Effect of triacontanol on photosynthesis, alkaloid content and growth in opium poppy (Papaver somniferum L.)


Srivastava, N.K.; Sharma, S.

Plant Growth Regulation 9(1): 65-72

1990


The influence of different foliar applications of Triacontanol (Trio.) on growth, CO2 exchange, capsule development and alkaloid accumulation in opium poppy was studied in glasshouse conditions. Plant height, capsule number and weight, morphine content, CO2 exchange rate, total chlorophyll and fresh and dry weight of the shoot were significantly maximum at 0.01 mg/l Tria. At the highest concentration (4 mg/l) total chlorophyll, CO2 exchange rate and plant height were significantly inhibited. Thebaine and codeine contents remained unaffected at all the concentrations. The concentration of Fe, Mn, Cu in shoots were maximum at .01 and Zn at 0.1 mg/l Tria. Increase in shoot weight, leaf area ratio and chlorophyll content were significantly correlated with morphine content.

Plant
Growth
Regulation
9:
65-71,
1990.
©
1990
Kluwer
Academic
Publishers.
Printed
in
the
Netherlands.
Effect
of
Triacontanol
on
photosynthesis,
alkaloid
content
and
growth
in
opium
poppy
(Papaver
Somniferum
L.)*
N.K.
SRIVASTAVA**
&
SRIKANT
SHARMA
Central
Institute
of
Medicinal
and
Aromatic
Plants,
Post
Bag
No.
1,
P.O.
RSM
Nagar,
Lucknow-226
016
India.
**Author
for
correspondence
(Submitted
7
February
1989;
accepted
5
August
1989)
Abstract.
The
influence
of
different
foliar
applications
of
Triacontanol
(Trio.)
on
growth,
CO,
exchange,
capsule
development
and
alkaloid
accumulation
in
opium
poppy
was
studied
in
glasshouse
conditions.
Plant
height,
capsule
number
and
weight,
morphine
content,
CO,
exchange
rate,
total
chlorophyll
and
fresh
and
dry
weight
of
the
shoot
were
significantly
maximum
at
0.01
mg/1
Tria.
At
the
highest
concentration
(4
mg/1)
total
chlorophyll,
CO,
exchange
rate
and
plant
height
were
significantly
inhibited.
Thebaine
and
codeine
contents
remained
unaffected
at
all
the
concentrations.
The
concentration
of
Fe,
Mn,
Cu
in
shoots
were
maximum
at
.01
and
Zn
at
0.1
mg/1
Tria.
Increase
in
shoot
weight,
leaf
area
ratio
and
chlorophyll
content
were
significantly
correlated
with
morphine
content.
Key
words:
alkaloids,
CO,
exchange,
growth
regulator,
opium
poppy,
Papaver
somniferum
L.,
Triacontanol
1.
Introduction
Growth
promoting
effects
of
Triacontanol
(Tria.)
on
vegetables
[18],
cereals
[19,
5]
and
horticultural
crops
[8]
are
well
documented.
These
are
associated
with
increased
protein
content,
water
uptake
[10],
uptake
of
elements
[17]
and
photosynthetic
CO
2
fi
xation
[8].
Most
previous
studies
on
Tria.
have
concentrated
on
physiological
aspects
of
crops
leading
to
effects
on
com-
ponents
of
yield.
The
opium
poppy
(Papaver
somniferum
L.)
is
a
rich
source
of
opium
alkaloids
used
frequently
as
analgesic,
antitissive
and
antispasmodic
agents
in
modern
medicine.
It
is
also
grown
as
a
source
of
edible
seeds
and
oil
in
tropics
and
subtropical
countries.
The
capsule
is
the
most
important
plant
organ
being
the
major
deposition
site
of
latex
which
is
the
raw
material
for
subsequent
biosynthesis.
However,
poppy
straw
has
also
been
used
for
the
extraction
of
alkaloids
[13].
The
influence
of
growth
regulators
on
alkaloid
biosynthesis
and
ac-
cumulation
has
received
little
attention.
Nevertheless,
the
effect
of
some
*
CIMAP
Communication
No.
839.
66
growth
regulators
including
chlormequat
chloride,
dimethyl
sulphoxide
[6],
maleic
hydrazide
[22]
and
ethephon
[16]
on
alkaloid
constituents
in
poppy
have
been
attributed
to
changes
in
membrane
permeability,
affects
on
seed
development
due
to
suppression
of
embryogenesis
and
enhancement
of
crude
opium
yield.
Since
Tria.
has
been
reported
to
increase
dry
matter
production
in
several
species,
it
was
decided
to
investigate
its
possible
affect
on
the
inter
-relation-
ship
between
primary
and
secondary
metabolism
in
opium
poppy.
This
is
an
area
of
active
research
as
evidenced
by
recent
publications
on
crops
having
secondary
metabolites
of
economic
value
[9,
4,
21,
7,
20].
The
present
study
examines
the
effect
of
Tria.
on
alkaloid
biosynthesis
in
opium
poppy
and
the
relationship
between
alkaloid
production
(capsule
and
constituents)
and
physiological
parameters
related
to
productivity
including
CO
2
exchange
rate,
chlorophyll
production,
leaf
area
ratio,
biomass
production
and
ion
uptake
using
a
range
of
concentrations
under
controlled
conditions
in
a
glasshouse.
2.
Material
and
methods
Opium
poppy
plants
(Papaver
somniferum
L.
Cv.
Shyama)
were
raised
in
pots
fi
lled
with
purified
acid
-washed
silica
sand
by
the
method
of
Hewitt
[11]
modified
by
Agarwala
and
Sharma
[1].
Plants
were
given
between
200
and
500
ml
of
complete
Hogland
solution
[12]
periodically
and
were
maintained
in
a
glasshouse
at
ambient
temperature
(30-35
°C)
and
light
intensity
(800-
1000
µE/m
2
/sec),
the
latter
measured
by
a
LICOR
light
meter
(Model
188B).
Sprays
of
Tria.
at
0.001,
0.01,
0.1,
1.0,
2.0,
and
4.0
mg/1
were
applied
on
three
occasions
at
intervals
of
four
weeks
using
a
hand
held
sprayer.
Tween
20
(0.1%
w/v)
was
added
as
a
surfactant.
Care
was
taken
to
wet
both
leaf
surfaces
to
ensure
maximum
application.
Solutions
were
applied
to
groups
of
six
plants
in
three
replicates
with
one
set
serving
as
control
(no
spray).
Measurements
of
plant
height
and
fresh
and
dry
matter
were
recorded
three
weeks
after
the
last
spray.
Total
Chlorophyll
(Chl.)
content
was
measured
according
to
the
method
of
Arnon
[3]
using
80%
acetone
with
absorption
measured
on
a
Pye
Unichem
Spectrophotometer
model
SP
550
and
computed
for
Chl.
a
and
b.
CO
2
exchange
rate
(CER)
was
measured
by
the
CO
2
depletion
technique
using
the
third
leaf
and
was
analysed
by
a
LICOR
portable
photosynthesis
system
model
LI
6000
at
a
natural
light
intensity
of
1500
pE/m
2
/sec.
CER
was
computed
as
mg
CO,/m
2
/sec
and
converted
to
mg
CO
2
/dm
2
/hr.
Leaf
area
ratio
(LAR)
was
determined
ac-
cording
to
the
method
of
Radford
[15]
as
L/A
where
L
is
the
total
leaf
area
and
A,
the
dry
matter
of
shoot.
Leaves
were
separated
and
the
cumulative
67
area
measured
on
a
LICOR
model
LI
3000
area
meter.
Shoot
samples
were
dried
at
60-70°C,
milled
and
digested
in
HNO
3
-Hc10
4
mixture
[14]
for
determination
of
elemental
content.
Fe,
Mn,
Cu,
and
Zn
were
determined
on
a
Pye
Unichem
model
SP
2900
atomic
absorption
spectrophotometer.
Data
on
capsule
parameters
which
included
number/plant
and
weight
were
recorded
at
the
ripe
stage.
The
alkaloid
constituents
morphine,
codeine
and
thebaine
were
extracted
from
capsules
and
analysed
on
a
Water's
HPLC
[2].
The
individual
peaks
were
identified
by
coinjection
of
authentic
compounds.
The
experiment
was
carried
out
using
the
layout
of
a
randomised
com-
plete
block
design
of
three
replications.
Mean
values
of
each
character
were
statistically
analysed
for
significance
by
means
of
a
Least
Significant
Dif-
ference
(LSD)
test.
Linear
correlation
coefficients
were
computed
to
deter-
mine
the
inter
-relationships
among
various
characters.
3.
Results
and
discussion
Foliar
applications
of
Tria.
at
lower
concentrations
(0.01
and
0.1
mg/1)
stimulated
overall
growth
and
significantly
enhanced
morphine
content
and
capsule
weight
but
was
inhibitory
at
the
highest
concentration
(4
mg/1).
Plant
height
and
shoot
fresh
and
dry
weight
were
significantly
increased
at
all
concentrations
of
Tria.
application
(Figure
1),
the
association
among
200
-
180
--
160
E
1
-
1
140
120
6
L-
=
cc
100
g
0
'^
a
0
60
0
I
40
20
0
C
0
.1
1.0
2.0
4.0
mg
/
I
TR
IACONTANOL
20
18
8
0
E
.,
c
6
TI
)
/..)
4
Og
2
0
Fig.
1.
Effect
of
Tria.
on
plant
height,
shoot
fresh
wt.,
shoot
dry
wt.,
and
capsule
number.
LSD
values
at
5%
and
1%
are
4.8,
6.7
(plant
height);
21.0,
29.5
(fresh
wt.);
4.3,
5.9
(dry
wt.);
1.1,
1.8
(capsule
number),
respectively.
68
these
characters
being
significantly
positive.
However,
at
concentrations
higher
than
0.1
mg/1
these
characters
exhibited
a
decreasing
trend.
Among
the
physiological
characters,
total
Chl.,
LAR
and
CER
increased
significantly
till
0.01
mg/1
and
then
decreased.
However,
Chl.
a/b
ratio
showed
an
inverse
relation
indicating
the
greater
influence
of
Tria.
On
Chl.
a
(Figure
2).
Both
CER
and
LAR
were
positively
correlated
with
shoot
fresh
weight
(r
=
.816,
.674)
and
dry
weight
(r
=
.945, .690).
This
provides
a
physiological
basis
for
overall
enhancement
in
growth
by
Tria.
application.
Elemental
contents
of
Fe,
Mn,
Cu
were
significantly
maximum
at
0.1
mg/1
and
Zn
at
0.01
mg/1
(Figure
3).
Above
these
Tria.
concentrations
Fe
and
Mn
concents
decreased
while
that
of
Cu
and
Zn
remained
unaffected.
This
indicated
the
differential
affects
of
Tria.
on
uptake
and
distribution
of
micronutrients
in
poppy.
Tria.
application
at
0.01
mg/1
increased
capsule
number
(Figure
1),
weight
and
morphine
content
(Figure
4)
significantly,
whereas
thebaine
and
codeine
contents
remained
unaffected
at
different
concentrations.
The
positive
cor-
relation
between
capsule
number
and
weight
(r
=
.794)
suggested
that
increasing
number
of
capsules
is
not
a
limiting
factor
for
capsule
growth.
This
is
possibly
because
of
significant
positive
association
of
both
these
capsule
characters
with
CER
(r
=
.881,
.868)
and
total
Chl.
(r
=
.931,
.821)
100
---
95
.
.E
E
90
-.......
8
1.0
o'
.--:-.
u
0.,
45
84
-9
4
3
E
0
W
cn
uj
0
4
0
75
13
-
---...
cre
e—
=e
a
35
E
cc
70`
1
300
.7.,
cc
z
30
4
65w
a
25er
.65
X
60
,..,.
4
20
-
a-
5
:71
u
x
e—
ILI
x
0
"
20
55
—1
1
5
u
.4
t--.
0
50
0
1
-
31
o
1
.,
1
5
la
10
45
-5
0
C
-00
-01
1
-0
2-0
40
mgp
TRIACONTANOL
Fig.
2.
Effect
of
Tria.
on
CER,
total
Chl.,
LAR
and
Chl.
a/b
ratio.
LSD
values
at
5%
and
1%
are
4.8,
6.7
(CER);
0.15,
0.21
(total
Chl.);
28.4,
39.8
(LAR);
0.6,
0.8
(Chl.
a/b
ratio)
respective-
ly.
69
500
475
450
425
400
375
O
35.
rn
100
75
50
25
0
C
-001
•01
'1
1.0
2-0
4.0
mg/1
TR1ACONTONOL
Fig.
3.
Influence
of
Tria.
on
total
elemental
concentrations
in
shoots.
LSD
values
at
5%
and
1% are
19.3,
27.1
(Fe);
4.2,
5.9
(Mn);
4.4,
6.2
(Cu);
6.3,
8.9
(Zn)
respectively.
19
18-
AMorphine
16
0
Codein
17-
15
Thebaine
16-
x
Capsule
wt.
-
14
7
:0
15-
•13
w
14-
-12
5-
-11
_1
4
-
10
cL
3-
9
2-
8
1
I I
T
O
C
.001
-01
.1
1-0
20
40
mg/I
TR1ACONT
ANOL
Fig.
4.
Influence
of
Tria.
on
alkaloid
constituents
and
capsule
wt.
LSD
values
at
5%
and
1%
are
0.8,
1.1
(Morphine);
0.5,
0.7
(Codeine);
0.3,
0.5
(Thebaine);
0.7,
1.0
(capsule
wt.)
respec-
tively.
which
contribute
to
the
sink
(capsule).
The
positive
association
of
morphine
content
with
LAR
(r
=
.963),
CER
(r
=
.740),
total
Chl.
(r
=
.792)
and
shoot
fresh
weight
(r
=
.767)
and
dry
weight
(r
=
.740)
suggest
the
in-
creased
supply
of
latex
from
shoot
to
capsule
where
it
is
then
biosyntheti-
cally
transformed.
70
The
present
investigation
reveals
that
Tria.
at
concentrations
upto
0.1
mg/
1
significantly
enhances
various
processes
related
to
production
physiology
in
opium
poppy.
The
primary
processes
in
turn
contribute
significantly
in
increasing
overall
yield
of
straw,
capsule
and
morphine
content.
Acknowledgment
The
authors
are
grateful
to
the
Director,
CIMAP
for
encouragement
and
necessary
facilities
during
investigation
and
to
the
Director,
CFTRI,
Mys-
ore
for
providing
Triacontanol.
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