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Evaluation of Fusarium Head Blight resistances in Italianwheat cvs. by phenotypic and molecular analyses
Francesconi S., Mazzaglia A., Balestra G.M.
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Dipartimento di Scienze Agrarie e Forestali (DAFNE), Università degli Studi della Tuscia, Via San Camillo de Lellis, snc, 01100, Viterbo
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INTRODUCTION – The wheat crop
• One of the most cultivated crop;
• High economic importance;
• Intense cytogenetic and genetic studies;
• Its domestication caused genetic erosion:
Increased SUSCEPTIBILITY to diseases.
Wheat land in «Agabiti» farm, Amelia (PG), Italy. Creditsto Prof. G.M. Balestra, University of Tuscia, Viterbo (Italy).
INTRODUCTION – Fusarium Head Blight
• The genus Fusarium has a global distribution;
• Many of them are phytopathogenic fungi andmycotoxins producers (DON);
• The main causal agent is F. graminearum;
• The infection is highly related to phenological stage andweather conditions;
• The infection biology is yet to be fully understood;
• Resistance to FHB is of a quantitative and multigenicnature TYPE I and TYPE II;
• Today, no genotype is completely immune;
• AIM: Evaluating Type I and Type IIresistances in three Italian wheat cvs.
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3
F. graminearum disease cycle (Virginia State University,https://pubs.ext.vt.edu/3102/3102-1535/3102-1535.html).
Infected wheat spike (Triticumdurum cv. Marco Aurelio). Creditsto Prof. G. M. Balestra, University ofTuscia, Viterbo (Italy).
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Materials and Methods
• Wheat Italian cvs.: Palesio, Claudio and Marco Aurelio;
• Fungal Reference strain: F. graminearum WT 3824;
• Two inoculation methods: Spray and Point inoculation;
• Three conidial concentrations: 500 c/s, 1500 c/s, 2500 c/s;
• Disease assessment: % of symptomatic spikelets;
• F. graminearum biomass quantification with Real-Time qPCR from chaff and rachis;
• Standard curves for wheat (Actin) and F. graminearum (Tri6) biomass: data reported as ng of fungal DNA/ng of plant DNA.
• TKW for yield impact evaluation;
• Analysis of variance and Tukey’s test.
Wheat plant grown in greenhouse (University of Tuscia,Viterbo, Italy). Credits to Sara Francesconi.
Gene and Accession Number Pair of primers (5'-3') bp Primers source
Actin (TaACT) - AB181991TCCTGTGTTGCTGACTGAGG
350 Tundo et al., 2016GGTCCAAACGAAGGATAGCA
Trichothecene Transcriptional Regulator (Tri6) -
AB017495, AF359361, AY102605, AB060689
TCTTTGTGAGCGGACGGGACTTTA245 Horevaj et al., 2011
ATCTCGCATGTTATCCACCCTGCT
Genes and primers for F. graminearum and wheat DNA quantification.
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Results and Discussion – Symptoms development by different conidial concentrations
0
10
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100
3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
% o
f in
fect
ed s
pik
elet
s
Palesio - Spray Inoculation
3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
Claudio - Spray Inoculation
*
3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
M. Aurelio - Spray Inoculation
*
**
0102030405060708090
100
3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
% o
f in
fect
ed s
pik
elet
s
days post infection
Palesio - Point Inoculation
500 c/s 1500 c/s 2500 c/s
**
** ** **
3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
days post infection
Claudio - Point Inoculation
500 c/s 1500 c/s 2500 c/s
3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
days post infection
M. Aurelio - Point Inoculation
500 c/s 1500 c/s 2500 c/s
**
**** **
(*) p < 0,05 and (**) p < 0,0,1
Results and Discussion – Symptoms development by different inoculation methods
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0
20
40
60
80
100
3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21% o
f in
fect
ed s
pik
elet
s
days post infection
Palesio
Spray inoculation Point inoculation
3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
days post infection
Claudio
Spray inoculation Point inoculation
*
3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
days post infection
M. Aurelio
Spray inoculation Point inoculation
**
**
**
****
**
• M. Aurelio showed Type Iresistance;
• The importance of both Sprayand Point methods in testing FHBresistances Advantages andDisadvantages.
(*) p < 0,05 and (**) p < 0,0,1
Results and Discussion – Fungal biomass quantification
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0.000
0.200
0.400
0.600
0.800
1.000
1.200
1.400
1.600
1.800
2.000
Spray Inoculation Point Inoculation
ng
fun
gal D
NA
/ng
pla
nt
DN
A
Palesio
**
0.000
0.100
0.200
0.300
0.400
0.500
0.600
0.700
Spray Inoculation Point Inoculation
ng
fun
gal D
NA
/ng
pla
nt
DN
A
Claudio
**
-0.050
0.000
0.050
0.100
0.150
0.200
0.250
0.300
Spray Inoculation Point Inoculation
ng
fun
gal D
NA
/ng
pla
nt
DN
A
M. Aurelio
*
• Point Inoculation resulted in a higherfungal DNA concentration;
• Genotypes have different ability toconfine the pathogen spread.
(*) p < 0,05 and (**) p < 0,0,1
Amplificaton curves and Melting curves of Tri6 gene.
Results and Discussion – TKW
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0
10
20
30
40
50
60
70
80
90
Spray Inoculation Point Inoculation
% o
f yi
eld
loss
Palesio
*
0
10
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50
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80
90
100
Spray Inoculation Point Inoculation
% o
f yi
eld
loss
Claudio
*
0
10
20
30
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50
60
70
80
90
100
Spary Inoculation Point Inoculation
% o
f yi
eld
loss
M. Aurelio
Uninfected seed (left) andinfected seed (right). Credit toSara Francesconi.
• % of yield loss ishigher with Point Inoculation.(*) p < 0,05
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Conclusions and Future Perspectives
• No differences in symptoms progress between different conidial concentration;
• Marco Aurelio is the most promising wheat cv. for FHB Type I resistance.
• Natural compounds as fungicides;
• How stomatal conductance is envolved in FHB resistance? Transcriptomic and Phenomic approaches.
Thank YouUniversità degli Sudi della Tuscia, Dipartimento di Scienze Agrarie e Forestali (DAFNE), Via San Camillo de Lellis, snc, 01100, Viterbo, Italy
+39 388183991Sara Francesconi
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