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Adjustments in the Tomato
Spray Program in Tennessee
Steve Bost
Professor and Extension Plant Pathologist
University of Tennessee
Control of early blight and bacterial spot two of the most
important diseases of tomato in Tennessee was less than
desirable Pathogen resistance to control products may be
responsible
Early blight drives the
tomato spray program
which has featured QoIrsquos
since Quadris was
labeled in 1997
Early Blight of Tomato2013 Field Trial Nashville
This trial was a search for replacements for the Group 11 fungicides ndash Quadris
Cabrio and Tanos Plants were sprayed 3 times at weekly intervals and evaluated
1 week after the 3rd spray
c
c
c
c
b
b
b
a
a
0 10 20 30
Untreated check
Quadris 62 fl oz
Tanos 8 oz
Cabrio 12 oz
Bravo 275 pt
Manzate 3 lb
Switch 14 oz
Inspire Superhellip
Fontelis 20 fl oz
Leaf area affected ()
Pro
du
ct
amp r
ate
75 g
al
The results were used to guide the development of non-strobilurin-based spray
programs
Early Blight Spray Programs2013 Trial Highland Rim REC
Four spray programs were evaluated on trellised lsquoMountain Gloryrsquo tomatoes
sprayed every 7-10 days beginning 3 wks after planting total of 10 sprays
Program Product
Early blight
leaf area
affected
(final)
Check none 880 a
1 Manzate alt with Quadris 575 b
2 Manzate every application 233 c
3 Fontelis 10 fl oz + Manzate + Kocidealt Manzate + Kocide
03 d
4 Inspire Super 1 pt then Manzate then Fontelis 1 pt then Manzate then repeat
01 d
The minimum labeled rate of Fontelis is 1 ptacre
Early Blight Fungicides2014 Field Trial Nashville
This trial was a search for candidates for inclusion in a spray program Plants were
sprayed 3 times at weekly intervals and evaluated 1 week after the 3rd spray
d
d
c
c
b
b
a
a
a
a
a
0 10 20 30 40 50
Untreated check
Quadris 62 fl oz
Double Nickel 1 qt
PhD 62 oz
Manzate 3 lb
Bravo 275 pt
Switch 14 oz
Endura 35 oz
Inspire Super 20 fl oz
Priaxor 8 fl oz
Fontelis 20 fl oz
Leaf area affected ()
Pro
du
ct
amp r
ate
75 g
al
7
7
39
7
912
M
M
19
NC
11
Do I really need
to use anything
more expensive
than chlorothalonil
or mancozeb
Early Blight Spray Programs2014 Trial Highland Rim REC
Four spray programs were evaluated on trellised lsquoMountain Gloryrsquo tomatoes
sprayed every 7-10 days beginning 5 wks after planting total of 7 sprays
Program Product
Early blight
leaf area
affected
(mid-harvest)
Check none 913 a
1 Manzate alt with Quadris every 3rd application 663 b
2 Manzate every application 513 c
3 Fontelis 10 fl oz + Manzate + Kocide every application
29 d
4 Inspire Super 1 pt then Manzate then Fontelis 1 pt then Manzate then repeat
43 d
The minimum labeled rate of Fontelis is 1 ptacre
Unsprayed check
Program 4
Program 4Unsprayed check
Excellent control of early blight was provided by spray programs
containing Inspire Super or Fontelis
Early Blight Resistant Varieties2014 Field Trial Cheatham County TSU ARECDeterminate varieties with resistance to early blight were compared
with the susceptible variety Mountain Glory Quadris was applied once
c
bc
bc
bc
bc
bc
b
a
0 20 40 60 80
Plum Regal
Mountain Merit
Plum Crimson
Mountain Fresh +
Iron Lady
BHN 964
Defiant
Mountain Glory
Leaf area affected at mid-harvest ()
Va
rie
ty
Spray program for
an early-blight resistant varietyMountain Merit
Plateau REC 2015
Spray program
Early blight severity ()
Mountain Glory Mountain Merit
Unsprayed control 563 a 150 a
Manzate each application 50 b 14 b
Fontelis then Manzate then
Inspire Super then Manzate then
repeat sequence
06 b 02 b
Ratings were taken just prior to harvest Early blight was later occluded by bacterial spot
1st spray 2nd spray 3rd spray 4th spray
Fontelis
1 pt
chlorothalonil
or
mancozeb
Inspire Super
1 pt
chlorothalonil
or
mancozeb
Repeat
sequence
until end
of season
We are looking at the prevalence of QoI resistance
Prevalence of Copper Resistance among Foliar
Bacterial Pathogens of Tomato in TennesseeJonathan Mixon
Xanthomonas spp (Xs)
Pseudomonas syringae
pv tomato (Pst)
Clavibacter michiganensis
pv michigansis (Cmm)
Needed an accurate method of determining copper
resistance that was practical for a large-scale study
Literature inconsistent on in vitro testing methods
In vitro copper resistance determination
Variables tested(Chose a CuSO4 concentration of 200 microgml)
pH level
choice of culture medium
buffer concentration
method of application of bacteria to medium
culture incubation time
effect of medium storage time
For Xs and Pst used bioassays as guides
For Cmm used sensitivity as the standard
0
10
20
30
40
50
60
70
80
90
100
451b
(HR)
943
(MR)
1254
(MR)
BL10
(HR)
MA11-1
(S)
MA11-2
(HR)
HA11
(HR)
FL87-2
(MR)
8-5
(S)
6-3
(S)
3-3
(MR)
3-20 a
(MR)
GR
OW
TH
(
OF
CO
NT
RO
L)
ISOLATE (BIOASSAY RATING)
Xanthomonas isolates pH 62
pH 65
pH 68
pH 7o
Effect of medium pH on copper sensitivity for several
Xanthomonas spp isolates
Because of its known low-complexing activity 2-(N-morpholino) ethanesulfonic acid
(20 mM) was used to counteract the depressing effect of copper on medium pH
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
Control of early blight and bacterial spot two of the most
important diseases of tomato in Tennessee was less than
desirable Pathogen resistance to control products may be
responsible
Early blight drives the
tomato spray program
which has featured QoIrsquos
since Quadris was
labeled in 1997
Early Blight of Tomato2013 Field Trial Nashville
This trial was a search for replacements for the Group 11 fungicides ndash Quadris
Cabrio and Tanos Plants were sprayed 3 times at weekly intervals and evaluated
1 week after the 3rd spray
c
c
c
c
b
b
b
a
a
0 10 20 30
Untreated check
Quadris 62 fl oz
Tanos 8 oz
Cabrio 12 oz
Bravo 275 pt
Manzate 3 lb
Switch 14 oz
Inspire Superhellip
Fontelis 20 fl oz
Leaf area affected ()
Pro
du
ct
amp r
ate
75 g
al
The results were used to guide the development of non-strobilurin-based spray
programs
Early Blight Spray Programs2013 Trial Highland Rim REC
Four spray programs were evaluated on trellised lsquoMountain Gloryrsquo tomatoes
sprayed every 7-10 days beginning 3 wks after planting total of 10 sprays
Program Product
Early blight
leaf area
affected
(final)
Check none 880 a
1 Manzate alt with Quadris 575 b
2 Manzate every application 233 c
3 Fontelis 10 fl oz + Manzate + Kocidealt Manzate + Kocide
03 d
4 Inspire Super 1 pt then Manzate then Fontelis 1 pt then Manzate then repeat
01 d
The minimum labeled rate of Fontelis is 1 ptacre
Early Blight Fungicides2014 Field Trial Nashville
This trial was a search for candidates for inclusion in a spray program Plants were
sprayed 3 times at weekly intervals and evaluated 1 week after the 3rd spray
d
d
c
c
b
b
a
a
a
a
a
0 10 20 30 40 50
Untreated check
Quadris 62 fl oz
Double Nickel 1 qt
PhD 62 oz
Manzate 3 lb
Bravo 275 pt
Switch 14 oz
Endura 35 oz
Inspire Super 20 fl oz
Priaxor 8 fl oz
Fontelis 20 fl oz
Leaf area affected ()
Pro
du
ct
amp r
ate
75 g
al
7
7
39
7
912
M
M
19
NC
11
Do I really need
to use anything
more expensive
than chlorothalonil
or mancozeb
Early Blight Spray Programs2014 Trial Highland Rim REC
Four spray programs were evaluated on trellised lsquoMountain Gloryrsquo tomatoes
sprayed every 7-10 days beginning 5 wks after planting total of 7 sprays
Program Product
Early blight
leaf area
affected
(mid-harvest)
Check none 913 a
1 Manzate alt with Quadris every 3rd application 663 b
2 Manzate every application 513 c
3 Fontelis 10 fl oz + Manzate + Kocide every application
29 d
4 Inspire Super 1 pt then Manzate then Fontelis 1 pt then Manzate then repeat
43 d
The minimum labeled rate of Fontelis is 1 ptacre
Unsprayed check
Program 4
Program 4Unsprayed check
Excellent control of early blight was provided by spray programs
containing Inspire Super or Fontelis
Early Blight Resistant Varieties2014 Field Trial Cheatham County TSU ARECDeterminate varieties with resistance to early blight were compared
with the susceptible variety Mountain Glory Quadris was applied once
c
bc
bc
bc
bc
bc
b
a
0 20 40 60 80
Plum Regal
Mountain Merit
Plum Crimson
Mountain Fresh +
Iron Lady
BHN 964
Defiant
Mountain Glory
Leaf area affected at mid-harvest ()
Va
rie
ty
Spray program for
an early-blight resistant varietyMountain Merit
Plateau REC 2015
Spray program
Early blight severity ()
Mountain Glory Mountain Merit
Unsprayed control 563 a 150 a
Manzate each application 50 b 14 b
Fontelis then Manzate then
Inspire Super then Manzate then
repeat sequence
06 b 02 b
Ratings were taken just prior to harvest Early blight was later occluded by bacterial spot
1st spray 2nd spray 3rd spray 4th spray
Fontelis
1 pt
chlorothalonil
or
mancozeb
Inspire Super
1 pt
chlorothalonil
or
mancozeb
Repeat
sequence
until end
of season
We are looking at the prevalence of QoI resistance
Prevalence of Copper Resistance among Foliar
Bacterial Pathogens of Tomato in TennesseeJonathan Mixon
Xanthomonas spp (Xs)
Pseudomonas syringae
pv tomato (Pst)
Clavibacter michiganensis
pv michigansis (Cmm)
Needed an accurate method of determining copper
resistance that was practical for a large-scale study
Literature inconsistent on in vitro testing methods
In vitro copper resistance determination
Variables tested(Chose a CuSO4 concentration of 200 microgml)
pH level
choice of culture medium
buffer concentration
method of application of bacteria to medium
culture incubation time
effect of medium storage time
For Xs and Pst used bioassays as guides
For Cmm used sensitivity as the standard
0
10
20
30
40
50
60
70
80
90
100
451b
(HR)
943
(MR)
1254
(MR)
BL10
(HR)
MA11-1
(S)
MA11-2
(HR)
HA11
(HR)
FL87-2
(MR)
8-5
(S)
6-3
(S)
3-3
(MR)
3-20 a
(MR)
GR
OW
TH
(
OF
CO
NT
RO
L)
ISOLATE (BIOASSAY RATING)
Xanthomonas isolates pH 62
pH 65
pH 68
pH 7o
Effect of medium pH on copper sensitivity for several
Xanthomonas spp isolates
Because of its known low-complexing activity 2-(N-morpholino) ethanesulfonic acid
(20 mM) was used to counteract the depressing effect of copper on medium pH
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
Early blight drives the
tomato spray program
which has featured QoIrsquos
since Quadris was
labeled in 1997
Early Blight of Tomato2013 Field Trial Nashville
This trial was a search for replacements for the Group 11 fungicides ndash Quadris
Cabrio and Tanos Plants were sprayed 3 times at weekly intervals and evaluated
1 week after the 3rd spray
c
c
c
c
b
b
b
a
a
0 10 20 30
Untreated check
Quadris 62 fl oz
Tanos 8 oz
Cabrio 12 oz
Bravo 275 pt
Manzate 3 lb
Switch 14 oz
Inspire Superhellip
Fontelis 20 fl oz
Leaf area affected ()
Pro
du
ct
amp r
ate
75 g
al
The results were used to guide the development of non-strobilurin-based spray
programs
Early Blight Spray Programs2013 Trial Highland Rim REC
Four spray programs were evaluated on trellised lsquoMountain Gloryrsquo tomatoes
sprayed every 7-10 days beginning 3 wks after planting total of 10 sprays
Program Product
Early blight
leaf area
affected
(final)
Check none 880 a
1 Manzate alt with Quadris 575 b
2 Manzate every application 233 c
3 Fontelis 10 fl oz + Manzate + Kocidealt Manzate + Kocide
03 d
4 Inspire Super 1 pt then Manzate then Fontelis 1 pt then Manzate then repeat
01 d
The minimum labeled rate of Fontelis is 1 ptacre
Early Blight Fungicides2014 Field Trial Nashville
This trial was a search for candidates for inclusion in a spray program Plants were
sprayed 3 times at weekly intervals and evaluated 1 week after the 3rd spray
d
d
c
c
b
b
a
a
a
a
a
0 10 20 30 40 50
Untreated check
Quadris 62 fl oz
Double Nickel 1 qt
PhD 62 oz
Manzate 3 lb
Bravo 275 pt
Switch 14 oz
Endura 35 oz
Inspire Super 20 fl oz
Priaxor 8 fl oz
Fontelis 20 fl oz
Leaf area affected ()
Pro
du
ct
amp r
ate
75 g
al
7
7
39
7
912
M
M
19
NC
11
Do I really need
to use anything
more expensive
than chlorothalonil
or mancozeb
Early Blight Spray Programs2014 Trial Highland Rim REC
Four spray programs were evaluated on trellised lsquoMountain Gloryrsquo tomatoes
sprayed every 7-10 days beginning 5 wks after planting total of 7 sprays
Program Product
Early blight
leaf area
affected
(mid-harvest)
Check none 913 a
1 Manzate alt with Quadris every 3rd application 663 b
2 Manzate every application 513 c
3 Fontelis 10 fl oz + Manzate + Kocide every application
29 d
4 Inspire Super 1 pt then Manzate then Fontelis 1 pt then Manzate then repeat
43 d
The minimum labeled rate of Fontelis is 1 ptacre
Unsprayed check
Program 4
Program 4Unsprayed check
Excellent control of early blight was provided by spray programs
containing Inspire Super or Fontelis
Early Blight Resistant Varieties2014 Field Trial Cheatham County TSU ARECDeterminate varieties with resistance to early blight were compared
with the susceptible variety Mountain Glory Quadris was applied once
c
bc
bc
bc
bc
bc
b
a
0 20 40 60 80
Plum Regal
Mountain Merit
Plum Crimson
Mountain Fresh +
Iron Lady
BHN 964
Defiant
Mountain Glory
Leaf area affected at mid-harvest ()
Va
rie
ty
Spray program for
an early-blight resistant varietyMountain Merit
Plateau REC 2015
Spray program
Early blight severity ()
Mountain Glory Mountain Merit
Unsprayed control 563 a 150 a
Manzate each application 50 b 14 b
Fontelis then Manzate then
Inspire Super then Manzate then
repeat sequence
06 b 02 b
Ratings were taken just prior to harvest Early blight was later occluded by bacterial spot
1st spray 2nd spray 3rd spray 4th spray
Fontelis
1 pt
chlorothalonil
or
mancozeb
Inspire Super
1 pt
chlorothalonil
or
mancozeb
Repeat
sequence
until end
of season
We are looking at the prevalence of QoI resistance
Prevalence of Copper Resistance among Foliar
Bacterial Pathogens of Tomato in TennesseeJonathan Mixon
Xanthomonas spp (Xs)
Pseudomonas syringae
pv tomato (Pst)
Clavibacter michiganensis
pv michigansis (Cmm)
Needed an accurate method of determining copper
resistance that was practical for a large-scale study
Literature inconsistent on in vitro testing methods
In vitro copper resistance determination
Variables tested(Chose a CuSO4 concentration of 200 microgml)
pH level
choice of culture medium
buffer concentration
method of application of bacteria to medium
culture incubation time
effect of medium storage time
For Xs and Pst used bioassays as guides
For Cmm used sensitivity as the standard
0
10
20
30
40
50
60
70
80
90
100
451b
(HR)
943
(MR)
1254
(MR)
BL10
(HR)
MA11-1
(S)
MA11-2
(HR)
HA11
(HR)
FL87-2
(MR)
8-5
(S)
6-3
(S)
3-3
(MR)
3-20 a
(MR)
GR
OW
TH
(
OF
CO
NT
RO
L)
ISOLATE (BIOASSAY RATING)
Xanthomonas isolates pH 62
pH 65
pH 68
pH 7o
Effect of medium pH on copper sensitivity for several
Xanthomonas spp isolates
Because of its known low-complexing activity 2-(N-morpholino) ethanesulfonic acid
(20 mM) was used to counteract the depressing effect of copper on medium pH
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
Early Blight of Tomato2013 Field Trial Nashville
This trial was a search for replacements for the Group 11 fungicides ndash Quadris
Cabrio and Tanos Plants were sprayed 3 times at weekly intervals and evaluated
1 week after the 3rd spray
c
c
c
c
b
b
b
a
a
0 10 20 30
Untreated check
Quadris 62 fl oz
Tanos 8 oz
Cabrio 12 oz
Bravo 275 pt
Manzate 3 lb
Switch 14 oz
Inspire Superhellip
Fontelis 20 fl oz
Leaf area affected ()
Pro
du
ct
amp r
ate
75 g
al
The results were used to guide the development of non-strobilurin-based spray
programs
Early Blight Spray Programs2013 Trial Highland Rim REC
Four spray programs were evaluated on trellised lsquoMountain Gloryrsquo tomatoes
sprayed every 7-10 days beginning 3 wks after planting total of 10 sprays
Program Product
Early blight
leaf area
affected
(final)
Check none 880 a
1 Manzate alt with Quadris 575 b
2 Manzate every application 233 c
3 Fontelis 10 fl oz + Manzate + Kocidealt Manzate + Kocide
03 d
4 Inspire Super 1 pt then Manzate then Fontelis 1 pt then Manzate then repeat
01 d
The minimum labeled rate of Fontelis is 1 ptacre
Early Blight Fungicides2014 Field Trial Nashville
This trial was a search for candidates for inclusion in a spray program Plants were
sprayed 3 times at weekly intervals and evaluated 1 week after the 3rd spray
d
d
c
c
b
b
a
a
a
a
a
0 10 20 30 40 50
Untreated check
Quadris 62 fl oz
Double Nickel 1 qt
PhD 62 oz
Manzate 3 lb
Bravo 275 pt
Switch 14 oz
Endura 35 oz
Inspire Super 20 fl oz
Priaxor 8 fl oz
Fontelis 20 fl oz
Leaf area affected ()
Pro
du
ct
amp r
ate
75 g
al
7
7
39
7
912
M
M
19
NC
11
Do I really need
to use anything
more expensive
than chlorothalonil
or mancozeb
Early Blight Spray Programs2014 Trial Highland Rim REC
Four spray programs were evaluated on trellised lsquoMountain Gloryrsquo tomatoes
sprayed every 7-10 days beginning 5 wks after planting total of 7 sprays
Program Product
Early blight
leaf area
affected
(mid-harvest)
Check none 913 a
1 Manzate alt with Quadris every 3rd application 663 b
2 Manzate every application 513 c
3 Fontelis 10 fl oz + Manzate + Kocide every application
29 d
4 Inspire Super 1 pt then Manzate then Fontelis 1 pt then Manzate then repeat
43 d
The minimum labeled rate of Fontelis is 1 ptacre
Unsprayed check
Program 4
Program 4Unsprayed check
Excellent control of early blight was provided by spray programs
containing Inspire Super or Fontelis
Early Blight Resistant Varieties2014 Field Trial Cheatham County TSU ARECDeterminate varieties with resistance to early blight were compared
with the susceptible variety Mountain Glory Quadris was applied once
c
bc
bc
bc
bc
bc
b
a
0 20 40 60 80
Plum Regal
Mountain Merit
Plum Crimson
Mountain Fresh +
Iron Lady
BHN 964
Defiant
Mountain Glory
Leaf area affected at mid-harvest ()
Va
rie
ty
Spray program for
an early-blight resistant varietyMountain Merit
Plateau REC 2015
Spray program
Early blight severity ()
Mountain Glory Mountain Merit
Unsprayed control 563 a 150 a
Manzate each application 50 b 14 b
Fontelis then Manzate then
Inspire Super then Manzate then
repeat sequence
06 b 02 b
Ratings were taken just prior to harvest Early blight was later occluded by bacterial spot
1st spray 2nd spray 3rd spray 4th spray
Fontelis
1 pt
chlorothalonil
or
mancozeb
Inspire Super
1 pt
chlorothalonil
or
mancozeb
Repeat
sequence
until end
of season
We are looking at the prevalence of QoI resistance
Prevalence of Copper Resistance among Foliar
Bacterial Pathogens of Tomato in TennesseeJonathan Mixon
Xanthomonas spp (Xs)
Pseudomonas syringae
pv tomato (Pst)
Clavibacter michiganensis
pv michigansis (Cmm)
Needed an accurate method of determining copper
resistance that was practical for a large-scale study
Literature inconsistent on in vitro testing methods
In vitro copper resistance determination
Variables tested(Chose a CuSO4 concentration of 200 microgml)
pH level
choice of culture medium
buffer concentration
method of application of bacteria to medium
culture incubation time
effect of medium storage time
For Xs and Pst used bioassays as guides
For Cmm used sensitivity as the standard
0
10
20
30
40
50
60
70
80
90
100
451b
(HR)
943
(MR)
1254
(MR)
BL10
(HR)
MA11-1
(S)
MA11-2
(HR)
HA11
(HR)
FL87-2
(MR)
8-5
(S)
6-3
(S)
3-3
(MR)
3-20 a
(MR)
GR
OW
TH
(
OF
CO
NT
RO
L)
ISOLATE (BIOASSAY RATING)
Xanthomonas isolates pH 62
pH 65
pH 68
pH 7o
Effect of medium pH on copper sensitivity for several
Xanthomonas spp isolates
Because of its known low-complexing activity 2-(N-morpholino) ethanesulfonic acid
(20 mM) was used to counteract the depressing effect of copper on medium pH
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
Early Blight Spray Programs2013 Trial Highland Rim REC
Four spray programs were evaluated on trellised lsquoMountain Gloryrsquo tomatoes
sprayed every 7-10 days beginning 3 wks after planting total of 10 sprays
Program Product
Early blight
leaf area
affected
(final)
Check none 880 a
1 Manzate alt with Quadris 575 b
2 Manzate every application 233 c
3 Fontelis 10 fl oz + Manzate + Kocidealt Manzate + Kocide
03 d
4 Inspire Super 1 pt then Manzate then Fontelis 1 pt then Manzate then repeat
01 d
The minimum labeled rate of Fontelis is 1 ptacre
Early Blight Fungicides2014 Field Trial Nashville
This trial was a search for candidates for inclusion in a spray program Plants were
sprayed 3 times at weekly intervals and evaluated 1 week after the 3rd spray
d
d
c
c
b
b
a
a
a
a
a
0 10 20 30 40 50
Untreated check
Quadris 62 fl oz
Double Nickel 1 qt
PhD 62 oz
Manzate 3 lb
Bravo 275 pt
Switch 14 oz
Endura 35 oz
Inspire Super 20 fl oz
Priaxor 8 fl oz
Fontelis 20 fl oz
Leaf area affected ()
Pro
du
ct
amp r
ate
75 g
al
7
7
39
7
912
M
M
19
NC
11
Do I really need
to use anything
more expensive
than chlorothalonil
or mancozeb
Early Blight Spray Programs2014 Trial Highland Rim REC
Four spray programs were evaluated on trellised lsquoMountain Gloryrsquo tomatoes
sprayed every 7-10 days beginning 5 wks after planting total of 7 sprays
Program Product
Early blight
leaf area
affected
(mid-harvest)
Check none 913 a
1 Manzate alt with Quadris every 3rd application 663 b
2 Manzate every application 513 c
3 Fontelis 10 fl oz + Manzate + Kocide every application
29 d
4 Inspire Super 1 pt then Manzate then Fontelis 1 pt then Manzate then repeat
43 d
The minimum labeled rate of Fontelis is 1 ptacre
Unsprayed check
Program 4
Program 4Unsprayed check
Excellent control of early blight was provided by spray programs
containing Inspire Super or Fontelis
Early Blight Resistant Varieties2014 Field Trial Cheatham County TSU ARECDeterminate varieties with resistance to early blight were compared
with the susceptible variety Mountain Glory Quadris was applied once
c
bc
bc
bc
bc
bc
b
a
0 20 40 60 80
Plum Regal
Mountain Merit
Plum Crimson
Mountain Fresh +
Iron Lady
BHN 964
Defiant
Mountain Glory
Leaf area affected at mid-harvest ()
Va
rie
ty
Spray program for
an early-blight resistant varietyMountain Merit
Plateau REC 2015
Spray program
Early blight severity ()
Mountain Glory Mountain Merit
Unsprayed control 563 a 150 a
Manzate each application 50 b 14 b
Fontelis then Manzate then
Inspire Super then Manzate then
repeat sequence
06 b 02 b
Ratings were taken just prior to harvest Early blight was later occluded by bacterial spot
1st spray 2nd spray 3rd spray 4th spray
Fontelis
1 pt
chlorothalonil
or
mancozeb
Inspire Super
1 pt
chlorothalonil
or
mancozeb
Repeat
sequence
until end
of season
We are looking at the prevalence of QoI resistance
Prevalence of Copper Resistance among Foliar
Bacterial Pathogens of Tomato in TennesseeJonathan Mixon
Xanthomonas spp (Xs)
Pseudomonas syringae
pv tomato (Pst)
Clavibacter michiganensis
pv michigansis (Cmm)
Needed an accurate method of determining copper
resistance that was practical for a large-scale study
Literature inconsistent on in vitro testing methods
In vitro copper resistance determination
Variables tested(Chose a CuSO4 concentration of 200 microgml)
pH level
choice of culture medium
buffer concentration
method of application of bacteria to medium
culture incubation time
effect of medium storage time
For Xs and Pst used bioassays as guides
For Cmm used sensitivity as the standard
0
10
20
30
40
50
60
70
80
90
100
451b
(HR)
943
(MR)
1254
(MR)
BL10
(HR)
MA11-1
(S)
MA11-2
(HR)
HA11
(HR)
FL87-2
(MR)
8-5
(S)
6-3
(S)
3-3
(MR)
3-20 a
(MR)
GR
OW
TH
(
OF
CO
NT
RO
L)
ISOLATE (BIOASSAY RATING)
Xanthomonas isolates pH 62
pH 65
pH 68
pH 7o
Effect of medium pH on copper sensitivity for several
Xanthomonas spp isolates
Because of its known low-complexing activity 2-(N-morpholino) ethanesulfonic acid
(20 mM) was used to counteract the depressing effect of copper on medium pH
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
Early Blight Fungicides2014 Field Trial Nashville
This trial was a search for candidates for inclusion in a spray program Plants were
sprayed 3 times at weekly intervals and evaluated 1 week after the 3rd spray
d
d
c
c
b
b
a
a
a
a
a
0 10 20 30 40 50
Untreated check
Quadris 62 fl oz
Double Nickel 1 qt
PhD 62 oz
Manzate 3 lb
Bravo 275 pt
Switch 14 oz
Endura 35 oz
Inspire Super 20 fl oz
Priaxor 8 fl oz
Fontelis 20 fl oz
Leaf area affected ()
Pro
du
ct
amp r
ate
75 g
al
7
7
39
7
912
M
M
19
NC
11
Do I really need
to use anything
more expensive
than chlorothalonil
or mancozeb
Early Blight Spray Programs2014 Trial Highland Rim REC
Four spray programs were evaluated on trellised lsquoMountain Gloryrsquo tomatoes
sprayed every 7-10 days beginning 5 wks after planting total of 7 sprays
Program Product
Early blight
leaf area
affected
(mid-harvest)
Check none 913 a
1 Manzate alt with Quadris every 3rd application 663 b
2 Manzate every application 513 c
3 Fontelis 10 fl oz + Manzate + Kocide every application
29 d
4 Inspire Super 1 pt then Manzate then Fontelis 1 pt then Manzate then repeat
43 d
The minimum labeled rate of Fontelis is 1 ptacre
Unsprayed check
Program 4
Program 4Unsprayed check
Excellent control of early blight was provided by spray programs
containing Inspire Super or Fontelis
Early Blight Resistant Varieties2014 Field Trial Cheatham County TSU ARECDeterminate varieties with resistance to early blight were compared
with the susceptible variety Mountain Glory Quadris was applied once
c
bc
bc
bc
bc
bc
b
a
0 20 40 60 80
Plum Regal
Mountain Merit
Plum Crimson
Mountain Fresh +
Iron Lady
BHN 964
Defiant
Mountain Glory
Leaf area affected at mid-harvest ()
Va
rie
ty
Spray program for
an early-blight resistant varietyMountain Merit
Plateau REC 2015
Spray program
Early blight severity ()
Mountain Glory Mountain Merit
Unsprayed control 563 a 150 a
Manzate each application 50 b 14 b
Fontelis then Manzate then
Inspire Super then Manzate then
repeat sequence
06 b 02 b
Ratings were taken just prior to harvest Early blight was later occluded by bacterial spot
1st spray 2nd spray 3rd spray 4th spray
Fontelis
1 pt
chlorothalonil
or
mancozeb
Inspire Super
1 pt
chlorothalonil
or
mancozeb
Repeat
sequence
until end
of season
We are looking at the prevalence of QoI resistance
Prevalence of Copper Resistance among Foliar
Bacterial Pathogens of Tomato in TennesseeJonathan Mixon
Xanthomonas spp (Xs)
Pseudomonas syringae
pv tomato (Pst)
Clavibacter michiganensis
pv michigansis (Cmm)
Needed an accurate method of determining copper
resistance that was practical for a large-scale study
Literature inconsistent on in vitro testing methods
In vitro copper resistance determination
Variables tested(Chose a CuSO4 concentration of 200 microgml)
pH level
choice of culture medium
buffer concentration
method of application of bacteria to medium
culture incubation time
effect of medium storage time
For Xs and Pst used bioassays as guides
For Cmm used sensitivity as the standard
0
10
20
30
40
50
60
70
80
90
100
451b
(HR)
943
(MR)
1254
(MR)
BL10
(HR)
MA11-1
(S)
MA11-2
(HR)
HA11
(HR)
FL87-2
(MR)
8-5
(S)
6-3
(S)
3-3
(MR)
3-20 a
(MR)
GR
OW
TH
(
OF
CO
NT
RO
L)
ISOLATE (BIOASSAY RATING)
Xanthomonas isolates pH 62
pH 65
pH 68
pH 7o
Effect of medium pH on copper sensitivity for several
Xanthomonas spp isolates
Because of its known low-complexing activity 2-(N-morpholino) ethanesulfonic acid
(20 mM) was used to counteract the depressing effect of copper on medium pH
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
Early Blight Spray Programs2014 Trial Highland Rim REC
Four spray programs were evaluated on trellised lsquoMountain Gloryrsquo tomatoes
sprayed every 7-10 days beginning 5 wks after planting total of 7 sprays
Program Product
Early blight
leaf area
affected
(mid-harvest)
Check none 913 a
1 Manzate alt with Quadris every 3rd application 663 b
2 Manzate every application 513 c
3 Fontelis 10 fl oz + Manzate + Kocide every application
29 d
4 Inspire Super 1 pt then Manzate then Fontelis 1 pt then Manzate then repeat
43 d
The minimum labeled rate of Fontelis is 1 ptacre
Unsprayed check
Program 4
Program 4Unsprayed check
Excellent control of early blight was provided by spray programs
containing Inspire Super or Fontelis
Early Blight Resistant Varieties2014 Field Trial Cheatham County TSU ARECDeterminate varieties with resistance to early blight were compared
with the susceptible variety Mountain Glory Quadris was applied once
c
bc
bc
bc
bc
bc
b
a
0 20 40 60 80
Plum Regal
Mountain Merit
Plum Crimson
Mountain Fresh +
Iron Lady
BHN 964
Defiant
Mountain Glory
Leaf area affected at mid-harvest ()
Va
rie
ty
Spray program for
an early-blight resistant varietyMountain Merit
Plateau REC 2015
Spray program
Early blight severity ()
Mountain Glory Mountain Merit
Unsprayed control 563 a 150 a
Manzate each application 50 b 14 b
Fontelis then Manzate then
Inspire Super then Manzate then
repeat sequence
06 b 02 b
Ratings were taken just prior to harvest Early blight was later occluded by bacterial spot
1st spray 2nd spray 3rd spray 4th spray
Fontelis
1 pt
chlorothalonil
or
mancozeb
Inspire Super
1 pt
chlorothalonil
or
mancozeb
Repeat
sequence
until end
of season
We are looking at the prevalence of QoI resistance
Prevalence of Copper Resistance among Foliar
Bacterial Pathogens of Tomato in TennesseeJonathan Mixon
Xanthomonas spp (Xs)
Pseudomonas syringae
pv tomato (Pst)
Clavibacter michiganensis
pv michigansis (Cmm)
Needed an accurate method of determining copper
resistance that was practical for a large-scale study
Literature inconsistent on in vitro testing methods
In vitro copper resistance determination
Variables tested(Chose a CuSO4 concentration of 200 microgml)
pH level
choice of culture medium
buffer concentration
method of application of bacteria to medium
culture incubation time
effect of medium storage time
For Xs and Pst used bioassays as guides
For Cmm used sensitivity as the standard
0
10
20
30
40
50
60
70
80
90
100
451b
(HR)
943
(MR)
1254
(MR)
BL10
(HR)
MA11-1
(S)
MA11-2
(HR)
HA11
(HR)
FL87-2
(MR)
8-5
(S)
6-3
(S)
3-3
(MR)
3-20 a
(MR)
GR
OW
TH
(
OF
CO
NT
RO
L)
ISOLATE (BIOASSAY RATING)
Xanthomonas isolates pH 62
pH 65
pH 68
pH 7o
Effect of medium pH on copper sensitivity for several
Xanthomonas spp isolates
Because of its known low-complexing activity 2-(N-morpholino) ethanesulfonic acid
(20 mM) was used to counteract the depressing effect of copper on medium pH
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
Unsprayed check
Program 4
Program 4Unsprayed check
Excellent control of early blight was provided by spray programs
containing Inspire Super or Fontelis
Early Blight Resistant Varieties2014 Field Trial Cheatham County TSU ARECDeterminate varieties with resistance to early blight were compared
with the susceptible variety Mountain Glory Quadris was applied once
c
bc
bc
bc
bc
bc
b
a
0 20 40 60 80
Plum Regal
Mountain Merit
Plum Crimson
Mountain Fresh +
Iron Lady
BHN 964
Defiant
Mountain Glory
Leaf area affected at mid-harvest ()
Va
rie
ty
Spray program for
an early-blight resistant varietyMountain Merit
Plateau REC 2015
Spray program
Early blight severity ()
Mountain Glory Mountain Merit
Unsprayed control 563 a 150 a
Manzate each application 50 b 14 b
Fontelis then Manzate then
Inspire Super then Manzate then
repeat sequence
06 b 02 b
Ratings were taken just prior to harvest Early blight was later occluded by bacterial spot
1st spray 2nd spray 3rd spray 4th spray
Fontelis
1 pt
chlorothalonil
or
mancozeb
Inspire Super
1 pt
chlorothalonil
or
mancozeb
Repeat
sequence
until end
of season
We are looking at the prevalence of QoI resistance
Prevalence of Copper Resistance among Foliar
Bacterial Pathogens of Tomato in TennesseeJonathan Mixon
Xanthomonas spp (Xs)
Pseudomonas syringae
pv tomato (Pst)
Clavibacter michiganensis
pv michigansis (Cmm)
Needed an accurate method of determining copper
resistance that was practical for a large-scale study
Literature inconsistent on in vitro testing methods
In vitro copper resistance determination
Variables tested(Chose a CuSO4 concentration of 200 microgml)
pH level
choice of culture medium
buffer concentration
method of application of bacteria to medium
culture incubation time
effect of medium storage time
For Xs and Pst used bioassays as guides
For Cmm used sensitivity as the standard
0
10
20
30
40
50
60
70
80
90
100
451b
(HR)
943
(MR)
1254
(MR)
BL10
(HR)
MA11-1
(S)
MA11-2
(HR)
HA11
(HR)
FL87-2
(MR)
8-5
(S)
6-3
(S)
3-3
(MR)
3-20 a
(MR)
GR
OW
TH
(
OF
CO
NT
RO
L)
ISOLATE (BIOASSAY RATING)
Xanthomonas isolates pH 62
pH 65
pH 68
pH 7o
Effect of medium pH on copper sensitivity for several
Xanthomonas spp isolates
Because of its known low-complexing activity 2-(N-morpholino) ethanesulfonic acid
(20 mM) was used to counteract the depressing effect of copper on medium pH
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
Early Blight Resistant Varieties2014 Field Trial Cheatham County TSU ARECDeterminate varieties with resistance to early blight were compared
with the susceptible variety Mountain Glory Quadris was applied once
c
bc
bc
bc
bc
bc
b
a
0 20 40 60 80
Plum Regal
Mountain Merit
Plum Crimson
Mountain Fresh +
Iron Lady
BHN 964
Defiant
Mountain Glory
Leaf area affected at mid-harvest ()
Va
rie
ty
Spray program for
an early-blight resistant varietyMountain Merit
Plateau REC 2015
Spray program
Early blight severity ()
Mountain Glory Mountain Merit
Unsprayed control 563 a 150 a
Manzate each application 50 b 14 b
Fontelis then Manzate then
Inspire Super then Manzate then
repeat sequence
06 b 02 b
Ratings were taken just prior to harvest Early blight was later occluded by bacterial spot
1st spray 2nd spray 3rd spray 4th spray
Fontelis
1 pt
chlorothalonil
or
mancozeb
Inspire Super
1 pt
chlorothalonil
or
mancozeb
Repeat
sequence
until end
of season
We are looking at the prevalence of QoI resistance
Prevalence of Copper Resistance among Foliar
Bacterial Pathogens of Tomato in TennesseeJonathan Mixon
Xanthomonas spp (Xs)
Pseudomonas syringae
pv tomato (Pst)
Clavibacter michiganensis
pv michigansis (Cmm)
Needed an accurate method of determining copper
resistance that was practical for a large-scale study
Literature inconsistent on in vitro testing methods
In vitro copper resistance determination
Variables tested(Chose a CuSO4 concentration of 200 microgml)
pH level
choice of culture medium
buffer concentration
method of application of bacteria to medium
culture incubation time
effect of medium storage time
For Xs and Pst used bioassays as guides
For Cmm used sensitivity as the standard
0
10
20
30
40
50
60
70
80
90
100
451b
(HR)
943
(MR)
1254
(MR)
BL10
(HR)
MA11-1
(S)
MA11-2
(HR)
HA11
(HR)
FL87-2
(MR)
8-5
(S)
6-3
(S)
3-3
(MR)
3-20 a
(MR)
GR
OW
TH
(
OF
CO
NT
RO
L)
ISOLATE (BIOASSAY RATING)
Xanthomonas isolates pH 62
pH 65
pH 68
pH 7o
Effect of medium pH on copper sensitivity for several
Xanthomonas spp isolates
Because of its known low-complexing activity 2-(N-morpholino) ethanesulfonic acid
(20 mM) was used to counteract the depressing effect of copper on medium pH
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
Spray program for
an early-blight resistant varietyMountain Merit
Plateau REC 2015
Spray program
Early blight severity ()
Mountain Glory Mountain Merit
Unsprayed control 563 a 150 a
Manzate each application 50 b 14 b
Fontelis then Manzate then
Inspire Super then Manzate then
repeat sequence
06 b 02 b
Ratings were taken just prior to harvest Early blight was later occluded by bacterial spot
1st spray 2nd spray 3rd spray 4th spray
Fontelis
1 pt
chlorothalonil
or
mancozeb
Inspire Super
1 pt
chlorothalonil
or
mancozeb
Repeat
sequence
until end
of season
We are looking at the prevalence of QoI resistance
Prevalence of Copper Resistance among Foliar
Bacterial Pathogens of Tomato in TennesseeJonathan Mixon
Xanthomonas spp (Xs)
Pseudomonas syringae
pv tomato (Pst)
Clavibacter michiganensis
pv michigansis (Cmm)
Needed an accurate method of determining copper
resistance that was practical for a large-scale study
Literature inconsistent on in vitro testing methods
In vitro copper resistance determination
Variables tested(Chose a CuSO4 concentration of 200 microgml)
pH level
choice of culture medium
buffer concentration
method of application of bacteria to medium
culture incubation time
effect of medium storage time
For Xs and Pst used bioassays as guides
For Cmm used sensitivity as the standard
0
10
20
30
40
50
60
70
80
90
100
451b
(HR)
943
(MR)
1254
(MR)
BL10
(HR)
MA11-1
(S)
MA11-2
(HR)
HA11
(HR)
FL87-2
(MR)
8-5
(S)
6-3
(S)
3-3
(MR)
3-20 a
(MR)
GR
OW
TH
(
OF
CO
NT
RO
L)
ISOLATE (BIOASSAY RATING)
Xanthomonas isolates pH 62
pH 65
pH 68
pH 7o
Effect of medium pH on copper sensitivity for several
Xanthomonas spp isolates
Because of its known low-complexing activity 2-(N-morpholino) ethanesulfonic acid
(20 mM) was used to counteract the depressing effect of copper on medium pH
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
1st spray 2nd spray 3rd spray 4th spray
Fontelis
1 pt
chlorothalonil
or
mancozeb
Inspire Super
1 pt
chlorothalonil
or
mancozeb
Repeat
sequence
until end
of season
We are looking at the prevalence of QoI resistance
Prevalence of Copper Resistance among Foliar
Bacterial Pathogens of Tomato in TennesseeJonathan Mixon
Xanthomonas spp (Xs)
Pseudomonas syringae
pv tomato (Pst)
Clavibacter michiganensis
pv michigansis (Cmm)
Needed an accurate method of determining copper
resistance that was practical for a large-scale study
Literature inconsistent on in vitro testing methods
In vitro copper resistance determination
Variables tested(Chose a CuSO4 concentration of 200 microgml)
pH level
choice of culture medium
buffer concentration
method of application of bacteria to medium
culture incubation time
effect of medium storage time
For Xs and Pst used bioassays as guides
For Cmm used sensitivity as the standard
0
10
20
30
40
50
60
70
80
90
100
451b
(HR)
943
(MR)
1254
(MR)
BL10
(HR)
MA11-1
(S)
MA11-2
(HR)
HA11
(HR)
FL87-2
(MR)
8-5
(S)
6-3
(S)
3-3
(MR)
3-20 a
(MR)
GR
OW
TH
(
OF
CO
NT
RO
L)
ISOLATE (BIOASSAY RATING)
Xanthomonas isolates pH 62
pH 65
pH 68
pH 7o
Effect of medium pH on copper sensitivity for several
Xanthomonas spp isolates
Because of its known low-complexing activity 2-(N-morpholino) ethanesulfonic acid
(20 mM) was used to counteract the depressing effect of copper on medium pH
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
We are looking at the prevalence of QoI resistance
Prevalence of Copper Resistance among Foliar
Bacterial Pathogens of Tomato in TennesseeJonathan Mixon
Xanthomonas spp (Xs)
Pseudomonas syringae
pv tomato (Pst)
Clavibacter michiganensis
pv michigansis (Cmm)
Needed an accurate method of determining copper
resistance that was practical for a large-scale study
Literature inconsistent on in vitro testing methods
In vitro copper resistance determination
Variables tested(Chose a CuSO4 concentration of 200 microgml)
pH level
choice of culture medium
buffer concentration
method of application of bacteria to medium
culture incubation time
effect of medium storage time
For Xs and Pst used bioassays as guides
For Cmm used sensitivity as the standard
0
10
20
30
40
50
60
70
80
90
100
451b
(HR)
943
(MR)
1254
(MR)
BL10
(HR)
MA11-1
(S)
MA11-2
(HR)
HA11
(HR)
FL87-2
(MR)
8-5
(S)
6-3
(S)
3-3
(MR)
3-20 a
(MR)
GR
OW
TH
(
OF
CO
NT
RO
L)
ISOLATE (BIOASSAY RATING)
Xanthomonas isolates pH 62
pH 65
pH 68
pH 7o
Effect of medium pH on copper sensitivity for several
Xanthomonas spp isolates
Because of its known low-complexing activity 2-(N-morpholino) ethanesulfonic acid
(20 mM) was used to counteract the depressing effect of copper on medium pH
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
Prevalence of Copper Resistance among Foliar
Bacterial Pathogens of Tomato in TennesseeJonathan Mixon
Xanthomonas spp (Xs)
Pseudomonas syringae
pv tomato (Pst)
Clavibacter michiganensis
pv michigansis (Cmm)
Needed an accurate method of determining copper
resistance that was practical for a large-scale study
Literature inconsistent on in vitro testing methods
In vitro copper resistance determination
Variables tested(Chose a CuSO4 concentration of 200 microgml)
pH level
choice of culture medium
buffer concentration
method of application of bacteria to medium
culture incubation time
effect of medium storage time
For Xs and Pst used bioassays as guides
For Cmm used sensitivity as the standard
0
10
20
30
40
50
60
70
80
90
100
451b
(HR)
943
(MR)
1254
(MR)
BL10
(HR)
MA11-1
(S)
MA11-2
(HR)
HA11
(HR)
FL87-2
(MR)
8-5
(S)
6-3
(S)
3-3
(MR)
3-20 a
(MR)
GR
OW
TH
(
OF
CO
NT
RO
L)
ISOLATE (BIOASSAY RATING)
Xanthomonas isolates pH 62
pH 65
pH 68
pH 7o
Effect of medium pH on copper sensitivity for several
Xanthomonas spp isolates
Because of its known low-complexing activity 2-(N-morpholino) ethanesulfonic acid
(20 mM) was used to counteract the depressing effect of copper on medium pH
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
Needed an accurate method of determining copper
resistance that was practical for a large-scale study
Literature inconsistent on in vitro testing methods
In vitro copper resistance determination
Variables tested(Chose a CuSO4 concentration of 200 microgml)
pH level
choice of culture medium
buffer concentration
method of application of bacteria to medium
culture incubation time
effect of medium storage time
For Xs and Pst used bioassays as guides
For Cmm used sensitivity as the standard
0
10
20
30
40
50
60
70
80
90
100
451b
(HR)
943
(MR)
1254
(MR)
BL10
(HR)
MA11-1
(S)
MA11-2
(HR)
HA11
(HR)
FL87-2
(MR)
8-5
(S)
6-3
(S)
3-3
(MR)
3-20 a
(MR)
GR
OW
TH
(
OF
CO
NT
RO
L)
ISOLATE (BIOASSAY RATING)
Xanthomonas isolates pH 62
pH 65
pH 68
pH 7o
Effect of medium pH on copper sensitivity for several
Xanthomonas spp isolates
Because of its known low-complexing activity 2-(N-morpholino) ethanesulfonic acid
(20 mM) was used to counteract the depressing effect of copper on medium pH
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
In vitro copper resistance determination
Variables tested(Chose a CuSO4 concentration of 200 microgml)
pH level
choice of culture medium
buffer concentration
method of application of bacteria to medium
culture incubation time
effect of medium storage time
For Xs and Pst used bioassays as guides
For Cmm used sensitivity as the standard
0
10
20
30
40
50
60
70
80
90
100
451b
(HR)
943
(MR)
1254
(MR)
BL10
(HR)
MA11-1
(S)
MA11-2
(HR)
HA11
(HR)
FL87-2
(MR)
8-5
(S)
6-3
(S)
3-3
(MR)
3-20 a
(MR)
GR
OW
TH
(
OF
CO
NT
RO
L)
ISOLATE (BIOASSAY RATING)
Xanthomonas isolates pH 62
pH 65
pH 68
pH 7o
Effect of medium pH on copper sensitivity for several
Xanthomonas spp isolates
Because of its known low-complexing activity 2-(N-morpholino) ethanesulfonic acid
(20 mM) was used to counteract the depressing effect of copper on medium pH
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
For Xs and Pst used bioassays as guides
For Cmm used sensitivity as the standard
0
10
20
30
40
50
60
70
80
90
100
451b
(HR)
943
(MR)
1254
(MR)
BL10
(HR)
MA11-1
(S)
MA11-2
(HR)
HA11
(HR)
FL87-2
(MR)
8-5
(S)
6-3
(S)
3-3
(MR)
3-20 a
(MR)
GR
OW
TH
(
OF
CO
NT
RO
L)
ISOLATE (BIOASSAY RATING)
Xanthomonas isolates pH 62
pH 65
pH 68
pH 7o
Effect of medium pH on copper sensitivity for several
Xanthomonas spp isolates
Because of its known low-complexing activity 2-(N-morpholino) ethanesulfonic acid
(20 mM) was used to counteract the depressing effect of copper on medium pH
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
0
10
20
30
40
50
60
70
80
90
100
451b
(HR)
943
(MR)
1254
(MR)
BL10
(HR)
MA11-1
(S)
MA11-2
(HR)
HA11
(HR)
FL87-2
(MR)
8-5
(S)
6-3
(S)
3-3
(MR)
3-20 a
(MR)
GR
OW
TH
(
OF
CO
NT
RO
L)
ISOLATE (BIOASSAY RATING)
Xanthomonas isolates pH 62
pH 65
pH 68
pH 7o
Effect of medium pH on copper sensitivity for several
Xanthomonas spp isolates
Because of its known low-complexing activity 2-(N-morpholino) ethanesulfonic acid
(20 mM) was used to counteract the depressing effect of copper on medium pH
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
Because of its known low-complexing activity 2-(N-morpholino) ethanesulfonic acid
(20 mM) was used to counteract the depressing effect of copper on medium pH
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
Results
A protocol was developed that included colony growth
measurement on solid medium amended with CuSO45H2O (200
microgml) expressed as a percent of that on a non-amended
control
Plates were streaked by touching the edge of a sterile
disposable loop one time to a culture on solid medium and
making four streaks onto the test medium
Parameters were modified to produce copper sensitivity
reactions that approximated in-vivo disease-control levels
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
Results
Factors causing greatest variability in cell resistance to
copper were medium composition medium pH and age
of prepared
Freshly-prepared sucrose peptone agar adjusted to pH 68 (Xs and Cmm) or 62 (Pst) provided copper sensitivity ratings that corresponded well with bioassay ratings
The potential sources of error in resistance determinations identified in this study underscore the need for attention to these parameters in all studies of resistance to copper
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
195 acres were sampled
19 fields 11 farms 6 counties
(Bledsoe Grainger Hamblen
Greene Unicoi Lauderdale)
pathogens recovered from 166
samples (1 sample per acre)
162 were b-spot pathogens
4 were b-canker
no b-speck (hot dry seasons)
4 home gardens sampled
3 isolates were b-spot
1 isolate was b-canker
Results Pathogen Presence
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
B-spot
154 resistant
8 sensitive (Lauderdale Co)
B-canker
All 4 sensitive
Home garden isolates
2 b-spot resistant
1 sensitive
1 b-canker was sensitive
Results Copper Resistance
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
Jonathan also worked on developing a multiplex PCR
protocol for simultaneous detection of all three
pathogens
Thank you
Thank you