In Vitro Propagation of Banana: Ezaul Arim Alek Ajia Ahman MIN AND Uhul MIN
In Vitro Propagation of Banana: Ezaul Arim Alek Ajia Ahman MIN AND Uhul MIN
In Vitro Propagation of Banana: Ezaul Arim Alek Ajia Ahman MIN AND Uhul MIN
Introduction
The banana and plantains (Musa spp.) belonging to the family Musaceae is one
of the world’s most important subsistence crops. It is originated in Malaysia
through a complex hybridization process (Novak, 1992). It is widely grown in the
tropics and subtropic regions of the world. In Bangladesh, banana is popular for
its year round availability, abundant production as well as high acceptability to
the consumers. It is a rich source of carbohydrate and also rich in some minerals,
notably phosphorus, calcium, and potassium. Banana is particularly rich in
vitamin C and also contains significant amounts of several other vitamins
(INIBAP, 1987). In addition, it has importance for tannin, latex and fiber
production. Banana occupies an area of 45 thousand hectares of land with total
production of 909 thousand metric tones with an average yield of 20.20 t/ha
(BBS, 2006). This yield is quite low compared to other banana growing countries
of the world like Argentina (34 t/ha) and Costa Rica (33 t/ha) (FAO, 2002). The
low yield and production of banana is influenced by many biotic and abiotic
stresses. Among the biotic stress, virus disease is one of the important factors.
The traditional clonal propagation method appears unable to satisfy the increase
in demand for disease free and healthy planting materials of banana. The
productivity of vegetative propagated crops, such as banana and plantain is
greatly reduced by virus disease (Lepoivre, 2000).
1&4
Biotechnology Divn., BARI, Joydebpur, Gazipur, 2&3Plant Genetic Resources Centre,
BARI, Joydebpur, Gazipur, 1701 5Shere-Bangla Agricultural University, Dhaka- 1207,
Bangladesh.
270 KARIM et al.
more effective in producing ball like structure. Similar results were also obtained
by Habib (1994) and Ali (1996) in their experiments. They observed that some
ball like structures formed from the base of the shoot during shoot multiplication.
These balls like structures are suitable for in vitro germplasm conservation.
Plant regeneration and subsequent shoot multiplication from meristem
derived plantlets of banana cv. BARI-1 were observed in MS medium
supplemented with different concentrations of BAP and NAA. The results
obtained from this study are presented in Table 1-4 and discussed under the
following sub-heads:
Table 1. Effect of different concentrations of BAP and NAA on shoot multiplication
of banana at different days after inoculation.
Treatments Shoot number Shoot length (cm)
BAP NAA 10 DAI 20 DAI 30 DAI 10 DAI 20 DAI 30 DAI
(mgL-1) (mgL-1)
0 0.0b 1.00c 1.00f 0.00i 0.00j 0.00j
0.5 1.0a 1.50bc 2.25def 0.30gh 1.03ghi 1.88defgh
0 1.0 0.5 ab 1.25 bc 2.50 de 0.30 gh 1.28 defgh 1.95 bcdefg
1.5 1.0 a 1.50 bc 1.50 ef 0.23 h 1.68 bcde 2.33 b
2.0 0.75 a 1.25 bc 2.00 def 0.20h 1.40 cdefg 1.78 fgh
0 1.0 a 1.00c l.50ef 0.23h 1.23 efgh l.9ocdefg
0.5 1.0 a 1.25 bc 1.75 ef 0.30 gh 1.48 cdefg 2.08 bcdefg
2.5 1.0 a 1.0 a 1.25 bc 2.50 de 0.45 feg 1.28 defgh 1.80 efgh
1.5 1.0a 1.75abc 1.75ef 0.35fgh 1.lofghi 1.83efgh
2.0 l.0a 1.50bc 2.25def 0.45efg 0.70i l.50hi
0 1.0a 1.25bc 1.50ef 0.25h 0.88hi 2.28bc
0.5 1.0 a 1.75 abc 2.50 de 0.48 ef 2.43 a 3.13 a
5.0 1.0 1.0a 2.25ab 2.50de 0.88abc 2.05ab 2.l3bcdef
1.5 1.0 a 1.50 bc 2.50 de 0.58 de 1.53 cdef 1.78 fgh
2.0 0.75a 1.50bc l.75ef 0.55e 1.28defgh 1.70gh
0 0.75 a 1.75 abc 2.50 de 0.58 de 1.20 efgh 1.25 i
0.5 0.75 a 2.75 a 6.25 a 1.03 a 2.45 a 3.38 a
7.5 1.0 0.75 a 2.75 a 5.25 ab 0.95 ab 1.75 bcd 2.23 bcd
1.5 1.0a 1.75 abc 4.25bc 0.88 abc 1.88bc 2.l0bcdef
2.0 l.0a 1.75 abc 2.25 def 0.78c 1.3odefgh 2.05 bcdefg
0 1.0a 1.25bc 2.25def 0.75c 1.05 fghi 1.93 cdefg
0.5 0.75a 2.00abc 2.50de 0.73cd 1.60bcde 2.l8bcde
10.0 1.0 0.75 a 2.00 abc 2.50 de 0.80 bc 1.48 cdefg 2.28 bc
1.5 0.5ab 1.50bc 3.25cd 0.73cd 1.35defgh 1.98bcdefg
2.0 0.75 a 1.50 bc 2.50 de 0.73 cd 1.23 efgh 1.90 cdefg
LSD value (0.Ol) 0.61 1.05 1.26 0.16 050 0.39
CV (%) 38.88 34.55 26.71 16.06 19.27 10.49
272 KARIM et al.
(1996) obtained same results from 5.0 mgL-1 BAP. The lowest number of leaves
(0.00, 0.50, and 2.25 at 10, 20, and 30 DAI leaves per explant) were obtained
from control treatment (Table 2).
Table 2. Effect of different concentrations of BAP and NAA on leaf number and leaf
length of banana at different days after inoculation.
Treatment Shoot number Shoot length (cm)
BAP NAA 10 DAI 20 DAI 30 DAI 10 DAI 20 DAI 30 DAI
(mgL-1) (mgL-1)
0 0.00 e 0.5 c 2.25 d 0.50 defg 0.55 h 0.95 i
0.5 1.50 cd 2.25 b 4.75 c 0.58 cdef 1.65 fg 2.03 gh
0 1.0 1.75bcd 2.75ab 4.50c 0.60bcde 1.70efg l.95h
1.5 1.75bcd 3.00ab 5.50bc 0.70abc 1.8obcdefg 2.00gb
2.0 1.50cd 2.25b 5.50bc 0.46defg 1.60g 2.08gh
0 1.50cd 2.50b 4.75c 0.5odefg 1.55g l.95h
0.5 1.75 bcd 3.00 ab 5.00 c 0.63 bcd 2.03 bcd 2.58 efg
2.5 1.0 2.25 abc 3.25 ab 5.50bc 0.600bcdc 1.95 bcdef 2.53 efgh
1.5 1.50cd 2.75ab 5.75abc 0.45efg 2.075bc 2.43fgh
2.0 2.00 abcd 3.25 ab 5.50 bc 0.45 efg 2.05 bcd 2.28 gh
0 1.25d 2.5b 4.75c 0.35g 1.68fg 2.15gh
0.5 2,50 ab 3.25 ab 5.25 c 0.75 ab 2.60 a 3.60 bc
5.0 1.0 2.75 a 4.00 a 6.75 ab 0.70 abc 2.05 bcd 3.10 cde
1.5 2.25 abc 3.25 ab 5.50 bc 0.50 defg 2.03 bcd 2.93 def
2.0 1.75 bcd 2.75 ab 5.25 c 0.50 defg 2.00 bcde 2.98 def
0 2.00 abcd 3.25 ab 5.25 c 0.43 fg 1.65 fg 2.23 gh
0.5 2.5 ab 3.25 ab 7.00 a 0.85 a 2.70 a 4.23 a
7.5 1.0 1.75 bcd 2.75 ab 5.25 c 0.70 abc 2.10 b 3.70 ab
1.5 2.00abcd 3.00ab 5.75abc 0.S5cdef 1.95bcdef 2.58efg
2.0 2.00 abcd 2.5 b 4.50 c 0.55 cdef 1.76 cdefg 2.28 gh
0 1.50cd 3.25ab 4,75c 0.S3def 1.53g 2.15gh
0.5 2.00 abcd 2.5 b 4.50 c 0.50 defg 1.75 defg 2.98 def
10.0 1.0 1.50cd 3.25 ab 5.25 c 0.43 fg 1.93 bcdef 3.23 bcd
1.5 2.25 abc 2.75 ab 4.75 c 0.45 efg 1.95 bcdef 3.23 bcd
2.0 1.75 bcd 2.75 ab 4.75 c 0.45 efg 1.60 g 3.08 cde
L.SD value (0.01) 0.97 1.28 1.39 0.16 0.31 0.58
CV (%) 28.71 24,22 14.46 15.12 8.86 11.86
274 KARIM et al.
The MS medium supplemented with BAP and NAA showed different results
for leaf length, which was significantly influenced by different concentrations.
The longest leaves were produced by the concentration of 7.5 mgL-1 BAP + 0.5
mgL-1 NAA (0.85, 2.70, and 4.23 cm at 10, 20, and 30 DAI, respectively, which
was statistically significant. Statistically identical results was observed in 7.5
mgL-1 BAP + 1.0 mgL-1 NAA at 20 DAI (2.10 cm) and 30 DAI (3.70 cm).
Rahaman et al. (2004) observed the similar result. They obtained longest leaf in
the treatment 5.0 mgL-1 BAP (3.62 cm) followed by 1.5 mgL-1 NAA and 4.0
mgL-1 BAP (3.40 cm) using BARI Banana-I. They also found shortest leaf in 2.0
mgL-1 BAP. The results of present experiment agree with the findings of Khanam
et al. (1996) who obtained longest leaves in banana on MS medium
supplemented with 25 µM BAP treatments.
Table 3. Effect of different concentrations of IAA and IBA on root number of
multiple shoot of banana at different days after inoculation.
Treatment Vigor of Root number
-1
IAA (mgL ) IBA (mgL ) -1 regenerated 10 DAI 20 DAI 30DAI
root
0 + 0.00 e 0.00 e 0.00 f
0.5 + 1.50 cd 2.00 d 3.25 e
0
1.0 ++ 2.25 bc 2.25 d 3.50 de
1.5 ++ 2.25 bc 2.50 cd 3.50 de
0 +++ 2.75 ab 2.25 d 3.25 e
0.5 +++ 3.50 a 4.50 a 6.50 a
0.5
1.0 ++ 3.25 ab 3.50 abc 6.00 ab
1.5 ++ 3.25 ab 4.00 ab 5.00 bc
0 + 1.00 de 2.50 d 3.25 e
0.5 + 2.75 ab 2.75 cd 3.75 de
1.0
1.0 ++ 1.SOcd 3.O0bcd 4.O0cde
1.5 + 1.25 cd 3.50 abc 4.50 cd
LSD value (0.01) 1.03 1.16 1.16
CV(%) 25.36 22.19 15.51
+ = Less vigorous growth, ++ = Good growth and vigor, +++ = Best growth and vigor
other treatments (Table 5). The treatment 0.5 mgL-1 IAA + 1.0 mgL-1 IBA
produced 6.0 roots per explant at 30 DAI, but at 20 DAI, 3.50 roots was
produced per explant. Vigorous roots of in vitro grown plantlet on MS media
supplemented with 0.5 mgL-1 IAA + 0.5 mgL-1 IBA was shown in plate 10 (a &
b). The present results are similar with the findings of Gubbuk and Pekmezci
(2001). Molla et al. (2004) obtained 8.28 roots per plantlet on 0.5 mgL-1 IBA
followed by 6.33 roots in 0.6 mgL-1 BAP that was similar to the present work.
The results were also similar with the findings of Khanam et al. (1996) but
Rahman et al. (2004) obtained only 2.88 roots per explant at 30
DAI by 3.0 mgL-1 NAA.
Table 4. Effect of different concentrations of IAA and IBA on root length of
multiple shoot of banana at different days after inoculation.
Treatment Root number
-1 -1
IAA (mgL ) IBA (mgL ) 10 DAI 20 DAI 30DAI
0 0.00 e 2.00 ef 2.00 f
0.5 1.08 d 1.88 f 2.30 e
0
1.0 1.08d 2.3Ode 3.15d
1.5 1.13d 2.45d 3.08d
0 l.23d 1.60f 2.08e
0.5 2.93 a 4.63 a 5.88 a
05
1.0 2.55 ab 3.88 b 4.83 b
1.5 3.03 a 3.85 b 4.88 b
0 1.80 c 2.33 de 3.45 cd
0.5 1.80 c 2.35 de 3.48 cd
10
1.0 2.O8bc 3.15c 3.75c
1.5 2.lSbc 2.70d 3.70c
LSD value (0.01) 0.49 0.42 0.55
CV(%) 16.28 7.95 7.57
statistically significant. The second highest result (3.03, 3.85, and 4.88 cm at 10,
20, and 30 DAI) was observed with 0.5 mgL-1 IAA and 1.5 mgL-1 IBA. A similar
result was obtained by Molla et al. (2004) where they got 2.60-5.67 cm root
length in 0.5 mgL-1 IBA. Habiba (2002), Khanam et al. (1996) and Ali (1996)
also got more or less same observation.
Establishment of plantlet
Meristem derived plantlets were transferred to poly bags containing ground
soil:cow dung mixture (1:1) for acclimatization and hardening in room
temperature (28-30°C) and a good number of established plant (95%) is ready for
planting in the field.
C. Multiple shoots with 7.5 D. Multiple shoots with 7.5 mgL-1 BAP+0.5
mgL-1 BAP+0.5 mgL-1 NAA mgL-1 NAA at 2nd subculture.
at 30 days after inoculation.
IN VITRO PROPAGATION OF BANANA 277
E. Multiple shoots with 7.5 F. Multiple shoots with 7.5 mgL-1 BAP+0.5
mgL-1 BAP+0.5 mgL-1 mgL-1 NAA at 4th subculture.
rd
NAA at 3 subculture.
Fig. 1. Shoot regeneration from meristem derived of banana cv. BARI-1
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