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