American Journal of Plant Sciences, 2011, 2, 467-475
doi:10.4236/ajps.2011.23055 Published Online September 2011 (http://www.SciRP.org/journal/ajps)
467
Conversion of Banana Synseed Influenced by the
Bead Type and Seed Coat
Ahmed Hassanein1, Ibrahim A. Ibraheim2, Abel-Naser Galal1, Jehan Salem1
1Central Laboratory of Genetic Engineering, Faculty of Science, Sohag University, Sohag, Egypt; 2Genetic Engineering and Biotech-
nical Laboratory, Sadat City, Monofia University, Monofia, Egypt.
Received April 26th, 2011; revised June 10th, 2011; accepted June 23rd, 2011.
ABSTRACT
Banana shoot tips extracted fro m in vitro grown plant materials were encapsu lated in different types of alginate beads.
The results obtained after conservation for one month at 4˚C indicated that single layered synseeds (where the encap-
sulated shoot tip was co vered by one la yer o f the artificia l en dosp erm) containing activated charcoal were the best con-
struction in stimulating conversion of synseeds, where they expressed the highest; conversion frequency, number of
shoots/synseed, length of shoot and fresh weight/shoot cluster. These parameters decreased when the shoot tips were
encapsulated within two layers of endosperm, except the ex vitro conversion. Conversion of banana synseeds was in-
fluenced by explant orientation inside the alginate beads, the highest conversion frequency was obtained when the shoot
tip was directed upward inside the synthetic endosperm. Synthesis of seed coats around the artificial endosperm im-
proved the conversion frequency, number of shoots/synseed and shoot length.
Keywords: Artificial Seeds, Banana, Conservation, Synsee d C onversion, Tissue Culture
1. Introduction
Edible and cultivated banana cultivars are highly sterile,
triploid and producing seedless fruits through partheno-
cary. These features prevent the implementation of breed-
ing strategies to improve banana cultivars. The materials
used for conventional propagation include: corms, large
and small suckers, sward suckers [1,2], transfer weevils,
fungal pathogens, nematodes and viruses [2,3]. Com-
pared to suckers, the use of plantlets obtained via tissue
culture techniques (TC plantlets) express many addi-
tional advantages [4], where TC plantlets are cheaper and
easier to propagate and transport. The uniformity of
growth of in vitro obtained plants makes it possible to
control the flowering time and harvesting. Also, they
express higher survival rate in the field, and give a sig-
nificant increase in yield and fruit quality [5,6], but su-
maclonal variation should be avoided [7,8].
In fact, micropropagated plantlet production needs
high investment [9] because the final products require
specific and expensive management until commercializa-
tion and final delivery [10,11]. Development of synthetic
seed production technology is currently considered an
effective and efficient alternative method of propagation
in several commercial important agronomic and hor-
ticultured crops [12,13]. Synthetic seeds can be also
called artificial seeds or synseeds. Somatic embryogene-
sis is considered the most appropriate technique for
rapid-large scale micropropagation and synseed technol-
ogy [14-16], but somatic embryogenesis technique is not
well established in many plant species. In such cases, the
encapsulation of shoot tips and/or axillary buds provides
an alternative plant material to produce synseeds [13,17].
Only few reports described successful encapsulation of
non-embryogenic in vitro-derived vegetative organs such
as maxillary buds or shoot tips [13,18].
Synseed consists of somatic embryo or alternative
plant materials encapsulated within a synthetic endos-
perm which containing nutrient reserves and other addi-
tives [15,19-21]. These structures could also be encapsu-
lated further with a synseed coats to suit mechanical
handling and planting [22]. These synseeds can be stored
for a longer period of time even up to 6 months without
losing viability, especially when stored at 4˚C. In addi-
tion, to prevent embryo desiccation and mechanical in-
jury, a number of useful materials such as fungicides,
pesticides, antibiotics and microorganisms (e.g., rhizobia)
can be included into the encapsulation matrix. Incorpora-
tion of activated charcoal improves the conversion and
the vigor of encapsulated somatic embryos.

Conversion of Banana Synseed Influenced by the Bead Type and Seed Coat
468
Conversion ability of synseeds would be very neces-
sary when ex vitro sowing is used [23]. In some species,
such as banana, cardamom, mulberry and raspberry, en-
capsulated microcuttings demonstrated high adventitious
rooting capacity after sowing [14,24,25], while other
species did not respond [23,26]. There are many factors
affect the conversion of synseeds after cold storage pe-
riod. While, some factors need further studies, some oth-
ers have not yet been studied, such as orientation of ex-
plants inside the artificial endosperm. Therefore, the aim
of this work was to study factors affecting conversion of
synseeds after cold storage on agar-solidified medium.
2. Material and Methods
2.1. Explant Preparation and Culture Initiation
Field grown plants (1.5 m - 2.5 m high) of Hindi cultivar
were obtained from Sohag Governorate and used as pri-
mary explants. The upper part of their shoots were cut
back to approximately 30 cm; roots and dry leaf sheaths
were removed and transferred to the laboratory. Succes-
sive leaf bases of corms were cut away and the shoot tips
were excised by making four angled cuts into the sub-
tending tissue. The explants (60 mm × 20 mm cylinder)
were surface sterilized in 5.25% NaOCl solution (80%
v/v commercial Clorox bleach) for 20 min followed by 5
min dip in 0.2% (w/v) mercuric chloride (HgCl2), and
rinsed three times in sterile distilled water. After disin-
fection, the excessive tissues were removed to get shoot
tip explants of approximately 7 mm × 6 mm, including a
basal corm tissue. These shoot tips were cut one time
vertically in such a way to keep the base of the explants
intact, and cultured on solid multiplication medium (MS
medium supplemented with 5 mg/l BAP). The vegetative
apices were transferred to fresh medium three times at
two weeks intervals to avoid phenolic accumulation sur-
rounding the cultured explants. When shoot multiplica-
tion began to form and continued growth warranted, they
were subdivided into single shoots and subcultured on
fresh multiplication media.
In vitro shoot tips (4 mm - 5 mm height and 2 mm - 3
mm at base) of banana cultivars (Hindi) derived from the
initial stage were monthly subcultured on the multiplica-
tion medium supplemented with 5 mg/l BAP and 200
mg/l ascorbic acid (ASA), or 0.2% activated charcoal as
antioxidants. Ascorbic acid was added to the medium
after autoclaving. In all experiments, MS [27] medium
supplemented with 3% sucrose was used. Media were
solidified with 4 gm/l agar (Difco Bacto agar) at pH 5.8
before autoclaving. Vitamins (mg/l) were: myo-inositol
(100), vitamin B1-hydrochloride (4), nicotinic acid (4),
pyridoxal hydrochloride (0.7), biotin (0.04) and folic acid
(0.5). Media were autoclaved at 121˚C for 20 minutes. In
vitro cultured plant materials were incubated under light
condition of a tissue culture room (25˚C ± 2˚C with 16 h
photoperiod at 100 μmol·m–2s–1).
2.2. Synseed Formation
2.2.1. Single Layered Beads
Shoot multiplication medium with or without 2 gm/l ac-
tivated charcoal was used as artificial endosperm com-
ponent. Sodium alginate (4%) and CaCl2 (75 mM) solu-
tion were used as gel matrix and complexing agent, re-
spectively. Both the gel matrix and complexing agent
were autoclaved at 120˚C for 20 min and stored at room
temperature. Single layered beads were obtained by dip-
ping the shoot tips in glass jar or Petri dish containing the
gel matrix. Then, alginate-covered explants were picked
up individually and dropped into the complexing agent
for 30 minutes. Transfer time from gel matrix to com-
plexing agent must not exceed five minutes. After hard-
ening, the calcium chloride solution was decanted and
the constructed alginate beads were rinsed three times
with autoclaved liquid multiplication medium for 15 min,
5 min/each, to wash away calcium chloride residues. The
obtained beads (synseeds) were 5 mm - 6 mm (Figure 1)
in diameter and could be easily handled under sterile
conditions without dehydration of the explant.
2.2.2. Double Layered Beads with Explant in Bead
Center
Single layered synseeds (with or without 2 gm/l activated
charcoal) were completely immersed in sodium alginate
medium (with or without 2 gm/l activated charcoal).
They were picked up and dropped into the complexing
agent (CaCl2 solution) again. The explants became at the
center and covered with two layers of artificial en-
dosperm. After 30 min, calcium chloride was decanted
and the beads were rinsed 3 times with autoclaved liquid
multiplication medium for 15 min, 5 min/each, to wash
away calcium chloride residues. The obtained beads were
7 mm - 8 mm in diameter (Figure 2).
6 m
m
Figure 1. Single layered synseed.
8 mm
Figure 2. Double layered synseed with explant in the center
of two layers of artificial endosperm.

Conversion of Banana Synseed Influenced by the Bead Type and Seed Coat469
2.2.3. Douple Layered Beads with Lateral Explant
These type of beads were constructed by the method de-
scribed by Kinoshita and Saito [36], where shoot tip ex-
plants and single layered beads (with or without 0.2%
(w/v) activated charcoal) were completely immersed in
sodium alginate medium with or without 0.2% (w/v) ac-
tivated charcoal. Then, they were together picked up and
dropped into the complexing agent. The explant was in-
cluded outside the inner layer but covered with the outer
layer only. After 30 min, calcium chloride solution was
decanted and the beads were rinsed 3 times in sterilized
multiplication medium to wash away CaCl2 residues. The
obtained beads were about 9 mm in diameter (Figure 3).
2.3. Conversion of Synseeds Influenced by Bead
Type
Shoot tip explants of Hindi cultivar encapsulated into ten
types of synseeds were tested to determine the effect of
bead type on synseed conversion. Single layered type
synseed was with or without activated charcoal (2 gm/l).
Two groups of double-layered synseeds were used: the
first group with explant in the center of bead (Figure 2)
and covered with two layers of artificial endosperm, and
the second group with explants included outside the inner
layer but covered only by the outer layer (Figure 3). In
double-layered type, both two layers were with or with-
out activated charcoal or one of them was supplemented
with activated charcoal. All types of artificial seeds were
conserved for one month as submerged beads in liquid
multiplication medium at 4˚C in refrigerator. The syn-
seeds were converted on semisolid MS medium supple-
mented with 5 mg/l BAP (conversion medium). A group
of thirty synseeds of each type was cultured in six glass
jar, five synseeds/each, and considered as a replicate for
each type. After four weeks, percentage of synseed con-
version, number of shoots per synseed, length of shoot
and fresh weight per shoot cluster were determined.
2.4. Effect of Explants Orientation on the
Conversion of Synseeds
In this experiment, one layer synseeds without charcoal
were sued. A set of 90 encapsulated shoot tips was made
and classified into three groups. Each group was cultured
in six glass jars containing conversion medium. The first
group was cultured where the orientation of shoot tip
inside the beads was upward. The second group was
9 mm
Figure 3. Double layered synseed with explants outside the
inner layer but inside the outer layer.
cultured where the shoot tip inside the bead was hori-
zontally oriented. The third group was randomly cultured.
After preservation for one month at 4˚C and incubation
for more four weeks on conversion medium, percentage
of converted synseeds, number of shoots per synseed,
length of shoot and fresh weight per cluster were deter-
mined. After 4 days, peroxidase activity of explant inside
the beads under conversion conditions was determined.
2.5. Formation of Synseed Coat
For synseed coating, the protocol proposed by Levy and
Edwards-Levy [28] was used. The protocol consisted of
the passage of shoot tip explants in artificial endosperm,
which was prepared as previously described. The artifi-
cial endosperm solution was supplemented with 2.5%
sodium alginate and after autoclaving, enriched with 2%
PGA (Propylene glycole alginate, Kelcoloid S®, Kelco
International, 80% - 85% esterification grade) and 5 ml/l
of a patented antimicrobic PPM (Plant Preservative
Mixture, Plant Cell Tech. Laboratories, USA). The solu-
tion was stirred for 24 h at room temperature to dissolve
PGA. The complexing solution was made of the artificial
endosperm solution with CaCl2 (75 mM). The coating
solution was made of 50 ml of artificial endosperm solu-
tion supplemented after autoclaving with 5 gm egg al-
bumin (10% w/v final concentration). Encapsulation was
made by immersing the explant with forceps in the first
alginate-PGA solution and then dropping it in the stirred
CaCl2 complexing solution. The encapsulated explants
were kept stirred for 25 minutes and then transferred to a
first rinse solution made of the simple artificial endosperm
solution. After rinsing, the capsules were dropped in the
coating solution, where they were left stirred for 5 min,
in order to allow penetration of the protein into the outer
layer of the capsules. At this point, 2 ml of NaOH (1 M)
solution were added to the stirring solution with the cap-
sules to start a transacylation reaction. The capsules were
continuously stirred for a further 15 min, then the solu-
tion was brought to pH 5.5 with 2.5 ml HCl (1 M), and
stirred for another 15 min. After this, the capsules were
finally rinsed in the artificial endosperm solution. In this
experiment, all artificial endosperm solutions were made
of conversion medium, and supplemented with 5 ml/l
PPM®. Forty coated synseeds were cultured in eight jars
(100 ml) contained semisolid multiplication medium 5
synseeds each After four weeks culture, percentage of
synseed conversion, number of shoots per synseed,
length of shoot and fresh weight per shoot cluster were
determined.
2.6. Determination of Peroxidase Activity
Estimation of relative peroxidase activity was measured
(O.D./gm fresh weight/h) and calculated according to

Conversion of Banana Synseed Influenced by the Bead Type and Seed Coat
470
Wakamatsu and Takahama [29]. For peroxidase analysis,
1 gm of tissue was ground at 4˚C in a mortar in 1 ml.
extraction buffer consisting of 0.1 M Tris-base pH 7.0
and containing 0.002 M cysteine. The homogenate was
centrifuged at 14000 rpm for 15 min. The supernatant
were collected for immediate peroxidase activity deter-
mination. The reaction mixture consisted of 5 mM guai-
col, 40 mM potassium phosphate buffer, pH 7.2, 0.1 mM
EDTA, 0.3 mM H2O2 and enzyme preparation (50 µl of
each soluble peroxidase) in a final volume of 5 ml. The
reaction was measured by the absorbance at 470 nm at
the room temperature.
3. Results and Discussion
Sucker shoot tips were used for in vitro multiplication of
banana on MS medium supplemented with 5 mg/l BAP.
In order to save large number of plant materials to pro-
duce synseeds in large scale, six subcultures of in vitro
obtained shoot tips were established. Banana shoot tips
extracted from in vitro grown plant materials were big
enough to facilitate the encapsulation procedure; conse-
quently, great number of synseed could be formed in one
hour (180 synseeds). The obtained capsules of synseeds
were spherical and hard enough to be handled easily.
Two types of synseeds, single layered and double layered
synseeds were made. Single layered synseed was 5 mm -
6 mm in diameter (Figure 1), however double layered
one was 8 mm - 9 mm in diameter (Figures 2 and 3).
Preliminary experiments showed that solution of 4%
sodium alginate with 75 mM CaCl2 produced firm and
clear uniform capsules within ion exchange duration in
30 min. On the other side, higher concentrations of so-
dium alginate (5%) were not suitable for synseed forma-
tion because the resulted beads were too hard to cause
considerable delay in conversion and reduce the conver-
sion frequency. Beads obtained from lower concentra-
tions of sodium alginate (3%) were too fragile to be han-
dled. Ghosh and Sen [30] found that the use of high or
low level of sodium alginate reduced the conversion fre-
quency.
Conversion of banana synseeds influenced by bead
type was studied by constructing several types of single
and double layered synseeds with or without activated
charcoal (Figures 4 and 5). They were cultured on con-
version medium in order to determine the best synseed
type. Induction of synseed conversion from encapsulated
shoot tip explants was commenced after 3 days when
explants were encapsulated in single layered synseeds
with 0.2% activated charcoal. In case of single layered
synseeds without charcoal and double layered synseeds
(with explant covered with the outer layer and the inner
layer with charcoal), the conversion of synseeds com-
menced in 4 days. In double layered synseeds with the
explant in the outer layer and both layers either with or
without charcoal, or only the outer layer with charcoal,
the conversion of synseeds commenced in 5 days. On the
other side, conversion of all types of double layered syn-
seeds with explant in the center (covered with 2 en-
dosperm layers) was delayed for 5 more days, where
conversion commenced in 10 days.
(a) (b) (c)
(d) (e) (f)
Figure 4. Different ty pes of banana (Hnidi cultivar) synseeds with different types of artificial endosperm. (a) Single layer en-
dosperm. (b) Single layer endosperm with charcoal. (c)-(f) Double layer synseed, explants were in the centers of the beads: (c)
double layer endosperm, (d) double layer endosperm with charcoal, (e) double layer endosperm and the inner layer with
charcoal, and (f) double layer endosperm and the outer layer with charcoal.

Conversion of Banana Synseed Influenced by the Bead Type and Seed Coat471
(a) (b)
(c) (d)
Figure 5. Double layer synse eds of banana (Hnidi cultivar) but the explant was coated with only one layer of endosperm, (a)
without charcoal in both layers, (b) with charcoal in both layers, (c) with charcoal in the inner layer, and (d) with charcoal in
the outer layer.
The results obtained after four weeks incubation on
conversion medium indicated that the conversion of syn-
seeds was affected by the number of endosperm layers,
activated charcoal and the explant position inside the gel
matrix. Single layered synseed with activated charcoal
was the best construction in stimulating conversion of
synseeds (Figure 6), where it expressed the highest per-
centage of synseed conversion, number of shoots, length
of shoot and fresh weight/shoot cluster (Table s 1 and 2).
Double layered synseeds with the explant in outer en-
dosperm layer were better than those with the explant in
the center and coated with two endosperm layers. The
most effective type of double layered synseeds were
those with explant in the outer layer and charcoal in both
two layers (Table 2), where they expressed high per-
centage of synseed conversion, high shoot number, high
shoot length and high fresh weight/shoot cluster.
Encapsulation of banana shoot tips in single layer was
better than encapsulation in double layered synseeds.
However, double layered synseeds with the explant in the
outer endosperm layer with charcoal in both two layers
were the most effective type where it expressed high
conversion frequency especially under ex vitro condition
(data not shown). This may be due to the ease of penetra-
tion and emergence of new shoots from one layer than
from two layers. Micheli et al. [31] reported that sprout-
ing of double layered synseeds was lower than that of
single layered ones. Maruyama et al. [23] found that dou-
ble layered synseeds were much better than single lay-
ered synseeds of Cidrela odorata L., Guazuma crinita
Mart., and Jacaranda mimosaefolia D. Don.
Figure 6. Conversion of banana single layer synseeds with
charcoal. Synseeds were cultured on conversion medium for
four weeks.

Conversion of Banana Synseed Influenced by the Bead Type and Seed Coat
472
Table 1. Conversion of encapsulated shoot-tip explants on conversion medium under the influence of explant position, acti-
vated charcoal, and number of endosperm layers. In case of double layered synseeds, the explant was encapsulated within
two endosperm layers. Values are mean ± SD.
Structure of synseed Percentage of synseed
conversion (%) No. of shoots / synseed Length of shoot (cm) F.wt. (gm)
Single layer
Single layer with charcoal
Double layers
Double layers with charcoal
Double layers and the inner one with chacoal
Double layers and the outer one with charcoal
82.57
98.34
75.00
52.17
31.81
32.00
1.00 ± 0.00
2.17 ± 0.38
1.50 ± 0.54
1.50 ± 0.54
1.0 ± 0.00
1.00 ± 0.00
3.85 ± 0.17
5.33 ± 0.76
2.27 ± 0.25
3.07 ± 0.11
0.98 ± 0.08
2.43 ± 0.40
0.68 ± 0.09
0.87 ± 0.04
0.38 ± 0.04
0.28 ± 0.08
0.12 ± 0.05
0.11 ± 0.01
Table 2. Conversion of encapsulated shoot-tips on conversion medium under the influence of position of explants, activated
charcoal, and number of endosperm layers. In case of double layered synseeds, the explant was encapsulated within one en-
dosperm layer. Values are mean ± SD.
Structure of synseed Percentage of synseed
conversion (%) No. of shoots / synseed Length of shoot (cm) F.wt. (gm)
Single layer
Single layer with charcoal
Double layers
Double layers with charcoal
Double layers and the inner one with chacoal
Double layers and the outer one with chacoal
82.57
98.34
79.16
90.00
80.00
72.00
1.00 ± 0.00
2.17 ± 0.38
1.00 ± 0.00
1.50 ± 0.54
1.33 ± 0.48
1.00 ± 0.00
3.85 ± 0.17
5.33 ± 0.76
3.08 ± 0.15
3.43 ± 0.40
2.75 ± 0.20
2.95 ± 0.19
0.68 ± 0.09
0.87 ± 0.04
0.36 ± 0.05
0.66 ± 0.05
0.26 ± 0.04
0.25 ± 0.04
The data in this work indicated that addition of acti-
vated charcoal into the gel matrix was beneficial because
it improved the conversion as well as the number of
shoots/synseed and shoot fresh weight. Vigorous growth
was detected when the synthetic endosperm contained
activated charcoal, where the shoot length, number of
roots and root length were higher than those of synseeds
without activated charcoal in synthetic endosperm. The
beneficial effect of activated charcoal on synseeds con-
version is attributed to their ability to absorb undesirable
exudates, such as 5-hydroxymethylfurfural (a toxic break-
down product of sucrose formed during autoclaving) and
other harmful phenolic oxidation products [23]. Also, it
has been suggested that charcoal breaks up the alginate
bead and thus increases respiration of somatic embryos,
as well as it retains nutrients within the hydrogel capsule
and slowly releases them to the growing embryo [32].
Furthermore, the addition of activated charcoal to the
alginate beads matrix significantly enhanced root devel-
opment and germination of encapsulated somatic em-
bryos [33]. Our results showed that single layered syn-
seed with 0.2% activated charcoal was the most effective
synseed type for banana plants.
The previous reports indicated that conversion of syn-
seeds depends on several factors where the conversion
took place under in vitro or ex vitro conditions [6,13,20].
In this work, conversion of synseeds on a sterilized mix-
ture of peat and sand (2:1 v/v), and incubated under tis-
sue culture conditions indicated that douple layered
beads (especially where the explants covered only with
one layer of artificial endosperm) were better than one
layered beads (data not shown) but still it needs further
studies to improve the ex vitro conversion of banana
synseeds.
The effect of orientation of encapsulated shoot tip ex-
plants of Hindi cultivar on synseed conversion was
shown in Table 3. Shoot tip explants were encapsulated
in gel matrix without charcoal to determine the orienta-
tion of explant on medium. The highest conversion fre-
quency was obtained when the direction of the shoot tip
explant inside synthetic endosperm was upward and its
base was in contact with the medium surface. It was ac-
companied by high increase in peroxidase activity. The
lowest conversion frequency was obtained when the

Conversion of Banana Synseed Influenced by the Bead Type and Seed Coat473
shoot tip explants inside the synthetic endosperm was in
horizontal position on conversion medium. The polarity
of auxin transport is basipetal, i.e., auxin moves from
apex to base region. When the encapsulated explants
were placed in upward position on conversion medium,
auxins transported from the apex of explant to its base,
and in the same time the explant absorbed the cytokinin
from the conversion medium. Consequently, right cyto-
kinin/auxin ratio was established to induce cell division
and led to conversion of encapsulated explants. So, when
the encapsulated explant was in horizontal position, the
portion of auxins that was flowed towards lower side of
the explant under the influence of gravity and diffused to
the medium [34]. In this case, the remained auxin in the
explant became low than that needed to establish the
right auxin/cytokinin ratio resulting in repressing the
synseed conversion. In random culture, some of the en-
capsulated explants may be in upward direction and oth-
ers may be in horizontal direction or in between, thus
their conversion frequency was better than horizontal
culture but lower than upward culture.
Single layered synseeds were subjected for seed coat-
ing procedure. Synthesis of seed coats did not change the
size of beads or their appearance as well as their consis-
tence. Consequently, it was difficult to distinguish be-
tween synseeds with or without seed coats (Figure 7).
Charcoal was not included in single layered synseed
when seed coats were synthesized to cover the artificial
endosperm. Culture of synseeds with or without coat on
conversion medium for four weeks indicated that there
was a big difference between the data of uncoated and
coated synseeds. Coated synseeds expressed higher con-
version frequency, shoot length and fresh weight of shoot
cluster than those obtained by uncoated ones (Table 4).
It’s worthy to mention that synthesis of seed coat stimu-
late not only shoot but also root formation (Figure 8) and
the resulted plantlets transferred to ex vitro conditions
with limited success.
Table 3. Effect of orientation of encapsulated explants cultured on conversion for four weeks on conversion and peroxidase
activity of encapsulated explants. Values are mean ± SD.
Orientation Conversion frequency
(%) No. of shoots/synseed Length of shoo t (cm) F.wt. (gm) POX activity (%)
Random
Upward
Horizontal
51.42
67.65
42.85
1.82 ± 0.79
3.50 ± 0.58
1.25 ± 0.5
3.91 ± 0.39
4.26 ± 0.25
3.73 ± 0.05
0.48 ± 0.07
0.65 ± 0.04
0.46 ± 0.08
100
348.12 ± 11.69
92.11 ±6.91
Figure 7. Conversion of coated single layer synseeds of Hindi
cultivar cultured for four weeks on conversi on med ium.
Figure 8. Conversion of coated synseeds in Petri dish at
5˚C. 2

Conversion of Banana Synseed Influenced by the Bead Type and Seed Coat
474
Table 4. Effect of coating on the conversion of synseeds of banana. Synseeds were cultured for four weeks on conversion me-
dium. Values are mean ± SD.
Type of synseed Conversion frequency (%) Shoot no./synseed Shoot length (cm) F.wt. (gm)
Synseed without coat
Synseed with seed coat
81.81
96.77
1.25 ± 0.50
4.00 ± 0.89
3.00 ± 0.39
2.03 ± 0.11
0.37 ± 0.05
0.65 ± 0.19
Hydrophobic thin coat over a gel surface plays an im-
portant role of preventing the rupture and rapid desicca-
tion of synseeds [35]. It also expected that coat acts as a
shield against microbial and chemical contamination of
synseeds. Consequently, coating of the encapsulated ba-
nana shoot tips was established to determine the advan-
tages of coating on banana synseed conversion. Our data
indicated that, the conversion frequency, number of shoots/
synseed and shoot length increased when the gel matrix
was coated. This indicated that the used method for for-
mation of synseed coat did not affect negatively on the
studied parameters. Redenbaugh et al. [36] reported that
coating did not reduce the efficiency of shoot emergence
from alfalfa somatic embryos.
This work described efficient procedure to produce ar-
tificial synseed in large scale in banana where six sub-
cultures of in vitro obtained shoot tips were established.
Encapsulation of shoot tips provided reliable method to
produce synseeds. Single layered synseeds with charcoal
expressed the highest conversion frequency. Creation of
seed coat around the artificial endosperm was recom-
mended for synseed conversion. In addition, to improve
synseed conversion on conversion substratum shoot tips
should be oriented to be upward.
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