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

Cuppels Biology 4218a

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)

Family: still unnamed

Phytoplasma (Yellows diseases, decline diseases in trees, proliferation disease)

** Please note that Firmicutes are Gram-positive

Cuppels Biology 4218a

BACTERIA
DIVISION: GRACILICUTES

CLASS: PROTEOBACTERIA

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.

Cuppels Biology 4218a

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.

Cuppels Biology 4218a

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

Cuppels Biology 4218a

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
DIVISION: GRACILICUTES

CLASS: PROTEOBACTERIA

Family: Pseudomonadaceae (Gram - )

*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
Cuppels Biology 4218a

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?

Cuppels Biology 4218a

BACTERIA

Ralstonia
solanacearum:
DIVISION:
GRACILICUTES
CLASS: PROTEOBACTERIA
Family: Pseudomonadaceae (Gram - )
*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

Cuppels Biology 4218a

BACTERIA
DIVISION: GRACILICUTES

CLASS: PROTEOBACTERIA

Family: Pseudomonadaceae (Gram - )

*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

Cuppels Biology 4218a

X. axonopodis pv. citri

citrus canker pathogen

BACTERIA
DIVISION: GRACILICUTES

CLASS: PROTEOBACTERIA

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

Cuppels Biology 4218a

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)

Cuppels Biology 4218a

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

Cuppels Biology 4218a

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:

Cuppels Biology 4218a

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

Cuppels Biology 4218a

T- DNA expression in plant cells

resulting in
plant hormone and opine production at the wound site

Cuppels Biology 4218a

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
Cuppels Biology 4218a

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
Cuppels Biology 4218a

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

Cuppels Biology 4218a

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

Cuppels Biology 4218a

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

Cuppels Biology 4218a

www.apsnet.org

MOLLICUTES
DIVISION: TENERICUTES

CLASS: MOLLICUTES

Family: still unnamed

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

Cuppels Biology 4218a

MOLLICUTES
DIVISION: TENERICUTES

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

Cuppels Biology 4218a

www.apsnet.org