New Findings Relating to Bacterial Wilt

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New Findings Relating to Bacterial Wilt
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Research Team
Dave Norman Jeanne Yuen Arianna Novo Drew Byrne
Patty Ramos Loren Walsh Rosa Resendiz Ryan D
onahoo
Joe Boswell
J. Chen Jake Henny Dean W. Gabriel

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Research Objectives

• Differentiation of Populations
• Chemical Control
• Varietal Differences in Geranium Susceptibility
• Expanded Study on Freeze Survival

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Research Objectives

• Differentiation of Populations
• Chemical Control
• Varietal Differences in Geranium Susceptibility
• Expanded Study on Freeze Survival

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• Infrasubspecific Groups
• 16S RNA gene sequence
• HrpB gene sequence
• Endoglucanase gene sequence

• Clonal Lines
• RAPDS
• AFLP
• rep-PCR

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Digital Photo Taken with Kodak Electrophoresis
Documentation System
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Using BioNumerics Rep-PCR Data can be Analyzed
Using Single or Multiple Primers
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Similarity coefficient
6 strains, Kenya geranium UW551
ATCC 33193
16 strains, Costa Rica geranium
A
Biovar 2
13 strains, Guatemala geranium
8 strains, LMG Controls
10 strains, Caribbean anthurium
B
Biovar 1, 3
C
6 strains, Caribbean vegetables
D
30 strains, Florida, Puerto Rico, Guadeloupe,
Martinique vegetable ATCC 33192
Biovar 1
27 strains, Costa Rica Pothos
E
Biovar 1
F
21 strains, Costa Rica Pothos, anthurium
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Research Objectives

• Differentiation of Populations
• Chemical Control
• Varietal Differences in Geranium Susceptibility
• Expanded Study on Freeze Survival

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Fosphite, Biophos, Agri-Fos, ProPhyt
K-Phite Plant Food Systems
Mono- and di-potassium salts of Phosphorous acid
(53)
Phosphonate fungcide
Phosphorus acid H3PO3 Potash K2O
Current Uses Fertilizer Pythium/ Phytophthora
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Testing Protocol
Tomato transplants
Inoculate
6 weeks
Soil isolation
4 weeks
Reisolation
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Control of Bacterial Wilt of Geranium With K-
Phite
R3B2, UW 551 3 X 108 cfu/g soil
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Concentration of RS in soil after test R3b2
Log10 cfu/g soil
One application
14 day interval ( 2 applications)
7 day interval ( 4 applications)
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Activity of Phosphorous Compounds
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Foliar analysis of geraniums with five
applications of K-Phite on a 7 - day cycle
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Water Control
K-Phite 1
K-Phite 2

• Treatments
• Water Control
• K-Phite 0.5
• K-Phite 0.75
• K-Phite 1
• K-Phite 2

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Research Objectives

• Differentiation of Populations
• Chemical Control
• Varietal Differences in Geranium Susceptibility
• Expanded Study on Freeze Survival

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Approximately 200 different Pelargonium species
used over a 350 year period
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Zonal - Pelargonium X hortorum
Ivy Pelargonium peltatum
Regal - Pelargonium X domesticum
Scented Many species mostly Pelargonium
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AFLP on 64 cultivars provided by Ball, Fisher,
Goldsmith, Oglevee, Shady Hill Garden, and Yoders
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Inoculated with 1000 to 5000 cfu
Reisoaltions done 60 days after inoculation
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Research Objectives

• Differentiation of Populations
• Chemical Control
• Varietal Differences in Geranium Susceptibility
• Expanded Study on Freeze Survival

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Cold tolerance defined as ability to survive
cold temperatures. Cold treatments 10º C ov
ernight, then -20º C for 20 hrs. followed by 30º
C for 4 hrs.
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UW551 cspD3 conferred cold tolerance to GMI1000.
cspD3 was recloned into wide host range plasmid
pUFJ05 and transformed into GMI1000. 12 cycles
of daily cold treatments of - 8º C for 20 hrs
followed by 30 º C for 3-hrs) in nutrient broth
indicated that cspD3 conferred cold tolerance to
GMI1000.