Improvement of biological control against Cydia pomonella by sucrose

SUCROSE: A PLANT RESISTANCE
INDUCER AGAINST Cydia pomonella L
Authors
E. Ferré *, H. Galy *, F. Moulin **, G. Clément ***, S. Derridj ****
Project conducted in collaboration with:
* ANADIAG SA
** ENSP – Le Potager du Roi
*** INRA – Plant Nitrogen Nutrition
**** INRA – Research unit 1272 - Insect Physiology: Signalling and Communication
Outline
• Soluble carbohydrates and sugar alcohols:
• Presence onto the plant surface and plant signalisation for
insect oviposition
• Induction of modifications of plant signalisation by
sugar applications at low concentrations:
• Field experiments conducted through Europe:
Efficacy of sucrose and fructose applications against C. pomonella
• Effect of sucrose and fructose applications on leaf surface
composition:
Analyse of the metabolite composition and plant signals for C. pomonella
Soluble carbohydrates: presence on the plant and signals for
C. pomonella oviposition
Host plant selection process for oviposition
by insects
Contact with legs and leaf
surface scanning with the
ovipositor
Soluble carbohydrates
Primary metabolites
Diffusion by transcuticular
Photosynthetic origin
pathways
Quantities (ng/cm²) are present at cuticle surface
Soluble-carbohydrates and sugar-alcohols
ratios influences C. pomonella egg laying
Female site acceptance
Number of eggs layed
Behavioural events leading a generalized insect to
lay eggs on host plant
2) Moving with displacement delimited by
responses to non contact cues
1) Moving not influenced
by plantcues
3) Attraction by emission
source
4)Arrestment on (at) plant
5) Scanning via sensory
apparatus (legs, ovipositor)
6) Site acceptance
7) Egg laying
Contact sensory cues increasing
Transcuticle pathways of nonpolar (A) and polar compounds (P)
Diffusion by Polysaccharidic pathways
(Dominguez (1999)
Non polar lipophile pathway
Polar acquous pathway
Epicuticule
Primary cuticle
A
A
P
Secondary cuticle
Mean lamina
Primary cell wall
Polar pathway
Cires
Epicuticle
Intracuticle
Area of formation of polar pores (hydrophyles)
Pectine
Mean lamina
Cellulose
Cellulose and Hemicellulose fibers
Menbrane
Jeffry, 1996
Membrane
C
U
T
I
C
L
E
Egg laying of Cydia pomonella on nylon cloth substrates
drench by a mixture of 6 leaf surface metabolites
Egg-laying of C. pomonella after 63 minutes (3 mn after darkness) on nylon cloth
substrates drench by a mixture of 6 leaf surface metabolites (f) of Golden Delicious Vs
same mixture without one of the metabolite.
metabolite
So : Sorbitol
Q: Quebraquitol
M: Myoinositol
14
12
N o m b re d 'o e u fs p a r fe m e lle
(m o y e n n e ± e .s .)
F : Fructose
10
a
f
f-F
8
Polyols
f - So
ab
f-Q
6
ab
4
b
2
b
0
Nature du substrat
f-M
Cydia pomonella L : biological cycle
2nd
generation
Juillet
2ème vol
ût
Ao
in
Ju
L1
L3
L5
L4
1er vol
Chenille L5
diapausante
Chrysalide
ril
Av
e
br
em
v
No
Ma
rs
C. pomonella development cycle
(example 2 generations)
Oc
tob
re
generation
Ponte
Mai
1st
bre
Septem
L2
Févri
er
Janvier
e
mbr
e
c
Dé
Possible 3rd
generation
Material and methods : Field trials
Sucrose or fructose applications
First application of sugars alone 20 days before forecasted oviposition period.
1st insecticide application at egg hatching.
Application timing : every 20 days.
Observations
Preliminary damage assessment (100 fruits/ plot) at the end of the 1st insect generation.
At each application assessment on safe and damaged fallen fruits.
At the end of the generation or harvest assessment on 250 fruits per elementary plot.
Statistical analysis
% of total attacked fruits and corresponding Abbott efficacy of treatments
Bartlett and Student Newman & Keuls mean comparison test
Field experiments conducted in Europe
Abbott Efficacy levels calculated in % vs UTC damages
Trials
06/FR EL
08/GR ML
08/ITA ML
07/FR RY
07/FR GA
07/GR11
07/ITA015
Standard references
(Ref)
Carpovirusine
Zolone Flo
Carpovirusine
Carpovirusine
Imidan 50WP
Imidan 50WP
Imidan 50WP
Dursban 480 EC
Imidan 50WP
Smart EW
100ml/100L
120g/100L
100ml/100L
100ml/100L
100g/100L
100g/100L
100g/100L
100ml/100L
100g/100L
215ml/100L
UTC %
damages
Suc.
10ppm
Suc.
100ppm
37,58
1,75
Open field experiment
locations
Fru.
Fru.
10ppm 100ppm
24,14 a
22,26
24,93
44,35
1,02
40,67
Suc.
1000ppm
31,34 a 21,23 a
63,33 b
38,1 b
61,9 ab
37,98 bc 30,66 c
33,25 a 46,81 a
60,04 b
19,52 d
36,01 a
Std Ref
Std Ref
+
Sugar
36,6 b
47,12 b
26,49 a
54,63 a
78,9 a
80,95 a
92,86 a
41,55 bc 61,34 a
46,53 abc 50,66 ab
30,42 cd
54,63 a
45,14 a
37,75 a
Mean Abbott efficacy level (%) for each type of
treatment for all experiments
100,00
92,86
90,00
80,95
80,00
Efficacy(%)
70,00
60,00
63,33
61,90
60,04
50,00
40,00
30,00
59,42
48,85
42,69
37,93
45,14
37,21
31,34
26,49
21,23
20,00
19,52
10,00
MAX.
MEAN
MIN.
Suc.
10ppm
Suc.
100ppm
Fru.
10ppm
Std Ref
Std Ref
+
Sugar
Leaf surface metabolite analysis: Material and methods
Bourse shoots sampling
3 samples of bourse shoots harvested in the upper part of 2 central trees per
elementary plot.
Timing of harvesting on several days of sugar treated and control plants together.
Surface chemical analysis
Leaf surface washings collected and analyzed on a GC-TOF chromatography.
(software AMDIS)
Metabolome result analysis
All results were delivered on to PCA performed on 119 metabolites to determine:
Wether the plant surface metabolite composition is modified by sugar treatment
Wether those known as influencing C. pomonella egg laying and neonate larval
behaviour are concerned
Effect of apple tree sucrose treatment (10 ppm) on the
metabolite composition of the under side of the bourse
shoot leaves
Mean centered (Principal components analysis)
Variety: Golden delicious
K‐mean cluster 2
9 NT+ 4suc
K‐mean cluster 1
6 suc
glycerol
sorbitol
quinate
1918/375
1 lactate
22 glycine
first two components 69%
pyroglu
quebraquitol
alanine
Myo‐inositol
inositol
Effect of apple tree fructose treatment (0,1ppm) on the
metabolite composition of the under side of the bourse
shoot leaves
Mean centered (Principal components analysis)
Variety: Golden delicious
K‐mean cluster 1
6NT + 1 Fru
K‐mean cluster 2
4 NT + 8 Fruc
pyroglu
quebraquitol
alaninelactate
inositol
quinateglycerol
mannitol
10 urée
22 glycine
sorbitol
first two components 67 %
Metabolites modified by applications of sucrose at
10ppm and fructose at 0,1ppm on leaf surface
Metabolites significantly modified by sucrose and fructose applications
vs. None sugars treated trees
Sucrose treated (10ppm)
Fructose treated (0.1ppm)
N°
Name
t-test
N°
Name
t-test
69
quebraquitol 5tms 1859.3/318
1380.5/184
scyllo-inositol 1949.3/318
glyceric acid 1335.5/292
GABA 1527.7/304
2-ketogluconate 1779.8/292
1145.1/235
myo-inositol 2086.7/318
succinic acid 1320.9/247
ethanolamine 1269.8/174
alanine 1109.1/116
decanoic acid 1459.2/229
glycerol 1278.1/205
raffinose 3355.5/361
1433.5/233
arabinose meox 1662.9/307
unk 1918.7/375
nonanoic acid 1368.3/215
beta-alanine 1430.7/248
beta-lactate ? 1148.8/219
erythronic acid 1538.6/292
C19 1.006 lysine 174
oxalic acid 1138.3 /147
betahydroxybutyrate [938] 1162
0,0009
30
69
0,0013
47
0,0014
14
0,0017
81
1380.5/184
quebraquitol 5tms 1859.3/318
GABA 1527.7/304
ethanolamine 1269.8/174
scyllo-inositol 1949.3/318
1145.1/235
myo-inositol 2086.7/318
alanine 1109.1/116
1530.2/314
beta-alanine 1430.7/248
erythronic acid 1538.6/292
Diethylene glycol ? 1245.8/117
1452.9/350
1247.7/127
0,0001
0,0010
30
81
26
47
63
4
88
23
14
2
39
17
115
37
57
78
29
36
5
49
77
3
6
0,0017
0,0025
0,0047
0,0047
0,0054
0,0097
0,0108
4
88
2
48
36
49
0,0154
11
0,0164
38
0,0188
12
0,0229
0,0248
0,0260
0,0270
0,0287
0,0323
0,0335
0,0353
0,0508
0,0002
0,0005
0,0011
0,0012
0,0027
0,0075
0,0095
0,0155
0,0200
0,0260
0,0363
0,0429
0,0464
Conclusions
Applications of 10ppm and 100ppm solutions of sugar
change the insect behaviour and significantly reduce
the damages vs. untreated control.
Sucrose or fructose applied in tank mix with an
insecticide enhance the efficacy of the treatment
Sugar applications modify leaf surface metabolome and
metabolites known as signals for C. pomonella
oviposition behaviour.
The Plant Resistance against Insect behavior
(antixenosis) is induced by the spraying of sucrose.
THANK YOU FOR YOUR ATTENTION!
Contacts:
Hubert Galy: [email protected]
Eric Ferré: [email protected]

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