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Tree Genetics and Molecular Breeding 2014, Vol.4, No.1, 1
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jackfruit (Roy et al., 1993; 1996). Acclimatization
procedures have been attempted to increase
ex vitro
survival of plantlets upon transplanting (Roy et al.,
1990; 1996; Amin and Jaiswal, 1993; Noweret et al.,
2005; Pati et al., 2013). The
in vitro
raised plants
showed profused growth when transplanted in soil.
The protocol establishes here is highly reproducible
for the production of plantlets which is not possible
with conventional methods. Plant growth regulators
and the physiological activity of the explants are very
important for successful regeneration. The plants that
were under further evaluation for fruiting and quality
characters. The plant regeneration protocol established
in this investigation may facilitate future research in
genetic transformation in lakoocha and related genus.
3 Materials andMethods
3.1 Establishment of culture
New shoots of 15~20 cm sizes from 8 year old tree of
Artocarpus lakoocha
Roxb were defoliated and
collected in an aqueous solution of ascorbic acid
(5.78 µM) and brought to the laboratory for further
processing. The nodal explants were washed
thoroughly under running tap water for 1h and treated
with Reidomyl 0.1%+Timentin 1.05 mM in 100 ml of
distilled water and 2 drops of Tween-20 for one hour
to avoid bacterial and fungal pathogens. The segments
were washed 3~4 times using sterile distilled water. A
pre-treatment with chilled antioxidant solution
comprising ascorbic acid (5.68 mM) and
Polyvinylpyrollidine (25 µM) for 1 h was also
followed in order to minimize
in vitro
oxidative
browning due to phenolic compounds. Then the
explants were rinsed with sterile double distilled water
and taken to laminar hood for surface sterilization.
Shoot buds were dipped in 70% ethanol for 30
seconds followed by air dried. The explants were
treated with 0.1% HgCl
2
for 8 minutes and then
washed (3~4 times) with sterile distilled water and
blotted dry on pre sterilized filter paper sheets.
Surface sterilized explants were placed vertically on
MS (Murashige and Skoog, 1962) medium supple-
mented with BAP (0, 2.22 µM, 4.44 µM, 8.88 µM and
13.33 µM), GA
3
(0.57 µM), ADS (2.71 µM) and IAA
(1.14 µM and 2.28 µM) for shoot bud induction and
multiplication. All the media were added with 0.7%
(w/v) agar and 3% (w/v) sucrose. The medium was
changed 2~3 times over the first 10~14 days of the
study to control phenolics exudates to establish
cultures. Observations were recorded on morph-
genetic characters such as days taken for bud break,
number of leaf per explant, length of axillary shoots
(cm) and average number of microshoots per explant
after 30 days of culture of the explants.
3.2
In vitro
rooting in regenerated shoots
Microshoots obtained
in vitro
were subjected to auxins
and cytokine such as IBA (0, 9.85 µM, 14.77 µM and
19.70 µM), IAA (1.14 µM) and BAP (0.89 µM) in half
and full strength of MS basal medium supplemented
with sucrose 3%, activated charcoal 500 mg/l and agar
0.7%. Data were recorded on per cent rooting, number
of roots, and length of roots. All cultures were
incubated at (25±2)
0
C and were exposed to a
photoperiod of 16/8 hours light and dark cycling in the
culture room by cool white fluorescent tubes
(80
mol m
2
s
-1
) with 70%±5% relative humidity.
3.3 Hardening of
in vitro
plantlets
Eight week old rooted shoots, were shifted to six inch
plastic cups for acclimatized filled with autoclaved
Sand:Soil:Peat moss (1:1:2) and drenched with ½
strength MS nutrients. The plastic cups was covered
with transparent polyethylene bags to prevent excess
water loss for 3 weeks in growth room at (25±2)
0
C
and then transferred to shade net house
(100
mol m
2
s
-1
; 50% shade 70%±5% RH) for
secondary hardening. Survival percent were
determined before field plantation. When 6~8 leaves
emerged in the plants, they were shifted to field.
References
Abd El-Zaher M.H., 2008, Studies on micro-propagation of jackfruit
1-behaviour of the jackfruit plants through the micropropagation stages,
World Journal of Agricultural Sciences, 4(2): 263-279
Ali N., Mulwa R.M.S., Norton M.A. et al., 2003, Micropropagation of
guava (
Psidium guajava
L.), J. Hort Sci. Biotechnol., 78: 739-741
Amin M.N., and Jaiswal V.S., 1993,
In vitro
response of apical bud explants
from mature trees of jackfruit
(Artocarpus heterophyllus
Lam.), Plant
Cell Tiss. Org. Cult. 33: 59-65
Ashrafuzzaman M., Sukarnakar, Dilafroza K. and Shamsul Haque P, 2012,
In vitro
regeneration and multiplication of Jack fruit (
Artocarpus
heterophyllus
L.), Res. J. Biol., 2(2): 59-65
Avato P., Bucci R., Tava A., et al., 2006, Antimicrobial activity of saponins