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Table 1. Response of subcultured calluses obtained
from internode explants of chickpea to Kinetin and
IAA in MS medium.
Growth regulators
KIN 1 + IAA 1
KIN 2 + IAA 1
KIN 3 + IAA 1
KIN 4 + IAA 2
KIN 1 + IAA 2
KIN 2 + IAA 2
KIN 3 + IAA 2
KIN 4 + IAA 2
% of calluses
formed
shoots
50
30
20
10
-
60
50
30
No. of
shoots
per culture
4
4
3
2
-
15
10
5
Shoot
length
(cm)
3.00
5.00
4.00
6.00
-
2.50
3.00
3.50
upper surface of calluses (Fig. 1) and morphological
changes were observed after 5 weeks in culture when the
globular structure grew into adventitious shoot buds.
The shoot buds developed normally when the explants
were subcultured in MS medium containing different
combinations of Kinetin and IAA (Table 1).
On transfer to MS medium without any growth
regulator or with only kinetin (1-4 mg L -1 ) light green
calluses become dark green or blackish and friable but
no further morphogenic changes were noticed. At the
end of the 5th week of subculture partial browning and
darkening of external portion of calluses started and
consequently they became necrotic. When the calluses
were transferred to a medium containing Kinetin (1-4
mg L -1 ) and IAA (1-2 mg L -1 ) shoot proliferation started
within 2 weeks of subculture. Highest percentage (60%)
of calluses producing shoots and maximum number (15)
of shoots per culture were obtained on medium containing
2 mg L -1 kinetin and 2 mg L -1 IAA (Fig. 1).
In the present investigation it was observed that 2, 4-D
without cytokinin could induce calluses but for better
proliferation a cytokinin in combination with 2, 4-D was
required. It was also observed that length of shoots
decreased with the increase in shoot number. Similar
observation was made earlier (Kartha et al. 1981, Rao
and Chopra 1989, and Islam and Riazuddin 1993).
References
Baker, CM., and Wetzstein, H.Y. 1992. Somatic
embryogenesis and plant regeneration from leaflet of
peanut, Arachis hypogaea. Plant Cell Report 11:71-75.
Najaj, Y.P.S., and Dhangu, M.S. 1979. Regenerations
of plants from apical meristem tips of some legumes.
Current Science 48:906-907.
Islam, R., and Riazuddin, S. 1993. Shoot organogenesis
in chickpea Cier arietinum L. from callus cultures of
hypocotyal explants. Journal of Biological Sciences 1:
1-5.
Kartha, K.K., Pahl, P., Leung, N.L., and Morginshi,
L.A. 1981. Plant regeneration from meristems of grain
legumes. Soybean, cowpea, peanut, chickpea, and bean.
Canadian Journal of Botany 59:1671-1679.
Murashig, T., and Skoog, F. 1962. A revised medium
for rapid growth and bioassay with tobacco tissue cultures.
Physiol Plant 15:473-497.
Rao, B.G., and Chopra, V.L. 1989. Regeneration of
apical meriston, stem nodes and cotyledons of chickpea.
Indian Institute of Pulses Research 2:20-21.
ICPN 7, 2000 29