Angers, France EPPO Conference on diagnostics for plant pests · S. endobioticum, and a test to discriminate between pathotype 1(D1) and non-pathotype 1(D1) isolates (Bonants et al.,

Angers, France December 2015

EPPO Conference on diagnostics for plant pests

Results of the Euphresco-SENDO project : improving diagnostics in Synchytrium endobioticum

Gerard van Leeuwen, NPPO-NL, Wageningen, the Netherlands

Contents of my presentation

3

Euphresco –SENDO project : January 2012-May 2015 -focus on potato wart disease (Synchytrium endobioticum)

1. Pathotypes/races, bio-assays

-Search for ‘new’ cultivars to replenish/replace the current set of differential cultivars -Test Performance Study with three pathotypes, 5-6 laboratories; Glynne-Lemmerzahl method.

Euphresco –SENDO project : January 2012-May 2015

2. Molecular tests - two types of PCR’s tested : to detect/identify S. endobioticum,

and a test to discriminate between pathotype 1(D1) and non-pathotype 1(D1) isolates (Bonants et al., 2015)

PJM Bonants, MPE van Gent-Pelzer, GCM van Leeuwen & TAJ van der Lee, 2015. A real-time Taqman PCR assay to discriminate between pathotype 1(D1) and non-pathotype 1(D1) isolates of Synchytrium endobioticum. European Journal of Plant Pathology 143, p 495-506

Consortium Euphresco-SENDO• FERA, UK• ILVO, Belgium• DAFF, Ireland• MOA, Lithuania • SASA, UK• VIZR, Russia (St Petersburg)• All-Russian Plant Quarantine Center, Russia (Moscow)• CLPQ, Bulgaria • JKI, Germany• IHAR, Poland • NPPO-NL, Plant Research International, Hilbrands laboratory (all

the Netherlands)

Kick-off meeting Wageningen, January 2013

Euphresco SENDOPathotypes/races in Synchytrium endobioticum

- Principle = bioassay used to identify pathotype, several cultivars included

Objective of the project :- search for ‘new’ cultivars to replenish/replace the current set of

differential cultivars - Test Performance Study with three pathotypes, 5-6 laboratories

(Glynne-Lemmerzahl method)

State of the art (differential cultivars)• EPPO Diagnostic Protocol-

PM 7/28 (2004) Cultivar 1(D1) 2(G1) 6(O1) 8(F1) 18(T1) Deodara S S S S S Tomensa S S S S S Eersteling S S S S S Producent R S S S S Combi R S S S S Saphir R S R R R Delcora R R R S S Miriam R R R R S Karolin R R R R R Ulme R R R R R Belita R R R R –

S = wart formation

R = no wart formation

Cultivar

1(D1)

2(G1)

6(O1)

8(F1)

18(T1)

Deodara

S

S

S

S

S

Tomensa

S

S

S

S

S

Eersteling

S

S

S

S

S

Producent

R

S

S

S

S

Combi

R

S

S

S

S

Saphir

R

S

R

R

R

Delcora

R

R

R

S

S

Miriam

R

R

R

R

S

Karolin

R

R

R

R

R

Ulme

R

R

R

R

R

Belita

R

R

R

R

–

EPPO Diagnostic protocol• Results in the table of PM 7/28 are mostly based on results

obtained with the Spieckermann method (NL)

EPPO Diagnostic protocol (Table 1)

• and based on wart formation : yes/no (S and R)• conclusion : … a Dutch table

• Which reaction types do we score when scoring the reaction of potato cultivars to infection with Synchytrium endobioticum ?

Reaction types in scoring results (5 classes)

1. Type ‘- ‘ 2. Type ‘P’


3. Type ‘F’ 4. Type ‘R’


5. Type ‘wart’….

Euphresco-SENDO• Initiative in Euphresco-SENDO :

rely on the Glynne-Lemmerzahl method (Test Performance Study)some differential cultivars proved to give non reliable/non-consistent results in interlaboratory tests (Flath et al., 2014)

1) Flath K, Przetakiewicz J, van Rijswick PCJ, Ristau V & GCM van Leeuwen, 2014. Interlaboratory tests for resistance to Synchytrium endobioticum in potato by the Glynne-Lemmerzahl method. EPPO Bulletin 44(3), p 510-517

EPPO Diagnostic Protocol Cultivar 1(D1) 2(G1) 6(O1) 8(F1) 18(T1) Deodara S S S S S Tomensa S S S S S Eersteling S S S S S Producent R S S S S Combi R S S S S Saphir R S R R R Delcora R R R S S Miriam R R R R S Karolin R R R R R Ulme R R R R R Belita R R R R –

Cultivar

1(D1)

2(G1)

6(O1)

8(F1)

18(T1)

Deodara

S

S

S

S

S

Tomensa

S

S

S

S

S

Eersteling

S

S

S

S

S

Producent

R

S

S

S

S

Combi

R

S

S

S

S

Saphir

R

S

R

R

R

Delcora

R

R

R

S

S

Miriam

R

R

R

R

S

Karolin

R

R

R

R

R

Ulme

R

R

R

R

R

Belita

R

R

R

R

–

New differentials New cultivars tested in the project : cv Logo, Talent and Transit (German origin), and cv Gawin (Polish origin) Logo, Talent : to replace Miriam Transit and Gawin : to replace Ulme, Belita

Results - pathotype 6(O1)

IHAR HLB JKI ILVO PPS-BG

c 4 c 5 c 4 c 5 c 4 c 5 c 4 c 5 c 4 c 5Talent rep 1 9 0 0 0 6 0 6 0 0 0

rep 2 10 0 0 0 8 0 4 0 0 0rep 3 10 0 1 1 5 0 2 0 0 0

Logo rep 1 5 0 0 0 0 0 0 0 0 0rep 2 6 0 0 0 0 0 0 0 0 0rep 3 7 0 0 0 0 0 0 0 0 0

Gawin rep 1 0 0 0 0 0 0 0 0 0 0rep 2 0 0 0 0 0 0 0 0 0 0rep 3 0 0 0 0 0 0 0 0 0 0

Transit rep 1 0 0 0 0 0 0 0 0 0 0rep 2 0 0 0 0 0 0 0 0 0 0rep 3 0 0 0 0 0 0 0 0 0 0

Blad1

pathotype6(O1)

IHARHLBJKIILVOPPS-BG

c 4c 5c 4c 5c 4c 5c 4c 5c 4c 5

Talentrep 19000606000

rep 210000804000

rep 310011502000

Logorep 15000000000

rep 26000000000

rep 37000000000

Gawinrep 10000000000

rep 20000000000

rep 30000000000

Transitrep 10000000000

rep 20000000000

rep 30000000000

Blad2

Blad3

Results – pathotype 18(T1)

IHAR HLB JKI ILVO PPS-BG

c 4 c 5 c 4 c 5 c 4 c 5 c 4 c 5 c 4 c 5Talent rep 1 1 9 2 2 0 9 3 7 2 5

rep 2 0 10 3 3 0 7 0 4 0 8rep 3 0 10 1 8 0 7 6 3 0 3

Logo rep 1 9 0 0 0 5 0 0 0 1 2rep 2 10 0 0 0 2 0 0 0rep 3 10 0 0 0 3 0 0 0 0 0

Gawin rep 1 0 0 0 0 0 0 0 0 0 0rep 2 0 0 0 0 0 0 0 0 0 0rep 3 1 0 0 0 0 0 0 0 0 0

Transit rep 1 4 0 0 0 0 0 0 0 0 0rep 2 4 0 0 0 1 0 0 0 0 0rep 3 6 0 0 0 1 0 0 0 0 0

Blad1

pathotype18(T1)

IHARHLBJKIILVOPPS-BG

c 4c 5c 4c 5c 4c 5c 4c 5c 4c 5

Talentrep 11922093725

rep 201033070408

rep 301018076303

Logorep 19000500012

rep 2100002000

rep 310000300000

Gawinrep 10000000000

rep 20000000000

rep 31000000000

Transitrep 14000000000

rep 24000100000

rep 36000100000

Blad2

Blad3

Conclusion and confusion …

Conclusions :• cv Talent to replace Miriam (distinction between path 6(O1) and

18(T1))• cv Gawin to replace Ulme/Belita (resistant to all pathotypes

mentioned in EPPO DP 7/28)

Confusion …• Identity of pathotype 8(F1) ?

Acknowledgements

I want to thank the partners making this work and related activities possible in the recent past :

- Patricia van Rijswick, André Tonk, Bart van de Vossenberg, Marcel Westenberg (NPPO-NL)

- Peter Bonants, Theo van der Lee, Marga van Gent (Plant Research International)

- Margriet Boerma, Ineke van Holst (Hilbrands laboratory)

Diapositive numéro 1Diapositive numéro 2Contents of my presentationDiapositive numéro 4Consortium Euphresco-SENDOKick-off meeting Wageningen, January 2013Euphresco SENDOState of the art (differential cultivars)EPPO Diagnostic protocolEPPO Diagnostic protocol (Table 1)�Diapositive numéro 11Reaction types in scoring results (5 classes)Reaction types in scoring results (5 classes) Reaction types in scoring results (5 classes) �Euphresco-SENDO EPPO Diagnostic Protocol New differentials Results - pathotype 6(O1) Results – pathotype 18(T1) Conclusion and confusion …Acknowledgements

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Angers, France EPPO Conference on diagnostics for plant pests · S. endobioticum, and a test to discriminate between pathotype 1(D1) and non-pathotype 1(D1) isolates (Bonants et al.,