Plant regeneration as affected by plant growth regulators PGR in mangosteen Garcinia mangostana
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African Journal of Biotechnology Vol. 7 (15), pp. 2693-2701, 4 August, 2008
Available online at http://www.academicjournals.org/AJB
ISSN 1684–5315 © 2008 Academic Journals
Full Length Research Paper
Plant regeneration as affected by plant growth
regulators (PGR) in mangosteen (Garcinia mangostana
L.)
Mohammad Hossein Torabi Sirchi*, M. A. Kadir, M. A. Aziz, A. A. Rashid, Arash Rafat and M.
B. Javadi
Department of Agriculture Technology, Faculty of Agriculture, Universiti Putra Malaysia, 43400 Serdang, Selangor Darul
Ehsan, Malaysia, C/p: +60176066149.
Accepted 20 June, 2008
A protocol was developed for in vitro plantlet regeneration of mangosteen (Garcinia Mangostana L.) from
shoot tip, stem and seed explants. Shoot tip explants produced the highest mean number of shoots per
explant on media supplemented with 1.0 mg/L 6-benzylaminopurin (BAP) and 0.05 mg/L kinetin (KIN)
(73.3). On medium containing 0.1 mg/L BAP and 0.05 mg/L KIN, the highest mean shoot height reached
1.2 cm. The highest percentage of callus formed (90%) was obtained in treatment containing 0.20 mg/L
KIN. The effect of BAP in combination with -naphthalene acetic acid (NAA) was also studied. On half
seed explants, the combination of 4 mg/L (w/v) of BAP with 0.2 mg/L (w/v) of (NAA) produced the highest
number of shoots per explant (75). In medium containing 2 mg/L (w/v) of BAP and 0.2 mg/L (w/v) of NAA,
the mean shoot height reached 1.79 cm also The highest percentage of callus formation (90%) was
obtained on treatment containing 0.5 mg/L (w/v) of NAA. For rooting, pretreatment in different indol-3-
butyric acid (IBA) and NAA concentrations and comparison of MS salt strengths were evaluated. Identical
pattern occurred on medium containing one-quarter strength MS salt and the highest mean number of
roots per explant (1.17) was produced in treatment with 2 mg/L IBA. The highest percentage of root
formation was obtained on one-quarter strength MS salt medium containing 0.1 mg/L NAA (90.4%).
Rooted plantlets were acclimatized on medium composed of soil + sand + organic matters + vermiculite
according to the ratio (2: 2: 1: 1).
Key words: Garcinia Mangostana L., 6-benzylaminopurine, -naphthalene acetic acid, Indole-3-butyric acid,
kinetin.
INTRODUCTION
Mangosteen (Garcinia mangostana Linn.) has been yield of seeds. Usually, it produces a maximum of two
hailed as the “Queen” of tropical fruits with promising seeds per fruit but some fruits have no seed. In addition,
economic value (Samson, 1980). The value of this fruit in the recalcitrant nature of the seeds causes difficulties in
2003 in Malaysia was about 2,447 tones with a value of producing planting material throughout the year.
RM 4.7 million that was mostly exported to Singapore Conventional vegetative propagation methods have been
and Hong Kong. This export value is the best value as commercially unsuccessful. Moreover, development of
compared to the value of other fruits export commodity. mangosteen tree is very slow, taking about 10-12 years
Apomictic seeds usually propagate mangosteen plant, before fruiting (Rehm and Espig, 1991). In vitro culture of
but propagation by seeds is insufficient due to the low mangosteen has been established using seeds (Goh et
al., 1988; Te-chato and Aengyong, 1988). The objectives
of this study are to determine the effect of different con-
centration of BAP either alone or in combination with
*Corresponding author. E-mail: hossein955@yahoo.com, kinetin on shoot multiplication from shoot tip and stem
mbjavadi2002@yahoo.com. Tel: 0389464148. Fax: segments, to determine the effect of different concen-
0389464115. tration of BAP either alone or in combination with NAA on2694 Afr. J. Biotechnol.
shoot induction from seed explants, to study the effect of Acclimatization
different auxins and their concentrations on in vitro
To determine the most suitable potting medium for transfer of the in
rooting of mangosteen, to determine the effect of different vitro rooted mangosteen plantlets to ex vitro conditions, rooted
MS salt strengths supplemented with the best auxin shoots, about ± 3 cm long, from in vitro conditions of medium were
treatment on in vitro rooting of mangosteen and esta- washed with sterile distilled water to remove traces of medium and
blishment of acclimatization protocol for mangosteen. placed in 5% benlat solution. The rooted plantlets were then
transferred into small pots (8 cm diameter and 7 cm high) which
contained (1) sand, (2) soil, (3) perlite, (4) sand : soil : organic
MATERIALS AND METHODS matter : vermiculite (2:2:1:1), (5) sand : soil : organic matters
(3:2:1), (6) sand : perlite (1:1), and (7) sand : soil (1:1). The Potted
plants were covered with perforated Zib blog plastic (10 cm in
Plant material and sterilization protocol
diameter) at 25±1oC, under a 16 h photoperiod with a light intensity
of 15.8-µmol m ²s ¹ and irradiance provided by cool white fluore-
Plant materials for this research which include the shoot tip, stem, scent tubes. These were watered with tap water twice a day. The
and seed explants were derived ex vitro from the mature trees. plastic covers were removed completely after 2 weeks. Each
After removing the outer leaves from shoot tip and stem explants treatment consisted of 25 plants and was repeated once.
and removing seeds from mature fresh fruits, they were immersed
under running tap water with 5% (v/v) Teepol solution. The explants
were next sterilized with 0.5% (v/v) Benlate solution for 15 min and Experimental design and statistical analysis
the following procedure was done in laminar airflow cabinet. The
explants were dipped in 70% ethanol for 3 min and were washed 3 The experiments were arranged in a Completely Randomized
times in sterile distilled water. Subsequently, they were disinfected Design (CRD) with ten replications and each replication per treat-
in 20% Clorox solution with two drops of Tween 20 for 15 min. After ment contained one explant. Data were analyzed using the analysis
that, they were rinsed with sterile distilled water for five times and of variance (ANOVA) and Duncan New Multiple Range Test
dried on a filter paper to minimizing infection. The nutrient medium (DNMRT) at =5% for comparison between treatment means.
used was modified MS (Murashige and Skoog, 1962) medium
supplemented with 30 g/L sucrose and 3.79 g/L gelrite agar.
Various plant growth regulators and additives, namely BAP, Indole-
3-butyric acid (IBA), kinetin (KIN) and NAA were incorporated at RESULTS
different concentrations in the culture media as required. The pH of
the medium was adjusted to 5.7 - 5.8 before autoclaving. The Multiple shoot induction from shoot tips
cultures were kept at 25 ± 2oC in the growth room with a daily fluo-
rescence lighting of 16 h providing an intensity of 15.8 molm-2s-1. It was observed that after eight weeks of culture, shoots
proliferated from the shoot tip (Plate 1A). The multiple
shoots developed further after 12 weeks of culture (Plate
Explant preparation and treatments 1B). Combinations of BAP and KIN tested affected the
After sterilization, the outer leaves were removed and 5 mm long,
mean number of shoots produced per explant, shoot
shoot tip and stem explants were excised. These explants were height and the percentage of explants that responded to
cultured on modified MS medium incorporated with different form callus after 12 weeks of culture. Significant diffe-
concentrations of BAP (0, 0.1, 1.0 and 2.0 mg/L) either alone or in rences were noted among the treatment combinations on
combination with KIN (0, 0.05, 0.1 and 0.2 mg/L). For seed mean number of shoots produced per explant (Figure 1),
explants, the concentrations of BAP (0, 2, 4, and 8 mg/L (w/v)),
mean shoot height (Figure 2) and the percentage of
either alone or in combination with NAA (0, 0.1, 0.2 and 0.5 mg/L
(w/v)) were used. For rooting study, MS medium containing different explants that formed callus (Figure 3). The combination
concentrations of IBA (0.0, 0.1, 1.0 and 2.0 mg/L) and NAA (0, 0.1, of 1 mg/L BAP with 0.05 mg/L KIN produced the highest
0.5 and 1 mg/L (w/v)) were applied. Medium of MS without any mean number of shoots per explant (73.3) and resulted in
growth regulator was considered as a control. a mean shoot height of 0.9 cm (Figures 1 and 2). On
medium containing 0.1 mg/L BAP and 0.05 mg/L KIN, the
mean shoot height reached 1.2 cm but the number of
Parameters observed
shoots produced was low (24.3). Treatments containing 0
Parameters observed in shoot tip and stem cultures were the mean to 0.2 mg/L KIN but without BAP produced less than two
number of shoots produced per explant, the mean shoot height shoots per explant (Figure 1). Increasing the BAP con-
(cm) and the percentage of explants that responded to form callus centration up to 1.0 mg/L resulted in increased number of
(%). Data were collected every two weeks until the twelfth week of shoots produced per explant. However, the number of
culture, while growth characteristics were observed every week. shoots produced per explant decreased when the BAP
Seed culture, the parameters observed were the percentage of
explants producing shoots (%), the mean number of shoots pro-
concentration was increased to 2.0 mg/L. The presence
duced per explant the mean shoot height (cm) and the percentage of KIN at higher concentrations (0.1 and 0.2 mg/L) either
of explants that responded to form callus. In rooting study the alone or in combination with BAP, caused significant
parameters recorded were the percentage of explants producing callus formation at the base of the explants (Figure 3). At
root (%), the mean number of roots per explant and the mean root 0.05 mg/L KIN and less, either alone or in combination
length (cm) attained. The data on rooting were collected until the 6 with BAP there were less or no callus formed. The
months of culture. In the acclimatization study, two parameters that
were recorded for 6 months were the plant height (cm) and the
highest percentage of callus formed (90%) was obtained
percentage of plant survival. in treatment containing 0.20 mg/L KIN without BAP. ThisSirchi et al. 2695
Number of shoots formed per explant
Callus formation (%)
Figure 3. Effect of BAP in combination with KIN on percentage of
callus formation from shoot tips after 12 weeks of culture.
Figure 1. Effect of BAP in combination with KIN on number of shoot
formation after twelve weeks of culture from shoot tips.
Plate 1. Shoot formation from shoot tip of mangosteen. (A) Effect of
1BAP in combination with0.05 KIN (mg/L) on shoot induction after
one week of culture. (B) Effect of 0.1 BAP in combination with 0.05
KIN (mg/L) on Shoot elongation after 4 weeks of culture. (C) Effect
of 0.2 KIN (mg/L) on callus formation on shoot tip explants after 12
weeks of culture.
Figure 2. Effect of BAP in combination with KIN on mean shoot aa
pe rce n tage of s h oots form e d t
bb
height (cm) after twelve weeks of culture from shoot tips.
c d c cd
(%)
was followed by treatment containing 0.20 mg/L KIN with
0.1 mg/L BAP at 60%. e e f
g g g
Shoot induction and callus formation from seed
explants
The seeds segments were cultured on MS medium
fortified with BAP (0, 2, 4, and 8 mg/L (w/v)) either alone
or in combination with NAA (0, 0.1, 0.2 and 0.5 mg/L
(w/v)). A culture medium (MSO) set without any growth
regulator was considered as the control. Statistically, in Figure 4. Effect of BAP in combination with NAA on percentage of
shoot formation per half seed explants after 12 weeks of culture.
the first week through the fifth week of culture, there were
no significant differences among all main effects (BAP
and NAA), The combination of 4 mg/L (w/v) of BAP with
0.2 mg/L (w/v) of NAA produced the highest number of medium containing 2 mg/L (w/v) of BAP and 0.2 mg/L
shoots per explant (75) (Figure 4) (Plate 2A) and resulted (w/v) of NAA, the mean shoot height reached 1.79 cm but
in mean shoot height of 1.3 cm (Figure 5) (Plate 2B). In the number of shoots produced was lower (24.3). Treat-2696 Afr. J. Biotechnol.
aa
number of shoots produced
bc
b
c d cd d
per explant
e e e
f ff f
Figure 7. Effect of different combinations of BAP and NAA
concentrations on percentage of callus formation from half seed
Figure 5. Effect of BAP in combination with NAA on mean number explants after 12 weeks of culture.
of shoots produced per half seed explants after 12 weeks of
culture.
between the height of shoot and the numbers of shoots
produced on a culture medium, the two parameters were
regressed. Figure 7 shows that the greater number of
shoots produced the lower height of shoot.
Shoot proliferation from stems
In general, the number of shoots produced by each stem
explants of mangosteen fast increase in number of
shoots produced was revealed on media with BAP at fifth
to twelfth week of culture. It appeared that both media of
BAP and Kinetin tended to produce more shoots from the
analysis of variance. All main effects between BAP and
kinetin were not significant in the first week through the
fourth week. The result also showed a non-significant
interaction between BAP and kinetin indicating that
kinetin effect did not differ significantly with concen-
trations BAP tested.
Figure 6. Effect of different combinations of BAP and NAA The highest percentage of shoot formation (60.2%)
concentrations on mean shoot height after 12 weeks of culture.
occurred on medium containing 1 mg/L (w/v) of BAP and
0.05 mg/L (w/v) Kin (Figure 9) (Plate 3D). The presence
of BAP in the media either alone or in combination with
ments containing 0 to 0.5 mg/L (w/v) of NAA without BAP Kin stimulated the explants to produce shoots. On me-
produced less than two shoots per explant (Figure 6) dium without BAP, shoots were not formed. The highest
(Plate 3A). The presence of NAA at higher concentrations mean number of shoots produced per explant (9.5) also
(0.2 and 0.5 mg/L (w/v)) either alone or in combination occurred on medium containing 0.1 mg/L (w/v) of BAP
with BAP, caused significant callus formation around the and 0.05 mg/L (w/v) Kin. This medium also produced in
explants (Figure 7) (Plate 3B). With 0.1 mg/L (w/v) of the highest mean shoot height (0.69 cm) followed by
NAA, either alone or in combination with BAP, there were medium containing 1.0 mg/L (w/v) of BAP and 0.05 mg/L
less or no callus formed. The highest percentage of (w/v) Kin (0.66 cm; Figure 8) (Plate 3C). Both treatments
callus formation (90%) was obtained on treatment showed significant difference in shoot height compared to
containing 0.5 mg/L (w/v) of NAA without BAP followed other treatments. In most treatments, the explants also
by treatment containing 0.5 mg/L (w/v) NAA with 2 mg/L produced callus and highest percentage of callus forma-
(w/v) BAP (60%). To clarify whether there is relationship tion (83.36%) occurred on medium containing 0.2 mg/L (w/v)Sirchi et al. 2697
Figure 9. Effect of BAP in combination with KIN on percentage of
stem explants producing shoots after 12 weeks of culture.
Plate 2. Shoot induction from seed of mangosteen. (A) Effect of 4
mg/L (w/v) BAP in combination with 0.2 NAA mg/L (w/v) on shoot
induction after one week of culture. (B) Effect of 4 mg/L (w/v) BAP
in combination with 0.2mg/L (w/v) NAA on shoot multiplication after
four weeks of culture. (C) Effect of 2 mg/L (w/v) of BAP in
combination with 0.2 mg/L (w/v) of NAA on shoot elongation after
four weeks of culture. (D) Effect of 0.5 mg/L (w/v) of NAA on
callus formation after 12 weeks of culture.
Shoot height (cm)
Plate 3. Shoot formation from stem of mangosteen. (A) Effect of
1mg/L (w/v) BAP in combination with 0.05 mg/L (w/v) KIN on shoot
induction (60.2%) (B). Effect of 1mg/L (w/v) BAP in combination
with 0.05mg/L (w/v) KIN on multiple shoot formation (9.5). (C)
Effect of 0.1 mg/L (w/v) BAP in combination with 0.05 mg/L (w/v)
KIN on shoot elongation (0.69cm). (D) Effect of 0.2 mg/L (w/v) KIN
Figure 8. Effect of BAP in combination with KIN on mean shoot on callus formation (83.36%).
height (cm) attained after 12 weeks of culture.
The shoots were excised and then transfer to quarter
Kin (Figure 11). However, callus was not formed in all strength, half strength and full strength of MS (Murashige
treatments without Kin. and Skoog, 1962). Either basal salt solution medium with
0.05% charcoal and 10 g/l sucrose and fortified with
Rooting various auxins, with indol-3-butyric acid (IBA), -
naphthalene acetic acid (NAA), IBA at the concentrations
The parameters observed in this study were percentage of 0, 0.5, 1 and 2 mg/L (w/v) and NAA at concentrations
of explants producing roots (%), mean number of roots of 0, 0.1, 0.5 and 1 mg/L (w/v). Medium of MS without
produced per shoot and mean root length attained (cm). any growth regulator was considered as a control. At the2698 Afr. J. Biotechnol.
DISCUSSION
The production of shoots from auxiliary bud and shoot tip
explant is the most applicable and reliable method of in-
vitro propagation. In this study the different concentration
and combinations of BAP and Kinetin tested affected
shoot development during culture. The presence of BAP
either alone or in combination with Kin in the medium
promoted or enhanced the number of shoots produced
Plate 4. Effect of IBA (mgL ¹) on root formation on mangosteen. (A) from shoot tips excised from in-vitro mangosteen plants.
Effect of 1.0 mgL ¹ (w/v) IBA on percentage of root formation
(63.34%) (Bar=0.33cm). (B) Effect of 1.0 mgL ¹ (w/v) IBA on
BAP may have broken the apical dominance and caused
highest mean root formation (3.43) (Bar= 0.45cm). shoot proliferation, while the presence of kin enhanced
meristem cells to elongate. The combination of 1 mg/L
(w/v) of BAP with 0.05 mg/L of Kin produced the highest
number of shoots per explant. Approximately 74 shoots
first month, the basal ends of explants began to swell and were obtained and reaching a mean height of 0, 85 cm in
callus was formed. From the second month to the Medium containing 0.1 mg/L (w/v) of BAP and 0.05 mg/L
seventh month of subculture, it was revealed that media (w/v) of Kin, the mean shoot height reached 1.18 cm but
of 1 mg/l IBA and 0.5 mg/l IBA gave the highest number the number of shoots produced was 24.3, which was low-
of roots (Plate 4B). These media were significantly wer than the produce in treatment 1.0 mg/L(w/v)
different from the other media at four, five, six, and 7 BAP+0.05 mg/L (w/v) Kin. Meanwhile, the treatments
month period, indicating that these media were the most containing 0 to 0.2 mg/L (w/v) Kin but without BAP
effective in inducing root formation. The maximum concentration up to 1.0 mg/L resulted in increase number
number of root yielded on this media was 3.43 and 2.87 of shoots produced per explant.
for 1 mg/l IBA and 0.5 mg/l IBA, respectively, at the In general, it appeared that both media of BAP and
seventh month of subculture. Based on statistical analy- Kinetin tended to produce more shoots from the analysis
sis there was an interaction between MS salt strength of variance. All main effects between BAP and Kinetin
and IBA concentrations on mean number of roots formed were not significant in the first week through the fourth
per explant. On medium containing full strength MS salt, week. The result also showed a non significant
mean number of roots formed per explant for treatment interaction between BAP and Kinetin, indicating that
without IBA (3.8) was significantly higher in comparison Kinetin effect did not differ significantly with concen-
to 1 mg/L IBA (3.43), 0.5 mg/L IBA (2.87) and 2 mg/L trations BAP tested. Referring to (Figure 9), 60.24% of
(1.23). There were significant differences in rooting stem explants produced shoots on medium containing
between treatment 1, 0.5 and 2 mg/L IBA (Table 1). 1.0 mg/L (w/v) of BAP and o.o5 mg/L (w/v) of Kin. The
Identical pattern occurred on medium containing one- highest number of shoots per explant (9.5) was obtained
quarter strength MS salt (Table 2); increase in IBA on the same medium (Figure 10). This indicated that 1.0
concentration in the culture medium increased the mean mg/L (w/v) of BAP and 0.05 mg/L (w/v) of Kin in the
number of roots formed per explant. The highest mean medium was the most suitable concentration to induce
number of roots formed per explant was produced in direct shoot regeneration from stem explant of mangos-
treatment with 2 mg/L IBA (1.17) (Table 1), while the teen. Whereas, all the media without BAP did not result in
lowest was produced in treatment without IBA (0.13). shoot formation (Figure 9). This phenomenon indicated
that BAP is important for shoot regeneration, medium
Root length and comparison between IBA and NAA containing 0.1 mg/L (w/v) BAP and 0.05 mg/L (w/v) Kin
on rooting produced the highest shoot height. However, it did not
differ significantly to treatment 1.0 mg/L (w/v) BAP and
0.05 mg/L (w/v) Kin, the ratio between BAP and Kin has
The maximum mean length of root was 7 cm and was
an effect on shoot height of mangosteen. Whereby an
obtained on medium of 1 mg/l IBA followed by 6.04 cm
increase in the concentration resulted in reduction of
on 0.1 mg/l NAA. Therefore, from the overall media
shoot length. Meanwhile treatments containing BAP
tested, medium containing 1 mg/l IBA was the best
without the presence of Kin produced shorter shoots
medium for root elongation particularly when compared to
compared to media containing Kin. This indicates that the
medium containing 1 mg/l NAA (4.57) or 0.5 mg/l NAA
Kin plays a role in rejuvenating the cell activity and shoot
(5.40) (Plate 4A). The highest percentage of root
elongation. The treatment containing 0.05 mg/L (w/v) of
formation was obtained on medium containing 0.1 mg/L
Kin in combination with 0.1-1.0 mg/L (w/v) of BAP in the
NAA (90.4%). Increasing levels of NAA decreased the
medium is suggested for shoot elongation.
percentage of root formation. There was no significant
In this study BAP was the only Plant Growth Regulator
difference on percentage of root formation between
(PGR) influencing shoots formation of seed explants in
control and 1 mg/L IBA.Sirchi et al. 2699
Table 1. Root length formation (cm) on different MS salt strength and IBA concentrations.
IBA concentration (mg/L) Full strength Half strength One-quarter strength
0 3.8±0.35a 2.27±0.25a 0.13±0.05d
0.5 2.87 ±0.12c 1.17 ±0.29c 0.47± 0.25c
1 3.43 ±0.40b 1.67±0.15b 0.90±0.10b
2 1.23±0.25b 0.57±0.11d 1.17±0.29a
CV (%) 8.55 17.5 36.27
Means with the same letter are not significantly different at 0.05 probability level according to Duncan New Multiple Range Test
(DNMRT) test.
Table 2. Percentage of root formation (%) on different MS salt strength and IBA Concentrations.
IBA concentration (mg/L) Full strength Half strength One-quarter strength
0 87.0±4.69a 56.7±3.33a 13.0 ±4.69c
0.5 26.5 ±3.84d 16.03±4.69d 23.2± 3.84ab
1 63.4 ±3.48b 53.4±3.33ab 26.5±3.84a
2 33.2±3.48c 23.2±3.84c 26.5±3.84a
CV (%) 6.25 10.67 27.35
Means with the same letter are not significantly different at 0.05 probability level according to Duncan New Multiple Range Test
(DNMRT) test.
Callus formation (%)
Mean number of shoots
h h h
Figure 10. Effect of BAP in combination with KIN on mean
number of shoots produced per stem explants after 12 weeks Figure 11. Effect of BAP in combination with KIN on
of culture. percentage of stem that responded to form callus (%) after 12
weeks of culture.
mangosteen while NAA Was only as a PGR supplement
improving the effect of BAP. There is a negative rela- et al. (2000). The combination of 4 mg/L (w/v) of BAP
tionship between the number produced and the high of with 0.2mg/L (w/v) of NAA produced the highest number
shoot. This result confirmed the study conducted that the of shoots per explant (75) and resulted in mean shoot
presence of BAP in media considerably enhanced the height of 1.3 cm. In medium containing 2 mg/L (w/v) of
level of caulogenic determination as shown by the BAP and 0.2 mg/L (w/v) of NAA, the mean shoot height
increased percentage of explants producing shoot buds reached 1.79 cm but the number of shoots produced was
as well as the number of shoots formed in half tissues of lower (24.3). Treatments containing 0 to 0.5 mg/L (w/v) of
mangosteen cultured in vitro. BAP is the most effective NAA without BAP produced less than two shoots per
when added singly in the medium for in vitro culture of explant. Among the concentrations of BAP in the basic
G.mangostana explants, which had been confirmed by MS medium 4 mg/L (w/v) BAP and 0.5 mg/L NAA con-
Lakshmanan et al. (1997), Normah et al. (1992), Huang sistently produced. The highest mean number of shoots2700 Afr. J. Biotechnol.
per explant and plant height
Number of leaves developed 8
7
6
5
4
3
2
1
0
sa so per sa+so+or+v sa+so+or sand+per sa+so
Figure 12. Effect of growth media on ex-vitro performance on number of leaves developed per explant and plant
height (cm). Sa = Sand, so = soil, per = perlite, org = organic matters, ver = vermiculite.
Table 3. Comparison between IBA and NAA on rooting.
Mean number of roots Percentage of root Root length Intensity of
Treatment formed per explant formation (%) (cm) callus formation
Control 4.23±0.25c 73.8±3.84b 5.57b -
1mg/L IBA 3.57±0.36cd 66.7±12.51bc 7.00a +
0.1 mg/l NAA 6.40±0.81b 90.04±12.11a 6.04ab +
0.5 mg/l NAA 7.87±0.53a 76.8±13.61b 5.40c ++
1 mg/L NAA 6.60±0.40b 43.3±15.94c 4.57d +++
Means with the same letter are not significantly different at 0.05 probability level according to Duncan New Multiple Range Test
(DNMRT) test.
- = Very slight callus, + = slight callus, ++ = moderate callus, +++ = heavy callus.
produced per half seeds (9.5) also occurred on medium roots per explant and percentage of root formation.
containing 20 mg/L (w/v) BAP and 10 mg/L (w/v) NAA. Medium containing one-quarter strength MS salt did not
NAA did not influence significantly on the shoot production. provide adequate nitrition for root induction and therefore,
Is a crucial and difficult stage in micropropagation of addition of IBA was needed (Arteca, 1996). In the experi-
mangosteen, especially to get roots with good charac- ment on “Comparison of IBA and NAA on Rooting” higher
teristics? The quality of roots produced plays an levels of IBA and NAA (>0.5 mg/L) induced moderate and
important role for successful transfer of plantlet to the soil heavy callus, while treatment at 0.1 mg/L NAA and 1
during acclimatization. The presence of auxin in rooting mg/L IBA induced slight callus. Callus formation should
medium is sometimes needed for root initiation. Also in be avoided during rooting, because if root develop from
absence of IBA and NAA and followed by the culture on callus–like areas the shoot, vascular connections are
basal medium (MSO) no root were formed. In the weak and can cause high mortality of plantlets during
experiment on “Effect of MS Salt Strength and IBA acclimatization (Goncalves et al., 1988).
concentrations on Rooting” on medium containing full, The highest percentage of plantlets survival was
half and quarter strength MS salts, the highest mean obtained in plants grown in sand+ soil+ organic maters
number of roots formed per explant and percentage of and vermiculite (Figure 12). This response may be due to
root formation were obtained on treatment without IBA. In the ability of the media to provide enough moisture for the
contrast, on medium containing one-quarter strength MS plants and for good root growth. In some of pots, the
salts treatment with 2 mg/L IBA produced the highest number of leaves produced per plant and plant height
mean number of roots formed per explant (Table 3) and were low which could be due to the low nutrient content
percentage of root formation. These response could be of perlite and sand. The compost containing a high
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