Avena genetic resources for quality in human consumption (AVEQ) WP5,Fusarium and Mycotoxins analysis. 1. Samples from artificial inoculation of Fusarium spp. in field tests of Germany, Czech Republic and Romania. 2. Natural infection in Northern Italy field test. 3. ELISA test for DON and T-2 detection 4. qPCR analysis on a subset of samples to quantify Fusarium CRA-GPG (ex Experimental Institute for Cereal Research), Fiorenzuola d’Arda, Italy Valeria Terzi, Caterina Morcia
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Bars with different colors are different at the Tukey’s HSD test (P=0.05) Avena genetic resources for quality in human consumption (AVEQ) WP5,Fusarium.
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Avena genetic resources for quality
in human consumption (AVEQ)
WP5,Fusarium and Mycotoxins analysis.
1. Samples from artificial inoculation of Fusarium spp. in field tests of Germany,
Czech Republic and Romania.
2. Natural infection in Northern Italy field test.
3. ELISA test for DON and T-2 detection
4. qPCR analysis on a subset of samples to quantify Fusarium
CRA-GPG (ex Experimental Institute for Cereal Research), Fiorenzuola d’Arda,
Italy
Valeria Terzi, Caterina Morcia
ELISA test for mycotoxin detection
127 samples from Fusarium Screening 2008 in Gross Lusewitz
ELISA tests for DON on these 127 samples are ongoing
ELISA tests for T2 on these 127 samples?
Starting from the results obtained we can select a set of genotypes grown in Northern Italy,
under natural infection, and evaluate them for DON
F.culmorum Primer Forward TCACCCAAGACGGGAATGAPrimer Reverse GAACGCTGCCCTCAAGCTTprobe CACTTGGATATATTTCC
Waalwijk C, van der Heide R, de Vries I, van der Lee T, Schoen C, Costel-de Corainville G, Hausen-Hahn I, Kastelein P, Kohl J, Lonnet P, Demarquet T, Kema GHJ (2004) Eur J Plant Pathol 110: 481-494
F.graminearum Primer Forward GGCGCTTCTCGTGAACACAPrimer ReverseGGCTAAACAGCACGAATGCprobe AGATATGTCTCTTCAAGTCT
Waalwijk C, van der Heide R, de Vries I, van der Lee T, Schoen C, Costel-de Corainville G, Hausen-Hahn I, Kastelein P, Kohl J, Lonnet P, Demarquet T, Kema GHJ (2004) Eur J Plant Pathol 110: 481-494
F.culmorum Primer Forward ATGGTGAACTCGTCGTGGCPrimer Reverse CCCTTCTTACGCCAATCTTCG
F. verticilloides: MPVP228Microchodium nivale: MPVP77
0
1
2
3
4
5
6
59 61 65 71 75 85
Growth stages (BBCH)
DN
A (
ln p
g)
F 2F 1
0
0.5
1
1.5
2
2.5
3
3.5
4
0 2 4 6 8 10
DNA (ln pg)
DO
N (
ln p
pm
)
F. culmorum
F. graminearum
3 bread wheat cv3 durum wheat cv2 yearsInoculation with F. graminearum (2 strains)
Inoculation with F. culmorum (3 strains) 6 developmental stages
anno di coltivazione (21%)
specie di Fusarium (6%)
stadio di crescita del frumento (32%)
varietà di frumento (4%)
anno x stadio di crescita (5%)
anno x varietà (8%)
specie di Fusarium x stadio di crescita
(4%)
varianza residua (20%)
Parametri di regressione 1 Campioni
N di casi
Coefficiente di
correlazione a b P2 R2
adj3
Totale dei campioni
44 0.93 458.3 10.11 < 0.001 0.86
con DON 500 μg/kg
28 0.91 997.7 9.77 < 0.001 0.83
Granella 11 0.97 -120.0 11.11 < 0.001 0.92
Farina integrale
11 0.89 -47.9 7.99 < 0.001 0.77
Farina 11 0.99 -49.6 12.60 < 0.001 0.98
Pane 11 0.87 -266.1 91.27 0.001 0.72
Cultivar Località Contenuto in DON (μg/kg)
A 30.217 B 29.630 C 15.685 D 2.288 E 247
Guadalupe
F 11 G 3.559 H 14.495 I 2.173 L 285
Serio
M 140
0 20 40 60 80 100
bread
flour
wholemeal
grainsqPCR based Fusarium
traceability in raw materials
and food.
Fusarium traceability for early diagnosis
Fun
gus
Hou
rs
Bila
ncia
Sag
ittar
io
Dui
lio
S.C
arlo
06
121824487214421628806
1218244872144216288
F.
culm
orum
F.
gram
inea
rum
2 bread wheat cv2 durum wheat cvIn potsInoculation with F. graminearum Inoculation with F. culmorumFlowering stage (65 BBCH)Samples from 6hours from inoculation to 12 days
Avena genetic resources for quality in human consumption (AVEQ)
qPCR based Fusarium traceability
1.We have checked our qPCR assay on strains fromCzech Republic and Germany. It works for F.culmorum and graminearum.
2.Preliminary qPCR analysis was done on a set of oat samples from Czech Republic.
Avena genetic resources for quality in human consumption (AVEQ)
qPCR based Fusarium traceability
1.2. We’ll apply the assay to 11 standard cv, inoculated and non
inoculated
3. PCR results will be gathered with ELISA results
4. Starting from the results, it will be evaluated to expand the PCR assays.