International Referred Reseach Journal,February,2011 ISSN-0975-3486 RNI: RAJBIL 2009/300097 VOL-I *ISSUE 17

Research PaperEnvironmental Science

EFFECT OF SALICYLIC ACID ON

TRIGONELLINE PRODUCTION IN TRIGONELLA
FOENUM- GRAECUM L. CELL
SUSPENSION CULTURE.
* Laxmi Mathur ** Rajesh Kr. Yadav

February, 2011

*-** Dept. of Environmental Science, S.S. Jain Subodh (PG) College, Jaipur
A B S T R A C T
Trigonelline (a pyridine alkaloid) was extracted from callus of Trigonella foenum-graecum. Cell suspension
culture of T. foenum-graecum was prepared to study the effect of elicitation. Two concentration doses of
salicylic acid (viz. 0.025 mM and 0.05 mM) were used as elicitor in the cell suspension culture. A two fold
increase in trigonelline production was observed at a concentration dose of 0.025 mM, while an increase
was also observed at 0.05mM concentration dose of salicylic acid.
Key words: Trigonelline, Trigonella foenum-graecum, Cell suspension culture, Elicitation, Salicylic acid.
Introduction:
Secondary metabolites are the compounds which play
an important role in plant defense mechanism.
Trigonelline, one of the pyridine alkaloid is widely
distributed in the plant Kingdom (Taguchi et. al., 1985).
Elicitation can enhance the efficiency of product
accumulation in cell cultures reducing the time required
to obtain a product (Wiznsama et. al., 1985).
Trigonella foenum-graecum L. commonly
known as fenugreek (Methis) is a rich source of
trigonelline. Enhancement in trigonelline content in
cell suspension culture of T. foenum-graecum using
salicylic acid as an elicitor showed a significant increase
at dose level of 0.025 mM and 0.05 mM concentrations
of salicylic acid as compared to normal callus.
Material and Methods:
Cell suspension cultures of T. foenum-graecum
were grown in liquid MS medium. Salicylic acid (Merck
India Ltd.) was used as an elicitor, two concentration
doses of salicylic acid (0.025 mM and 0.05 mm) were
added in both the sets of experiment, separately by
using erilsons micropipette.
The flasks were incubated on shaker for 24
hours of incubation; callus tissue was weighed and
separated from the medium by filtration through
Whatmann filter paper. Both tissue and culture medium
were used for extraction and estimation of their
trigonelline content.
The dried callus samples and culture medium
were extracted and analysed according to method of
Kogan et. al. (1953). The extracted samples were
R

I S

dissolved in 0.5 ml methanol and co-chromatographed

on tlc along with standard trigonelline.
Results:
The effect of low concentration doses (0.025
mM and 0.05 mM) of salicylic acid on the production
of trigonelline was observed with control. The callus
growth was not effected throughout the experiment,
while a very significant two fold increase (0.59%) in
trigonelline content was observed in 0.025 mM
treatment dose of salicylic acid over the control. The
trigonelline content was also increased (0.39%) in 0.05
mM (Table-1).
Table 1
Effect of different dose treatment of Salicylic acid on
trigonelline content in T. foenum-graceum cell
suspension culture.

Control

Salicylicaciddose
0.025mM
0.05mM

Trigonelline
(%)

0.27

0.59

0.39

Discussions:
Elicitor in cell cultures offer a novel approach
for rapid accumulation of certain secondary metabolites
by reducing time required to obtain the efficiency of
product (Kamal et. al. 1995). Rokem et. al. (1984) reported
70% increase in disogenin content from Dioscorea
deoltoidea cell cultures by using homogenate of
Rhizopous arrhizus.Salicylic alcohol and salicylic acid
are the basic compound of salicylate an important group
of pharmaceutical agents.
N

I O

137

International Referred Reseach Journal,February,2011 ISSN-0975-3486 RNI: RAJBIL 2009/300097 VOL-I *ISSUE 17

It has been observed that application of

exogenous salicylic acid induces expression of defense
related genes and provides partial protection against
pathogen.Ward (1991) reported salicylic acid plays an
important role in disease resistant in tobacco.In the
present study two months old callus culture of T.
foenum-graecum were fed with different treatment
doses of salicylic acid in the medium. Trigonelline

recovery in the tissue was highly significant in

comparison to control.This increase in trigonelline
content can also be co-related with the studies of Kamal
et. al. (1995), that the salicylic acid might be acting as
a signal compound in the induction of plant defense
mechanism thereby increasing the production of
secondary metabolites specifically trigonelline in the
present investigations on T. foenum-graecum.

R E F E R E N C E
Kamal, R., Moreno, P.R. H. and Verpoorte, R. 1995. Effect of elicitors on production of alkaloid and enzymes activities of
Catharanthus roseus suspension cultured cells. Plant Cell, Tissue and Organ Culture.Rokey, J.S., Schwarzberg, J. and Godbetry,
J. 1984. Autoclaved fungal mycelia increase disogenin production in cell suspension culture of Dioscorea delotoiedea Plant
Cell Rep. 3; 159-160.Taguchi, H., Muneto, S. and Shimabayushi, Y. 1986. Content of quinolic acid trigonelline and M. methyl
nicotinamide in various food and thermal conversion of acid and nicotinamide. Betamin 60(11); 537-546.Warel, E.R., Ukhers,
S.J. William, S.C. Diches, S.S., Wiederhol, D.L., Alexander, D.C., Ahil-evrg, P., Metrauo, J.P. and Ryeul, J.A. 1991. Plant Cell
3; 1085-1094.Warren, L. 1959. The Triobarbituric acid assay of sialic acids! J. Biol. Chem.,234; 1971.

138

I S

I O