ArticlePDF Available

Effects of nano silver on the growth of banana (Musa spp.) cultured in vitro

Authors:

Abstract and Figures

Nano silver has positive effects on the growth and development of in vitro plants. In this study, shoots of in vitro banana with 1 cm in length were cut off the tip which was cultured in Murashige and Skoog (MS) medium supplemented with 30 g.L-1 sucrose, 15% coconut water (v/v), 5 mg. L-1 6-Benzyladenine (BA), 8 g.L-1 agar and nano silver (1, 3, 5, 7 ppm). After twenty days cultured, shoots in the medium supplemented with 1 ppm nano silver have grown well with 8.4 times multiplication and total chlorophyll content (2.05 mg.g-1), three-fold higher than in the control experiment. All new shoot samples were cultivated in a new medium with same component and supplemented with 1 mg.L-1 active charcoal to study root formation. The results showed that the best medium for root development was the one with 3 ppm nano silver. In this treatment, the height of shoots, number of leaves, number of roots, root length, fresh weight, dry weight and total chlorophyll content were 2.90 cm, 4.40 leaves per explant, 7.10 roots per explant, 7.7 cm per explant, 1.47 g per explant, 0.136 g and 3.17 mg.g-1, respectively. In vitro healthy seedlings with equal size were grown in nurseries. Different concentrations of nano silver (5, 10, 15 ppm) were sprayed on once a week. After 30 days, the highest quality result was observed at concentration 5 ppm with shoot of 4.86 cm in length, 5.20 leaves per shoot, 4.60 roots per explant, root of 4.87 cm in length and 3.07 g per explant in fresh weight.
Content may be subject to copyright.
J. Viet. Env. 2018, 10(2):92-98
DOI: 10.13141/jve.vol10.no2.pp92-98
92
* Corresponding author
Email: dodanggiap@gmail.com
http://d x.doi.org/10.13141/JVE
ISSN 2193-6471
RESEARCH ARTICLE
Effects of nano silver on the growth of banana (Musa spp.)
cultured in vitro
nh hưởng ca nano bc lên s sinh trưởng ca cây chui (Musa spp.) nuôi cy in vitro
DO, Dang Giap*; DANG, Thi Kim Thuy; NGUYEN, Thi Huyen Trang; NGUYEN, Thi Duoc; TRAN,
Trong Tuan; DUONG, Duc Hieu
Institute of Tropical Biology, Vietnam Academy of Science and Technology. 9/621 Hanoi highway, Linh Trung ward, Thu Duc district, Ho Chi
Minh City, Vietnam
Nano silver has positive effects on the growth and development of in vitro plants. In this study, shoots of in vitro bana-
na with 1 cm in length were cut off the tip which was cultured in Murashige and Skoog (MS) medium supplemented
with 30 g.L-1 sucrose, 15% coconut water (v/v), 5 mg. L-1 6-Benzyladenine (BA), 8 g.L-1 agar and nano silver (1, 3, 5, 7
ppm). After twenty days cultured, shoots in the medium supplemented with 1 ppm nano silver have grown well with
8.4 times multiplication and total chlorophyll content (2.05 mg.g-1), three-fold higher than in the control experiment. All
new shoot samples were cultivated in a new medium with same component and supplemented with 1 mg.L-1 active
charcoal to study root formation. The results showed that the best medium for root development was the one with 3
ppm nano silver. In this treatment, the height of shoots, number of leaves, number of roots, root length, fresh weight,
dry weight and total chlorophyll content were 2.90 cm, 4.40 leaves per explant, 7.10 roots per explant, 7.7 cm per ex-
plant, 1.47 g per explant, 0.136 g and 3.17 mg.g-1, respectively. In vitro healthy seedlings with equal size were grown in
nurseries. Different concentrations of nano silver (5, 10, 15 ppm) were sprayed on once a week. After 30 days, the
highest quality result was observed at concentration 5 ppm with shoot of 4.86 cm in length, 5.20 leaves per shoot, 4.60
roots per explant, root of 4.87 cm in length and 3.07 g per explant in fresh weight.
Nano bc có tác động tích cc lên quá trình sinh trưởng và phát trin ca mt s loài thc vt nuôi cy in vitro. Trong nghiên
cu này, chi non in vitro có chiu cao 1 cm đã ct b đỉnh được nuôi cy trên môi trường MS b sung 30 g/L đường,
15% nước da (v/v), 5 mg/L BA, 8 g/L agar và nano bc nng độ 1, 3, 5, 7 ppm. Sau 20 ngày nuôi cy, mu cy phát trin
tt trên môi trường b sung nano bc nng độ 1 ppm vi h s nhân chi 8,4 ln, hàm lượng chlorophyll tng s đạt 2,05
mg/g, cao gp ba ln so vi mu đối chng. Mu cy được chuyn sang môi trường mi có thành phn tương t và b sung
thêm 1 mg/L than hot tính để kho sát s to r. Kết qu cho thy mu cy trên môi trường có nng độ nano bc 3 ppm
phát trin tt nht vi chiu cao cây 2,90 cm/mu; s 4,40/ mu; s r 7,10/ mu; chiu dài r 7,7 cm/mu; khi lượng
tươi 1,47 g/mu, khi lượng khô 0,136 g/mu hàm lượng chlorophyll tng đạt 3,17 mg/g. Nhng cây con in vitro phát
trin tt được trng ngoài vườn ươm. Các dung dch nano bc nng độ 5, 10, 15 ppm được dùng để phun lên cây 1 ln/
tun. Sau 30 ngày, kết qu cao nht ghi nhn dung dch nng độ 5 ppm vi chiu cao chi đạt 4,86 cm, 5,20 lá/chi,
4,60 r/mu, chiu dài r đạt 4,87 cm và khi lượng tươi đạt 3,07 g/mu.
Keywords:
chlorophyll content; development; banana; growth; Musa spp.; nano silver
1. Introduction
Banana (Musa spp.) belongs to the Musaceae family and
is the most widely exported fruit in the world, providing
millions of people living in the tropics and subtropics such
as a kind of staple food. In terms of gross value of pro-
duction, banana is ranked fourth, next to rice, wheat and
maize in terms of gross value of production. It is grown in
around 150 countries worldwide on an area of 4.84 mil-
lion ha with gross output of 95.6 million tons (Singh et al.,
2011). For high quality applications, the development of
uninfected plant material through tissue culture has been
one of the main research fields during recent years. In
vitro banana clonal propagation started in the early of
1970s (Ma and Shii, 1972) and became a normal tech-
nique for a wide range of cultivars by the middle of 1980s
(Vuylsteke, 1989; Smith and Drew, 1990). Nowadays, three
main procedures have been described for somatic em-
bryogenesis in banana and based on vegetable tissues
such as rhizome fragments or leaf bases (Novak et al.,
1989), in vitro proliferating meristems (Dhed’a et al., 1991)
and immature male inflorescences (Escalant et al., 1994).
However, microbial contaminants are the major challeng-
es of in vitro plant propagation during different stages of
culture processes such as initiation of callus or sub-
culturing (Omamor et al., 2007). It was reported that
J. Viet. Env. 2018, 10(2):92-98
93
about 40-60% of in vitro banana cultures were lost in
spite of using mainly reliable, aseptic procedures
(Msogoya et al., 2012). Nano silver is a non-toxic material
that shows high capabilities against microorganisms e.g.
fungus, bacteria, and viruses. This chemical can disadvan-
tageously affect more than 600 microorganisms (Abdi et
al., 2008). The recent years of development in nanotech-
nology have also increased the use of silver nanoparticles
(AgNPs) (Luoma, 2008). AgNPs is commonly used as coat-
ing for many products such as medical devices, food stor-
age containers, handrails etc. AgNPs is also spun down in
fabrics and in some cases as powder for use in shoes
(Luoma 2008). The optical and physical properties of
AgNPs make it also very useful in medical applications
(Winjhoven et al., 2009).
A lot of research on AgNPs has been documented on
microbial and animal cells; however, only a few studies
were done on plants (Krishnaraj et al., 2012; Monica and
Cremonini, 2009). AgNPs increased plants growth process
(shoot and root length, leaf area) and biochemical param-
eters (chlorophyll, carbohydrate and the content of pro-
tein, antioxidant enzymes) of Brassica juncea, common
bean and corn (Salama, 2012; Sharma et al., 2012). The
aim of this paper to evaluate effect of nano silver on ba-
nana growth and development.
2. Materials and methods
2.1 Materials
Shoots of 1.0 cm in length having one pair of leaf from in
vitro cultured banana cultured at Plant Cell Technology
Department, Institute of Tropical Biology were used in this
study.
PVP-coated Ag nanoparticles were purchased as a na-
nosilver colloid (Institute of Material Science, Ho Chi Minh
city). AgNP were roughly spherical. The size distribution
was as mean 25 30 nm. A dilution in deionized water
was prepared to make a 1000 ppm Ag stock solution.
2.2 Culture media
Shoot multiplication medium was MS basal medium (Mu-
rashige and Skoog, 1962) and was modified by Do et al.
(2012). MS basal medium was supplemented with 30 g/l
sucrose, 8 g/l agar, 5 mg/l 6-Benzyladenine (BA) (Duchefa
Biochemie, Netherlands), 15% coconut water (v/v) and
different concentrations of nano silver (0, 1, 3, 5, 7 ppm).
Rooting medium was the same composition of shoot mul-
tiplication medium and supplemented with 1 mg/l activat-
ed carbon. The pH of the two media was adjusted to 5.7
5.8 with 0.1 N HCl or KOH before adding 0.8% agar and
autoclaved at 121oC, 1 atm for 15 minutes.
2.3. Experimental design
2.3.1. Effect of nano silver concentrations on
shoot multiplication of in vitro banana
Shoot of in vitro banana with 1 cm in length was cut off
the tip and the rest of its size 3-5 mm. Then, explants
were cultured into the above shoot multiplication medi-
um. The study was carried out to evaluate the effect of
different concentrations of nano silver on shoot for-
mation and development of in vitro banana.
After 20 days of culture, shoot multiplication was evaluat-
ed by the number of shoots/explant, shoot length (cm),
number of leaves/shoot, fresh weight (g/explant), dry
weight (g/explant), and total chlorophyll content (mg/g).
2.3.2. Effect of nano silver concentrations on root
formation in vitro banana
After 20 days of culture, shoots were separated and sub-
cultured on rooting medium. The effect of various con-
centrations of nano silver on root formation and devel-
opment of in vitro banana was evaluated by number of
leaves/plantlet, number of roots/plantlet, root length (cm),
shoot length (cm), fresh weight (g/plantlet), dry weight (g/
plantlet), total chlorophyll content (mg/g). These parame-
ters were recorded after 20 days of culture.
2.3.3. Effect of nano silver concentrations on
growth and development of ex vitro banana
Rooted plantlets were removed from the medium,
washed under running tap water to remove the agar and
planted in a fiber coconut coal ashes mixture (1:2). The
plantlets were watered two times/day by nano silver solu-
tions with different concentrations (0, 5, 10, 15 ppm) and
watered one time/week. The aim of this experiment was
to evaluate the effect of nano silver on growth and devel-
opment of ex vitro banana.
The data was recorded after 30 days of cultivation with
parameters such as the number of leaves/plantlet, num-
ber of roots/plantlet, root length (cm), shoot length
(cm/plantlet), fresh weight (g/plantlet).
2.4. Culture condition
The cultures were maintained at 25 ± 2°C, 70 80% air
humidity under continuous illumination of 18.70 ± 1.00
µmol m-2 s-1 during 12h/day provided by fluorescent
lamps (Do et al., 2012).
2.5. Statistical analysis
The experiments had completely randomized design and
were repeated three times with each treatment contain-
ing ten explants. The data obtained from the experiments
were analysed, calculated and compared by using Dun-
J. Viet. Env. 2018, 10(2):92-98
94
can’s multiple-range test at a 5% level of significance by
SPSS software version 16.0 (Duncan, 1955).
3. Results and discussion
3.1. Effect of nano silver concentrations on
shoot multiplication of in vitro banana
When AgNPs was added into the cultivation medium, the
response was positive not only on shoot initiation, num-
ber and growth rate but also increased leaf number and
total chlorophyll content (Table 1). All shoot multiplication
media supplemented with AgNPs (except 7 ppm) led to
higher values than the control experiment. The explants
on medium with 1 ppm nano AgNPs gained the maximum
number of shoots (8.40), highest shoot length (2.45 cm),
maximum number of leaves (12.10) and highest rate fresh
weight/dry weight (2.24/0.20 g). These parameters were
1.5 2 folds higher than those of explants cultured on
medium without nano silver, especially, total chlorophyll
content was 3 folds higher (2.05/0.69 mg.g-1). When nano
silver solution was increased to 3, 5 and 7 ppm, the pa-
rameters began to decrease; particularly, the explants on
medium containing 7 ppm developed less than the con-
trol but the total chlorophyll content (1.24 mg.g-1) was
more than control (0.69 mg.g-1). In the medium with con-
centrations of 3 and 5 ppm, the number of shoots per
explant were 1.4 1.6 folds higher and the total chloro-
phyll content was 2.3 2.4 folds higher compared to con-
trol (without nano silver). However, the data recorded in
SH2 and SH3 treatment was no significantly different be-
tween experiments (p <0.05). In brief, in shoot multiplica-
tion medium, the concentration of 1 ppm nano silver was
found to be the optimum concentration required to in-
duce maximum shoot growth.
Table 1. Effect of nano silver on multiple shoot formation from cultured shoot tip explants for 20 days
Treatment
Concentration of
nano silver
solution (ppm)
No. of
shoots/
explant
Fresh
weight
(g/shoots)
Dry
weight
(g/shoots)
Total
chlorophyll
content (mg/g)
SH0
0
4.30c
1.42b
0.12b
0.69c
SH1
1
8.40a
2.24a
0.20a
2.05a
SH2
3
6.80b
1.73b
0.15b
1.60ab
SH3
5
6.03b
1.67b
0.14b
1.65ab
SH4
7
4.90c
1.51c
0.13b
1.24b
Mean values followed by the same letter(s) within a column are not significantly different (P <0.05)
Figure 1. Banana shoots on medium containing different concentrations of nano silver solution after 20 days incubated in
vitro (SH0: 0 ppm; SH1: 1 ppm; SH2: 3 ppm; SH3: 5 ppm; SH4: 7 ppm)
3.2. Effect of nano silver concentrations on
root formation of in vitro banana
Nano silver not only induced shoot multiplication but also
influenced root formation of explant (Table 2). More roots
were produced in explants inoculated on medium con-
taining nano silver solution in concentrations from 1 to 5
ppm. It was also observed that in vitro shoot cultures on
medium containing 3 ppm AgNPs produced maximum
number of roots (7.10) per explant with higher root length
(7.70 cm). The highest shoot length (2.90 cm) and number
of leaves per shoot (4.40) was obtained on the medium
containing of 3 ppm AgNPs. Fresh weight/dry weight
(1.47/0.136 g) with total chlorophyll content 3.17 mg.g-1
was also higher than other treatments. However, these
parameters were no significantly different between the R2
and R1 treatment experiments (p < 0.05). When concen-
tration of nano silver solution was increased to 7 ppm,
J. Viet. Env. 2018, 10(2):92-98
95
the growth of shoot was limited. The parameters of
growth were lower than control experiment. Neverthe-
less, the content of chlorophyll in the control experiment
(2.80 mg.g-1) was still higher than the treatment experi-
ment (2.62 mg.g-1).
Table 2. Effect of nano silver concentrations on rooting medium of in vitro banana after 20 days of incubation
Treat-
ment
Concentration
of nano silver
solution (ppm)
Shoot
length
(cm)
No. of
leaves/
plantlet
No. of
roots/
plantlet
Root
length
(cm)
Fresh
weight
(g/plantlet)
Dry
weight
(g/plantlet)
Total
chlorophyll con-
tent (mg/g)
R0
0
2.30bc
4.00ab
5.90bc
5.97bc
1.05b
0.093b
2.62bc
R1
1
2.68ab
3.90ab
6.60ab
6.90ab
1.15b
0.098b
2.91ab
R2
3
2.90a
4.40a
7.10a
7.70a
1.47a
0.136a
3.17a
R3
5
2.51abc
3.56b
5.40c
6.04bc
1.09b
0.094b
2.56c
R4
7
2.22c
3.41b
5.09c
5.10c
0.71c
0.061c
2.80bc
Mean values followed by the same letter(s) within a column are not significantly different (P <0.05)
Figure 2. Banana plantlets on rooting medium containing different concentrations of nano silver solution after 20 days incu-
bated in vitro (R0: 0 ppm; R1: 1 ppm; R2: 3 ppm; R3: 5 ppm; R4: 7 ppm)
3.3. Effect of nano silver concentrations on
growth and development of ex vitro banana
The plantlets in vitro were planted and tended in arbore-
tum. The results were recorded after 30 days’ cultivation
(Table 3). The plantlets were watered with concentrations
of AgNPs from 0 to 5 ppm. N1 treatment experiment ex-
posed effectively growth and development, and a signifi-
cant difference (p < 0.05) among the concentrations test-
ed. The plantlets watered with solution containing 5 ppm
AgNPs achieved the highest values of shoot length (4.86
cm), number of leaves per shoot (5.20), number of roots
(4.60), root length (4.87), fresh weight (3.07). When con-
centration of AgNPs was increased, the parameters de-
creased, at 15 ppm AgNPs being even lower than the con-
trol experiment.
Table 3. Effect of nano silver concentrations on growth and development of ex vitro banana at nursery garden after 30 days
of cultivation
Treatment
Concentration
of AgNPs
(ppm)
Shoot
length
(cm/plantlet)
No. of
leaves/
plantlet
No. of
roots/
plantlet
Root
length
(cm)
Fresh
weight
(g/plantlet)
N0
0
3.58b
3.50d
3.36c
3.64a
2.49bc
N1
5
4.86a
5.20a
4.60a
4.87c
3.07a
N2
10
3.60b
4.77b
4.06b
4.10b
2.65b
N3
15
3.40b
4.10c
3.53c
3.70a
2.39c
Mean values followed by the same letter(s) within a column are not significantly different (P <0.05)
J. Viet. Env. 2018, 10(2):92-98
96
Figure 3. Banana plants were watered with different concentrations of nano silver solution after 30 days at nursery garden
(N0: 0 ppm; N1: 5 ppm; N2: 10 ppm; N3: 15 ppm)
4. Discussion
AgNPs have positive effects on growth and development
of banana plants. The explants were cultured on shoot
multiplication medium and rooting medium supplement-
ed with 1 ppm and 3 ppm AgNPs, respectively. These me-
dia were optimum for formation and development of
shoot and root. Particularly, the chlorophyll content of
shoots cultured on medium containing AgNPs was higher
than the control experiment (without AgNPs). Salama
(2012) and Sharma et al. (2012) reported that AgNPs in-
creased plant growth process (shoot and root length,
area of the leaf) and biochemical parameters (chlorophyll,
carbohydrate and the content of protein, antioxidant en-
zymes) of common bean and corn.
Salama (2012) reported that low concentrations of silver
nanoparticles had a stimulating effect on the growth of
the plantlets, while the enhanced concentrations induced
an inhibitory effect. However, the increase in the concen-
tration of silver nanoparticles from 20 to 60 ppm led to an
increase in shoot and root lengths, leaf surface area, chlo-
rophyll, carbohydrate and protein contents of the two
tested crop plants (Phaseolus vulgaris L. and Zea mays L.).
Sharma et al. (2012) investigated the effect of silver nano
particles (at 0, 25, 50, 100, 200 and 400 ppm) on the
growth and antioxidant status of 7-day old Brassica juncea
seedlings. The results showed that fresh weight, root and
shoot length and vigor index of seedlings is positively af-
fected by silver nanoparticle. It induced a 326% increase
in root length and 133% increase in vigor index of the
treated seedlings. The photosynthetic quantum efficiency
is improved and the content of chlorophyll in leaves of
treated seedlings is recorded higher than the control
seedlings.
Nano silver was used with optimum concentration that
had a significant effect on the plant shoot, the number of
leaves, the height of the plant, plant dry weight, inflo-
rescence dry weight, seed yield, weight of one hundred
seeds, polyphenol and tannin content in shoot. The con-
centration of nano silver from 20 to 60 ppm has led to an
improvement in the seed yield (Seif et al., 2011). AgNPs
induce root growth by blocking ethylene signalling in Cro-
cus sativus (Rezvani et al. 2012). Silver ions can displace
copper ions from the receptor proteins and play an im-
portant role in ethylene binding upon receptors.
The effect of peptone and silver nitrate on in vitro shoot
formation in Hevea brasiliensis Muell Arg was studied by
Yupaporn and Sompong (2012). The addition of silver
nitrate in the induction medium at concentrations ranging
from 0.5 to 2 mg/l was successful in promoting shoot
multiplication in all the explants with an average number
of 5 shoots per explant. In addition, silver nitrate provides
silver ions which may interact with polyamines, leading to
the promotion of organogenesis and embryogenesis
(Zhang et al., 2001).
5. Conclusion
Culture medium supplemented with 1 ppm nano silver
was the best one for shoot multiplication. After twenty
days cultured, shoots in this medium have grown well
with 8.4 times multiplication and total chlorophyll content
J. Viet. Env. 2018, 10(2):92-98
97
(2.05 mg. g-1) was three-fold higher than in the control.
The suitable medium for rooting was the one added 3
ppm nano silver. In this treatment, the height of shoots,
number of leaves, number of roots, root length, fresh
weight, dry weight and total chlorophyll content were 2.90
cm, 4.40 leaves per explant, 7.10 roots per explant, 7.7
cm per explant, 1.47 g per explant, 0.136 g and 3.17
mg/g, respectively. When the plantlets were planted and
tended at the arboretum, nano silver also exposed effec-
tively on growth and development of plant when using 5
ppm of concentration. After 30 days, the plant achieved
the highest quality with shoot of 4.86 cm in length, 5.20
leaves per shoot, 4.60 roots per explant, root of 4.87 cm
in length and 3.07 g per explant in fresh weight.
Acknowledgement: The authors are grateful to the
Southern Region National Key Laboratory of Plant Cell
Technology - Institute of Tropical Biology, Vietnam Acad-
emy of Science and Technology for the financial support
to carry out the present experiment.
6. References
[1] Abdi, G., Salehi, H. and Khosh-khuri, M. (2008). Nano
silver: A novel nanomaterial for removal of Bacterial
contamination in Valerian (V. officinalis) tissue cul-
ture. Acta Physiol Plant. 30: 709-714.
[2] Do, D.G, Pham, N.V, Tran, T.T, Nguyen, T.H.T, Pham,
N.A.T, Thai, X.D. (2012). “High-frequency shoot multi-
plication of laba banana (Musa sp.) cultured in vitro
by using light, myo-inositol and adenin sulphate”.
Journal of Biology. 34(3SE): 180-187 (in Vietnamese
with English summary).
[3] Dhed' A, D., Dumortier, F., Panis, B., Vuylsteke, D. and
De Langhe, E. (1991). Plant regeneration in cell sus-
pension cultures of the cooking banana cultivar
'Bluggoe' (Musa spp. ABB group). Fruits. 46: 125-135.
[4] Duncan, D.B. (1955). Multiple range and multiple F
tests. Biometrics. 11(1): 1-5.
[5] Escalant, J.V., Teisson, C. and Cote, F. (1994). Ampli-
fied somatic embryogenesis from male flowers of
triploid banana and plantain cultivars (Musa spp.). In
vitro Cellular Developmental Biology. 30P: 181-186.
[6] Krishnaraj, C., Jagan, E.G., Ramachandran, R., Abira-
mi, S.M., Mohan, N. and Kalaichelvan, P.T. (2012). Ef-
fect of biologically synthesized silver nanoparticles
on Bacopa monnieri (Linn.) Wettst. Plant growth me-
tabolism. Process Biochem 47(4): 51658.
[7] Luoma, N.S. (2008). Silvernanotechnologies and the
environment: Old problems or new challenges? The
Project on Emerging Nanotechnologies. Vol. 15. Sep-
tember 2008.
[8] Ma, S.S. and Shii, C.T. (1972). In vitro formation of
adventitious buds in banana shoot apex following
decapitation. Journal of Chinese Society for Horticul-
tural Science. 18: 135-142.
[9] Monica, R.C. and Cremonini, R. (2009). Nanoparticles
and higher plants. Caryologia. 62(2): 161165.
[10] Msogoya, T., Kayagha, H., Mutigitu, J., Kulebelwa, M.
and Mamiro, M. (2012). Identification and manage-
ment of microbial contaminants of banana in vitro
cultures. J Appl Biol. 55: 3987-3994.
[11] Murashige, T. and Skoog F. (1962). A revised medium
for rapid growth and Bio-assays with tobacco tissue
cultures. Phys. Plant. 15: 473-497.
[12] Novak, F.J., Afza, R., Van Duren, M., Perea-Dallos, M.,
Conger, B.V. and Tang, X. (1989). Somatic embryo-
genesis and plant regeneration in suspension cul-
tures of dessert (AA or AAA) and cooking (ABB) ba-
nanas (Musa spp.). Nature Biotechnology. 7: 154-
159.
[13] Omamor, I.B., Asemota, A.O., Eke, C.R. and Eziashi,
E.I. (2007). Fungal contaminants of the oil palm tis-
sue culture in Nigerian Institute for oil palm Re-
search (NIFOR). Afr J Agric Res. 2(10): 534-537.
[14] Rezvani, N., Sorooshzadeh, A. and Farhadi, N. (2012).
Effect of nano-silver on growth of saffron in
ooding stress. World Acad. Sci. Eng. Technol. 1:
517522.
[15] Salama, H.M.H. (2012). Effects of silver nanoparticles
in some crop plants, common bean (Phaseolus vul-
garis L.) and corn (Zea mays L.). Int Res J Biotech.
3(10): 190197.
[16] Seif, S.M., Sorooshzadeh, A.H., Rezazadeh, S. and
Naghdibadi, H.A. (2011). Effect of nano-silver and sil-
ver nitrate on seed yield of borage. Journal of Medic-
inal Plants Research. 5(5): 706-710.
[17] Sharma, P., Bhatt, D., Zaidi, M.G., Saradhi, P.P., Khan-
na, P.K., Arora, S. (2012). Silver nanoparticlemediated
enhancement in growth and antioxidant status of
Brassica juncea. Appl Biochem Biotechnol. 167:
22252233.
[18] Singh, H., Uma, S., Selvarajan, R. and Karihaloo, J.
(2011). Micropropagation for production of quality
banana planting material in Asia-Pacific. Asia-Pacific
Consortium on Agricultural Biotechnology (APCoAB),
New Delhi, India, 92 p.
[19] Smith, M.K. and Drew, R.A. (1990). Current applica-
tions of tissue culture in plant propagation and im-
provement”. Australian Journal of Plant Physiology.
17: 267-289.
[20] Vuylsteke, D.K. (1989). Shoot - tip culture for the
propagation, conservation and exchange of Musa
germplasm. In: Withers LA, ed. “Practical Manuals for
Handling Crop Germplasm in vitro”. Vol. 2, Rome:
IBPGR: 1-56.
[21] Winjhoven, W.P S., Peinenburg, J.G.M W., Herberts, A.
C., Hagens I. W., Oomen, G. A., Heugens, H.W. E.,
Roszek, B., Bisshops, J., Gosens, I., Van De Meent, D.,
Dekkers, S., De Jong, H. W., Van Zijverden, M., Sips,
J. Viet. Env. 2018, 10(2):92-98
98
J.A.M A. and Geersma, E. R. (2009). Nano-Silver- a re-
view of available data and knowledge gaps in human
and environmental risk assessment. Nanotoxicology.
3(2): 109-138.
[22] Yupaporn, S. and Sompong, T. (2012). The effect of
peptone and silver nitrate on in vitro shoot for-
mation in Hevea brasiliensis Muell Arg. Journal of Ag-
ricultural Technology. 8(4): 1509-1516.
[23] Zhang, P., Phansiri, S. and Puonti, K.J. (2001). Im-
provements of cassava shoot organogenesis by the
use of silver nitrate in vitro. Plant Cell, Tissue and
Organ Culture. 67: 47-54.
... Clonal propagation of banana in vitro was the first report by Ma and Shii [8] in the early year 1972 and has been successfully developed for some last decades and eventuated a common technique for wide applications by the middle of 1980 [9][10]. Recently, three major in vitro banana procedures have been developed including somatic embryogenesis [11]; in vitro proliferating meristem and immature male inflorescences [12]. ...
... Among the various metallic nanoparticles, silver nanoparticles (AgNPs) are non-toxic substances and which were reported to unveil broad-spectrum suppression against the growth of over 600 harmful plant pathogens [16] and simultaneously ameliorate callus induction, organogenesis, somatic embryogenesis, somaclonal variation, genetic transformation and metabolite plant production [17][18]. Nowadays, AgNPs are extensively studied in various scientific fields and one of the most rapid-growing nanomaterials due to their unique chemical and physical properties [10,19]. Moreover, AgNPs were reported to use as the main factor to improve plant growth (leaf area, ...
... Moreover, the different parts of plants respond contrarily to AgNPs, the growth of plant tissue or organ may enhance by AgNPs while it curbs the growth of other organs [48]. Our current results have been inconsistent with the recent report of Giap et al. [10] who found that the optimal concentration at 1.0 ppm of AgNPs for banana shoot multiplication, 3.0 ppm and 5.0 ppm for rooting and nursery explants of banana in vitro. It can explain that due to using the different organ and medium, culture conditions which may give different results. ...
Article
Full-text available
The King banana (Musa ssp.) is a native specialty variety in Vietnam with distinctive features, delicious taste specialty and high economic value. The objectives of this study were to investigate the effects of different concentrations of silver nanoparticles (AgNPs) on the callus formation, shoot regeneration and multiplication, root induction and nursery phase during in vitro propagation. The results have shown that approximately 98.0% of pseudostems were formed in the callus in the MS medium supplemented by 8.0 ppm of AgNPs after 3 weeks of culture. The calluses were then examined on the basal MS medium added with different AgNPs concentrations (0.0, 2.0, 4.0 and 6.0 ppm). The maximum of shoot numbers was found in the medium supplemented by 4.0 ppm after 4 weeks cultured. Moreover, the highest shoot regeneration rate was found in the stem culture supplemented by 6.0 ppm by 93.33% with a shooting coefficient of 4.22 value. Similarly, the medium supplemented with 4.0 ppm of AgNPs has been the best choice for rooting in vitro with a rate of 98.33%, an average of 5.22 roots per shoot, root length of 4.26 cm, respectively. The plantlets had remarkable characteristics in the nursery stage with a height of 8.17 cm, 1.68 leaves per plant. Our results have revealed that AgNPs have exerted strikingly positive influences on in vitro morphogenesis of the King banana. We have also discussed the complex effects of AgNPs induced on plant growth in vitro which show not only AgNPs concentrations-culture medium dependent but also part of organ and plant species uses dependent, respectively. Our findings may open a new window for using AgNPs to conserve both the specialty fruits and endanger species in vitro in the future.
... Few studies have reported the phytotoxicity of copper [17,18] on Musa acuminata and on Philodendron selloum [5] plants, which confirmed that the high activity of the antioxidant enzymes could cause ROS detoxification, hence, decreasing the oxidative damage of plants. Many studies have been published about the different stresses on in vitro Musa sp., such as water stress [19], osmotic stress [20], cold stress [21], stress of nanoparticles [22][23][24], and salinity stress [4], but very few on copper stress [18,25]. ...
... As the chlorophyll content was reduced due to its degradation, the photosynthesis of banana plantlets also reduced, as confirmed by many studies. Due to the stress resulting from nano silver, different concentrations led to a decrease in the chlorophyll content of banana plantlets, such as for 7 and 200 mg L −1 [22,23], respectively. The chlorophyll content of banana plants was significantly influenced by the liquid MS basal medium, containing excess salt (NaCl up to 800 mM) or suffering under drought stress (mannitol up to 600 mM), as reported by [41]. ...
Article
Full-text available
Developing a successful protocol for banana in vitro culture is a guarantee for the mass propagation of pathogen-free, high-quality, true-to-type planting materials with low production costs. The current work aimed to investigate the influence of increasing copper levels in an MS medium on endophytic bacterial contamination; shoot multiplication; rooting and the acclimatization of in vitro cultured banana; minerals and chlorophyll content; antioxidant enzymes activity; electrolyte leakage; and the genetic stability of banana regenerants. Four different concentrations of copper sulphate (0.025 as a control, and 30, 60, and 120 mg L􀀀1) were examined. The growth of the endophytic bacteria was inhibited at 60 mg L􀀀1 of copper sulphate which recorded zero contamination, without a significant difference at 120 mg L􀀀1. However, 0.025 mg L􀀀1 of copper sulphate was optimal for the maximum shoot number and shoot length (10 shoots and 6 cm, respectively) without significant differences at 30 mg L􀀀1. The root length of banana plantlets was significantly enhanced at 30 mg L􀀀1 of copper sulphate but without significant differences to the control, regarding the number of roots (9.92 cm and 3.80 roots, respectively). In vitro plants were acclimatized successfully at 30 mg L􀀀1 of copper sulphate with 100% survival. The uptake of minerals, antioxidant enzyme activity and electrolyte leakage was improved because of the copper sulphate, but the chlorophyll level decreased. RAPD profiling showed polymorphism in only one plant treated with 60 mg L􀀀1 of copper sulphate, with an average of 1.8%. The genome template stability percentage was almost 100% for all treated plants.
... Ibrahim (2015) reported the green synthesis of silver NPs and tested their antimicrobial property against representative microorganisms. However, there still exists a report wherein the effect of silver NPs on growth and development of in vitro cultured banana was studied where the results showed that the best medium for shoot and root formation of Musa was 1 ppm and 3 ppm silver NPs, respectively (Do et al. 2018). On the other hand, Hasanin et al. (2021) illustrated that there was an increase in chlorophyll content, and several other growth parameters of banana like number of leaves, root length, and number of roots were observed in medium fortified with 9 mg/l St@CuONPs. ...
Article
Full-text available
Banana (Musa spp.), commonly known as ‘Adam fig’ and ‘Fruit of wise man’, is a commercial herbaceous tropical fruit, which governs its antiquity from ancient periods in the Indian and African subcontinent. All parts of the plant, i.e. stem, leaf, root, inflorescence, peel, fruit, and flower, have significant medicinal and nutritional values. Owing to its multitude of uses, it is known as ‘Kalpavriksha’ (plant of virtues). To combat multi-faceted issues related to traditional propagation, in vitro–based regeneration-cum-genetic improvement approaches become the trend of the hour. The present review illustrates various physico-chemical factors that are responsible for successful in vitro regeneration and acclimatization, protoplast culture, anther and microspore culture, cryopreservation and synthetic seed production, genetic transformation, mutagenesis, and nanotechnological and omics approaches. The key intent of this article is to present an insight on in vitro biotechnological research advances in the past decade, to identify the research gaps, unexplored areas, and major shortcomings associated with banana biotechnology and to highlight the potential approaches to mitigate them. Eventually, this review made salient conclusions and recommendations paving the way forward for the banana researchers to develop innovative ideas in order to enhance the propagation frequency and to ensure the genetic improvement of banana. Key points • This review addresses biotechnological interventions in Banana (Musa spp.) for enhanced propagation and quality improvement. • Highlights factors influencing in vitro regeneration, conservation, and genetic transformation. • Provides novel ideas to harness the qualitative and quantitative genetic improvement.
Article
Nano silver has positive effects on the growth and development of in vitro plants. The current study was carried out to investigate the effect of adding different concentrations of nano silver nitrate (0.0, 1.0, 2.0, 3.0 and 4.0 ml/l) to the culture medium in multiplication and rooting stage of the date palm cv. Hayani. The results showed that there was a significant difference of Ag NPs concentrations in regeneration stage as it increased the number and length of shoots at 3.0 ml/l concentrations compared to the control treatment. The outcome of shoots multiplication experiment indicated that Ag NPs concentration had a significant positive effect on studied traits, when the 3.0 ml/l of Ag NPs added to MS basal medium supplemented with BA at 2.0, GA3 at 0.5 and NAA at 0.1 (mg/l) gave the highest shoots and leaves number as it reached 10.74 shoots and 30.11 leaf/culture with length 6.60 cm, respectively in comparison with lowest results recorded 2.50 shoot/culture, 10.70 leaf/culture with length 2.00 cm, respectively with the control treatment. All shoots were cultivated in a new medium supplemented with NAA at 1.0 mg/l and various concentrations of Ag NPs to study root formation. The findings indicated that the optimal medium for root formation was one that included Ag NPs at 3.0 ml/l which resulted in the highest rooting percentage (85.4%), root number (8.4 root/culture) with length 6.3 cm. In concentration 3.0 ml/l of Ag NPs recorded that, chlorophyll A, chlorophyll B and carotenoids increases to 1.50, 1.10 and 2.33 (mg/g). Also, occurred increase in carbohydrate content to 26.33 mg/g, the proline was enhanced and increased gradually to 3.31 mg/g and protein content to 0.77 mg/g.
Article
Full-text available
Plant tissue culture plays an important role in plant biotechnology due to its potential for massive production of improved crop varieties and high yield of important secondary metabolites. Several efforts have been made to ameliorate the effectiveness and production of plant tissue culture, using biotic and abiotic factors. Nowadays, the addition of nanoparticles as elicitors has, for instance, gained worldwide interest because of its success in microbial decontamination and enhancement of secondary metabolites. Nanoparticles are entities in the nanometric dimension range: they possess unique physicochemical properties. Among all nanoparticles, silver-nanoparticles (AgNPs) are well-known for their antimicrobial and hormetic effects, which in appropriate doses, led to the improvement of plant biomass as well as secondary metabolite accumulation. This review is focused on the evaluation of the integration of nanotechnology with plant tissue culture. The highlight is especially conveyed on secondary metabolite enhancement, effects on plant growth and biomass accumulation as well as their possible mechanism of action. In addition, some perspectives of the use of nanomaterials as potential therapeutic agents are also discussed. Thus, the information provided will be a good tool for future research in plant improvement and the large-scale production of important secondary metabolites. Elicitation of silver-nanoparticles, as well as nanomaterials, function as therapeutic agents for animal well-being is expected to play a major role in the process. However, nanosized supramolecular aggregates have received an increased resonance also in other fields of application such as animal welfare. Therefore, the concluding section of this contribution is dedicated to the description and possible potential and usage of different nanoparticles that have been the object of work and expertise also in our laboratories.
Article
Full-text available
Background Banana bunchy top virus (BBTV) is a destructive viral disease in many countries including Egypt; it causes severe economic losses in banana crop. Recently, nanotechnology was used to generate resistance against plant viruses. The main purpose of this study was to use silver nanoparticles (AgNPs) as antiviral agents against BBTV. In this research, three different concentrations of AgNPs (40, 50 and 60 ppm) were applied by foliar spray post-BBTV inoculation. In addition, photopigments, oxidative enzymes, proline and phenolic compounds were determined. Besides, Random amplified polymorphic DNA (RAPD) and Sequence-related amplified polymorphism (SRAP) markers were used to evaluate the genotoxicity of AgNPs as antiviral factors against BBTV, compared with the control plants. Results In the current study, it was observed that banana plants infected with BBTV and treated with 50 ppm AgNPs have not shown any external symptoms where the rate of infection was 36%. On the other hand, banana plants treated with 50 ppm AgNPs after viral infection gave a significant increase in dry weight and leaf area, compared with BBTV infected banana plants (viral control). Our study showed that 50 ppm AgNPs treatment post-virus inoculation induced non-significantly and significant changes in chlorophyll (a and b) and carotenoids, respectively, compared with healthy and nano-controls. In contrast, phenol, proline and oxidative enzymes were significantly increased in all plants treated with 50 ppm AgNPs post-virus inoculation, compared with the healthy control. Our findings observed that the banana plants sprayed with 50 ppm AgNPs after BBTV infection induced a few changes at the genomic DNA level in the banana plants, whereas both RAPD and SRAP markers scored nearly the same polymorphism 36.99 and 37.5%, respectively. So, genotoxicity induced by banana plants treated with 50 ppm AgNPs post-BBTV inoculation was low. Conclusions It is evident from the study results the role of AgNPs as a novel, safe and effective antiviral agent against BBTV. These results should be taken into consideration in future for the use of AgNPs for plant viruses management.
Chapter
The use of agrochemicals has revolutionized agricultural production, modifying the availability of resources like nutrients, regulators, and biostimulants, modifying biotic relationships and physicochemical flows in agroecosystems. Nanoparticles (NPs) and nanomaterials (NMs) are a new class of physicochemical agents that promise to significantly improve the production capability of agricultural systems. In this sense, it is critical to understand the impacts that NPs and NMs can have in the short and medium-term, on the different physical, chemical and biological elements of agroecosystems: soil, water, plants, the soil microbiome and the plant itself. The understanding should emphasize the transfer of NPs and NMs in trophic chains, as well as to soil, water, and atmosphere. This manuscript presents updated information about the impact of NPs and NMs of essential and beneficial elements, metals, semimetals, and non-metals, carbon, and zeolites on the different components of the agricultural ecological system. The content highlights the sustainability-focused balance that should exist between pollution and trophic transfer vs. the positive impacts on crop productivity. Keywords: nanoparticles in ecosystems, trophic transfer, heavy metals, environmental pollution, soil health, food safety, agricultural systems, forestry systems.
Article
Full-text available
Borage plant (Borago officinalis L.) is an herbaceous annual plant. It was used as a medicinal plant since ancient times. Seed oil of this plant has been used to treat many skin disorders. It is the richest plant source of gamma-linoleic acid (GLA). However, seed production of borage is limited by flower and seed abscission. Ethylene is responsible for plant organs abscission which may be inhibited by silver ion (Ag+). Therefore, this experiment was designed to study the effect of nano silver and silver nitrate on abscission and yield of seed in borage. The study was carried out in a randomized block design with three replications. Four levels of either silver nitrate (0, 100, 200 and 300 ppm) or nano silver (0, 20, 40, and 60 ppm) were sprayed on borage plant at seed growth stage. The results showed that there was no significant difference between 100 ppm of silver nitrate and 60 ppm concentration of nano silver on the shoot silver concentration. However, increasing the concentration of silver nitrate from 100 ppm to 300 ppm caused a decrease in seed yield. On contrast, a raise in the concentration of nano silver from 20 ppm to 60 ppm has led to an improvement in the seed yield. Additionally, the lowest amount of seed yield was found with control plants. Finally, with increasing level of silver nitrate, the polyphenols compounds content were raised, but the enhancing level of nano silver resulting in the reduction of these components. In conclusion, nano silver can be used instead of other compounds of silver.
Article
Full-text available
Nanomaterials and nanotechnology have been widely applied in the world in this last decade. Nanotechnology provides the tool and the technological platforms for the study and transformation of biological systems. Few studies have focused on the effects and mechanisms of nanomaterials on plants. The results of these studies have been reported with the aim to provide further insight into connections between plants and nanomaterials.
Article
The present paper describes the effect of light intensity, myo-inositol and adenin sulphate on in vitro shoot formation of Laba banana (Musa sp.). In vitro shoots were cultured on MS medium supplemented with 5 mg l⁻¹ BA (6-benzylaminopurine) and different concentrations of myo-inositol (0, 100, 300, 500, 700 mg l⁻¹) or adenin sulphate (0, 80, 100, 130, 160 mg l⁻¹). After 4 weeks, the best multiple shoots were formed on MS medium with 5 mg l⁻¹ BA + 100 mg l-1 adenin sulphate (6.9 shoots per explant) and on MS medium containing 5 mg l⁻¹ BA + 100 mg l⁻¹ myo-inositol (8.0 shoots per explant). Explants on MS medium with suitable concentration of BA were maintained under 12-hphotoperiod with different intensities of light: Diffuse light, 18.70 ± 1.00, 26.20 ± 1.00 and 42 ± 1.00 µmol m⁻²s⁻¹. The best shoot formation rate (4.33 shoots per explant) was achieved on MS medium supplemented BA with a light intensity of 18.70 ± 1.00 µmol m⁻²s⁻¹.
Article
Plant tissue culture involves the culture of all types of plant cells, tissues and organs under aseptic conditions. This definition also extends to the culture of excised embryos and to protoplast culture. An overview of tissue culture techniques and their applications in plant propagation and genetic improvement of plants is presented. The areas under review include: (1) embyro culture, (2) meristem culture, (3) micropropagation, (4) somatic embryogenesis, (5) somaclonal variation, (6) in vitro selection, (7) anther culture and (8) protoplast culture. Problems and limitations of each of the techniques are also discussed. Examples are given of work that has been undertaken or that is currently in progress on the application of these techniques to the improvement of Queensland's subtropical horticultural industries. Key examples are: (1) embryo culture to facilitate incorporation of genes conferring disease-resistance from wild Cucurbita species into cultivated varieties, (2) meristem culture for virus elimination in strawberries (Fragaria × ananassa) and sweet potato (Ipomoea batatas), (3) micropropagation for rapid increase in new varieties of ginger (Zingiber officinale) and pineapple (Ananas comosus) to enable more rapid field evaluation and early release, (4) micropropagation of disease-free, genetically uniform planting material of superior female papaya (Carica papaya) selections and banana (Musa spp.) selections and (5) the use of somaclonal variation and gamma-irradiation for the genetic improvement of banana. Finally, future opportunities for the utilisation of tissue culture in plant propagation and improvement in Queensland's horticultural industries are summarised.