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Adv. Hort. Sci., 2018 32(2): 205-212 DOI: 10.13128/ahs-22019
Micropropagation of dwarf schefflera
[Schefflera arboricola (Hayata) Merr.]
via direct shoot regeneration
O. Rezaei Baghbidi, A. Jowkar (*)
Department of Horticultural Science, College of Agriculture, Shiraz
University, Shiraz, Iran.
Key words: In vitro propagation, NAA, Schefflera arboricola,shoot induction,
TDZ, variegated leaves.
Abstract: Schefflera [Schefflera arboricola (Hayata) Merr.] is one of the most
popular ornamental house plants conventionally propagated by seeds. Rapid
multiplication of elite clones is an important driving force for the pot plant’s
market. In this regard, In vitro clonal propagation of three schefflera cultivars,
‘Luseane’, ‘Charlotte’ and ‘Gold Capella’, was examined. Sterilization was done
by 70% ethanol for 2 min and 1% sodium hypochlorite solution for 15 min.
Shoot proliferation of the nodal segments was dependent on cytokinin supply.
The greatest number of shoots was obtained when nodal explants were cul-
tured on the MS medium with 0.5 mg l-1 TDZ for ‘Luseane’, or 8 mg l-1 BA for
‘Charlotte’ and ‘Gold Capella’. Subculture of nodal segments harvested from
the in vitro derived axillary shoots on the multiplication medium enabled con-
tinuous production of healthy shoots with similar frequency. Plantlets of
‘Luseane’ and ‘Gold Capella’ demonstrated 100% rooting using 2 mgl-1 NAA,
while ‘Charlotte’ showed 93.75% root induction by 1 mgl-1 NAA. Plantlets were
acclimatized successfully using peat moss and sand mixture (‘Luseane’), loam
soil, sand and leaf compost (‘Charlotte’) or peat moss and perlite mixture (‘Gold
Capella’).
1. Introduction
Dwarf Schefflera [Schefflera arboricola (Hayata) Merr.] is an evergreen
ornamental plant of the Araliaceae family, native to China and Taiwan
(Ohashi, 1993). It is mostly used indoors as a foliage pot plant because of
the attractiveness of the umbrella-like palmately compound leaves and
variegated cultivars (Gilman and Watson, 1994; Chen et al., 2002).
Schefflera’s ability to clean the air and its tolerance to harsh interior envi-
ronments has further increased its worldwide popularity (Yang et al.,
2009; Dela Cruz et al., 2014). To satisfy grower’s calls for potted plants of
schefflera, methods for rapid propagation of selected cultivars are crucial.
Multiplication of this house plant is mainly by seeds which results with
the segregation of the progeny traits and limited to its native plantation
in the tropics (Griffith, 1998; Chen at al., 2002) and only to cultivars of
(*) Corresponding author:
ajowkar@gmail.com
Citation:
REZAEI BAGHBIDI O., JOWKAR A., 2018 -
Micropropagation of dwarf schefflera [Schefflera
arboricola (Hayata) Merr.] via direct shoot rege-
neration. - Adv. Hort. Sci., 32(2): 205-212
Copyright:
© 2018 Rezaei Baghbidi O., Jowkar A. This is an
open access, peer reviewed article published by
Firenze University Press
(http://www.fupress.net/index.php/ahs/) and
distributed under the terms of the Creative
Commons Attribution License, which permits
unrestricted use, distribution, and reproduction
in any medium, provided the original author and
source are credited.
Data Availability Statement:
All relevant data are within the paper and its
Supporting Information files.
Competing Interests:
The authors declare no competing interests.
Received for publication 8 November 2017
Accepted for publication 24 January 2018
AHS
Advances in Horticultural Science
Adv. Hort. Sci., 2018 32(2): 205-212
206
schefflera which have non-variegated green leaves
(Marcotrigiano, 1997). Other propagation means are
leaf-bud cuttings (Hansen, 1986), air layering (Gilman
and Watson, 1994) and stem cuttings (Hansen, 1986).
These practices are hindered by difficulties such as
low number of propagules per plant, increased time
of production and the risk of disease spread from
several pathogens such as Pseudomonas cichorii,
Xanthomonas campestris, Phytophthora parasitica,
Pythium splendens and Alternaria panax (Chase and
Poole, 1986).
Micropropagation is a reasonably more efficient
way for schefflera production as a greater number of
plants can be produced faster compared to the tradi-
tional cuttings. Moreover, in vitro culture of tropical
ornamental plants has been recommended as a tool
to eradicate the diseases that are frequently wide-
spread in the mother plants (Hartmann and Kester,
2011). Tissue culture has furthermore resulted in
enhanced features compared to common propaga-
tions. It was revealed that micropropagated foliage
pot plants such as syngonium, spathiphyllum, dief-
fenbachia (Conover, 1992) and philodendron (Chen
et al., 2012) had a packed and denser plant forms in
co mpa ris on to the c onv ent ion al stem cut tin gs.
Consequently, small tissue culture-grown scheffleras
may be readily suitable for the limited space avail-
able in the terrariums or bonsai pots.
Despite the fact that micropropagation is exten-
sively exploited by the floriculture industry, their for-
mulas are not released to the general public. To our
knowledge, there is no tissue culture protocol avail-
able in the literature for S. arboricola, though a few
number of articles have been published so far for
other Araliaceae species with horticultural impor-
tance such as Cussonia paniculata (Tetyana and van
Staden, 2001), Fatsia japonica (Choi et al., 2005),
Eleutherococcus senticosus (Amin et al., 2003; You et
al., 2005), Panax quinquefolius (Uchendu et al., 2011)
Hedera helix (Sivanesan et al., 2011) Polyscias bal-
fouriana (Ilyas et al., 2013) and Polyscias fruticosa
(Sakr et al., 2014).
The current research developed, for the first time,
a micropropagation procedure for three cultivars of
shefflera with green, white and yellow variegated
leaves. In order to reach a high propagation rate and
get well plantlets, the effects of various sterilization
treatments, plant growth regulators’ type and con-
centration were studied on the survival rate, shoot
proliferation and rooting of scheffleras. Subsequent
acclimatization of the micropropagated plantlets was
also investigated using different potting media.
2. Materials and Methods
Plant material and culture medium
Three marketable cultivars with green, white and
yellow variegated leaves (i.e. ‘Luseane’, ‘Charlotte’
and ‘Gold Capella’, respectively) were used for micro-
propapagation. The explants were cultured on MS
(Murashige and Skoog, 1962) medium supplemented
with 3% (w/v) of sucrose, and 0.65% (w/v) of agar.
The pH of the medium was set to 5.7 before auto-
clave sterilization (i.e., 20 min, at 121°C). The plant
growth regulators were added to the medium later
by filter sterilization. The sterile nodal segments were
established in culture medium containing 2 mg l-1
benzyl adeni ne (BA). In order to proliferate the
shoots, the emerging buds were removed after 30
days and subcultured to media containing different
cytokinin treatments.
Sterilization of three Schefflera cultivars
The stock plants were sprayed with Ridomil fungi-
cide (0.3%) one week before explant excision. Stem
cuttings were soaked in 0.1% detergent solution for
10 min, then washed under running tap water for 30
min. The explants were moved to laminar flow cabi-
net and 1.5-cm long nodal segments were excised
fr om the s tem. F or surf ace ster ilizati on of the
explants, 70% EtOH was tested for 0, 30, 60, 90, 120,
150 and 180 seconds. Explants were then disinfected
using 0, 0.5, 0.75, 1 or 1.25% (v/v) sodium hypochlo-
rite containing 0.01% Tween-20 for 5, 10 or 15 min
on a shaker. After four rinses with sterile water and
excision of the damaged edges, the explants were
cultured on PDA medium (300 g potato extract, 30 g
l-1 of dextrose and 8 g l-1 of agar) for the evaluation of
the contamination. Five replicates were examined for
each treatment with three explants in each flask.
Contaminations and survival rates were recorded
after 1 month and those having noticeable infection
signs were instantly discarded.
Effect of cytokinin type and concentration on shoot
proliferation
Inducing new shoots was carried out using BA and
Kinetin (Kin) at 0, 1, 2, 4 and 8 mg l-1 concentrations
while thidiazuron (TDZ) was tested at 0, 0.125, 0.25,
0.5 and 1 mg l−1 concentrations. Five replicate flasks
were used for each treatment with three explants in
each flask. The number and length of the proliferated
shoots, and number of leaves produced per explant
were recorded after 30 days. The regenerated shoots
were later subcultured to the best cytokinin-contain-
ing MS medium.
Rezaei Baghbidi and Jowkar - Micropropagation of dwarf schefflera
207
Effect of auxin type and concentration on root induc-
tion
Shoots from the finest proliferation treatment
were put in the glass flasks of MS medium containing
0, 0.1, 0.5, 1, 2 or 4 mg l−1 of indole-3-butyric acid
(IBA) or naphthaleneacedic acid (NAA). After 15 days
the shoots were transferred to half-strength MS
medium devoid of auxins. Five replicate flasks were
tested in each treatment with three shoots in each
flask. The number and length of the induced roots
and rooting percentage of the shoots were recorded
15 days later.
Effect of potting media on plant acclimatization
Rooted shoots were carefully taken out from the
glass flasks and washed under distilled water to get
rid of attaching agar from the roots. The plantlets
were then transferred to 10-cm plastic pots filled
with different potting media: peat moss, peat moss
and perlite (1:1), peat moss and sand (1:1), loam soil,
sand and leaf compost (1:1:1). Ten replications were
tested for each of the treatments. Potted plants
were grown in a greenhouse with temperatures
ran g ing from 2 4 t o 2 8 °C, rel a tive humidi t y o f
between 70 and 90%, and light intensity of 35 µmol
m-2 s-1 under a 12 h photoperiod. After 45 days the
survival rate and plant height were analyzed.
Culture conditions, experimental design and data
analysis
The in vitro researches were carried out with an
environment temperature of 25±2°C and a 16/8 h
light/dark photoperiod delivered by cool fluorescent
lamps at 35 µmol m-2 s-1. All the tests were arranged
in a completely randomized design. Analysis of the
variances was done by SAS software (SAS Institute
Inc., 2002) and means were compared by LSD test at
5% probability level.
3. Results
Sterilization of three Schefflera cultivars
For the determination of the most effective sur-
face sterilization, explants were soaked in 70% EtOH
soluti on for 0-180 sec. As the soaking time w as
in crea sed to 180 sec, surf ace ste rili zati on was
increased gradually and eventually reached 100% in
all three cultivars, however viability was rapidly
decreased after 120 sec (Table 1). Thus, 120 sec, pro-
viding the highest disinfection (72.8%, 79.6% and
72.8% for ‘Luseane’, ‘Charlotte’ and ‘Gold Capella’,
respectively) and viability (72.8%, 74.6% and 72.8%
for Luseane’, ‘Charlotte’ and ‘Gold Capella’, respec-
tively) altogether, was selected. The results were not
significant between the cultivars.
The explants of ‘Luseane’ were effectively disin-
fected when 1.25% sodium hypochlorite was applied
for 15 min, however this application resulted with a
significant decrease of the viability to 13% (Table 2).
Th e most effect ive combin ati on of dis infectio n
(93.2%) and viability (86.4%) was obtained using 1%
sodium hypochlorite for 15 min. ‘Charlotte’ demon-
strated a similar pattern of sterilization. The utmost
co ntro l of con tami nat ion (93. 2%) and v iabi lity
(86.4%) was observed using 1% sodium hypochlorite
for 15 min or 1.25% sodium hypochlorite for 10 min.
Similarly, also for ‘Gold Capella’, the best percent-
ages of sterilization (93.2%) and viability (86.4%)
were obtained by using 1% sodium hypochlorite for
15 min.
Effects of cytokinin type and concentration on shoot
proliferation
Effects of benzyl adenine and kinetin treatments
on each schefflera cultivars. The increase of BA con-
centration to 8 mg l-1 significantly increased the num-
Table 1 - Effects of ethanol treatments on surface sterilization and viability of three schefflera cultivars
Data (±SE) are the mean values of five replicates with three explants in each.
For each cultivar, different lowercase letters indicate significant differences among treatments (LSD multiple range test, P≤0.05).
70% Ethanol (sec)
‘Luseane’ ‘Charlotte’ ‘Gold Capella’
Bacterial sterilization
(%)
Viability
(%)
Bacterial sterilization
(%)
Viability
(%)
Bacterial sterilization
(%)
Viability
(%)
0 0±0 e 0±0 e 0±0 d 0±0 d 0±0 e 0±0 d
30 19.8±8.08 d 19.8±0 d 13.2±8.08 d 13.2±0 d 13.2±8.08 de 13.2±0 cd
60 33±0 d 33±0 cd 46.2±8.08 c 46.2±0 b 26.4±6.60 d 26.4±0 c
90 52.8±8.08 c 52.8±0 b 53.2±8.25 c 53.2±0 b 59.6±6.65 c 59.6±0 a
120 72.8±6.80 b 72.8± 0 a 79.6±8.33 b 74.6±6.6 a 72.8±6.80 bc 72.8±0 a
150 93.2±6.80 a 39.6±6.60 c 100±0 a 33±0 c 86.4±8.33 ab 39.6±6.6 b
180 100±0 a 19.8±8.08 d 100±0 a 13.2±8.08 d 100±0 a 19.8±8.08 c
Adv. Hort. Sci., 2018 32(2): 205-212
208
ber of shoots per explant of ‘Luseane’ to 3.75 (Table
3). This concentration of BA also produced 12.5
leaves per explant. However, the largest shoot length
of 0.88 cm was obtained using 4 mg l-1 BA. On the
contrary, increasing Kin concentration to 4 mg l−1 and
above resulted with the decrease in the shoots num-
ber. Kin also produced significantly lesser leaves at all
the concentrations tested, and did not significantly
increase the shoot length.
Treatment of ‘Charlotte’ with cytokinins revealed
a similar pattern as ‘Luseane’. Indeed, the greatest
shoot proliferation of 3.75 shoots per explant was
obtained using 8 mg l−1 BA, which also produced 15.3
leaves per explant and maximum shoot length of
0.90 cm. Also Kin increased the shoots number at the
same concentration, though it was significantly lower
than what was obtained with BA. The maximum
shoot length and number of leaves observed with 8
mg l-1 Kin were 0.73 cm and 5.67, respectively, and
bot h we r e si gnifi c a ntly lowe r th a n wh at was
observed with BA.
Proliferation of the ‘Gold Capella’ with 8 mg l-1 BA
resulted with maximum of 1.75 shoots per explant,
the largest shoot length of 0.83 cm and 7.58 leaves
per explant, which were all significantly higher than
Kin treatments (Fig. 1). Adding 2 or 4 mg l-1 Kin to the
medium slightly increased the shoot length com-
pared to the control.
Table 2 - Effects of sodium hypochlorite treatments on sterilization and viability of three schefflera cultivars
Data (±SE) are the mean values of five replicates with three explants in each.
For each cultivar, different lowercase letters indicate significant differences among treatments (LSD multiple range test, P≤0.05).
Cultivar Minutes
Sodium hypochlorite (%)
Sterilization (%) Viability (%)
0 0.5 0.75 1 1.25 0 0.5 0.75 1 1.25
‘Luseane’ 5 13.2±8.08 g 24.75±6.39 fg 33±0 e-g 41.25±6.39 d-f 49.5±7.38 de 13.2±0 f 24.75±0 ef 33±0 d-f 41.25±0 c-e 49.5±0 cd
10 13.2±8.08 g 33±0 e-g 52.8±8.08 de 74.5±6.58 bc 86.4±8.33 ab 13.2±0 f 33±0 d-f 52.8±0 cd 74.5±0 ab 79.6±6.80 ab
15 13.2±8.08 g 49.5±7.38 de 59.6±12.51 cd 93.2±6.80 ab 100±0 a 13.2±0 f 49.5±0 cd 59.6±0 cd 86.4±6.80 a 13.2±8.08 f
‘Charlotte’ 5 19.8±8.08 f 33±10.44 ef 33±0 ef 41.25±6.39 de 49.5±7.38 de 19.8±0 ef 33±0 d-f 33±0 d-f 41.25±0 c-e 49.5±0 cd
10 19.8±8.08 f 41.25±6.39 de 52.8±8.08 de 74.5±6.58 bc 93.2±6.80 ab 19.8±0 ef 41.25±0 c-e 52.8±0 b-d 74.5±0 ab 86.4±6.80 a
15 19.8±8.08 f 49.5±7.38 de 59.6±12.51 cd 93.2±6.80 ab 100±0 a 19.8±0 ef 49.5±0 cd 59.6±0 bc 86.4±6.80 a 13.2±8.08 f
‘Gold Capella’ 5 13.2±8.08 g 19.8±8.08 fg 26.4±6.60 e-g 39.6±6.60 d-f 52.8±8.08 cd 13.2 0 f 19.8±0 ef 26.4±0 d-f 39.6±0 c-e 52.8±0 bc
10 13.2±8.08 g 26.4±6.60 e-g 52.8±8.08 cd 66±0 bc 86.4±8.33 ab 13.2±0 f 26.4±0 d-f 52.8±0 bc 66±0 ab 86.4±6.80 a
15 13.2±8.08 g 46.2±8.08 c-e 59.6±12.51 cd 93.2±6.8 0ab 100±0 a 13.2±0 f 46.2±0 b-d 59.6±0 bc 86.4±6.80 a 13.2±8.08 f
Cultivar Control BA (mgl-1) Kin (mgl-1)
1248 1 2 48
‘Luseane’
No. of shoots 0.83±0.1 f 1.25±0.05 de 1.5±0.07 c 2.75±0.08 b 3.75±0.25 a 1.42±0.03 cd 1.42±0.05 cd 1.17±0.06 e 0.92±0.08 f
Shoot length (cm) 0.24±0.03 f 0.50±0.02 c 0.39±0.01 d 0.88±0.01 a 0.73±0.02 b 0.27±0.03 ef 0.27±0.02 ef 0.31±0.005 e 0.31±0.01 e
No. of leaves 0.66±0.36 f 8±0.64 c 8.5±0.4 c 10.42±0.81 b 12.5±2.06 a 1.42±0.32 e 2±0.49 e 2.75±0.52 d 1.67±0.27 e
‘Charlotte’
No. of shoots 0.45±0.21 g 1.67±0.14 f 1.92±0.36 e 2.5±0.48 c 3.75±0.55 a 2.25±0.28 d 1.67±0.41 f 1.67±0.14 f 3±0.36 b
Shoot length (cm) 0.36±0.11 f 0.36±0.12 f 0.45±0.16 de 0.72±0.11 b 0.90±0.15 a 0.47±0.05 d 0.40±0.08 ef 0.38±0.05 f 0.73±0.13 b
No. of leaves 1.17±0.4 g 10.5±0.74 c 10.83±0.1 c 11.83±0.74 b 15.3±1.6 a 2.25±0.31 f 3.67±1.03 e 3.33±0.36 e 5.67±0.62 d
‘Gold Capella’
No. of shoots 0.42±0 g 0.75±0.04 de 0.83±0.1 cd 1.08±0.08 b 1.75±0.08 a 0.67±0.08 ef 0.75±0.08 de 0.92±0.08 c 0.58±0.08 f
Shoot length (cm) 0.19±0.05 e 0.59±0.04 b 0.47±0.04 c 0.76±0.06 a 0.83±0.03 a 0.25±0.02 e 0.39±0.02 cd 0.33±0.01 d 0.23±0.01 e
No. of leaves 0.58±0.08 e 4.41±0.64 c 5.66±0.56 b 5.83±0.52 b 7.58±0.75 a 0.75±0.21 e 0.83±0.22 e 2±0.38 d 1.66±0.14 d
Table 3 - Effects of Benzyl adenine and Kinetin on shoot proliferation of each schefflera cultivars
Data (±SE) are the mean values of five replicates with three explants in each.
For each cultivar, different lowercase letters indicate significant differences among treatments (LSD multiple range test, P≤0.05).
Fig. 1 - Effect of different cytokinin treatments on shoot prolife-
ration of schefflera cvs. Luseane (a), Charlotte (b) and
Go ld Cap ell a (c). Shoot s we re pro du ced us ing MS
medium supplemented with 0.5 mg l-1 TDZ for ‘Luseane’
and 8 mg l-1BA for the others.
Rezaei Baghbidi and Jowkar - Micropropagation of dwarf schefflera
209
Comparison of thidiazuron treatments on shoot
proliferation of schefflera cultivars. The highest shoot
proliferation (6.25 shoots per explant) was observed
in ‘Luseane’, using 0.5 mg l-1 TDZ (Table 4) (Fig. 1).
The maximum shoot proliferation of ‘Charlotte’ and
‘Gold Capella’ significantly lower than ‘Luseane’,
were 3.33 and 1.33 shoots per explant, using 0.5 and
1 mg l-1 TDZ, respectively. The largest shoot length of
2.33 cm was observed in the ‘Charlotte’ when 0.5 mg
l-1 TDZ was added to the medium. The other cultivars’
shoot length were significantly lower. Furthermore,
‘Ch arlo tte’ p r oduc ed the m aximu m leav es per
explant (i.e. 13.5 leaves) using 0.5 mg l-1 TDZ, while
other cultivars yielded significantly fewer leaves per
explant (8.75 for ‘Luseane’, 2.42 for ‘Gold Capella’) at
0.25 mg l-1 TDZ and 1 mg l-1 TDZ, respectively.
Effect of auxin type and concentration on root induc-
tion
Rooting of ‘Luseane’ was 100% when 2 or 4 mg l−1
IBA as well as 0.5, 2 or 4 mg l−1 NAA treatments were
applied (Table 5). The maximum number of roots per
explant (i.e. 18.67 roots) was observed using 2 mg l-1
NAA (Fig. 2). Moreover, the largest root length of
2.74 cm was produced by this treatment. However,
0.5 or 2 mg l-1 NAA appeared to have no significant
differences with the former treatment. Also, the
highest shoot length of 2.96 cm was recorded using 1
mg l-1 NAA, albeit no significant differences were
found with 0.5, 2 and 4 mg l-1 NAA treatments.
The rooting of ‘Charlotte’ raised to the maximum
93.75% using 2 mg l-1 IBA or 1 mg l-1 NAA. Other con-
TDZ (mgl-1)Luseane' Charlotte' ‘Gold Capella’
No. of shoots Shoot length (cm) No. of leaves No. of shoots Shoot length (cm) No. of leaves No. of shoots Shoot length (cm) No. of leaves
0 0.83±0.1 gh 0.24±0.03 I 0.66±0 36 j 0.45±0.21 I 0.36±0.11 h 1.17±0.4 I 0.42±0 I 0.19±0.05 I 0.58±0.08 j
0.125 1.65±0.02 e 0.71±0.01 ef 5.40±0.48 f 1.75±0.08 e 1.28±0.19 c 5.75±0.67 f 0.67±0h I 1.02±0.01 d 1.92±0.16 h
0.25 5.58±0.08 b 1.05±0.05 d 8.75±0.76 c 1.83±0.1 e 1.34±0.13 c 7.33±0.79 d 0.75±0.08 h 0.58±0.04 g 1.75±0.21 h
0.5 6.25±0.08 a 0.66±0.04 fg 6.25±0.82 e 3.33±0.47 c 2.33±0.25 a 13.5±2.58 a 1.08±0.08f g 0.36±0.02 h 1.75±0.16 h
1 5.75±0.25 b 0.80±0.02 e 5.75±0.61 f 2.33±0.24 d 1.76±0.15 b 9.5±1.31 b 1.33±0.02 f 0.41±0.02 h 2.42±0.08 g
Table 4 - Effects of thidiazuron on shoot proliferation of schefflera cultivars
Data (±SE) are the mean values of five replicates with three explants in each.
For each cultivar, different lowercase letters indicate significant differences among treatments (LSD multiple range test, P≤0.05).
Cultivar Control IBA (mgl-1) NAA (mgl-1)
0.25 0.5 12 4 0.25 0.5 124
‘Luseane’
Rooting (%) 8.25±6.39 d 33±0 c 66.25±10.59 b 74.5±6.58 b 100±0 a 100±0 a 33±10.44 c 100±0 a 91.75±6.39 a 100±0 a 100±0 a
No. of roots 0.25±0.11 h 2.58±0.26 g 7.43±0.24 f 11.24±0.35 d 13.83±0.56 c 10.38±0.37 de 2.75±0.12 g 9.75±0.54 e 15.08±0.34 b 18.67±0.33 a 13.83±0.44 c
Root length
(cm)
0.31±0.24 c 0.58±0.14 c 1.78±0.16 b 1.78±0.2 1.94±0.1 b 1.49±0.08 b 0.66±0.2 c 2.46±0.17 a 2.67±0.23 a 2.74±0.08 a 1.95±0.11 b
Shoot length
(cm)
1.85±0.1 c 1.5±0.05 d 1.95±0.08 c 2.03±0.14 bc 1.94±0.06 c 1.89±0.06 c 2.27±0.08 b 2.87±0.12 a 2.96±0.19 a 2.77±0.12 a 2.82±0.14 a
‘Charlotte’
Rooting (%) 58.25±12.44 c 62.5±7.76 bc 83.5±7.38 ab 87.5±5.59 a 93.75±4.84 a 85.5±6.61 a 62.5±7.76 bc 91.75±6.39 a 93.75±4.84 a 91.75±6.39 a 83.5±7.38 ab
No. of roots 3.75±0.58 e 5.02±0.33 d 9.49±0.36 c 12.25±0.45 b 12.28±0.33 b 10.18±0.56 c 6.33±0.86 d 15.06±0.46 a 16.46±0.69 a 13.33±1.21 b 9±0.32 c
Root length
(cm)
1.13±0.12 c 1.04±0.14 c 1.88±0.04 b 1.86±0.1 b 1.88±0.24 b 1.17±0.06 c 1.32±0.19 c 2.49±0.06 a 2.47±0.15 a 2.49±0.33 a 1.49±0.09 b
Shoot length
(cm)
2.41±0.26 b 1.53±0.07 c 1.99±0.07 b 2.28±0.11 b 2.19±0.13 b 1.52±0.1 c 2.28±0.1 b 2.92±0.1 a 3.34±0.16 a 3.21±0.18 a 2.26±0.14 b
‘Gold Capella’
Rooting (%) 0±0 41.75±12.44 b 85.5±6.61 a 100±0 a 100±0 a 100±0 a 50±16.70 b 100±0 a 100±0 a 100±0 a 100±0 a
No. of roots 0±0 0.72±0.14 d 6.99±0.3 c 6.86±0.25 c 9.39±0.46 b 7.14±0.35 c 1±0 d 8.69±0.62 b 8.88±0.26 b 11.83±0.76 a 9.13±0.32 b
Root length
(cm)
0±0 0.75±0.06 f 1.63±0.03 e 1.91±0.1 d 2.09±0.07 cd 1.92±0.02 c 0.90±0.08 f 2.15±0.04 c 2.53±0.14 b 2.78±0.09 a 2.55±0.03 b
Shoot length
(cm)
1.29±0.01 g 1.3±0.03 g 1.66±0.13 d-f ** 1.45±0.07 fg 1.92±0.14 cd 1.52±0.06 e-g 1.77±0.11 de 2.47±0.18 b 2.17±0.09 bc 2.83±0.19 a 2.26±0.08 b
Table 5 - Effects of auxins on rooting and young shoot growth of three schefflera cultivars
Data (±SE) are the mean values of five replicates with three explants in each.
For each cultivar, different lowercase letters indicate significant differences among treatments (LSD multiple range test, P≤0.05).
Fig. 2 - Root induction of schefflera cvs. Luseane (a), Charlotte
(b) and Gold Capella (c) after using 2, 1 and 2 mg l-1 of
NAA, respectively. Note: the roots were submerged in
distilled water for better visibility.
Adv. Hort. Sci., 2018 32(2): 205-212
210
centrations of IBA and NAA did not show significant
discrepancies. Supplementing the medium with 1 mg
l-1 NAA produced the greatest number of roots per
explant with an average of 16.46 roots (Fig. 2). On
the other hand, 0.5 mg l-1 NAA resulted with 15.06
roots which had no significant differences with the
former treatment. Furthermore, using 0.5 and 2 mg
l-1 NAA produced the largest root length of 2.49 cm,
though 1 mg l-1 NAA produced similar root length of
2.47 cm with no significant differences. The maxi-
mum shoot length of 3.34 cm was measured using 1
mg l-1 NAA in the medium, although 0.5, and 2 mg l-1
NAA treatments did not yield significant differences
with the former.
One hundred percent rooting was observed with
‘Gold Capella’ shoots when IBA was applied at 1, 2 or
4 mg l-1, and NAA at 0.5, 1, 2 or 4 mg l-1. The greatest
number of roots per plantlet (i.e. 16.46 roots) was
ob taine d using 2 mg l-1 NAA treatm ent (Fi g. 2).
Moreover, the highest shoot and root length (2.78
and 2.83 cm respectively), were measured by addi-
tion of 2 mg l-1 NAA, which showed significant differ-
ences compared to the other auxin treatments.
Effect of potting media on plant acclimatization
Schefflera’s survival rate was not significantly
affected by different potting media. The highest sur-
vival rate of 100% was obtained for ‘Luseane’ and
‘Charlotte’ using peat moss and sand (1:1) and loam
soil, sand and leaf compost (1:1:1), respectively
(Table 6). ‘Gold Capella’ showed a survival rate of
90% using peat moss or peat moss and perlite medi-
um (1:1).
4. Discussion and Conclusions
Infection of tissue cultures of ornamental pot
plants is a common phenomenon and an important
barrier for their mass production. Contaminations in
Agl a onema ( C h en a n d Yeh, 2 0 07), An t hurium
(Kunisaki, 1980), Dieffenbachia (Brunner et al., 1995),
Spathyphyllum and Syngonium (Kn e ifel and
Leonhardt, 1992), Zantesdeschia (Kritzinger et al.,
1998), as well as Philodendron (Fisse et al., 1987,
Chen et al., 2012) are commonly reported. Thus, the
current research was designed by setting up a dis-
ease-free shoot stock culture. Following surface ster-
ilization with 70% EtOH for 120 sec and disinfection
with 1% sodium hypochlorite for 15 min, more than
93% of the cultures were devoid of visible contami-
nations and they demonstrated 86.4% of viability for
all tested cultivars. These results are consistent with
the findings on Fatsia japonica Decne. (Choi et al.,
2005), Polyscias balfouriana (Ilyas et al., 2013) and
Po lys cia s fru ticosa (Sakr et al ., 2014) from th e
Araliaceae family.
The green schefflera cultivar ‘Luseane’, showed a
significantly greater number of shoot induction (6.25
shoots per explant) compared to the white- and yel-
low-variegated cultivars (Tables 3, 4). This phenome-
non could be ascribed to the fact that variegated
pla n ts have a decr e ased pro p agatio n r ate
(Marcotrigiano, 1997). Vitrification problem and fur-
ther necrosis of the new shoots was observed in
‘Charlotte’ and ‘Gold Capella’ when applying maxi-
mum concentration of 8 mg l-1 BA (Fig. 3). Thus, a
comparable 0.5 mg l-1 TDZ was suggested for in vitro
propagation of these cultivars instead of the former
treatment. Sivanesan et al. (2011) reported increased
number of shoots in Hedera helix ‘Mini’ by using a
combination of 0.5 mg l−1 TDZ and 0.1 mg l−1 NAA. The
mode of action of TDZ may be through alteration in
energy levels, nutrient uptake, nutrient assimilation,
or cell membranes of plants (Murthy et al., 1998).
However, Tetyana and van Staden (2001) demon-
strated that 2.5 mg l-1 BA supplement to the media in
Fig. 3 - Vitrification and necrosis of schefflera plantlets using 8
mg l-1 of BA in ‘Charlotte’ (a) and ‘Gold Capella’ (b).
Mixture Survival (%) Shoots length (cm)
‘Luseane’ ‘Charlotte’ ‘Gold Capella’ ‘Luseane’ ‘Charlotte’ ‘Gold Capella’
Peat moss 90±10 ab 90±10 ab * 90±10 ab 3.86±0.07 b 3.66±0.25 b 3.48±0.18 bc
Peat moss and perlite 90±10 ab 90±10 ab * 90±10 ab 4.81±0.22 a 4.49±0.2 a 4.92±0.17 a
Peat moss and sand 100±0 a 90±10 ab * 80±13.33 ab 3.60±0.1 bc 3.23±0.18 cd 2.94±0.23 d
Loam soil, sand and leaf compost 90±10 ab 100±0 a * 70±15.28 b 3.65±0.08 bc 3.72±0.24 bc 3.79±0.17 b
Table 6 - Effect of potting media on acclimatization of three schefflera cultivars
For each cultivar, different lowercase letters indicate significant differences among treatments (LSD multiple range test, P≤0.05).
Rezaei Baghbidi and Jowkar - Micropropagation of dwarf schefflera
211
Cussonia paniculata of the Araliaceae family, resulted
with the highest shoot induction of 3.5 shoots per
explant. On the other hand, the outcomings of You et
al. (2005) in Eleutherococcus senticosus, the other
member of the Araliaceae family, were different than
our findings. It was revealed that 2 mg l−1 BA prolifer-
ated shoots more effectively than TDZ, Kin and 2iP.
Various cytokinin types and concentrations affect
shoot proliferation of plant species, though all belong
to the same family. Therefore, for each plant species,
the shoot induction has to be newly investigated.
It was revealed that after the TDZ treatment,
shoots’ length of all cultivars were comparatively
longer than those of the Kin and BA treatments. The
white variegated schefflera ‘Charlotte’ showed the
greatest length (2.33 cm) using 0.5 mg l-1 TDZ. This
effect could be related to further increase of auxin
production by TDZ treatment (Murthy et al., 1998). It
is assumed that TDZ has the ability to affect the
amount of interna l plan t horm ones (Murch and
Saxena, 2001). Zaytseva et al. (2016) reported a simi-
lar high activity for low concentrations of TDZ in
Rhododendron.
Al thou gh the h ighe st numb er of leav es was
recorded by using 8 mg l-1 BA in all of the cultivars,
other treatments are suggested due to the afore-
mentioned vitrification problem. For ‘Charlotte’, 0.5
mg l-1 TDZ treatment and for the two other cultivars 4
mg l-1 BA produced maximum leaves without necro-
sis. Generally, cytokinins enhance the photosynthesis
and help translocate the nutrients to the leaves (Taiz
and Zeiger, 2006).
A sought-after plant tissue culture protocol essen-
tially depends on sufficient rooting along with a suc-
cessful acclimatiza tion of the young pl ants. Th e
results of the current study show that in all of the
cultivars NAA is more effective in rooting than IBA.
There are inconsistent reports on the rooting efficacy
of different auxins. The research of You et al. (2005)
indicated that 0.5 mg l−1 NAA is more effective than
IBA for rooting of E. senticosus shoots, while a con-
centration of 0.75 mg l−1 IBA or 1 mg l−1 NAA have
proven useful for C. paniculata (Tetyana and van
St aden , 2001 ). Auxi n trea tme nt of ‘Cha rlo tte’ ,
‘Luseane’ and ‘Gold Capella’ increased the rooting
percentage of the shoots, compared to the control by
35%, 92% and 100%, respectively. All shoots of
‘Charlotte’ and ‘Luseane’ were well acclimatized with
a survival rate of 100% using media of loam soil, sand
and leaf compost (1:1:1) and a mixture of peat moss
and sand (1:1), respectively, while ‘Gold Capella’
demonstrated a none-significant lower survival fre-
quency of 90% in the peat moss or peat moss and
sand media (1:1).
To our knowledge, the current research is the first
report on micropropagation of different cultivars of
schefflera. Results demonstrated that green cultivar
‘Luseane’ had a greater shoot proliferation than the
other two cultivars with average shoot number of
6.25 shoots per explant using 0.5 mg l−1TDZ. The
multiplicated shoots could lengthen on PGR-free MS
medium and later showed effective rooting on NAA-
contained media. Although the yellow variegated cul-
tivar ‘Gold Capella’ revealed a smaller proliferation
rate of shoots compared to the control, its rooting
was 100% improved. Subsequent relocation of the
‘Luseane’ and ‘Charlotte’ plantlets to the acclimatiza-
tion greenhouse resulted with 100% survival rate of
the rooted shoots. Generally, peat moss and perlite
media (1:1) had a more positive effect on the young
acclimatized plants of schefflera. The protocol devel-
oped in this research can be used for the aseptic pro-
duction of schefflera.
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