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Jan van der Wolf - EAPR · Collaborators in the Erwinia Deltaplan (H. Velvis, K. Kristelijn, D....
Transcript of Jan van der Wolf - EAPR · Collaborators in the Erwinia Deltaplan (H. Velvis, K. Kristelijn, D....
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Potato blackleg: remedies for a way out
Jan van der Wolf
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Outline presentation
Symptoms and causative agents
Initial introduction and spread of the pathogen
Control
● Clean seed (seed certification)
● Hygiene and cultural practices
● Resistance
● Tuber treatments
● Biocontrol
● Disinfectants
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Outline presentation
Symptoms and causative agents
Initial introduction and spread of the pathogen
Control
● Clean seed (seed certification)
● Hygiene and cultural practices
● Resistance
● Tuber treatments
● Biocontrol
● Disinfectants
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Symptoms Dickeya/Pectobacterium potato
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Important features causative agent
Enterobacteriaceae (not spore forming)
1-2 µm
Pectinolytic
Facultative anaerobes
Motile
0.8 µm
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Reason problems
Rotten tubers infectious smearing
Introductions difficult to avoid
Often latent infections
No chemicals to control blackleg
No resistant varieties
● Breeding for resistance hampered by absence of reliable bioassays
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“Erwinia” species of potato Pectobacterium
P. carotovorum subsp. carotovorum
P. wasabiae P. atrosepticum
P. carotovorum subsp. brasiliensis
Dickeya
D. dadantii
D. solani D. dianthicola
D. zeae
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Virulence in the field (NL, 2011)
d d
a
ab
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de
cd
ef
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a
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Outline presentation
Symptoms and causative agents
Initial introduction and spread of the pathogen
Control
● Clean seed (seed certification)
● Hygiene and cultural practices
● Resistance
● Tuber treatments
● Biocontrol
● Disinfectants
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Erwinia infections during multiplication
Yr 0: start minitubers (11 growers)
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Potential sources of introduction: machines
Planting Spraying (tyres): Infections of haulms, roots and tubers
Flailing (haulm topper): Infections of haulms (via flails), roots and tubers (via tyres or indirectly via infected haulms)
Harvesting Sorting
+ +
+
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Potential sources initial infection: “air –
and soilborne”
+?
Insects Rain/aerosols Irrigation water
Weeds/potato volunteers Soil
+/-?
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Haulm infections in first years minitubers
(19 fields, 100 leaves/field, 15 July 2012, E-TaqMan)
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Haulm infections can result in infected
progeny at 42 dpi (D. solani, 106 cfu/ml)
Inoculation point: 100%
Stem top: 40%
Progeny: 20%
Stolons: 40%
Abaxial side
Stem base: 20%
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Spread from rotten tubers via free water in
soil resulting in root and stem infections
control
Roots, 1
dpi
D. solani
stems, 15 dpi
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Spread via machines
Planting Spraying (tyres): Infections of haulms, roots and tubers
Flailing (haulm topper): Infections of haulms (via flails), roots and tubers (via tyres or indirectly via infected haulms)
Harvesting Sorting
+
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Outline presentation
Symptoms and causative agents
Initial introduction and spread of the pathogen
Control
● Clean seed (seed certification)
● Hygiene and cultural practices
● Resistance
● Tuber treatments
● Biocontrol
● Disinfectants
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Clean seed
Role of seed testing
● Field inspections
● Recommended protocol laboratory testing
● Test at least 200 tubers in multiple composite samples to calculate disease incidence I = {1 – [(N – p)/N]1/n}*100 I = incidence, N= number of
subsamples, p = number positives, n = number of tubers per
subsample (Plant Disease 86, 960 (2002))
● Test peel and heel ends
● Incubate in Pectate Enrichment Broth prior to testing (72 h, low oxygen conditions)
● Use TaqMan assays for final testing
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TaqMan detection Dickeya/Pectobacterium
Generic Pectobacterium/Dickeya (JHI, PRI)
Generic Dickeya (FERA)
P. atrosepticum (FERA, PRI)
P. wasabiae (PRI)
D. solani (JHI, ILVO) D. dianthicola (JHI, PRI) D. dadantii (PRI) D. dieffenbachiae (PRI) D. zeae (PRI) D. chrysanthemi (PRI) D. paradisiaca (PRI)
+
Internal amplification control
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Hygiene and cultural practices: plant
growth
Wash and disinfect machines and materials
Avoid water logging of soil drainage
Don’t use surface water for irrigation
Selection of diseased plants (roguing): only useful early in season
Full field spraying followed by flailing (after 5 days) is preferred above flailing followed by spraying
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Hygiene and cultural practices:
(post)harvest
Avoid wounding set machines carefully
Harvest manually as long as possible
Remove rotten tubers from harvesters and graders
Harvest under dry conditions
Dry, dry, dry ........
Store tubers in well-ventilated rooms at low temperatures
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Resistance breeding: petiole test
Test conditions
● 25 oC
● 16 h L/8 h D
● Ca. 100% RH
Bacterial strains
● Pcc 1948 (107 cfu/ml)
● Pw 1957 (107 cfu/ml)
● Ds 2222 (105 cfu/ml)
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340 Accessions 532 genotypes Nearly all potato species were included
The CBSG collection
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Results petiole assay
* Yungasensa, tolerance known from literature
Pcc Pw Ds Pcc Pw Ds
< 40% maceration
49 141 23 6 24 1*
40-80% maceration
182 209 188 48 128 18
> 80% maceration
301 182 321 178 380 513
2 DPI
3 DPI
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Preliminary conclusions
Tolerance against stem maceration (but also susceptibility) is present in Yungasensa (S. chacoense)
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Biocontrol with Serratia plymuthica A30
A30 is an endophyte isolated from potato tubers
Potato tubers inoculated with 106 cells/ml ‘D. solani’ (dsRed-tagged) and 1010 cells/ml S. plymuthica A30 (GFP-tagged)
Evaluation:
● Symptom expression
● Presence ‘D. solani’ and S. plymuthica A30 in roots and stems
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Greenhouse experiment: overview
T=28 dpi
S. plymuthica A30 water co-inoculation D. solani
Czajkowski et al., Plant Pathology 2012
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Effect disinfection of seed tubers on blackleg
development
Inoculation with 108 cfu/ml D. solani or with water (1 h)
drying tubers (15 min)
disinfection (30 min) and drying
planting in glasshouse in potting compost (28 °C,75% RH, 3-4 weeks)
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Effect disinfection seed tubers on blackleg
development
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Conclusions
The diversity of blackleg causing pathogens found in Europe is increasing
Initial infections are probably caused by air-borne infections and contaminated machines
Spread of the disease occurs mainly during mechanical harvesting
Use of pathogen-free seed in combination with hygiene and cultivation practices are still the most important management tools
Sources of resistance (tolerance) have been identified
There are perspectives for tuber treatments (biocontrol agents, disinfectants) able to reduce disease incidence
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Acknowledgement
Colleagues from PRI (P. Kastelein, L. de Haas, P. van der
Zouwen, M. Krijger, H. Rietman, R. Visser, T. Been, L. Leite)
Collaborators from NIOO (R. Czajkowski, H. van Veen)
Collaborators in the Erwinia Deltaplan (H. Velvis, K. Kristelijn,
D. Boomsma)
Collaborators from the EU Euphresco project
Collaborators in Canada and USA (S. de Boer, S. Li, A.
Charkowski)
Dutch potato growers (organizations)
Dutch Ministry of Economic Affairs
Dutch foundation for Technical Science (STW)