The effect of environmental factors on growth development and alkaloid production of poppy (Papaver somniferum L.) II. Interaction of light and temperature


Bernath, J.; Tetenyi, P.

Biochemie und Physiologie der Pflanzen 176(7): 599-605

1981


Optimum growth of Papaver somniferum cultivars of different origin was observed at 1.6 104 lux light intensity under "low temperature" programme (daily rhythm of 12.517.5°C increasing to 18.5/11.5°C during the vegetation). The maximum of dry matter was also produced in a "low temperature" programme, but at a higher light intensity of 3.2 104 lux. At the same time low temperature hindered the development by 10-15 days and modified the ratio of organs: proportion of leaves increased, that of the stems diminished. The alkaloid accumulation in dry capsules was determined by external conditions followed the generative differentiation of apex. The total amount of alkaloids (morphinane phthalideisoquinoline + benzylisoquinoline) reached a maximum value in a "high temperature" programme under the 3.2 104 lux illumination (18.31 mg/g). Under the same illumination level the accumulation was limited by the cold effect. Inside the morphinanes, the ratio of codeine was decreased below 10 %, even in individuals have been evaluated as a codeine-type, Thereby, the importance of light-temperature interaction in the demethylation processes of poppy alkaloids has been proved.

Biochem.
Physiol.
Pflanzen
176,
599-605
(1981)
The
Effect
of
Environmental
Factors
on
Growth,
Development
and
Alkaloid
Production
of
Poppy
(Papaver
somniferum
L.)
IL
Interaction
of
Light
and
Temperature
JEN5
BERNATH
and
PATER
TETENYI
Research
Institute
for
Medicinal
Plants,
Budakalasz,
Hungary
Key
Term
Index
morphinane-,
phthalideisoquinoline-,
benzylisoquinoline
alkaloids,
al
ka
biosynthesis,
effect
of
temperature,
light
-temperature
interaction;
Papaver
somniferum.
Summary
Optimum
growth
of
Papaver
somniferum
cultivars
of
different
origin
was
observed
at
1.6
10
4
lux
light
intensity
under
"low
temperature"
programme
(daily
rhythm
of
12.517.5
°C
increasing
to
18.5/11.5
°C
during
the
vegetation).
The
maximum
of
dry
matter
was
also
produced
in
a
"low
temperature"
programme,
but
at
a
higher
light
intensity
of
3.2
10
4
lux.
At
the
same
time
low
tem-
perature
hindered
the
development
by
10-15
days
and
modified
the
ratio
of
organs:
proportion
of
leaves
increased,
that
of
the
stems
diminished.
The
alkaloid
accumulation
in
dry
capsules
was
determined
by
external
conditions
followed
the
generative
differentiation
of
apex.
The
total
amount
of
alkaloids
(morphinane
phthalideisoquino-
line
+
benzylisoquinoline)
reached
a
maximum
value
in
a
"high
temperature"
programme
under
the
3.2
10
4
lux
illumination
(18.31
mg/g).
Under
the
same
illumination
level
the
accumulation
was
limited
by
the
cold
effect.
Inside
the
morphinanes,
the
ratio
of
codeine
was
decreased
below
10
%,
even
in
individuals
have
been
evaluated
as
a
codeine
-type,
Thereby,
the
importance
of
light
-tem-
perature
interaction
in
the
demethylation
processes
of
poppy
alkaloids
has
been
proved.
Introduction
In
the
first
part
of
our
paper,
the
effect
of
the
light
intensity
and
day
-length
were
discussed
on
the
growth,
development
and
alkaloid
formation
of
poppy
(BERNkTH
and
TETENYI
1979).
The
described
changes
in
alkaloid
formation
and
accumulation
complementing
the
results
of
NYMAN
and
HANSSON
(1978)
led
to
a
new
viewpoints
of
the
chemotaxonomic
evaluation.
By
our
results
the
overall
alkaloid
formation
and
the
stereospecific
enzyme
reaction
of
reticuline
and
the
demethylation
processes
of
morphinane
group
could
be
influenced
by
light.
According
to
the
experimental
proposal
of
the
U.N.
Narcotics
Laboratory
(1975)
the
influence
of
the
temperature
must
also
be
determinant.
The
intention
of
present
work
is
to
establish
alteration
of
growth,
development
and
alkaloid
formation
affecting
by
temperature
at
different
light
conditions.
40
Biochem.
Physiol.Pflanzen,
Bd
.176
600
J.
BE
and
P.
TETENY1
Material
and
Methods
Plant
material
Taking
into
the
consideration
the
earlier
establishment
cultivar
"Kompolti
M"
(with
morphine
character)
and
cultivar
"Reading"
(with
codeine
character)
differing
in
both
chemism
and
origin
were
taken.
The
genetical
homogeneity
was
insured
by
sowing
I
s
seeds.
Standard
growth
conditions
The
general
experimental
conditions
were
published
in
the
first
part
of
our
paper
(BERNATH
and
TkTENYT
1979).
In
the
phytotron
chambers
(Conviron
E-15
and
mofified
G
30)
plants
were
cultivated
in
sand-perlite
mixture
(v/v
1:
1)
using
KNOP
nutrient
solutions.
Temperature
conditions
Both
the
"high
temperature"
and
"low
temperature"
programmes
were
calculated
on
the
basis
of
vegetatvie
cycle
in
Hungary.
The
programmes
were
accomplished
by
12/12
h
day/night
rhythm
as
follows
"High
temperature"
programme
{day/night)
1—
5
weeks
12.5/
7.5
0
0
6-11
weeks
18.5/11.5
°C
12-17
weeks
22.0/13.0
°C
After
18
weeks
26.0/16.0
°C
"Low
temperature"
programme
1
5
weeks
6-11
weeks
12-17
weeks
After
18
weeks
12.5/
7.5
°C
1.60/10.0
°C
17.0/11.0
°C
18.5/11.5
0
0
High
-low
programme
transfers
were
taken
at
generative
apex
initiation,
flowering,
half
as
well
as
whole
sized
capsule
stages.
Light
conditions
The
light
intensity
was
chosen
with
consideration
of
the
results
of
earlier
experiments.
After
the
differentiation
of
apex
14
h
illumination
have
uniformly
be
set
by
General
Electric
F72T12
(CW)
VHO
fluorescent
lamps
supplemented
by
incandescent
bulbs.
Experimental
variations
of
light
and
temperature
treatments
are
shown
in
Fig.
1.
Morpho-phenological
investigation,
alkaloid
analysis
The
detailed
evaluation
of
25-50
individuals
(15
in
high
-low
transfer
variations)
was
accom-
plished
for
each
treatment
and
for
each
cultivar
in
course
of
the
investigations.
This
was
extended
to
the
morpho-phenological,
producting
and
alkaloid
forming
processes,
similarly
to
our
earlier
"
high
temperature'
programme
low
temperature
"
programme
01.
10
Luxi
1.6
.10
4
Lux
24.10
4
Luxj
32
.10
LUX
Fig.
1.
Light
-temperature
combinations
applied
in
the
course
of
experiments.
Papaver
Alkaloid
Production
Affected
by
Temperature
601
investigations.
The
quantitative
control
was
performed
in
a
Hewlett
Packard
HPLC
equipment,
in
addition
to
the
TLC
method
of
DINos
(1968).
Results
Changes
of
growth,
development
and
dry
-matter
production
The
characteristic
height
and
capsule
production
data
of
plants
cultivated
in
various
"low
and
high
temperature"
programmes
are
summarized
in
Table
1.
It
could
be
estab-
lished
on
the
data
that
both
cultivars
reached
the
maximum
height
under
"low
tem-
perature"
conditions.
This
was
the
result
of
a
longer
growth
periods
which
followed
the
stretched
sigmoid
curve
affecting
by
cold.
The
prolonged
growth
caused
a
delay
of
10-15
days
in
flowering
too.
The
number
of
capsules
was
less
at
a
"low
temperature".
1.15
on
the
"Kompolti
M"
and
about
1
on
the
cultivar
"Reading"
being
characteristic
feature
of
that
cultivar.
However,
the
capsules
thus
formed
were
larger
and
their
weight
was
by
about
10-53%
heavier
than
those
of
"high
temperature"
programme.
The
seed
formation
was
promoted
by
cold
temperature
too.
The
interaction
of
different
light
and
temperature
conditions
is
shown
in
Fig.
2.
The
greater
production
unambiguously
was
yielded
by
the
plants
grown
in
the
"low
temperature"
programme.
The
light
-dependence
was
indicated
by
the
fact
that
while
optimum
production
appeared
at
2.4
10
4
lux
in
the
"high
temperature"
programme,
in.
the
"low"
programme
further
significant
growth
could
be
realised
at
3.2
10
4
lux
ilumi-
nation.
Proportion
of
stem
rised
under
"high",
while
that
of
leaves
under
"low"
tempera-
ture
programme.
The
action
of
increasing
light
intensity
was
similar
to
warm
condi-
tions.
Quantitative
and
qualitative
changes
in
alkaloid
formation
The
alkaloid
content
of
dry
capsules
affected
by
temperature
is
summarized
in
Table
2.
The
whole
amount
of
alkaloids
increased
under
"high
temperature"
programme
in
Table
1.
The
Effect
of
"low"
and
"high"
temperature
on
height
and
capsule
production
of
poppy
height
mean
weight/one
dimension
(mm)
number
of
capsules
(g)
of
capsules
(mm)
capsules
per
plant
capsule
seed
primer
secondary
diam.
lenght
diam.
lenght
(piece)
"Kompolti
M"
"low
temperature"
1,302
1.15
0.91
0.59
24
31
13
19
"high
temperature"
1,182
1.38
0.69
0.53
22
27
5
6
"Reading"
"low
temperature"
1,171
1.03
1.58
1.13
37
37
21
16
"high
temperature"
1,038
1.62
1.11
0.07
31
33
7
6
S
Z
D
5
%
temperature
62
0.31 0.21
0.29
2
3
5
6
40*
602
J.
BERNATH
and
P.
TETiNYI
0.8x
1.6
x
2.4x
12
x
Q
1
PO
0.8x
1.6x
2.4x
3.2x
(10
4
Lux)
F
If
ig
h
terrp
e
raturi
programm
e
"Low
temperature"
IT
0.8
x
1.6
r
2.1
x
3.2
x
0.8x
14x
2.4x
3.2
x(10
4
Lux)
VVVVV
lull
01E11
40
A00'
CAPSULE
t;11:
1
4
LEAF
Fig.
2.
Effect
of
light
-temperature
iniercation
on
dry
-matter
production
of
poppy.
the
capsules,
but
the
cultivar
"Reading"
reacted
in
a
more
sensible
way.
The
strongest
change
was
observed
in
the
amount
and
ratio
of
codeine:
under
"high
temperature"
programme
its
amount
was
3
times
larger
in
cultivar
of
morphine
and
17
times
larger
in
cultivar
of
codeine
character.
Similarly
any
noticeable
thebaine
accumulation
appear-
ed
at
warm
conditions
only.
The
amount
of
benzylisoquinolines
was
unchanged
in
"Kompolti
IV,
and
increased
to
a
higher
level
in
the
Reading
at
the
"high
temperature"
programme.
The
quantity
of
phthalideisoquinolines
was
higher
in
that
later
case
equally.
Temperature
-light
interaction
is
demonstrated
in
Figs.
3.
and
4.
In
the
cultivar
"Kompolti
M"
the
increase
of
light
intensity
was
expressed
by
acceleration
of
mor-
phine
formation
according
to
the
following
saturation
curves:
Papaver
Alkaloid
Production
Affected
by
Temperature
603
Table
2.
The
effect
of
"low"
and
"high"
temperature
programme
on
quality
and
quantitative
composi-
tion
poppy
alkaloids
(Alkaloid
mg
/g
dry
matter)
main
alkaloid
groups
morphinane
phthalide-,
benzyl-,
isoquinoline
morphine
codeine
thebaine
narcotine
laudanine
mg/g
ratio
mgig
ratio
mg/g
ratio
narco-
+
papa-
toline
verine
mglg
mg/g
(
0
/0)
(%)
0/0)
Total
rngig
"Kompolti
M"
"low
temperature"
5.36
(95)
0.21
(4)
0.08
(1)
2.07
"high
temperature"
5.91
(86)
0.62
(9)
0.34
(5)
2.72
"Reading"
"low
temperature"
4.61
(95)
0.23
(5)
0.00
(0)
1.31
"high
temperature"
2.98
(42)
3.92
(56)
0.16
(2)
2.25
0.15
0.15
2.48
4.74
7.87
9.65
8.63
14.05
SZD5%
(temperature)
1.50
0.30
0.07
0.62
1.23
1.52
Y"low
temperature"
'"high
temperature"
7
.
7(1
e
1.0616-0.0001.5
X)
7
.
8(1
e
1.0357-0•00036
X)
Thus,
the
effect
of
"high
temperature"
appeared
in
the
more
rapid
accumulation
of
morphine
and
the
higher
level
of
its
saturation.
The
more
intensive
accumulation
of
alkaloid
mg/g
8
,Kompoliii
Low
temperature
programme
2.4
3.2
x
1.6x
0,8x
Th
High
temperature
programme
2.4
K
x
Th
Fig.
3.
Morphinane
spectrum
of
dry
capsules
changing
under
the
pressure
of
light
-temperature
inter-
action.
M,
morphinane;
Co,
codeine;
Th,
thebaine
604
J.
BERNATH
and
P.
TETENYI
alkaloid
mg/g
,Reading'
Low
temperature
programme
8
8-
4-
4
-
High
temperature
programme
Ux
Th
M
Co
Th
Fig.
4.
Morphinane
spectrum
of
dry
capsules
changed
under
the
pressure
of
light
-temperature
inter-
action.
M,
morphine;
Co,
codeine;
Th,
thebaine
codeine
and
thebaine
was
also
observed
in
that
later
case.
A
chemism
dependent
reac-
tion
of
the
cultivar
"Reading"
was
proved
by
the
saturation
curve
calculated
in
codeine
formation:
Y"iow
temperature"
Y"hi
g
h
temperature"
=
0.5(1
e0.69269-0400076
X)
6.0(1
0.92604-0.000129
X)
The
effect
of
light
intensity
promoting
the
accumulation
of
codeine
was
observed
only
under
warm
conditions.
In
the
cold
programme
the
demethylation
processes
remained
more
characteristic,
simultaneously
with
an
increased
accumulation
of
mor-
phine.
On
the
basis
of
our
"low
-high
temperature"
combinations
this
rule
of
alkaloid
accumulation
can
be
modified
only
in
the
case
when
the
temperature
acts
at
least
before
reaching
of
the
half
capsule
development
stage.
After
that
and
especially
during
maturation
no
further
accumulation
changes
occure
in
capsules.
Discussion
According
to
our
observations
the
characteristic
rhythm
of
growth
an
development
was
slowed
down
by
a
lower
temperature
under
the
given
light
conditions.
The
longer
vegetative
period
made
possible
a
higher
production
of
dry
-matter
and
appearance
a
quasi
more
vegetative
habit's.
The
"low
temperature"
showed
especially
favourable
stimulation
on
seed
production
of
cultivar
"Reading".
The
amount
and
composition
of
the
main
alkaloid
groups
were
substantially
deter-
mined
by
light
and
temperature
and
their
interaction
as
well.
The
alkaloid
formation
is
Papaver
Alkaloid
Production
Affected
by
Temperature
605
accelerated
by
the
increase
of
light
intensity
universally,
which
is
expressed
in
a
inten-
sive
accumulation
of
each
alkaloid
group
depending
on
the
chemism
of
the
cultivar
studied.
At
"low
temperature"
not
only
the
formation
of
alkaloids
is
significantly
di-
minished
but
the
ratio
of
main
groups
is
modifyed
also.
The
strenghtening
of
demethyla-
tion
processes
is
indicated,
as
under
the
pressure
of
"low
temperature"
the
total
amount
of
thebaine
and
large
amount
of
codeine
have
been
transformed
into
morphine.
The
di-
rection,
of
the
transformation
is
in
accord,
with
the
data
obtained
in
the
biosynthesis
of
poppy
alkaloid
(MoTHEs
and
SCHUTTE
1969)
and
conforms
to
the
metabolic
scheme
of
BILLS
(1978).
Taking
into
consideration
the
biosynthetic
pathway
the
established
cold
effect
could
promote
the
return
of
alkaloids
into
the
general
metbolism
of
the
plant
this
possibility
is
proved
by
FAIRBAIRN
et
al.
(1978).
References
BERNATH,
J.,
and
TEANYI,
P.:
The
effect
of
environmental
factors
on
growth,
development
and
alkaloid
production
of
poppy
(Papaver
somniferum
L.)
I.
Responses
to
day
-length
and
light
intensity.
Bioehem.
Physiol.
Pflanzen
174,
468-478
(1979).
BILLS,
D.
D.:
Biosynthesis
of
Phenanthrene
alkaloids.
Report
of
Working
Group
of
U.N.
Narcotics
Laboratory.
Oslo.
8-12
May
(1978).
DA.Nos,
D.:
Retegkromatografias
mOdszer
a
Papaver
somniferum
L.
alkaloidspektrumanak
nyo-
monkovetesere
II.
Herba
Hung.
7,
27-37
(1968).
FAIRBAIEN,
J.
W.,
HANDA,
S. S.,
GURKAN,
E.,
and
PHILLIPSON,
J.
D.:
In
vitro
conversation
of
mor-
phine
to
its
N
-oxide
in
Papaver
somniferum
latex.
Phytochemistry
17,
261-262
(1978).
MOTHES,
K.,
and
SCHUTTE,
H.
R.:
Biosynthese
der
Alkaloide.
pp.
367-419.
Berlin
1969.
NYMAN,
U.,
and
HANSSON,
B.:
Morphine
content
variation
in
Papaver
somniferum
L.
as
affected
by
the
presence
of
some
isoquinoline
alkaloids.
Hereditas
88,
17-26
(1978).
U.N.
Narcotics
Laboratory:
Relationship
between
environment
and
phena.nthrene
alkaloid
con-
tent.
Research
proposal.,
Geneve
1975.
Received
August
4,
1980;
in
revised
form
December
16,
1980.
Author's
address:
Prof.
Dr.
P.
TETENyi
and
Dr.
J.
BERNATH,
Research
Institute'
for
Medica
Plants,
2011
Budakalasz,
Hungary.