Phytopathogenic Bacteria

Disease Diagnosis and Identification of
Phytopathogenic Bacteria
Internal/external symptoms
Culture bacteria and characterize:
morphology
growth on selective media
chemical composition
nutritional tests
enzyme production
serological tests
molecular tests (PCR, etc)
pathogenicity tests
Bacterial
streaming
Cuppels Biology 4218a
Overview of the primary genera of phytopathogenic bacteria:
Kingdom Procaryotae
BACTERIA
DIVISION: GRACILICUTES
CLASS: PROTEOBACTERIA
Family: Enterobacteriaceae (Gram - )
Family: Pseudomonadaceae (Gram - )
*Erwinia species (Fireblight, soft rot)
Acidovorax species (Leaf spot of watermelon,

orchids and corn)
Pantoea species (Stewarts wilt of corn)
*Pseudomonas species (Leaf spots, fruit spots,

blights, vascular wilts, soft rots, galls)
Serratia species (Yellow vine disease of

cucurbits)
*Ralstonia species (Wilts of solanaceous crops,

banana, plantain, peanuts, geranium)
Sphingomonas species (Brown spot of

yellow Spanish melon)
Rhizobacter (Bacterial gall of carrot)

Rhizomonas (Corky root of lettuce)
*Xanthomonas species (Leaf spots, fruit spots,
blight, vascular wilts, citrus canker)
Family: Rhizobiaceae (Gram - )
Family: still unnamed
*Agrobacterium (Crown gall of dicots)
Xylella (Leaf scorch and dieback of trees and

vines)
Xylophilus species (Bacterial necrosis and canker
of grape)
* = The six most studied genera of phytopathogenic bacteria

BACTERIA
DIVISION: FIRMICUTES**
CLASS: FIRMIBACTERIA (single-celled)
Bacillus (Tuber rot)

Clostridium (Tuber rot, wetwood of elm and poplar)
DIVISION: FIRMICUTES**
CLASS: THALLOBACTERIA (branched)
Arthrobacter (Bacterial blight of holly)

*Clavibacter (Bacterial wilts in potato, tomato, alfalfa; ring rot of potato)
Leifsonia (Ratoon stunting of sugarcane)
Rhodococcus (Fasciation of ornamentals and sweet pea)
Streptomyces (Common scab of potato)
MOLLICUTES
DIVISION: TENERICUTES
CLASS: MOLLICUTES
Family: Spiroplasmataceae
Spiroplasma (Corn stunt disease, citrus stubborn disease)

Phytoplasma (Yellows diseases, decline diseases in trees, proliferation disease)
** Please note that Firmicutes are Gram-positive

BACTERIA
Family: Enterobacteriaceae (Gram -)

*Erwinia species
Fire blight, soft rot (most common on fleshy vegetables and fruit)
Pantoea species
Stewarts wilt of cornspread by corn flea beetles
Serratia species (fastidious, phloem-inhabiting)
Yellow Vine Disease of Cucurbitsspread by squash bug
Sphingomonas melonis
Brown spot of yellow Spanish melon fruit
Erwinia, Oregon State Univ.

Bacterial ooze from infected

apple, Van der Zwet, USDA
Erwinia carotovora supsp. atroseptica

Blackleg of potato,S. DeBoer, CFIA, PEI
Fire blight of apple in Michigan
This picture was taken in early June of 2000, 2 weeks after a hail storm and onset of widespread fire
blight symptoms in the region. New shoots have been killed and the leaves have died giving the trees a
brown appearance. Both 'Gala' and the M9 rootstock are very susceptible to fire blight.
photo by M. Longstroth, Mich. State Univ.
Disease cycle of fire blight of pear and apple

Bees carry
bacteria to
flowers
Bacteria penetrate flowers then

multiply and spread rapidly
Infe
shr cted fl
ivel
o
and wers
die
Infection
spreads
Infection of
young twigs
Intercellular
multiplication in bark
Bacteria in exudate
carried by insects
and rain
Infected bark
cells collapse
Bacteria travel
in xylem and
between cells
Bacteria overwinter on
margins of old cankers
Young tree heavily

infected with fire
blight
Twig killed
by fire blight
Formation of
new cankers
on branches
and stems
[from Plant Pathology, 5th ed, G. Agrios]
BACTERIA

*Pseudomonas species (Leaf spots, fruit spots, blights, vascular wilts, soft rots, galls)
Examples: marginalis and syringae
Fluorescent
-RNA Grp I
-PHB (-)
P. syringae pv. tomato

bacterial speck of tomato
fluorescent bacterial growth
biology.uwsp.edu
P. marginalis on lettuce
www.ctahr.hawaii.edu/nelsons
Phytopathogenic Pseudomonas sp. continued
Non-Fluorescent
- RNA Grp II
- PHB (+)
Examples: cepacia, gladioli
Now is Burkholderia cepacia

in Burkholderiaceae
Sour skin on onion

www.apsnet.org
http://www.apsnet.org/education/feature/BurkholderiaCepacia/
Friend or Foe?
BACTERIA
Ralstonia
solanacearum:
DIVISION:
GRACILICUTES
*Ralstonia species (Wilts of solanaceous crops, banana, plantain, peanuts, geranium)
Geranium wilt: race 3

www.agf.gov.bc.ca
Moko Disease, www.tpp.uq.edu.au
Potato brown rot

BACTERIA

*Xanthomonas species (Leaf spots, fruit spots, blight, vascular wilts, citrus canker)
Yellow pigment (brominated aryl-polyenes)

Xanthan gums
Monotrichous flagellation
Host-specific
Examples:
X. gardneri , tomato pathogen
on nutrient broth yeast ext. agar
X. axonopodis pv. citri

citrus canker pathogen
BACTERIA
Family: Rhizobiaceae (Gram - )

* Agrobacterium species (Crown gall, hairy root)
A. tumefaciens: Crown gall of apple rootstock
A. vitis: Crown gall of wine grapes
A. rhizogenes:
hairy root of carrot
A. rubi: galls on blackberry/rasberry
Agrobacterium infection process:

1. Chemotaxis to wound:
soil line
wound
bacteria
2. Plant cells become conditioned

and bacteria bind to them:
[ bacteria make cellulose fibrils
and Chv proteins ]
3. Induction of bacterial vir genes:
Vir A protein
Wound exudates
Ti
Ti plasmid (site of vir genes)
Ti Plasmid
(~150-250 kb !!)
vir genes
(about 35-45 kb)
H
F
E
D
C
G
BA
http://arabidopsis.info/students/paaras/t_dna.htm
4. T- DNA processing and transfer to nucleus of plant cell:

Plant nucleus
T-DNA
Ti
Vir B transporter core
T- DNA insertion into plant nuclear DNA:
Genes carried on T-DNA:

I. Genes affecting plant hormones:
(oncogenes)
tms 1: Tryptophan monooxygenase
(iaaM)
tms 2: Indoleacetamide hydrolase (iaaH)
ipt: Isopentenyl transferase
II. Genes involved in opine synthesis

(will be utilized as food by the bacteria)
Octopine: N2-(1-carboxyethyl)arginine
Nopaline: N2-(1,3-dicarboxypropyl)arginine
Agropine: N2-(1-mannose)glutamine
T- DNA expression in plant cells

resulting in
plant hormone and opine production at the wound site
BACTERIA
DIVISION: FIRMICUTES
CLASS: THALLOBACTERIA (branched)
*Clavibacter (Bacterial wilts in potato, tomato, alfalfa; ring rot of potato)

Streptomyces (Common scab of potato)
Clavibacter spp.
pleomorphic, mostly rod-like
diaminobutyric acid in cell wall
usually non-motile
C. michiganensis subsp. sepedonicum

Ring rot of potato
Streptomyces scabies
spore-forming
branched vegetative hyphae
soil inhabitants
attacks potato and other root vegetables
Common scab of potato
PLANT DISEASES CAUSED BY FASTIDIOUS

VASCULAR BACTERIA
Observed in 1972 in the phloem of clover and
periwinkle with clover club leaf disease and in the
phloem of citrus with citrus greening disease
Observed in 1973 in the xylem of grapes with Pierces

disease and the xylem of alfalfa with alfalfa dwarf
Citrus greening disease

www.apsnet.org
Pierces disease of grape

www-plb.ucdavis.edu
Fastidious Xylem-inhabiting Bacteria

Example: Xylella fastidiosa (Gram - )
Disease: Pierces disease of grape
Small* with rippled cell wall

No flagella
Grows on complex nutritional media
Vector: sharpshooter leafhoppers
entomology.ucdavis.edu
www.apsnet.org
* Cell size = 0.2 - 0.5 x 1 - 4 m

inventabrasilnet.t5.com.br
Fastidious Xylem-inhabiting Bacteria

Example: Leifsonia xyli (Gram +, branched thallobacteria)
Disease: Ratoon stunting of sugarcane
Small coryneform bacterium

No flagella
Grows on specialized nutritional media
Transmitted by seed cane, farm tools
pinkish discoloration at nodes

www.tpp.uq.edu.au
Fastidious Phloem-limited Bacteria

Symptoms: stunting of leaves, shoot proliferation, greening of floral parts
Example : Candidatus liberobacter asiaticus or africanus

Disease: Citrus greening or Huanglongbing (yellow dragon)
Small with rippled cell wall

No flagella
Has not been cultivated
Transmitted by insectspsyllid
Example : Serratia marcescens

Disease: Yellow vine disease of cucurbits
Watermelon, squash, pumpkin
Squash bug vector
Control: resistance genes
www.apsnet.org
MOLLICUTES
CLASS: MOLLICUTES

Phytoplasma (Yellows diseases, decline diseases in trees, proliferation disease)
Cannot be cultivated
Small genome (500-1000 kb)
Greater than 200 diseases
Pleomorphic
Found in phloem sap, insect organs
Leafhopper-transmitted
Aster yellows
www.apsnet.org
MOLLICUTES
CLASS: MOLLICUTES
Family: Spiroplasmataceae
Spiroplasma (Corn stunt disease, citrus stubborn disease)
Can be cultivated
Small genome (500-1000 kb)
Relatively few diseases
Helical to branched filaments
Motile but no flagella
In phloem sap
Leafhopper-transmitted
Can live saprophytically
www.zarzy.com
Corn stunt spiroplasma
www.apsnet.org

Phytopathogenic Bacteria

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Phytopathogenic Bacteria

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Disease Diagnosis and Identification of

Cuppels Biology 4218a

Overview of the primary genera of phytopathogenic bacteria:

Family: Enterobacteriaceae (Gram - )

Family: Pseudomonadaceae (Gram - )

*Erwinia species (Fireblight, soft rot)

Acidovorax species (Leaf spot of watermelon,

Pantoea species (Stewarts wilt of corn)

*Pseudomonas species (Leaf spots, fruit spots,

Serratia species (Yellow vine disease of

*Ralstonia species (Wilts of solanaceous crops,

Sphingomonas species (Brown spot of

Rhizobacter (Bacterial gall of carrot)

Family: Rhizobiaceae (Gram - )

Family: still unnamed

*Agrobacterium (Crown gall of dicots)

Xylella (Leaf scorch and dieback of trees and

* = The six most studied genera of phytopathogenic bacteria

CLASS: FIRMIBACTERIA (single-celled)

Bacillus (Tuber rot)

CLASS: THALLOBACTERIA (branched)

Arthrobacter (Bacterial blight of holly)

Family: still unnamed

** Please note that Firmicutes are Gram-positive

Family: Enterobacteriaceae (Gram -)

Erwinia, Oregon State Univ.

Bacterial ooze from infected

Erwinia carotovora supsp. atroseptica

Fire blight of apple in Michigan

Cuppels Biology 4218a

photo by M. Longstroth, Mich. State Univ.

Disease cycle of fire blight of pear and apple

Bacteria penetrate flowers then

Cuppels Biology 4218a

Young tree heavily

Family: Pseudomonadaceae (Gram - )

Examples: marginalis and syringae

P. syringae pv. tomato

Phytopathogenic Pseudomonas sp. continued

Now is Burkholderia cepacia

Sour skin on onion

Cuppels Biology 4218a

Geranium wilt: race 3

Moko Disease, www.tpp.uq.edu.au

Potato brown rot

Family: Pseudomonadaceae (Gram - )

Yellow pigment (brominated aryl-polyenes)

Cuppels Biology 4218a

X. axonopodis pv. citri

Family: Rhizobiaceae (Gram - )

A. tumefaciens: Crown gall of apple rootstock

A. vitis: Crown gall of wine grapes

Cuppels Biology 4218a

A. rubi: galls on blackberry/rasberry

Agrobacterium infection process:

2. Plant cells become conditioned

3. Induction of bacterial vir genes:

Ti plasmid (site of vir genes)

Cuppels Biology 4218a