROC

A R T I C LE I N FO A B S T R A C T

Keywords: Some species of Kalanchoe, including those of sections Kalanchoe and Bryophyllum do not flower naturally. This
Section barrier to flowering might hinder hybridization and breeding. The role of gibberellin acid (GA3) in promoting
Kalanchoe flowering was investigated, and the effects of GA3 treatment on flowering under different temperatures were
Section determined. The findings indicated that flowering was promoted in 100% of the K. sexangularis, K. nyikae, K.
Bryophyllum
marnieriana, K. longiflora, and K. × richaudii plants treated with different GA3 gradients. The percentage of
Breeding
Temperature
flowering in K. marnieriana and K. nyikae increased with increasing concentrations of applied GA3. The number
Percentage of flowering of days taken to flower in K. nyikae and K. sexangularis decreased with increasing concentrations of applied GA3.
Days to flowering In K. × richaudii treated with 100 mg L−1 GA3 flowers appeared the earliest. The number of flowers of K.
sexangularis and K. × richaudii increased, while that of K. nyikae decreased, with increasing GA3 concentrations.
GA3-treated K. nyikae, K. poincarei, and K. sexangularis plants showed higher percentages of flowering at 25/20 °C
(day/night) than at higher temperatures. The findings from this study confirmed that GA3, applied in a broad
range of concentrations, promotes floral induction in the studied species, affecting mainly the number of days
taken to flower and the number of flowers produced. However, the GA3-treated plants had reduced flowering at
higher temperatures. These findings can be applied in the regulation of flowering and breeding of Kalanchoe in
future.

1. Introduction
Photoperiod, temperature and the juvenile phase of plants are be-
lieved to affect the induction of flowering in Kalanchoe. The genus
Kalanchoe can be divided into two sections: Kalanchoe and Bryophyllum
(Descoings, 2003). Species belonging to the section Bryophyllum usually
exhibit longer juvenile phases, e.g., six years in B. proliferum (Zeevaart,
1985). Currey and Erwin (2011) showed that Kalanchoe beauverdii, K.
beharensis, K. fedtschenkoi, K. longiflora, K. marmorata, K. marnieriana, K.
streptantha, K. tomentosa, and K. vigueridoi did not flower under a
photoperiod induction treatment. Therefore, the juvenile phase is one
of the important factors influencing flowering in Kalanchoe species.
Gibberellin acids (GAs) can effectively shorten the length of the juvenile
phase (Sgamma, 2017). GAs are known as environmental signals that
induce stem elongation and flowering (Hisamatsu et al., 2000; Mutasa-
Göttgens and Hedden, 2009; Wilkie et al., 2008). Several reports have
suggested that GAs play an important role in the promotion of flow-
ering in plants, including Arabidopsis thaliana (Wilson et al., 1992),
Brunonia (Wahyuni et al., 2011), Bryophyllum (Huang, 2007), Henckelia
humboldtianus (Sumanasiri et al., 2013), Helleborus niger, Helleborus ×
ericsmithii ‘Pink Beauty’ (Christiaens et al., 2012), Lolium temulentum
(King et al., 2003), Philodendron ‘Black Cardinal＇(Chen et al., 2003),
and Iris nigricans (Al-Khassawneh et al., 2006). Previous studies have
confirmed that GAs contents significantly increased in the long-short
day plant B. daigremontianum, when transferred from under long-day to
short-day condition, and floral primordia were formed in the plants
(Skene and Lang, 1964; Zeevaart, 1969a). Huang (2007) showed that
flowering was induced in 90% of the treated two-month-old K. pinnata
and K. poincarei plants when 5 mg L−1 GA3 was applied on them.
Therefore, exogenous GAs appears to promote the transition from ve-
getative growth to reproductive growth (King et al., 2003; Wilson et al.,
1992).
However, some species belonging to the sections Kalanchoe and
Bryophyllum did not flower under any of the studied photoperiods, e.g.,
K. longiflora and K. marnieriana (Currey and Erwin, 2011). The contents
of endogenous GAs were found to increase at low temperatures, and
flowering was simultaneously induced (Su et al., 2001; Xu et al., 1997).
Chen (2007) showed that flowering was not induced in Lavandula het-
erophylla by applying GA3 at high temperature conditions. Flower de-
velopment in the GA3-treated Phalaenopsis also was prevented at high
temperature conditions (Chen et al., 1994; 1997). Flowering in K.
nyikae and K. sexangularis is unstable and unpredictable under the
short-day environmental conditions of tropical regions. Therefore, it
appears that flowering in Kalanchoe species may be affected by

⁎
Corresponding author.
E-mail address: huangch@mail.npust.edu.tw (C.-H. Huang).

https://doi.org/10.1016/j.scienta.2018.04.001
Received 2 November 2017; Received in revised form 1 April 2018; Accepted 2 April 2018
Available online 24 April 2018
0304-4238/ © 2018 Elsevier B.V. All rights reserved.
M.-Z. Chang, C.-H. Huang
environmental temperature after GA treatment.
A barrier to flowering may hinder hybridization and prolong the
breeding time. As such, the present study was undertaken with an ob-
jective to investigate (1) the effects of exogenous GA3 concentrations on
the promotion of flowering in Kalanchoe species, and (2) the effects of
different temperatures on flowering following treatment with GA3. The
findings of this study are expected to address the problem of flowering
barrier in Kalanchoe species and regulate flowering to improve its
breeding efficiency, in future.
2. Materials and methods
2.1. Plant materials and cultivation
K. longiflora, K. nyikae, K. sexangularis (section Kalanchoe), K. mar-
nieriana, K. × richaudii, and K. poincarei (section Bryophyllum) were
used as study plants. Stem cuttings with three leaf-pairs were planted in
6-cm plastic pots filled with a 2:0.5:1 mixture (v/v) of peat moss
(DSM05W, Kekkilä, Finland), perlite (No. 4, South Sea Co., Ltd.,
Taiwan) and coconut fiber (CB53, Da-Yi Agritechnology Co., Ltd.,
Taiwan). After four weeks of rooting, the plants were potted in 9-cm
pots and grown in a shaded greenhouse with a maximum photo-
synthetic active radiation of 320 μmol m−2 s-1. The potted plants were
grown in the experimental farm of Pingtung University of Science and
Technology, under average day and night temperatures of 33 and 27 °C,
respectively. A slow-release fertilizer, containing 14% N, 14% P2O5,
and 14% K2O (Osmocote®, Everris International B.V., Netherlands), was
applied at the rate of 0.5 g per container. The plants were overhead
irrigated as required and fertilizers (20 N-8.1 P2O5-16.9 K2O, Peters®,
Everris NA Inc., USA) with N at 200 ppm were applied once a week.
They were grown under long-day conditions and the long-day treat-
ment was carried out with 23-W energy-saving bulbs (East Asia, China
Electric Co., Ltd., Taiwan) from 10:00–14:00 h (the shoot apex re-
ceived > 150 lx) from September 21 to November 21. Thereafter, the
experiments were carried out from November 2014 until March 2015.
2.2. Effects of GA3 concentration on flowering
In the experiment involving a short-day environment (11-h to 11.5-
h photoperiod), 2.5, 5, 10, and 20 mg L−1 GA3 (Sigma Co., Mo., USA)
were applied on three-month-old K. marnieriana, K. nyikae, and K.
sexangularis. In addition, K. longiflora and K. × richaudii were treated
with 50, 100, and 200 mg L−1 GA3 solution. Twenty milliliters of GA3
solution was applied on the leaves of each plant. The control group was
treated with deionized water. GA3 treatment was applied once every
seven days, for three weeks. The conditions of plant fertilizer applica-
tion and irrigation were the same as that described above. After flow-
ering, the vegetative and reproductive characteristics of each plant
were recorded.
2.3. Effects of different temperatures and GA3 application on flowering
K. nyikae, K. poincarei, and K. sexangularis were used for this study.
Table 1
Effects of GA3 concentration on flowering of Kalanchoe nyikae.
Concentration (mg Percentage of Plant height Nodes Branches
L−1) flowering (%) (cm)
0 0 cy 67.2 c 8.0 b 0.0 a -z
2.5 50 b 91.3 ab 11.0 a 0.2 a 78.8
5.0 50 b 87.3 ab 11.8 a 0.0 a 75.0
10.0 83 ab 94.1 a 11.8 a 0.0 a 71.0
20.0 100 a 84.1 b 11.7 a 0.0 a 70.0
y
Means with the same letter in a column are not significantly different by LSD test at P < 0.05 level.
z
Plants did not flower until end of experiment.
Scientia Horticulturae 238 (2018) 7–13
The afore-mentioned cutting propagation method and growth medium
were used. The rooted plants were transferred to 9-cm plastic pots and
maintained under long-day conditions. Two months after cultivation,
the plants were transferred to two controlled-environment cabinets
(EH-1800, Juxing Chemical Instrument Co., Ltd., Taiwan) with tem-
peratures set at 30/25 °C and 25/20 °C (day/night temperature; varia-
tion from set points ± 1 °C), respectively. Light was provided by white
fluorescent tubes and the photoperiod was set at 11 h:13 h (light:dark).
A photon flux density of 100 μmol m−2 s−1 was measured on the young
leaves of the plants. The plants were treated with 20 mg L−1 GA3, which
was applied once every seven days, for a total of three times. The
control group was treated with deionized water. After the first florets
opened, the vegetative and reproductive characteristics of the plants
were recorded.
2.4. Growth measurements
The percentages of flowering, plant height, number of nodes,
number of branches, number of flowers, days to flowering, flower
diameter, and length and diameter of the flower tubes were recorded.
Plant height was measured from the base of the stem to the shoot apex
or the top of the first floret. The percentage of flowering was defined as
the proportion of plants with open florets to the total number of plants
used in the study. The number of days to flowering was defined as the
number of days from the GA3 treatment to the opening of the first floret.
Flower diameter was recorded by measuring the diameter of the first
open floret. Floret diameter, and the length and width of the flower
tube were measured using a digital slide caliper.
2.5. Statistical analysis
All experiments were performed using a completely randomized
design. Data representing means of five replications were analyzed by
the SAS software (SAS Institute Inc., North Carolina, USA). Analysis of
variance (ANOVA) and the least significant difference (LSD) were used
to compare the treatment means at the 5% significance level.
3. Results
3.1. Effects of GA3 concentration on flowering
Application of 2.5 and 5 mg L−1 GA3 led to flowering in 50% of the
treated K. nyikae plants, while flowering occurred in 100% of the
treated plants upon treatment with 20 mg L−1 GA3. The percentage of
flowering and the number of nodes generated in the K. nyikae plants
subjected to different GA3 treatments were significantly different from
that in the control group plants. The number of days to flowering in K.
nyikae plants under 10 and 20 mg L−1 GA3 treatments were 7.8–8.8
days earlier than that in the plants under 2.5 mg L−1 GA3 treatment.
The highest number of K. nyikae flowers was observed in the plants
treated with 2.5 mg L−1 GA3 (Table 1). K. sexangularis plants in all GA3
treatments showed 100% flowering, and application of GA3 sig-
nificantly reduced the days taken to flower in this species (Table 2). The
Days to Number of Flower diameter Length of flower Width of flower
flower flowers (mm) tubes (mm) tubes (mm)
– – – –
a 182.3 a 22.3 a 19.5 a 4.4 a
ab 140.8 b 21.2 a 19.7 a 4.4 a
b 135.2 b 21.0 a 19.5 a 4.4 a
b 138.8 b 21.4 a 19.5 a 4.3 a
8
M.-Z. Chang, C.-H. Huang Scientia Horticulturae 238 (2018) 7–13

Table 2
Effects of GA3 concentration on flowering of Kalanchoe sexangularis.
Concentration (mg Percentage of Plant height Nodes Branches Days to Number of Flower diameter Length of flower Width of flower
L−1) flowering (%) (cm) flower flowers (mm) tubes (mm) tubes (mm)

0 0 by 32.7 b 12.0 a 1.3 a -z – – – –

2.5 100 a 66.3 a 10.2 bc 0.2 b 85.8 a 389.5 b 8.3 b 15.5 a 3.4 a
5.0 100 a 69.8 a 9.5 c 0.0 b 83.7 a 477.7 a 9.0 ab 15.1 ab 3.4 a
10.0 100 a 68.3 a 11.5 ab 0.0 b 71.2 b 496.0 a 9.3 a 14.7 bc 3.5 a
20.0 100 a 62.8 a 10.8 abc 0.0 b 62.5 c 515.5 a 8.3 b 14.2 c 2.7 b

y
Means with the same letter in a column are not significantly different by LSD test at P < 0.05 level.
z
Plants did not flower until end of experiment.

number of days to flowering in K. sexangularis plants under the
20 mg L−1 GA3 treatment was effectively reduced by 23.3 days from
that taken by the 2.5 mg L−1 GA3-treated K. sexangularis plants. The
numbers of flowers in the K. sexangularis plants subjected to 5, 10, and
20 mg L−1 GA3 treatments were higher than that in the K. sexangularis
plants under 2.5 mg L−1 GA3 treatment. The number of flowers pro-
duced per plant increased with an increase in GA3 concentration
(Table 2).
The percentage of flowering in K. marnieriana was enhanced with
the increase in GA3 concentration. Flowering occurred in 100% of the
plants treated with 20 mg L−1 GA3. However, the number of days to
flowering was not significantly different among the K. marnieriana
plants subjected to different GA3 treatments. The plants under the 5 and
10 mg L−1 treatments had the highest number of flowers (Table 3).
Treatment with 50 mg L−1 GA3 resulted in the flowering of 83% of the
treated K. × richaudii plants, while flowering occurred in 100% of the
plants treated with 100 and 200 mg L−1 GA3. The number of nodes in
all the GA3 treated K. × richaudii plants was significantly decreased as
compared with that in the control group plants. The number of days to
flowering in K. × richaudii decreased in the plants under 100 mg L−1
GA3 treatment by 11.8 days as compared with that in the plants under
50 mg L−1 GA3 treatment (Table 4). Flowering occurred in 100% of all
GA3-treated K. longiflora plants. The number of days to flowering and
the number of flowers in the K. longiflora plants under different GA3
treatments were not significantly different (Table 5).
3.2. Effects of different temperatures and GA3 application on flowering
Flowering occurred in 50 and 67% of the GA3-treated K. nyikae and
K. sexangularis plants, respectively, at 25/20 °C. The plants did not
flower under the high day and night temperature conditions, regardless
of the concentration of the applied GA3 (Tables 6 and 7). Flowering
occurred in 67 and 17% of the GA3-treated K. poincarei plants at 25/
20 °C and 30/25 °C, respectively. The untreated control plants did not
flower (Table 8).
4. Discussion
GAs are tetracyclic diterpenoid compounds that are associated with
the promotion of seed germination, stem and root growth, flower bud
differentiation and flowering, pollen development, pollen tube growth,
parthenocarpy, shortening of the juvenile period, and determination of
the sex of flowers (Buchanan et al., 2015; Matsuoka, 2003; Pharis and
King, 1985; Taiz and Zeiger et al., 2010). Wilson et al. (1992) confirmed
that GA4 was produced in the leaves and transported to the shoot apex,
where it causes flowering under short-day conditions in Arabidopsis.
Kulikowska-Gulewska et al. (2000) suggested that GAs are involved in
the control of phytochrome levels during vegetative growth until
flowering, and play a promotive role in the early process of flowering in
Pharbits nil (Wijayanti et al., 1997). Exogenous GAs have been found to
induce or promote flowering in many species (Pharis and King, 1985;
Wilson et al., 1992; Zeevaart, 1985). In the family Cupressaceae, GAs
promote early maturation and flowering (Pharis and Ross, 1985). The
percentage of flowering increased with increase in GA3 concentration in
Philodendron ‘Black Cardinal’ (Chen et al., 2003). B. daigremontianum
required 5 μg GA3 per plant to induce flowering (Zeevaart, 1969b).
Flowering in B. crenatum was promoted by the application 0.2 μg GA3,
GA4, or GA7 per plant (Michniewicz and Lang, 1962). Huang (2007)
showed that flowering was induced in 100% of the treated six-month-
old and two-month-old K. pinnata plants by spraying 25 and 5 mg L−1
GA3, respectively. Applying 25 mg L−1 GA3 in K. luciae and K. mar-
morata resulted in the flowering of 100% of the treated plants (Lu,
2013). In the present study, the leaves treated with GA3 could effec-
tively induce flowering in species belonging to the sections Kalanchoe
and Bryophyllum (Tables 1–5). Flowering was induced in 100% of the
treated K. sexangularis, K. nyikae, and K. × richaudii plants by applying
2.5, 20, and 100 mg L−1 GA3, respectively (Tables 1, 2, and 4). Currey
and Erwin (2011) showed that K. marnieriana and K. longiflora did not
flower under flowering-inducing photoperiods. However, we observed
flowering in 100% of the plants of these species treated with 20 and
50 mg L−1 GA3 (Tables 3 and 5). We also observed that the percentage
of flowering in K. marnieriana and K. nyikae was promoted with an
increase in the concentration of applied GA3. These findings indicate
that the application of a broad range of GA3 concentration can induce
flowering in the studied species.
GAs can effectively shorten the length of the juvenile phase
(Sgamma, 2017) and play a very important role in the regulation of
flowering (Pharis and King, 1985). The juvenile phase represents the
early period of plant development when flower initiation does not occur
even under suitable conditions (Erwin, 2002; Ison and Humphreys,

Table 3
Effects of GA3 concentration on flowering of Kalanchoe marnieriana.
Concentration (mg Percentage of Plant height Nodes Branches Days to Number of Flower diameter Length of flower Width of flower
L−1) flowering (%) (cm) flower flowers (mm) tubes (mm) tubes (mm)

0 0 by 45.2 b 26.5 b 6.8 a -z – – – –

2.5 0b 51.8 a 29.3 ab 6.8 a 106.0 b 89.3 b 9.6 a 26.6 a 8.2 a
5.0 33 b 48.2 ab 32.5 a 8.0 a 106.0 b 114.5 a 8.9 a 26.1 ab 8.1 a
10.0 83 a 48.2 ab 29.5 ab 7.2 a 105.3 b 111.3 a 8.8 a 26.8 a 8.1 a
20.0 100 a 45.7 b 30.2 ab 6.2 a 106.7 b 66.3 c 7.8 b 25.1 b 7.5 b

y
Means with the same letter in a column are not significantly different by LSD test at P < 0.05 level.
z
Plants did not flower until end of experiment.

9
M.-Z. Chang, C.-H. Huang Scientia Horticulturae 238 (2018) 7–13

Table 4
Effects of GA3 concentration on flowering of Kalanchoe×richaudii.
Concentration (mg Percentage of Plant height Nodes Branches Days to Number of Flower diameter Length of flower Width of flower
L−1) flowering (%) (cm) flower flowers (mm) tubes (mm) tubes(mm)

0 0 by 42.3 b 38.2 a 2.2 a -z – – – –

50 83 a 47.3 ab 25.8 bc 0.7 b 85.8 a 155.2 a 15.6 a 26.9 a 6.4 a
100 100 a 50.0 a 25.0 c 1.2 ab 74.0 c 174.0 a 15.9 a 26.4 a 6.9 a
200 100 a 52.8 a 27.7 b 1.0 b 80.2 b 180.3 a 16.1 a 27.6 a 7.1 a

y
Means with the same letter in a column are not significantly different by LSD test at P < 0.05 level.
z
Plants did not flower until end of experiment.

1984; Sgamma, 2017; Wadhi and Ram, 1967). Wadhi and Ram (1967)
found that the endogenous GA content in juvenile plants was low,
which was insufficient to induce flowering. Low endogenous GA con-
tents delayed floral induction and flowering in Raphanus sativus
(Nishijima et al., 1997). In the B. daigremontianum plants transferred
from under long-day to short-day environmental conditions, large
amounts of GAs were found to be produced, which stimulated flow-
ering. GAs are the limiting factor controlling the production of floral
stimulus in Bryophyllum under short-day conditions (Zeevaart and Lang,
1962; Zeevaart, 1969a). Huang (2007) also found that flowering in two-
month-old K. pinnata plants required a five-times higher GA3 con-
centration than the six-month-old plants. The juvenile phase may last
from only a few days in annual plants to several years in perennials
(Sgamma, 2017), and depends on the age of the plant, number of
leaves, or size of meristem. For instance, the juvenile phases in Stylo-
santhes guianensis ‘Schofield’ and ‘Cook,’ a long-short day plant, are 45
days and 61 days, respectively (Ison and Humphreys 1984). It lasts for
two years in B. daigremontianum and K. pinnata (Wadhi and Ram, 1967;
Zeevaart, 1962), six years in K. prolifera (Zeevaart, 1985), and five–ten
years in Hedera helix (Sgamma, 2017). K. marnieriana, K. longiflora and
K. × richaudii do not bloom naturally after one year of cultivation
(personal observation). However, in this study, flowering in the studied
species were induced depending on GA3 concentration (Tables 1–5).
Therefore, the juvenile phase also appears to be one of the limiting
factors influencing flowering in Kalanchoe species. The application of
GA3 has a promotive effect on flowering, and effectively shortens the
juvenile phase in Kalanchoe.
Increase in endogenous GA3 was shown to accelerate flowering time
(Domagalska et al., 2010). Plants treated with increasing concentra-
tions of GA3 or GA4 + 7 showed a decrease in the time taken to flower
(Al-Khassawneh et al., 2006; Harkess and Lyons, 1994; Sumanasiri
et al., 2013). Application of 100 mg L−1 GA3 to Belamcanda chinensis
reduced the number of days to flowering and increased plant height,
number of leaves, inflorescence length, flower length, and flower
number (Bhuj et al., 1998). Application of GA3 promoted early flow-
ering in chrysanthemum (Sajid et al., 2016; Sharifuzzaman et al.,
2011). Phengphachanh et al. (2012) has also suggested that GA3 pro-
moted flower initiation and early flowering in Rhynchostylis gigantea.
Application of exogenous GA3 reduced the time taken to flowering in
the non-cold treated plants under short day conditions, suggesting that
endogenous GAs act as limiting factors of spring rape (Brassica napus
var. annua) (Dahanayake and Galwey, 1999). The present study also
showed that GA3 treatment significantly reduced the number of days
taken to flower in K. nyikae and K. sexangularis (Tables 1 and 2).
GA3 treatment was shown to increase the number of flowers in
Pharbits nil (Kulikowska-Gulewska et al., 2000). K. pinnata treated with
a high concentration GA3 produced more flowers than the plants
treated with lower concentrations of GA3 (Wadhi and Ram, 1967).
Application of GA3 enhanced early flowering, and increased the number
of flowers and flower diameter in Helleborus niger and Helleborus ×
ericsmithii ‘Pink Beauty’ (Christiaens et al., 2012). GA3-treated Brunonia
flowered early and showed 30% more inflorescences per plant under
short-day conditions (Wahyuni et al., 2011). Henckelia humboldtianus
treated with 200 mg L−1 GA3 exhibited early flowering, increased
number of inflorescences and numbers of flowers per inflorescence, and
improved vegetative growth (Sumanasiri et al., 2013). The percentage
of flowering and flower number increased with increase in GA3 con-
centration in Philodendron ‘Black Cardinal’ (Chen et al., 2003). Chen
(2007) showed that treatment with 100 mg L−1 GA3 significantly re-
duced the number of flowers in L. heterophylla. In the present study, the
number of flowers per plant increased in K. sexangularis and K. × ri-
chaudii with an increase in applied GA3 concentration (Table 2), al-
though, K. marnieriana treated with 20 mg L−1 GA3 showed a decreased
number of flowers (Table 3).
Abscisic acid(ABA) antagonizes GAs during many plant develop-
mental processes (Kinet et al., 1985). The role of ABA has been de-
monstrated in delaying or inhibiting flowering (Bernler et al., 1981;
Domagalska et al., 2010). GAs and ABA may act at the same site during
flowering, but in an opposite manner (Wijayanti et al., 1997).
Domagalska et al. (2010) suggested that hormonal regulation of the
transition from vegetative to reproductive development in Arabidopsis
thaliana depends on the balance between GAs and ABA. GA3 was also
found to promote flower initiation and early flowering by decreasing
the concentration of ABA in Rhynchostylis gigantea (Phengphachanh
et al., 2012). Badr et al. (1970) suggested that application of ABA in-
hibited flowering in olive exposed to inductive winter chilling tem-
peratures. Flowering in Pharbitis nil was inhibited by applying ABA
under short day conditions (Wilmowicz et al., 2008) and the inhibitory
effects of ABA were reversed by application of GA3 (Wijayanti et al.,
1997). Therefore, the balance between endogenous inhibitors and GAs
appears to be a likely candidate controlling flower induction. High
temperatures typically promote ABA synthesis, consequently inhibiting
GA synthesis and signaling through the action of ABA (Seo et al., 2006).
Other reports suggest that plant exposure to low temperature can

Table 5
Effects of GA3 concentration on flowering of Kalanchoe longiflora.
Concentration (mg Percentage of Plant height Nodes Branches Days to Number of Flower diameter Length of flower Width of flower
L−1) flowering (%) (cm) flower flowers (mm) tubes (mm) tubes (mm)

0 0 by 26.3 b 8.3 b 4.0 a -z – – – –

50 100 a 41.7 a 10.7 a 0.5 b 80.7 a 94.5 a 12.7 a 14.7 a 3.1 a
100 100 a 44.5 a 10.3 a 0.3 b 81.8 a 93.3 a 13.1 a 17.1 a 3.1 a
200 100 a 45.0 a 10.7 a 0.3 b 82.3 a 103.0 a 13.2 a 17.2 a 3.1 a

y
Means with the same letter in a column are not significantly different by LSD test at P < 0.05 level.
z
Plants did not flower until end of experiment.

10
M.-Z. Chang, C.-H. Huang Scientia Horticulturae 238 (2018) 7–13

Table 6
Effects of GA3 application on flowering of Kalanchoe nyikae in different temperature conditions.
Day/ night GA3 concentration (mg Percentage of Plant height Nodes Branches Number of Flower diameter Length of flower Width of flower
temperature (℃) L−1) flowering (%) (cm) flowers (mm) tubes (mm) tubes (mm)

25/20 0 0 bx 48.8 bc 11.2 b 0a -y – – –

20 50 a 82.7 a 17.7 a 0a 37.7 20.8 17.8 4.2
30/25 0 0b 43.5 c 10.0 b 0a – – – –
20 0b 55.8 c 11.0 b 0a – – – –
Tz ** *** *** ns
C ** *** *** ns
T*C ** ** ** ns

x
Means with the same letter in a column are not significantly different by LSD test at P < 0.05 level.
y
Plants did not flower until end of experiment.
z
Levels of significant are represented by *: P < 0.05, **: P < 0.01, ***: P < 0.001 and ns: not significant.

increase biosynthesis of GA and endogenous GA concentrations in
Arabidopsis thaliana (Derkx et al., 1994), Pisum sativum (Grindal et al.,
1998), Aster savatieri (Ishida and Takano, 1971), Phalaenopsis hybrida
(Su et al., 1994; Su et al., 2001), Thlaspi arvense (Hazebroek and
Metzger, 1990; Hazebroek et al., 1993), wheat (Lin and Stafford, 1987),
canola (Zanewich and Rood, 1995), and lily (Takayama et al., 1993).
The findings of Sawhney (1983) indicated that endogenous GA3 con-
centrations of Lycopersicon esculentum was increased at low tempera-
tures. Furthermore, the petals, stamens, carpels, and ventricles in a low
temperature environment (18/15 °C) were significantly larger than
those in a high temperature environment (28/23 °C). Higher en-
dogenous GA levels were detected following treatment with GA3 in
Phalaenopsis hybrida (‘Minho Princess’× Phalaenopsis equestris) exposed
to low temperature (25/20 °C) than in plants exposed to high tem-
peratures or in untreated plants (Su et al., 2001). Application of GA4+7
induced flowering in Aquilegia × hybrida at lower temperatures than at
higher temperatures (Gianfagna and Merritt, 1998). Flowering was
induced in Lavandula heterophylla by applying 500 mg L−1 GA3 or
GA4+7 at 15/12 °C and 20/15 °C, not, however, under high temperature
conditions. Evidently, flowering was affected by the type of GAs used
and the post-GA treatment temperature conditions (Chen, 2007). GA
substitutes were required to induce flowering in Gypsophila panicalata at
high night temperatures under long-day conditions (Shlomo et al.,
1985). Hisamatsu et al. (2004) showed that ent-kaurene biosynthesis,
which is stimulated under low temperature conditions, leads to stem
elongation in Eustoma grandiflorum. The plants treated with GA4 and
GA9 had rapidly elongating stems and earlier flower formation under
short-day conditions (Xu et al., 1997). GA3 contents in plants, therefore,
increase by metabolic processes at low temperatures or by exogenous
GA3 application on the plants (Hazebroek and Metzger, 1990;
Hazebroek et al., 1993; Su et al., 2001). In addition, the GA metabolism
may be important in promotion of flowering. Su et al. (2001) found that
floral induction and development of Phalaenopsis hybrida was induced
by cold temperature or application of GA3 owing to reduced conversion
of active GA1 to its inactive forms. The active GA1 level decreased,
resulting in delayed flowering of Raphanus sativus in a long day en-
vironment after cold induction (Nishijima et al., 1997). Several reports
indicated that activity of the enzymes involved in GAs biosynthesis
were changed by cold temperature (Metzger, 1990; Hazebroek et al.,
1993; Zanewich and Rood, 1995). The bioactive GA in imbibed seeds of
Arabidopsis thaliana stay at low levels under high temperature condition
(Toh et al., 2008). Yamauchi et al. (2004) suggested that imbibed seeds
of Arabidopsis thaliana had increased levels of bioactive GA under low
temperature conditions. GA3 or GA1 may provide the signal for the
induction of flowering in aster, Solidago, Phlox, and Hypericum under
long day or low temperature condition (Yossi, 2000). K. nyikae, K.
poincarei, and K. sexangularis in the present study flowered after ap-
plication of GA3 at 25/20 °C, but not at higher temperatures. It ap-
peared that the decrease in flowering might be associated with in-
creased ABA content or conversion of active forms of GA as compared
with that of the inactive forms under high temperature conditions.
Therefore, the findings of this study reveal that promotion of flowering
by GA3 is inhibited at high temperatures (8).
5. Conclusion
Application of GA3 could promote flowering in species of sections
Kalanchoe and Bryophyllum. The GA3 concentration required depended
on the species of plant. The present study indicates that flowering oc-
curred in 100% of the treated K. sexangularis, K. nyikae, K. marnieriana,
K. longiflora, and K. × richaudii plants upon application of 2.5, 20, 20,
50, and 100 mg L−1 of GA3, respectively. The number of days taken to
flower in K. nyikae and K. sexangularis was significantly reduced fol-
lowing GA3 treatment. The number of flowers of K. sexangularis and
K. × richaudii increased with increasing GA3 concentrations. The pro-
moting effect of GA3 on flowering was inhibited at high temperature.
This study clearly elucidated the optimal concentration of GA3 required
for flowering induction in the studied plants. These findings can be
applied for regulation of flower, breeding, and hybridization of
Kalanchoe in future.

Table 7
Effects of GA3 application on flowering of Kalanchoe sexangularis in different temperature conditions.
Day/ night GA3 concentration Percentage of Plant height Nodes Branches Number of Flower diameter Length of flower Width of flower
temperature (℃) (mg L−1) flowering (%) (cm) flowers (mm) tubes (mm) tubes (mm)

25/20 0 0 bx 31.5 c 10.7 b 1.3 a -y – – –

20 67 a 46.3 b 11.7 ab 0.8 a 128.5 8.7 15.7 2.1
30/25 0 0b 33.3 c 11.8 ab 0b – – – –
20 0b 54.7 a 18.3 a 0b – – – –
Tz ** ** ** ***
C ** *** ** –
T*C ** ** ns ns

x
Means with the same letter in a column are not significantly different by LSD test at P < 0.05 level.
y
Plants did not flower until end of experiment.
z
Levels of significant are represented by *: P < 0.05, **: P < 0.01, ***: P < 0.001 and ns: not significant.

11
M.-Z. Chang, C.-H. Huang Scientia Horticulturae 238 (2018) 7–13

Table 8
Effects of GA3 application on flowering of Kalanchoe poincarei in different temperature conditions.
Day/ night GA3Concentration (mg Percentage of Plant Height Nodes Branches Number of Flower Length of Width of flower
temperature (℃) L−1) flowering (%) (cm) flowers diameter (mm) flower tubes tubes (mm)
(mm)

25/20 0 0 bx 25.0 bc 14.7 b 0a -y – – –

20 67 a 45.3 a 16.8 a 0a 69.8 4.5 29.5 6.9
30/25 0 0b 15.8 c 9.8 c 0a – – – –
20 17 b 28.3 b 16.7 a 0a 72.0 4.4 16.2 6.1
Tz – ** ** ns
C – *** *** ns
T*C – ** ** ns

x
Means with the same letter in a column are not significantly different by LSD test at P < 0.05 level.
y
Plants did not flower until end of experiment.
z
Levels of significant are represented by *: P < 0.05, **: P < 0.01, ***: P < 0.001 and ns: not significant.

Acknowledgment
This study was supported by the Agriculture and Food Agency,
Council of Agriculture of Taiwan (grant number 103AS-9.2.5-FD-Z1).
We declare the absence of any conflicts of interest with government
agencies and authors.
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