IJSAT: ISSN 1815-1272, Volume 10 Issue 8 2014

 
Title code: IJSAT/140801
Int. J. Sustain. Agril. Tech. 10(8): 01-04, August 2014
An online Journal of “G-Science Implementation and Publication”, website: www.gurpukur.com or www.gscience.net

KINETIN (KIN) AND INDOLE-3- ACETIC ACID (IAA) ON IN VITRO SHOOT AND ROOT
INITIATION OF TUBEROSE
M. R. ALI, H. MEHRAJ and A. F. M. JAMAL UDDIN*

Department of Horticulture, Sher-e-Bangla Agricultural University, Dhaka-1207, Bangladesh

*Corresponding author’s Email: jamal4@yahoo.com.
ABSTRACT
An experiment was conducted at the Tissue Culture Laboratory, Department of Biotechnology, Sher-e-
Bangla Agricultural University, Dhaka Bangladesh from July 2011 to September 2012 to evaluate the
effect of different concentrations of KIN and IAA on In vitro shoot and root initiation of tuberose.
Experiment consisted seven treatments viz. T1: Simple MS, T2: MS+0.5 mg/L, T3: MS+1.0 mg/L, T4:
MS+1.5 mg/L, T5: MS+2.0 mg/L and T6: MS+2.5 mg/L for both KIN and IAA following Completely
Randomized Design with five replications. Early shoot initiation (10.1 days), leaf initiation (12.2 days),
root initiation (14.92 days) was found from T3. On the other hand, maximum number of leaves
(3.00/plantlet), longest leaves (5.48 cm) and maximum number of roots (3.05/plantlet) was found from T3
at 60 DAI.

Keywords: KIN, IAA, Shoot, Root and Tuberose.

INTRODUCTION
Tuberose (Polianthes tuberosa L.) is a bulbous plant belongs to Amaryllidaceae family normally
propagated by bulb. The smaller the bulb sizes take more time for plant to flower (Roostika et al.,
2005). Now a day, plant tissue culture techniques as alternative to conventional propagation are being
applied for clonal propagation of tuberose which required nutrients and plant growth regulators
(BadrDin and Pierre, 1995). In culturing of plant cells, plant growth regulators are used to produce
callus growth, multiplication and rooting. Four broad classes of growth regulators, namely Auxin,
Cytokinin, Gibberellin and Abscisic acid are important in plant tissue culture (Razdan, 2002). Plant
growth regulators are the critical medium components in determining the development pathway of plant
cells. It is well known that several factors can affect in vitro Micropropagation (George and Debergh,
2008). Most important of these parameters are the plant growth regulators content in the culture media
(Gomes and Canhoto, 2003). Plant growth regulators act like signals to stimulate, inhibit or regulate
growth in the developmental programs of plants (Mercier et al., 1997). Thus, experiment was aimed to
study the effect of KIN and IAA on in vitro shoot and root initiation of tuberose.
MATERIALS AND METHODS
An experiment on in vitro regeneration of tuberose was conducted in the Tissue Culture Laboratory,
Department of Biotechnology, Sher-e-Bangla Agricultural University, Dhaka, Bangladesh during July
2011 to March 2012. Disease free bulbs of tuberose were collected from Bangladesh Agricultural
Research Institute (BARI), Gazipur for explants. Murashige and Skoog (MS) (1962) medium were used
with different hormonal supplements as culture medium for shoot and root initiation. The culture tube
containing medium was sterilized in an autoclave at temperature of 121°C for 20 minutes at l5 psi
pressure. The medium was then cooled at room temperature before use. The lead of cabinet then was
closed well and UV was switched on while turning off the air flow. The UV light of cabinet was left on
for 30 minutes and the surface area is wiped with 70% ethanol. The temperature of culture room was
maintained within 25±1°C with the help of air conditioner with 16 hours photoperiod and 3000 lux light
intensity by using white florescent lights. Buds were first sterilized with 70% (v/v) ethanol for few
seconds after that surface sterilized by immersing in 0.1% HgCl2 solution for 4-5 minutes and then was
washed several times with sterilized distilled water. Then the bud was cut aseptically into small pieces
with the help of sharp aseptic knife and the small segments of bud were used as explants. Three to five
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 IJSAT: ISSN 1815-1272, Volume 10 Issue 8 2014
 
Title code: IJSAT/140801
segments of explants were directly inoculated into each test tube containing 10 ml of MS medium
supplemented with different callus induction hormonal combinations of IAA and KIN. Single factorial
experiment consisted seven treatments viz. T1: Simple MS, T2: MS+0.5 mg/L, T3: MS+1.0 mg/L, T4:
MS+1.5 mg/L, T5: MS+2.0 mg/L and T6: MS+2.5 mg/L for both KIN and IAA following Completely
Randomized Design with five replications. Data were taken on days to shoot initiation, days to leaf
initiation, days to root initiation, number of leaves/plantlet, length of the leaves/plantlet, number of
roots/plantlet and length of roots/plantlet (cm). The collected data on different parameters were
analyzed using a MSTAT-C package computer program. The analysis of variance was performed and
means were compared by Duncan Multiple Range Test (DMRT) at 5% level of significance.
RESULTS AND DISCUSSION
Effect of KIN on in vitro shoots initiation in tuberose
Days to shoot initiation and leaf initiation: Early shoot and leaf initiation was recorded on T3 (10.1 days
for shoot initiation and 12.2 days for leaf initiation) while late from T6 (14.5 days for shoot initiation
and 27.1 days for leaf initiation) (Table 1). It was observed that no shoot or leaf initiated in simple MS
medium (Plate 1a). Initiated shoot on 15 days on MS medium supplemented with 1.0 mg/l was shown
on plate 1b.
Number of leaves/plantlet: Maximum number of leaves was found from T3 (3.0/plantlet) while
minimum from T2 (1.54/plantlet) at 60 DAI (Table 1). Maximum numbers of initiated leaves were
shown on Plate 1c.
Length of the leaves/plantlet (cm): Longest leaves was found from T3 (5.48 cm) while shortest from T6
(0.25 cm) at 60 DAI (Table 1). Longest leaves were shown on plate 1c.

(a) (b) (c)

Plate 1. (a) No shoot initiation on T1, (b) Initiated shoot at 15 DAI on T3 and (c) Maximum number with
longest leaf at 60 DAI on T3.
Effect of IAA on in vitro root initiation in tuberose
Days to root initiation: Early root initiation was found from T3 (14.92 days) whereas late from T6 (34.16
days) (Table 2). Root initiation was not found from T1 (Plate 2a).
Number of roots/plantlet: Maximum numbers of roots were found from T3 (3.05/plantlet) while
minimum from T6 (1.52/plantlet) at 60 DAI (Table 2). Maximum number of initiated roots at 60 DAI
was showed in plate 2b.
Length of roots (cm): Longest root was found from T2 (1.69 cm) followed by T3 (1.03 cm) while
minimum from T6 (0.28 cm) at 60 DAI (Table 2).

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 IJSAT: ISSN 1815-1272, Volume 10 Issue 8 2014
 
Title code: IJSAT/140801

(a) (b)
Plate 2. (a) Root initiation was not noticed at 60 DAI in T1 and (b) Maximum number
of roots initiated at 60 DAI T3..
Table 1. Effect of different concentrated KIN on shooting of tuberose.
Days to Number of leaves/plantlet at different Length of leaves (cm) at different
Days to leaf day after inoculation (DAI) days after inoculation (DAI)
KIN (mg/L) shoot
initiation
initiation 15 30 45 60 15 30 45 60
T1 - - - - - - - - - -
T2 11.7c 24.7b 0.10e 0.53d 0.88d 1.54e 0.30b 0.77b 1.55b 2.23b
T3 10.1d 12.2d 0.55a 0.96a 1.37a 3.00a 0.77a 1.75a 4.54a 5.48a
T4 11.9c 22.4c 0.17d 0.66c 1.12c 1.75d 0.12c 0.32c 0.74c 1.14c
T5 13.4b 24.9b 0.35b 0.75b 1.17b 2.23b 0.00d 0.12d 0.27d 0.52d
T6 14.5a 27.1a 0.38c 0.70b 1.15bc 2.02c 0.00d 0.00e 0.11e 0.25e
LSD(0.05) 1.1 1.3 0.04 0.04 0.04 0.08 0.04 0.04 0.07 0.09
CV (%) 7.9 5.5 12.28 4.47 2.41 3.48 6.54 3.90 4.68 4.21
In column, figure with same letter(s) do not differ significantly at 5% level of significance according to DMRT.

Table 2. Effect of different concentrated IAA on rotting characteristics of tuberose.

Number of roots/plantlet at different days Length of roots (cm) at different days after
Days to root
IAA (mg/L) after inoculation (DAI) inoculation (DAI)
initiation
15 30 45 60 15 30 45 60
T1 - - - - - - - - -
T2 22.40d 0.33b 1.04b 1.98b 2.75b 0.90a 0.25a 0.77a 1.69a
T3 14.92e 0.47a 1.26a 2.21a 3.05a 0.50a 0.22a 0.52b 1.03b
T4 31.60b 0.00c 0.47c 1.09c 2.02c 0.00b 0.11b 0.32c 0.74c
T5 24.57c 0.00c 0.43c 0.90d 1.71d 0.00b 0.12b 0.24d 0.52d
T6 34.16a 0.00c 0.43c 0.75e 1.52e 0.00b 0.70b 0.12e 0.28e
LSD(0.05) 1.32 0.04 0.05 0.06 0.15 0.04 0.04 0.04 0.07
CV (%) 4.77 8.65 7.65 4.03 6.17 13.24 12.67 9.65 7.81
In column, figure with same letter(s) do not differ significantly at 5% level of significance according to DMRT.

CONCLUSION
It can be concluded that addition of 1.0 mg/L KIN and IAA with MS medium was best for the shoot and
root initiation respectively in tuberose.

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Title code: IJSAT/140801

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