critical components of bacterial canker surveillance...
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CRITICAL COMPONENTS OF BACTERIAL CANKER SURVEILLANCE AND MANAGEMENT IN FIELD AND GREENHOUSE TOMATOES Sally A. Miller1, Melanie L. Lewis Ivey1, Fulya Baysal-Gurel1, Xiulan Xu1, Warren Arinaitwe2 and Michael E. Bledsoe3 1The Ohio State University, OARDC, Wooster, OH 2Makerere University, Kampala, Uganda 3Village Farms International, Inc., Heathrow, FL
Bacterial Canker • High consequence bacterial disease in temperate climates and greenhouses • Clavibacter
michiganensis subsp. michiganensis (Cmm) • Seedborne • Long-lived in environment • Mechanically transmitted • 1°, 2° canker in open fields
Midwestern US outbreak 2011
GH tomato outbreak, 2010
Why Identify and Track Cmm Strains?
• Understand the movement of Cmm
• Identify the source of outbreaks
• Identify critical control points
• Improve management practices
Production Flow Overview – GH Tomatoes
Germination*
Seedling*production
Grafting
Training*Dropping*
Trans6planting
Topping
De6lea7ing*
Seeding
Harvest*
Propa6gation
Crop*Work
Post6harvest*Handling***
Field Tomato Production Overview
Germination*
Seedling*production Pruning*
Staking*Spraying***
Trans6planting
Seeding
Harvest*
Propa6gation
Crop*Work
Post6harvest*Handling***
Development of a Multivariate Matrix for Cmm Surveillance • Rep-PCR fingerprinting
• Historical Cmm types: ABCD • Field tomatoes, Midwestern US, 1998 (Louws et al.)
• Field tomato haplotypes: ABCD - Israel, 2008 (Kleitman et al.) • Greenhouse tomato haplotypes: ABCD – Japan 2010 (Kawaguchi
et al.)
• Gene sequence analysis – dnaA Replication Initiation Factor (RIF) (Schneider et al. 2011)
• This study • 40 strains collected from North American GHs between 2006 and 2011
• BOX-PCR and RIF analyses • 63 strains from processing field tomatoes in OH in 2011 – BOX PCR
RIF Sequence Analysis vs. Box-PCR: Clavibacter Strains
1 C 1 CG1 11 CG3 9 CG5
1 B 2 CG1 13 CG2 1 CG3 1 CG4 2 CG5 1 A 1 CG2
BOX PCR Clonal Groups
dnaA RIF
Cmm Strain Matrix GH 2006-2011 Loca%on' State'or'
Province' Country' Tomato'Type'' Scion'' Rootstock'' 'Phase' GH'ID' House' Row'
Number' Post' Week' Year' Strain' RepBPCR'Profile'
A' TX' USA' TOV' Grandella' Maxifort' 3' '' '' '' '' '' 2009' 09B158' CG1'A' TX' USA' TOV' Grandella' Maxifort' 3' '' '' '' '' '' 2009' 09B159'' CG1'A' TX' USA' TOV' '' '' 3' '' '' '' '' 37' 2008' 08B687' CG1''A' TX' USA' TOV' Trisha' Maxifort' 1' '' '' '' '' '' 2007' VF3B70B08' CG2'A' TX' USA' TOV' Trisha' Maxifort' 2' '' '' '' '' '' 2007' VF3B71B08' CG2'A' TX' USA' TOV' Grandella' Maxifort' '' S' 22' '' 8' '' 2009' VFB22SB09' CG2'
A' TX' USA' TOV' Grandella' Maxifort' '' S' 50' '' 19' '' 2009' VFB50SAB09' CG2'A' TX' USA' TOV' Grandella' Maxifort' '' S' 50' '' 19' '' 2009' VFB50SBB09' CG2'A' TX' USA' TOV' Grandella' Maxifort' '' S' 50' '' 19' '' 2009' VFB50SCB09' CG2'A' TX' USA' TOV' Grandella' Maxifort' 3' B' 72' 2' 10' 14' 2009' 09B165'' CG2'A' TX' USA' TOV' Grandella' Maxifort' 3' '' 72' 2' 12' 14' 2009' 09B166'' CG2'A' TX' USA' TOV' Grandella' Maxifort' '' N' 21' '' 4' '' 2009' VFB21NB09' CG2'B' TX' USA' TOV' Trisha' Maxifort' 1' '' '' '' '' '' 2007' VF6B7B08' CG2'B' TX' USA' TOV' Tricia' Maxifort' 2' 13' 3' 31' 2' 42' 2009' 42B3B1' CG2'B' TX' USA' Roma' Savantas' Maxifort' '' '' '' '' '' '' 2009' 09B198A' CG2'B' TX' USA' TOV' Komeet' Maxifort' 2' 16' 10' 100' 2' 12' 2010' SM157B10' CG2'A' TX' USA' TOV' Grandella' Maxifort' '' '' '' '' '' '' 2006' VF3B1B08' CG2''A' TX' USA' TOV' Grandella Maxifort 3 B 7 2008' VF6B8B08' CG3'A' TX USA TOV Grandella Maxifort S 8 8 17 2009 VFB8SB09' CG3 A' TX' USA' Beef' Starbuck' Maxifort' '' '' 2' '' '' 15' 2009' 09B177'' CG3'B' TX' USA TOV Tricia Maxifort 2 23 46 468 5 36 2009 SM101B09' CG3'
B' TX' USA' Roma' Savantas' Maxifort' '' '' '' '' '' '' 2009' 09B197A' CG3'B' TX' USA' Roma' Savantas' Maxifort' '' '' '' 242' '' '' 2009' 09B190A' CG3'C' BC' Canada' TOV' Idol' Maxifort' '' '' '' '' '' '' 2008' B6B78' CG3'C' BC' Canada' TOV' Idol' Maxifort' '' '' '' '' '' '' 2008' B6B79' CG3'C' BC' Canada' TOV' Trisha' Maxifort' '' '' '' '' '' '' 2009' 09B192B' CG3'
D' PA' USA' TOV' Castella'or'Grandella' Maxifort' '' '' '' '' '' '' 2008' 08B568' CG3'
D' PA' USA' TOV' Castella'or'Grandella' Maxifort' '' '' '' '' '' '' 2008' 08B567' CG3'
A' TX' USA' Beef' Starbuck' Maxifort' '' '' 2' '' '' 15' 2009' 09B176'' CG3''A' TX' USA' TOV' Grandella' Maxifort' 3' '' '' '' '' '' 2009' 09B157' CG4''E' ME' USA' TOV' Success' Maxifort' 1' '' '' 227' 1' 3' 2011' SM287B11' CG5'E' ME' USA' TOV' Success' Maxifort' 1' '' '' 227' 1' 3' 2011' SM288B11' CG5'E' ME' USA' TOV' Success' Maxifort' 1' R' '' 381' 12' 3' 2011' SM290B11' CG5'E' ME' USA' TOV' Success' Maxifort' 1' L' '' 286' 11' 3' 2011' SM291B11' CG5'E' ME' USA' TOV' Success' Maxifort' 1' L' '' 286' 11' 3' 2011' SM292B11' CG5'E' ME' USA' TOV' Success' Maxifort' 1' L' '' 286' 11' 3' 2011' SM293B11' CG5'E' ME' USA' TOV' Success' Maxifort' 1' R' '' 132' 11' 5' 2011' SM295B11' CG5'E' ME' USA' TOV' Success' Maxifort' 1' L' '' 103' 1' 4' 2011' SM297B11' CG5'E' ME' USA' TOV' Success' Maxifort' 1' R' '' 132' 16' 5' 2011' SM298B11' CG5'E' ME' USA' TOV' Success' Maxifort' 1' R' '' 381' 12' 3' 2011' SMB289B11' CG5'E' ME' USA' TOV' Success' Maxifort' 1' R' '' 132' 11' 5' 2011' SM296B11' CG5'
Location E - 2011
Loca%on' State'or'Province' Country' Tomato'
Type'' Scion*'' Rootstock*'' 'Phase' GH'ID' House' Row'Number' Post' Week' Year' Strain' RepBPCR'
Profile'
E' ME' USA' TOV' Success' Maxifort' 1' '' '' 227' 1' 3' 2011' SM287B11' CG5'
E' ME' USA' TOV' Success' Maxifort' 1' '' '' 227' 1' 3' 2011' SM288B11' CG5'
E' ME' USA' TOV' Success' Maxifort' 1' R' '' 381' 12' 3' 2011' SM290B11' CG5'
E' ME' USA' TOV' Success' Maxifort' 1' L' '' 286' 11' 3' 2011' SM291B11' CG5'
E' ME' USA' TOV' Success' Maxifort' 1' L' '' 286' 11' 3' 2011' SM292B11' CG5'
E' ME' USA' TOV' Success' Maxifort' 1' L' '' 286' 11' 3' 2011' SM293B11' CG5'
E' ME' USA' TOV' Success' Maxifort' 1' R' '' 132' 11' 5' 2011' SM295B11' CG5'
E' ME' USA' TOV' Success' Maxifort' 1' L' '' 103' 1' 4' 2011' SM297B11' CG5'
E' ME' USA' TOV' Success' Maxifort' 1' R' '' 132' 16' 5' 2011' SM298B11' CG5'
E' ME' USA' TOV' Success' Maxifort' 1' R' '' 381' 12' 3' 2011' SMB289B11' CG5'
E' ME' USA' TOV' Success' Maxifort' 1' R' '' 132' 11' 5' 2011' SM296B11' CG5'
Only Cmm CG5 was detected throughout large greenhouse 3-5 weeks after transplanting *Does not infer source of Cmm
Village Farms Ft Davis
CG2 and CG3 detected between 2007 and 2010 CG2 Detected in GH 10 in both 2009 and 2010 CG3 introduced in a specific variety in 2009 but affected many other varieties *Does not infer source of Cmm
Grower'or'Company'
Grower'Facility'' Region,'City'State,'Region,'or'Province' Country' Tomato'
Type'' 'Scion*' Rootstock*' Field'or'Phase' A/B'' House' Row'
Number' Post' Week' Year' CG''
Village'Farms'Ft'Davis' Fort'Davis' Texas' USA' '' Trisha' Maxifort' 1''' '' '' '' '' 2007' CG2'Village'Farms'Ft'Davis' Fort'Davis' Texas' USA' Roma' Savantas' Maxifort' '' '' '' '' '' '' 2009' CG3'Village'Farms'Ft'Davis' Fort'Davis' Texas' USA' Roma' Savantas' Maxifort' '' '' '' 242''' '' 2009' CG3'Village'Farms'Ft'Davis' Fort'Davis' Texas' USA' Roma' Savantas' Maxifort' '' '' '' '' '' '' 2009' CG2'Village'Farms'Ft'Davis' Fort'Davis' Texas' USA' Roma' Savantas' Maxifort' '' '' '' '' '' '' 2009' CG2'Village'Farms'Ft'Davis' Fort'Davis' Texas' USA' TOV' Tricia' Maxifort' 2' 15' 10' 101' 5' 38' 2009' CG2'Village'Farms'Ft'Davis' Fort'Davis' Texas' USA' TOV' Tricia' Maxifort' 2' 13' 3' 31' 2' 42' 2009' CG2'Village'Farms'Ft'Davis' Fort'Davis' Texas' USA' TOV' Tricia' Maxifort' 2' 23' 46' 468' 5' 36' 2009' CG3'Village'Farms'Ft'Davis' Fort'Davis' Texas' USA' TOV' Tricia' Maxifort' 2' 14' 2' 26' 16' 42' 2009' CG3'Village'Farms'Ft'Davis' Fort'Davis' Texas' USA' TOV' Komeet' Maxifort' 2' 16' 10' 100' 2' 12' 2010' CG2'Village'Farms'Ft'Davis' Fort'Davis' Texas' USA' TOV' Komeet' Maxifort' 2' 16' 12' 128' 3' 12' 2010' CG2'Village'Farms'Ft'Davis' Fort'Davis' Texas' USA' TOV' Komeet' Maxifort' 2' 13' 9' 97' 9' 12' 2010' CG2'
Cmm Matrix (partial) GROWER
INFORMATION Crop Information PROPAGATOR SAMPLE INFORMATION
Farm State Year GPS Variety* Type Date
Received CG Source
Lab Source Lab Identifier
OSU Sample Name
A OH 2011 46 Processing 2 23#Aug' 8 OSU 59A#1' SM502#11'A OH 2011 46 Processing 2 23#Aug' 8 OSU 59A#2' SM516#11'A OH 2011 46 Processing 2 23#Aug' 8 OSU 59A#3' SM513#11'A OH 2011 46 Processing 2 23#Aug' 8 OSU 59B#1' SM559#11'A OH 2011 46 Processing 2 23#Aug' 8 OSU 59B#2' SM525#11'A OH 2011 46 Processing 2 23#Aug' 8 OSU 59C#1' SM555#11'A OH 2011 46 Processing 2 23#Aug' 8 OSU 59C#2' SM569#11'A OH 2011 46 Processing 2 23#Aug' 8 OSU 59C#3' ''A OH 2011 331 Processing 1 23#Aug' 8 OSU 60A#1' SM551#11'A OH 2011 331 Processing 1 23#Aug' 8 OSU 60A#2' SM570#11'A OH 2011 331 Processing 1 23#Aug' 8 OSU 60A#3' SM571#11'A OH 2011 331 Processing 1 23#Aug' 8 OSU 60B#3' SM542#11'A OH 2011 9704 Processing 1 23#Aug' 8 OSU 61A#2' SM539#11'A OH 2011 9704 Processing 1 23#Aug' 8 OSU 61A#3' SM541#11'A OH 2011 9704 Processing 1 23#Aug' 8 OSU 61B#2' SM552#11'A OH 2011 9704 Processing 1 23#Aug' 8 OSU 61C#2' ''B OH 2011 46 Processing 1 23#Aug' 8 OSU 65A#2' ''B OH 2011 46 Processing 1 23#Aug' 8 OSU 65A#3' SM553#11'B OH 2011 46 Processing 1 23#Aug' 8 OSU 65A#4' ''B OH 2011 9704 Processing 1 23#Aug' 8 OSU 64#2' SM524#11'B OH 2011 9704 Processing 1 23#Aug' 8 OSU 64#3' SM573#11'B OH 2011 9704 Processing 1 23#Aug' 9 OSU 64#1' SM524#11'C OH 2011 331 Processing 1 23#Aug' 8 OSU 58A#2' SM511#11'C OH 2011 331 Processing 1 23#Aug' 8 OSU 58A#3' SM509#11'C OH 2011 331 Processing 1 23#Aug' 8 OSU 58B#1' SM505#11'C OH 2011 331 Processing 1 23#Aug' 8 OSU 58B#4' SM510#11'C OH 2011 331 Processing 1 23#Aug' 8 OSU 58C#3' SM507#11'C OH 2011 331 Processing 1 23#Aug' 8 OSU 58D#1' SM499#11'C OH 2011 331 Processing 1 23#Aug' 9 OSU 58A#1' ''C OH 2011 331 Processing 1 23#Aug' 9 OSU 58B#2' SM498#11'C OH 2011 331 Processing 1 23#Aug' 9 OSU 58C#4' SM508#11'C OH 2011 9704 Processing 1 23#Aug' 9 OSU 61C#3' ''
*Does not infer source of Cmm
Distribution of Cmm Clonal Group 9 in Contiguous Tomato Fields in OH
WEST EAST Variety/Field 46 4007 331 401 9704
Greenhouse
2
2
1
2
1
Canker Incidence
5% 0 80% 0 85%
Canker Severity
1-2% 0 30% 0 50%
CG 9 - 9 - 9
August 23, 2011
Summary: Cmm Clonal Groups in Multivariate Matrices • Recently isolated strains different from historical groups
ABCD
• Strains/CGs from greenhouse and field outbreaks are different • GH, field fresh market and processing varieties are produced by
different companies in many different locations
• Matrices detailing substantial information on GH and field outbreaks can provide clues to pathogen source and movement
Bacterial Canker Management • Early intervention is critical – GH and field systems
• Seed • Prevention • Testing • Treatment
• Propagation • Environment • Sanitation • Bactericidal treatments – Copper, streptomycin • Testing?
• Production • Field – not much; long rotations necessary • GH – sanitation, sanitation, sanitation
After 26 Years, What Have We Accomplished in Management of Bacterial Canker? • No resistant varieties • No highly effective bactericides • A new GH tomato industry threatened • Seeds are still a source of Cmm
• We know a lot about the epidemiology of this disease • But not the frequency and mechanism of seed infection
• Cmm testing methods have improved significantly • Standardization is still needed • Effective sanitizing seed treatments are known but are generally not
adopted • Field production practices have changed
• The industry moved away from bare-root transplants but canker did not go away
• Greenhouse environment for seedling production highly conducive to bacterial disease development
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