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Effect of endomycorrhizal infection and biocides on phytoalexin accumulation in soybean roots
303
Agriculture, Ecosystems and Environment, 29 (1989) 303-305 Elsevler Science Publishers B.V., Amsterdam Printed in Czechoslovakia
EFFECT OF EMDOMYCORRHIZAL INFKCTIONANDBIOCIDKSONPHYTOALBXIN ACCUMULATXGfIIN SO~BOOT6 D. MORANDI INRA, L a b , r a t , i r e de P h y t o p a r a s t t o l o g t e , BV 1 ~ 0 ,
S t a t i o n d ' h m 6 1 t o r a t i o n des P l a n t e s ,
21034 DIJON C~dex, France AB6TRACT
Two b i o c t d e a arbuscular
were
mycorrhtzal
(AOA). E f f e c t s
to
fungus
soybeans inoculated : Altette
or not with a vesicular
(Fosetyl-hl),
and
omtnooxyacettc acid
on VAM t n £ e c t t o n were p o s i t i v e w i t h A l i e t t e and n e s a t t v e w i t h AOA.
Concentrations
of the lso£1avonoide
stEnt£tcantly
81yceolltns,
c o u m e s t r o l and d a t d z e i n were
d e c r e a s e d by AOA b u t remained i n c h a n g e d w i t h A l i e t t e .
augEest that that
applied
phytoalexins
t h e u s e o£ b t o c i d e s
These r e s u l t s
do n o t c o n t r o l V A M d e v e l o p m e n t a n d emphasize t h e £act c a n m o d i f y t h e p h y s i o l o g i c a l s t a t e o£ t h e r o o t system
and t h e plant-VAJ4 £ungus i n t e r a c t i o n . INTROmgCTION Beside
their
mycorrhtzal
on p h o s p h a t e n u t r i t i o n ,
vesicular
arbuscular
(VAM) £ u r ~ i have been shown t o i n c r e a s e p l a n t r o o t c o n c e n t r a t i o n s o£
antimtcrobial
compounds l i k e p h y t o a l e x i n s (Morasdi e t e l . ,
could contribute
t o t h e i m p r o v e m e n t o£ p l a n t
VAM, e s p e c i a l l y (Morandt,
main effect
disease
1984). T h i s phenomenon r e s i s t a n c e o b s e r v e d with
a g a i n s t s o i l b o r n e p a t h o g e n s , and r e s u l t s o b t a i n e d w i t h nematodes
1987) s t r e n g t h e n t h i s h y p o t h e s i s . On t h e o t h e r h a n d , t h e s e 8 n t i m l c r o b t a l
compounds p r o d u c e d by t h e h o s t p l a n t could have an i n h i b i t i n g e f f e c t on t h e VAM £unEus i t s e l f test
this
in£ectlon the
and c o n s e q u e n t l y
hypothesis,
btocides
known t o i n t e r f e r e
I n o r d e r to w i t h VAN
o r w i t h p h e n y l p r o p a n o l d metabolism t o s o y b e a n s , and t r i e d t o e v a l u a t e
relation
infection.
c o n t r o l i t s progression in the root.
we h a v e a p p l i e d
between In
this
amtnooxyacetic
acid
and a funEtcide, (JabaJi-hare~nd
biocide
paper
effect
we r e p o r t
on p h y t o a l e x i n results
(AOA), known t o i n h i b i t PAL a c t i v i t y £osetyl-A1
H e n d r i c k , 1985).
(Aliette),
production
obtained
known to
with
a n d on VAN
a herbicide,
(hmrhein e t e l . , increase
1976),
VAN i n f e c t i o n
304
IIAlI~IALSAlmlaTSOi~S S o y b e a n s (AMSOY 71) were grown i n p o t s c o n t a i n i n g c a l c i n e d c l a y , i n a growth c h a m b e r ( d a Y / n i & h t : 22 / 1 8 r C , 70~/90~ rh ; 16h day, 1 8 k l u x ) . They r e c e i v e d 40ml m o d i f i e d Long Aehton s o l u t i o n (-N : KC1 i n s t e a d o£KNO 3 ; 18.4 m ~ l NaH 2 PO 4 H 2 O) t h r e e japonicum
t i m e s a week. A l l p l a n t s
and h a l f
by P h i l l i p s
w i t h GlomuB i n t r a r a d i ~ e a
w e r e i n o c u l a t j d w i t h Brad~rhizobium (LPA 8 ) .
VAM i n f e c t i o n
and Haymans (1970) method m o d i f i e d as f o l l o w s
: I L =
i n f e c t i o n o f r o o t system by l e n g t h I T = ~ o f VAN i n f e c t i o n IL ~ IT. d i a m e t e r and o v e r a l l i n f e c t i o n : I M = 100 Three ieoflavonotde daidzetn.
were a n a l y s e d
E x t r a c t i o n and q u a n t i f i c a t i o n
in roots
II,
III,
knowing that
glyceolltn
%
o f
V A M
o f i n f e c t e d r o o t s , by
: glyceolltn,
c o u m e a t r o l and
were as d e s c r i b e d p r e v i o u s l y (Norondi and
O ~ a n i n a z z t - P e a r s o n , 1986). For g l y o e o l l i n , I,
was e s t i m a t e d
r e s u l t s a r e t h e sum o f t h r e e i s o m e r s :
I i s t h e major isomer i n r o o t s . Methods f o r
a p p l y i n g t h e two b i o c i d e s a r e g i v e n i n t h e T a b l e s .
iJ~JLIS~DIS~J~ION .VAM i n f e c t i o n 2).
was i n c r e a s e d by A l i e t t e
Root c o n c e n t r a t i o n s o f a l l
(apart
i e o f l a v o n o i d s were s i g n i f i c a n t l y
from d a i d z e i n i n non , ~ / c o r r h l z a l r o o t s ) ,
The d e p r e s s i v e
t o be a PAL i n h i b i t o r
with the decrease a central
strengthened
d e c r e a s e d by AOA,
b u t were n o t a f f e c t e d by A l i e t t e .
e f f e c t o£ AOA on I s o f l a v o n o i d s i s c o n s i s t e n t w i t h l i t e r a t u r e
AOA i s r e p o r t e d play
( ~ a ~ . a 1) and d e c r e a s e d by AOA ( T a b l e
(Amrhein e t e l . ,
1976) ; t h i s ,
where
together
i n VAN t n £ e o t i o n due t o AOA, s u u g e a t s t h a t p h y t o a l e x i n s do n o t
role
in controlling
by t h e f a c t
that
VAN d e v e l o p m e n t .
t h e VAN i n f e c t i o n
This Interpretation
increased
by A l i e t t e
is
is not
a s s o c i a t e d w i t h a change i n r o o t i a o f l a v o n o i d c o n c e n t r a t i o n s . Futher involved
research
is necessary
in the biooide
effects
in order to better
u n d e r s t a n d t h e mechanisms
on VAN development which seem t o be i n d e p e n d e n t
o f t h e i r i n f l u e n c e on r o o t p h y t o a l e x i n a c c u m u l a t i o n . Table 1 - Aliette
a p p l i c a t i o n - a e r i a l spray with 2 g / l A l i e t t e ,
2 . 5 m l / p l a n t on
2 and 6 wef,k o l d soybeans ; s o y b e a n s h a r v e s t e d a f t e r 10 weeks g r o w t h . Shoot
Infection
Gl~ceolline
Coumestrol
f.wt.(g)
level
( 1 ~ / ~ Fresh
(Mg/R Fresh
roots) 4.80 a
-
Daidzein (~/R
roots)
too ts )
4.02 a
415 e
325 a
5.67 •
IOI+AL
4.62 a
436 •
295 a
M
7.81 c
51 a
27.7
b
290 b
200 b
N+AL
7.85 c
62 b
22.3
b
270 b
176 b
-
fresh
305 Table
2 -
solution
AOA a p p l i c a t i o n
-
3 day old
o f O, 10, o r I O 0 ~ M AOA b e f o r e Infection
Shoot f.wt.
soybean
planting
were
immersed
for
and h a r v e s t e d a f t e r
Glyceollins
48h
in
Coumestrol
Daidzein
NM 0
4.81 a
-
3.78 c
392 b
205 a
NM 10
3.92 b
-
2.79 b
202 a
215 a
NM 100
3.05 c
M 0
5.33 d
34.8 a
M 10
3.95 b
32.5 s
M 100
2.76 c
24.2 b
- For
each
column
-
in
each
1.68 a
267 a
196 a
672 c
509 b
8.28 d
324 ab
288 a
1,66 a
468 b
221 a
12.6
table
reans
e
(4 replicates)
a
7 weeks,
non f o l l o w e d
b y a same
letter are slEnlflcantly different at P = 0.05. BEFEIE~CES AMRHEIN
N.
, GODEKE
K.H.
and KEFELI
V.I.
: The estimation
of relative
intracellular phenylalanime ammonialysse (PAL) activities and the modulation in and by
vivo
in
competitive
vitro
inhibitous
D e u t a c h , Gea. 8 9 : 2 4 7 - 2 5 9
- Bet,
1976. JABAJI-HARE
S.H.
and KENDRICK W.B.
on c o m p o s i t i o n
of
J. Pl~mt Pathol.
7 : 118-126,
NORANDI D . , soybean
extracts
BAILEY J.A.
roots
infected
of
mycorrhizal
nutrition
Physiological
of Fosetyl-A1
and non m y c o r r h i z a l
and GIANINAZZI-P~ARSON V. with
on r o o t
exudation
leek roots
and
- Can.
1985. :Isoflavonnoid
vesicular-arbuscular
P l a n t P n t h o l . 24 : 3 5 7 - 3 6 4 , 1984. MORANDI D. a n d GIANZNAZZI-PEARSON phosphate
: Effects
on s e c o n d a r y
and g e n e t l e a l
V.
mycorrhizal
: Influence
metabolite
accumulation funEi-
of mycorrhizse
contents
of
in
Pb~ya$ol.
infection
soybean roots
and
- In
:
a s p e c t s o f myeorrhlxal. GIANINAZZI-PEARSON,
V.,GIANINAZZI, S./ eds/, INRA-PrQsa p. 217-222, 1986. MORANDI D° : VA Mycorrhizae, Nematodes, Phosphorus and Phytoalexins on Soybean In
:
Nyeerrhizae
Priorities.
in
the
next
decade.
SYLVIA, D.M, HUNG, L . L . ,
PHILLIPS
J.M.
staining
pacasitio
assessement of
and
HAYMAN D , S . and
infection-
Practical
GRAHAM, J . M .
:Improved
vesicular-arbuscular
application
-
and Research
/ e d s / - p. 212, 1987.
procedures
For clearing
mycorrhizal
T r a n s . B r . I ~ y c o l , S o c . 55 : 1 5 8 - 1 6 1 ,
funEi
roots for
1970.
Morandi, D., 1989: E f f e c t of e n d o m y c o r r h i z a l i n f e c t i o n and biocides on phytoalexins accumulation in s o y b e a n roots. Agric. E c o s y s t e m s Environ., 29: 303-305.
and rapid
Agriculture, Ecosystems and Environment, 29 (1989) 303-305 Elsevler Science Publishers B.V., Amsterdam Printed in Czechoslovakia
EFFECT OF EMDOMYCORRHIZAL INFKCTIONANDBIOCIDKSONPHYTOALBXIN ACCUMULATXGfIIN SO~BOOT6 D. MORANDI INRA, L a b , r a t , i r e de P h y t o p a r a s t t o l o g t e , BV 1 ~ 0 ,
S t a t i o n d ' h m 6 1 t o r a t i o n des P l a n t e s ,
21034 DIJON C~dex, France AB6TRACT
Two b i o c t d e a arbuscular
were
mycorrhtzal
(AOA). E f f e c t s
to
fungus
soybeans inoculated : Altette
or not with a vesicular
(Fosetyl-hl),
and
omtnooxyacettc acid
on VAM t n £ e c t t o n were p o s i t i v e w i t h A l i e t t e and n e s a t t v e w i t h AOA.
Concentrations
of the lso£1avonoide
stEnt£tcantly
81yceolltns,
c o u m e s t r o l and d a t d z e i n were
d e c r e a s e d by AOA b u t remained i n c h a n g e d w i t h A l i e t t e .
augEest that that
applied
phytoalexins
t h e u s e o£ b t o c i d e s
These r e s u l t s
do n o t c o n t r o l V A M d e v e l o p m e n t a n d emphasize t h e £act c a n m o d i f y t h e p h y s i o l o g i c a l s t a t e o£ t h e r o o t system
and t h e plant-VAJ4 £ungus i n t e r a c t i o n . INTROmgCTION Beside
their
mycorrhtzal
on p h o s p h a t e n u t r i t i o n ,
vesicular
arbuscular
(VAM) £ u r ~ i have been shown t o i n c r e a s e p l a n t r o o t c o n c e n t r a t i o n s o£
antimtcrobial
compounds l i k e p h y t o a l e x i n s (Morasdi e t e l . ,
could contribute
t o t h e i m p r o v e m e n t o£ p l a n t
VAM, e s p e c i a l l y (Morandt,
main effect
disease
1984). T h i s phenomenon r e s i s t a n c e o b s e r v e d with
a g a i n s t s o i l b o r n e p a t h o g e n s , and r e s u l t s o b t a i n e d w i t h nematodes
1987) s t r e n g t h e n t h i s h y p o t h e s i s . On t h e o t h e r h a n d , t h e s e 8 n t i m l c r o b t a l
compounds p r o d u c e d by t h e h o s t p l a n t could have an i n h i b i t i n g e f f e c t on t h e VAM £unEus i t s e l f test
this
in£ectlon the
and c o n s e q u e n t l y
hypothesis,
btocides
known t o i n t e r f e r e
I n o r d e r to w i t h VAN
o r w i t h p h e n y l p r o p a n o l d metabolism t o s o y b e a n s , and t r i e d t o e v a l u a t e
relation
infection.
c o n t r o l i t s progression in the root.
we h a v e a p p l i e d
between In
this
amtnooxyacetic
acid
and a funEtcide, (JabaJi-hare~nd
biocide
paper
effect
we r e p o r t
on p h y t o a l e x i n results
(AOA), known t o i n h i b i t PAL a c t i v i t y £osetyl-A1
H e n d r i c k , 1985).
(Aliette),
production
obtained
known to
with
a n d on VAN
a herbicide,
(hmrhein e t e l . , increase
1976),
VAN i n f e c t i o n
304
IIAlI~IALSAlmlaTSOi~S S o y b e a n s (AMSOY 71) were grown i n p o t s c o n t a i n i n g c a l c i n e d c l a y , i n a growth c h a m b e r ( d a Y / n i & h t : 22 / 1 8 r C , 70~/90~ rh ; 16h day, 1 8 k l u x ) . They r e c e i v e d 40ml m o d i f i e d Long Aehton s o l u t i o n (-N : KC1 i n s t e a d o£KNO 3 ; 18.4 m ~ l NaH 2 PO 4 H 2 O) t h r e e japonicum
t i m e s a week. A l l p l a n t s
and h a l f
by P h i l l i p s
w i t h GlomuB i n t r a r a d i ~ e a
w e r e i n o c u l a t j d w i t h Brad~rhizobium (LPA 8 ) .
VAM i n f e c t i o n
and Haymans (1970) method m o d i f i e d as f o l l o w s
: I L =
i n f e c t i o n o f r o o t system by l e n g t h I T = ~ o f VAN i n f e c t i o n IL ~ IT. d i a m e t e r and o v e r a l l i n f e c t i o n : I M = 100 Three ieoflavonotde daidzetn.
were a n a l y s e d
E x t r a c t i o n and q u a n t i f i c a t i o n
in roots
II,
III,
knowing that
glyceolltn
%
o f
V A M
o f i n f e c t e d r o o t s , by
: glyceolltn,
c o u m e a t r o l and
were as d e s c r i b e d p r e v i o u s l y (Norondi and
O ~ a n i n a z z t - P e a r s o n , 1986). For g l y o e o l l i n , I,
was e s t i m a t e d
r e s u l t s a r e t h e sum o f t h r e e i s o m e r s :
I i s t h e major isomer i n r o o t s . Methods f o r
a p p l y i n g t h e two b i o c i d e s a r e g i v e n i n t h e T a b l e s .
iJ~JLIS~DIS~J~ION .VAM i n f e c t i o n 2).
was i n c r e a s e d by A l i e t t e
Root c o n c e n t r a t i o n s o f a l l
(apart
i e o f l a v o n o i d s were s i g n i f i c a n t l y
from d a i d z e i n i n non , ~ / c o r r h l z a l r o o t s ) ,
The d e p r e s s i v e
t o be a PAL i n h i b i t o r
with the decrease a central
strengthened
d e c r e a s e d by AOA,
b u t were n o t a f f e c t e d by A l i e t t e .
e f f e c t o£ AOA on I s o f l a v o n o i d s i s c o n s i s t e n t w i t h l i t e r a t u r e
AOA i s r e p o r t e d play
( ~ a ~ . a 1) and d e c r e a s e d by AOA ( T a b l e
(Amrhein e t e l . ,
1976) ; t h i s ,
where
together
i n VAN t n £ e o t i o n due t o AOA, s u u g e a t s t h a t p h y t o a l e x i n s do n o t
role
in controlling
by t h e f a c t
that
VAN d e v e l o p m e n t .
t h e VAN i n f e c t i o n
This Interpretation
increased
by A l i e t t e
is
is not
a s s o c i a t e d w i t h a change i n r o o t i a o f l a v o n o i d c o n c e n t r a t i o n s . Futher involved
research
is necessary
in the biooide
effects
in order to better
u n d e r s t a n d t h e mechanisms
on VAN development which seem t o be i n d e p e n d e n t
o f t h e i r i n f l u e n c e on r o o t p h y t o a l e x i n a c c u m u l a t i o n . Table 1 - Aliette
a p p l i c a t i o n - a e r i a l spray with 2 g / l A l i e t t e ,
2 . 5 m l / p l a n t on
2 and 6 wef,k o l d soybeans ; s o y b e a n s h a r v e s t e d a f t e r 10 weeks g r o w t h . Shoot
Infection
Gl~ceolline
Coumestrol
f.wt.(g)
level
( 1 ~ / ~ Fresh
(Mg/R Fresh
roots) 4.80 a
-
Daidzein (~/R
roots)
too ts )
4.02 a
415 e
325 a
5.67 •
IOI+AL
4.62 a
436 •
295 a
M
7.81 c
51 a
27.7
b
290 b
200 b
N+AL
7.85 c
62 b
22.3
b
270 b
176 b
-
fresh
305 Table
2 -
solution
AOA a p p l i c a t i o n
-
3 day old
o f O, 10, o r I O 0 ~ M AOA b e f o r e Infection
Shoot f.wt.
soybean
planting
were
immersed
for
and h a r v e s t e d a f t e r
Glyceollins
48h
in
Coumestrol
Daidzein
NM 0
4.81 a
-
3.78 c
392 b
205 a
NM 10
3.92 b
-
2.79 b
202 a
215 a
NM 100
3.05 c
M 0
5.33 d
34.8 a
M 10
3.95 b
32.5 s
M 100
2.76 c
24.2 b
- For
each
column
-
in
each
1.68 a
267 a
196 a
672 c
509 b
8.28 d
324 ab
288 a
1,66 a
468 b
221 a
12.6
table
reans
e
(4 replicates)
a
7 weeks,
non f o l l o w e d
b y a same
letter are slEnlflcantly different at P = 0.05. BEFEIE~CES AMRHEIN
N.
, GODEKE
K.H.
and KEFELI
V.I.
: The estimation
of relative
intracellular phenylalanime ammonialysse (PAL) activities and the modulation in and by
vivo
in
competitive
vitro
inhibitous
D e u t a c h , Gea. 8 9 : 2 4 7 - 2 5 9
- Bet,
1976. JABAJI-HARE
S.H.
and KENDRICK W.B.
on c o m p o s i t i o n
of
J. Pl~mt Pathol.
7 : 118-126,
NORANDI D . , soybean
extracts
BAILEY J.A.
roots
infected
of
mycorrhizal
nutrition
Physiological
of Fosetyl-A1
and non m y c o r r h i z a l
and GIANINAZZI-P~ARSON V. with
on r o o t
exudation
leek roots
and
- Can.
1985. :Isoflavonnoid
vesicular-arbuscular
P l a n t P n t h o l . 24 : 3 5 7 - 3 6 4 , 1984. MORANDI D. a n d GIANZNAZZI-PEARSON phosphate
: Effects
on s e c o n d a r y
and g e n e t l e a l
V.
mycorrhizal
: Influence
metabolite
accumulation funEi-
of mycorrhizse
contents
of
in
Pb~ya$ol.
infection
soybean roots
and
- In
:
a s p e c t s o f myeorrhlxal. GIANINAZZI-PEARSON,
V.,GIANINAZZI, S./ eds/, INRA-PrQsa p. 217-222, 1986. MORANDI D° : VA Mycorrhizae, Nematodes, Phosphorus and Phytoalexins on Soybean In
:
Nyeerrhizae
Priorities.
in
the
next
decade.
SYLVIA, D.M, HUNG, L . L . ,
PHILLIPS
J.M.
staining
pacasitio
assessement of
and
HAYMAN D , S . and
infection-
Practical
GRAHAM, J . M .
:Improved
vesicular-arbuscular
application
-
and Research
/ e d s / - p. 212, 1987.
procedures
For clearing
mycorrhizal
T r a n s . B r . I ~ y c o l , S o c . 55 : 1 5 8 - 1 6 1 ,
funEi
roots for
1970.
Morandi, D., 1989: E f f e c t of e n d o m y c o r r h i z a l i n f e c t i o n and biocides on phytoalexins accumulation in s o y b e a n roots. Agric. E c o s y s t e m s Environ., 29: 303-305.
and rapid