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International Journal of Research in Plant Science

Universal Research Publications. All rights reserved

ISSN 2249-9717
Original Article
In vitro propagation of Curcuma longa (Turmeric)
22. Gomathy1, M. Anbazhagan2* and K. Arumugam2
1. Research and Development Center, Bharathiar University, Coimbatore-641046.
2. Department of Botany, Annamalai University, Annamalainagar-608002, Tamil Nadu, India.
*Corresponding author E-mail: anbungm@yahoo.co.in
Received 24 January 2014; accepted 03 February 2014
Abstract
Experiments were conducted for the standardization of in vitro culture technique for the mass propagation of Curcuma
longaLinn.(Syn. C. domesticaValet.) is one of the important spice crops. It is a rhizomatous perennial belonging to family
Zingiberaceae. Scooped rhizomatous buds were used as explants and they were cultured on MS medium supplemented
with different concentrations of BAP, Kn and IAA both in individual and in combined form for shoot inductions and the
best results were obtained from MS medium supplemented with BAP+IAA at the concentration of 2.0mg/l and 0.5mg/l
respectively. Best root formation of in vitro developed shoots could be achieved on halt strength MS medium supplemented
with IBA at concentration 1.0mg/l. The in vitro developed plantlets were transferred to pot and they were grown in
greenhouse for hardening and finally they were planted in the open field. Around 90% of plants were successfully
established in natural field condition.
2011 Universal Research Publications. All rights reserved
Key words: Curcuma longa Linn.;in vitroPropagation; MS medium; BAP; Kn; IAA; IBA; NAA.
turmeric is being used in propagation has limitation
Introduction Indian system of medicine as on account of smaller size
Turmeric (Curcuma longa Linn. Syn. C. domesticaValet.) isstomachic,carminative, ofrhizomes in the first
one of the important spicecrops. It is a rhizomatous perennial blood purifier, vermicide and generation of plantlets and
belonging to family Zingiberaceae. Underground antiseptic. Wound healing high cost [7]. Still,
modifiedstem, the rhizomes are processed and used for antiseptic property thistechnique can be used
various purposes such as condiment, dye, drug and cosmetic ofturmeric is well known to for crop improvement
in addition to its use in religious ceremonies. India is a Indians since long. studies. There is need to
Curcumin is the main standardize therequirements
leading producer and exporter of turmeric in the world.
biologically of in vitro techniques for
Andhra Pradesh, Tamil Nadu, Orissa, Karnataka, West
activephytochemical local varieties.
Bengal, Gujarat, Meghalaya, Maharashtra, Assam are some
compound of turmeric with M
of the important states cultivates turmeric, of which, turmeric
wide range of therapeutic a
can be grown in diverse tropical conditions from sea level to effects. In recent t
1500m ASL, at a temperature range of 20-35C with an yearspharmacological e
annual rainfall of 1500mm or more, under rain fed or properties and actions of r
irrigated conditions. Though it can be grown on different curcumin have been widely i
types of soils, it thrives best in well drained sandy or clay researched and itsbeneficial a
loam soils with a pH range of 4.5-7.5 with good organic effects have been well l
s
status [1]. established [3].
As a spice, turmeric is used for its colouring Advent of in vitro a
properties and aroma. It is an importantingredient of curries techniques has opened new n
and curry powder. Important constituents of turmeric avenues in improvement of d
rhizomes arecurcumin (2-8%) and volatile oil (5-6%) [2,4,5]. crop plants.Tissue culture M
Turmeric is used for dying wool and cotton fabrics.It is also techniques have been e
employed as a colouring material in pharmacy, confectionarystandardized for many t
and food industry, aswell as in paints and varnishes. In horticultural crops. The in h
cosmetics, turmeric is an inexpensive and indigenousbeauty vitropropagation of turmeric o
d
aid.Traditionally was first attempted by [6]. s
However,
commercialapplication of S
this technique for o
urce of Explants fromrhizomatous buds for After excision, the surface
For this study, the field grown Curcuma longawere collectedshoot initiation. sterilization, the explants
from the Department of Agriculture, Annamalai University,Surface sterilization were
Annamalainagar and South India. Explants were collected
 International Journal of Research in Plant Science 2014; 4(1): 30-33
30
 Figure 1. The successive stages of in vitro propagation of Curcuma longa
A Initiation of shoot formation, B Initiationof multiple shoot formation, C Development of more number of multiple soot,
D Root formation from regenerated shoot, E Hardening in plastic pot, F Establishmentsregenerated plants
was served as control. Table 1. Effect of plant
After 35-40 days of growth regulator on shoot
primarily rinsed in tap water for 20 minutes culturing, the multiple induction
followed gently rinsed with 70% ethanol for shoots were separated
60 seconds and with 5% sodium hypochloride into pieces and the Sl. No. Plant Growth Regulator S
solution for 10 minutes. After each step of separation was done at (mg/l) (M
sterilization, the explants were washed with the base of multiple 1 Nil 0
sterile double distilled water for three times. shoots and they were 2 BAP 0.5 2
Further sterilization procedures were carried transferred to 500ml 3 BAP 1.0 3
out in laminar air flow chamber by using culture bottle containing 4 BAP 1.5 5
0.1% HgCl2 for 5 minutes. The explants were 50ml of the same kind of 5 BAP 2.0 9
then rinsed five times with sterile distilled medium 6 BAP 2.5 7
water. Finally, in the laminar air flow 7 BAP 3.0 5
chamber, the explants were cut into small 8 Kn 0.5 1
pieces. 9 Kn 1.0 3
Inoculations 10 Kn 2.0 4
11 Kn 3.0 4
After complete sterilization and slicing, the
12 BAP 1.0 + Kn 0.2 3
explants were inoculated in MS medium [8],
supplemented with cytokinins and auxins 13 BAP 1.0 + Kn 0.5 5
used either singly or in combination. The pH 14 BAP 2.0 + Kn 0.2 6
of the medium was adjusted to 5.8 and prior 15 BAP 2.0 + Kn 0.5 10
to autoclaving, 0.7% agar (Himedia, 16 BAP 1.0 + IAA 0.2 8
Mumbai) was added to the medium. Then, 17 BAP 1.0 + IAA 0.5 11
autoclaving was done at 121C and 15psi for 18 BAP 2.0 + IAA 0.2 16
20 minutes. 19 BAP 2.0 + IAA 0.5 9
Establishment of culture to get a more number of
After inoculation, the cultures were shoots. For root initiation,
maintained at a temperature of 252C with a the shoots were
photoperiod of 16hrs per day. Lighting of transferred to half
strength MS medium
80Em-2s-1was supplied by using cool and supplemented with IAA,
white fluorescent tubes (Philips India Ltd.). IBA and NAA.
Various types of plant growth regulator Hardening
(PGR), viz. BAP, Kn and IAA were added
Healthy plantlets with 4
with MS medium either alone or in
combination for better shoot induction (Table- to 5cm long roots were
1). The MS medium without adding of PGR individually removed
from the culture tubes.
After washing their roots carefully with tapsand, soil and greenhouse for hardening.
water, plantlets were transplanted into 10cm vermicompost (1:1:1) and The plants
diameter plasticpots containing a mixture of they were placed in the
International Journal of Research in Plant Science 2014; 4(1): 30-33
31
Table 2. Effect of different concentrations of IAA, IBA and NAA on root formation
Sl. Concentration of Percentage of root No. of roots per Average length of root per shoot
No. auxins (mg/l) per shoot (%) shoot (Mean SE) (cm, Mean SE)
IAA
1. 0.2 68 5.00 0.31 2.40 0.18
2. 0.5 80 6.00 0.44 3.00 0.16
3. 1.0 85 10.40 0.74 4.10 0.31
4. 2.0 70 4.60 0.24 2.40 0.20
IBA
5. 0.2 72 5.20 0.37 3.00 0.17
6. 0.5 87 6.30 0.37 3.80 0.26
7. 1.0 95 12.10 0.80 4.90 0.31
8. 2.0 75 4.20 0.37 3.20 0.13
NAA
9. 0.2 55 3.40 0.50 1.94 0.16
10. 0.5 64 4.70 0.37 2.60 0.15
11. 1.0 72 6.40 0.37 3.50 0.20
12. 2.0 52 4.00 0.54 2.10 0.14
Kn 0.5mg/l worked
well for the shoot
were watered with half-strength MS salts solution proliferation and
every week and covered with a polythene bag for 2 elongation from the
weeks. Afterwards, the hardened plants were gradually same explants.
transferred to 20cmpots containing pure garden soil Root induction
and kept in the field for developing into mature plants. Themicro cuttingof
Statistical Analysis in vitroproliferated
All experiments were repeated five times. The effects shoots were
of different treatments were quantified and the data implanted on half
subjected to statistical analysis using SPSS software strength MS
[9]. medium
supplemented
Results and Discussion individually with
Explants selection and shoot induction IAA, IBA and NAA
concentration of
In this study, rhizomatous buds were excised from the 1.0, 2.0, and
plant species of Curcuma longaas explants sources, 3.0mg/l for root
and the rhizomatous buds were found best explants for initiation. Among
shoot induction on MS medium supplemented with them, maximum
different concentrations of BAP like 0.5, 1.0, 1.5, 2.0, number of root
2.5, and 3.0mg/l or Kn like 0.5, 1.0, 2.0, 3.0mg/l formation (95%)
individually. In another way, the MS medium was observed in
supplemented with the combined form of BAP+Kn medium
and BAP+IAA. In individual form, the BAP usingsupplementedstandardwith(1.
concentration above 3.0mg/l level caused poor shoot 0mg/l)IBA (12error.100.80,
induction in two explants studied. The more number Tableof-2) .the m The root
of shoot induction was observed in two explants induction was gradually
studied on MS medium supplemented with BAP decreased with increasing
concentrations of auxin types.
2.0mg/l + IAA 0.5mg/l. When observations were No root formation was
made on the cultures of 35 days period, development
observed auxin free
of 16 (16.200.37, Table-1) shoots were observed basal medium.
from a single rhizomatous bud explants, while 14 Similar types of
(14.000.31, Table-1). results were found
by earlier workers
This in vitro propagation studies confirmed in the same species
the importance of PGR in the initiations of callus, Acclimatization of
shoot, root and on the whole the regeneration of plant. regenerated plants
In this way, the two cytokinins namely, BAP and Kn
For acclimatization,
used in this study, the BAP proved better one than Kn
the plantlets were
in shoot induction from rhizomatous bud. On the other
taken out from the
hand, mineral nutrients are being as the basic
component of culture media play a vital role in rapid culture tubes when
growth of tissue and the quality of morphogenesis of the roots were
tissue [10]. In this study, the synergistic effect of both partially brown in
BAP + IAA at concentration 2.0 and 0.5mg/l colour and root
respectively, were found best in regenerating shoot tip portion was washed
explants. Similarly the combination of BAP 2.0mg/l with tap water to
and remove the attached
medium. Then they
were transferred to pots containing sand, of Botany, production in
soil and vermicompost in the ratio 1:1:1 Annamalai turmeric. Plant
and they were placed in greenhouse for University for Cell, Tissue
hardening for the period of four weeks. having and Organ
rendered the Culture.64: 5-
The use of sufficiently porous substratum necessary
that allows adequate drainage and aeration 11.
facilities.
has been recommended for fast References 2. Panda, M.K., S.
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and R.S. Assessment of
transferred to natural field conditions for Nadgauda genetic stability
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. 2001. micropropagated
study, around 85% of plants were thrived Factors
well in natural situation. affecting plants of Curcuma
Acknowledgements in vitro longa by
The authors are also grateful to Dr. R. microrhiz
Paneerselvam, the Professor, Department ome
 International Journal of Research in Plant Science 2014; 4(1): 30-33
32
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o t: Nil;
S f Conflic
o s t of
u u interes
r p t: None
c p declare
e o d 33 International