Diagnosing Plant Problems-Plant Diseases

National Plant Diagnostic Network
Diagnosing Plant Problems: Plant Diseases

National Plant Diagnostic Network First Detector Training
Williamson, Riley, and Maloy. December 2006.
Update, August 2008.
NPDN Publication No. 0013
MODULE 4 Diagnosing plant problems: Plant Diseases
Adapted from Plant Disease Diagnosis by M. B. Riley, M. R. Williamson and O. Maloy.

In: The Plant Health Instructor. DOI:10.1094/PHI-I-2002-1021-01
Used by permission of the American Phytopathological Society
Plant Problem Diagnosis

Diagnosis process used for the identification of problems. Tools:
Plant sciences, especially plant pathology and entomology Arts of investigation and detective work
Plant Problem Diagnosis: Preliminary Considerations

Know Normal Appearance of Plant
Proper plant identification is critical.

Scientific vs. common names Variety or cultivar Whats normal for specific plant? Compare with healthy plants of same genus, species and cultivar.
Recognize healthy plant appearance.

Plant Problem Diagnosis Steps

1. Consider the Possible Agents:
Abiotic non-living agent
Damage from chemicals, weather, mechanical Nutritional problems
Biotic living agent

Pathogens - parasitic microorganisms that cause diseases Pests insects or mammals feeding on or damaging plants

2. Consult literature resources for possible diseases and disorders.
Indexes listing hosts and their pathogens Websites providing information Books with background info and host/pathogen lists Compendia for information on specific crops

3. Investigate symptom progression.
Biotic disease symptoms progress and nearby plants become infected. Abiotic disease generally a lack of symptom progression. Does not spread. Exception nutritional symptoms progress slowly.

4. Ask questions
When was the problem noticed?
Was the damage sudden or gradual? How old are affected plants? Percentage of plants affected? What is the degree of injury?

5. Observe patterns.
Determine prevalence of problem Large area/All plants generally abiotic Scattered, localized generally biotic Check for distribution of symptoms Uniform generally abiotic Random generally biotic
Observation of Patterns
Scattered symptom distribution Random pattern of damage
Photo: R. Billings, Texas Forest Service, http://www.forestryimages.org
Loblolly pine killed by southern pine beetle
Observation of Patterns
Symptoms distributed in a large area. Damage pattern is uniform.
Used by permission of M. Williamson
Cotton field with chemical damage

6. Review cultural practices.
Proper planting technique Fertilizer and pesticide application Irrigation frequency
7. Review environmental conditions.

Temperature extremes Drought or excess rain Soil type and conditions
8. Check host specificity many hosts may mean an abiotic problem

9. Check for symptoms and signs.
Symptoms - plant reactions or alterations of a plants appearance due to a disease or disorder Signs - actual pathogen, parts or byproducts seen on a diseased host plant
Symptoms
Identify and classify symptoms
Underdevelopment Overdevelopment Necrosis or death Alteration of normal appearance Wilting
Identify plant parts affected May need to identify source of symptoms within plant system
Symptoms
Underdevelopment
Stunting of plants, leaves Shortened internodes Inadequate chlorophyll production Caused by many types of pathogens
Photo: J. Dunez, www.forestryimages.org
Chrysanthemum stunt viroid infection of chrysanthemum showing stunting and earlier blooming of affected plants; healthy plants in middle.
Symptoms Overdevelopment
Photo: Joseph O'Brien, USDA Forest Service, www.ipmimages.org
Photo: Edward L. Barnard, Florida Department of Agriculture and Consumer Services, www.ipmimages.org
Oak leaf blister, caused by Taphrina caerulescens.
Gall on stem of eastern redcedar caused by Agrobacterium tumefaciens.
Overgrowth of leaf tissue causes thickening and distortion.
Galls form on all plant parts, caused by many pathogens.
Symptoms
Over-development - more biotic causes
Root knot nematodes - root galls Callus formation around cankers Tissue proliferation -some downy mildews and phytoplasmas Some insects and mites also cause galls.
Symptoms
Over-development Abiotic causes
Oedema Fasciation can be physiological or due to biotic causes. Some tree burls
Used by permission of M. Willliamson
Oedema of camellia, a physiological disorder
Symptoms and Signs

Tissue Necrosis
Fungal leaf spots Usually round, not veinlimited Elongated on narrow leaves or stems May have alternating zones of light and dark tissue Sporulation or mycelia may be evident
Photo: Paul Bertrand, University of Georgia, www.ipmimages.org
Target spot of tobacco, caused by Thanatephorus cucumeris.
Symptoms and Signs

Tissue Necrosis Bacterial leaf spots Often dark and watersoaked Often vein-limited, giving angular shape Bacterial flow observed under microscope
Photo: Volcani Center Archives, Agricultural Research Organization, www.ipmimages.org
Bacterial leaf spot of sweet pepper caused by Xanthomonas campestris pv. vesicatoria
Symptoms and Signs

Tissue Necrosis Fruit rots
Firm or soft and watery Colors vary Fungal sporulation may be present Fungal and bacterial causes
Photo: Clemson University - USDA Cooperative Extension Slide Series, , www.ipmimages.org
Brown rot of peach, caused by Monilinia fruticola
Symptoms
Tissue Necrosis Cankers = localized necrotic lesions
Sunken or swollen or both Mainly caused by fungi and bacteria Mechanical injury can cause
Photo: Joseph O'Brien, USDA Forest Service, www.ipmimages.org
Sugar maple canker caused by Nectria spp.
Symptoms
Tissue Necrosis Blight = rapid death or dieback.
Also from coalescing leaf spots, e.g. early blight of tomato Mainly fungal and bacterial causes
Photo: Robert L. Anderson, USDA Forest Service, www.ipmimages.org
Fire blight caused by Erwinia amylovora
Symptoms
Tissue Necrosis Dieback - many causes
Girdling cankers. Follow dieback to detect Dieback pathogens w/out cankers Root problems Mechanical, chemical or cold damage
Photo: Edward L. Barnard, Florida Department of Agriculture and Consumer Services, www.ipmimages.org
Pitch canker of Virginia pine, caused by Fusarium subglutinans
Symptoms
Tissue Necrosis Root rots
Root lesions Darkening and softening of roots Sloughing off of outer tissues Yellowing of foliage and stunting of plants Fungal and bacterial causes
Root rot of boxwood, caused by Pythium sp.
Symptoms
Tissue Necrosis Damping off
Rapid death of seedlings and cuttings Stem infected at soil line, seedling topples Spreads rapidly under crowded conditions Mainly fungal agents
Photo: Clemson University - USDA Cooperative Extension Slide Series, www.ipmimages.org
Damping off of tobacco, caused by Rhizoctonia spp. and Pythium spp.
Symptoms
Wilting
Biotic factors
Root, crown or stem rots Vascular wilts Root crown or stem damage from insects or animals Mainly fungal and bacterial causes
Used by permission of M.Williamson
Stem wilt of Exacum from INSV infection
Symptoms
Wilting
Abiotic factors
Damage from weather extremes Dry or flooded soil Mechanical damage to roots, crown or stem
Cold damage on azalea stem. Shoots wilt above damage.
Symptoms
Alteration of normal appearance Mosaic
Irregular patches of discolored tissue. Often with distortion Viruses mainly
Photo: David B. Langston, University of Georgia, www.ipmimages.org
Mosaic symptoms on cucumber due to a viral infection
Symptoms
Alteration of normal appearance Ringspot
On leaves and fruits. Distinct ring shaped lesions, often in concentric zones Viruses
Photo: Dr. Backhaus, Biologische Bundesanstalt fr Land- und Forstwirtschaft, www.ipmimages.org
Tomato spotted wilt virus (TSWV) on geranium leaf
Symptoms
Alteration of normal appearance
Abiotic causes Chemical damage Nutritional deficiencies
Cotton damage by the herbicide 2,4-D
Symptoms
Alteration of normal appearance
Yellowing Often nutritional, cultural or environmental Root malfunction biotic or abiotic Nematode infestation Yellows phytoplasma diseases
Burford hollies yellowing due to root malfunction. Soil is poorly drained.
Symptoms
Be aware of symptom variability.
May have more than one problem. More than one pathogen may be involved. Pathogens have varying levels of virulence. Environmental conditions can affect symptom expression. Host genetics and physiology can effect symptom expression.
Symptom variability due to multiple pathogens

Peach seedling on left infected with both Prune dwarf virus (PDV) and Prunus necrotic ringspot virus (PNRSV) Seedling in middle infected with PDV alone Seedling on right infected with PNRSV alone
Used by permission of S. Scott
Peach seedlings infected by single or multiple viruses show symptom variability.
Signs
Fungal signs
Evidence of pathogen on tissue. Spores, mycelia or fruiting bodies. Use hand lens or knife for field detection.
Photo: USDA Forest Service Archives, USDA Forest Service, www.ipmimages.org
White mycelia of Armillaria sp. on tree trunk affected by Armillaria root and stem rot.
Signs
Fungal signs
Fruiting bodies
Shape of fruiting body aids in fungal identification. Tissue location may help differentiate between species.
Photo: Central Science Laboratory, Harpenden Archives, British Crown, www.ipmimages.org
White rust of chrysanthemum, fruiting bodies of Puccinia horiana
Signs
Fungal signs
Powdery mildew fungi form mycelia and spores on tissue surface. Powdery material rubs off. Leaves often distorted, discolored Dark, round fruiting bodies form in fall.
Oidium sp. causing powdery mildew on Viburnum suspensum.
Symptoms and Signs

Downy mildews often produce angular leaf spots.

Downy mildew of Buddleia caused by Peronospora harotii.
Downy mildews develop sporulation on leaf undersides only.

Downy mildew of veronica, caused by Peronospora sordida
Signs
Bacterial signs
Streaming from freshly cut stem in water. Stringing from cut stems pushed together, then pulled apart. Bacteria flow observed from tissue mount on compound microscope. Bacterial ooze can be observed on-site with some diseases.
Bacterial streaming from tomato infected by Ralstonia solanacearum.

12. Laboratory Examination and Testing
Dissecting microscope Compound microscope Electron microscope Moist chamber incubation Culturing Additional tests for biotic agents Tests for abiotic agents
Laboratory Examination Examination using dissecting microscope

Observe sporulation thats invisible to naked eye. Closer view of structures seen by naked eye Nematologist can ID nematodes to genus.
Laboratory Examination
Examination using microscope of higher magnification
ID fungi to genus or to species. View bacterial flow; observe morphology at highest magnification. View virus inclusion bodies. Using electron microscope, view viruses, phytoplasmas
Rust spores, Puccinia sp.
Laboratory tests
Moist Incubation
Goal to induce sporulation Important for obligate fungal pathogens Avoid overly moist conditions. Can surface sterilize
Moist chamber
Laboratory tests
Fungal isolation

Fungal culture.
Surface sterilize. Use margin of diseased area. Nutritive agar media, can be selective Diagnostician can often ID to genus from appearance of culture
Isolate identification
Fungal identification Other methods Examination and comparison with drawings in literature Utilizing keys in literature Literature sources for host diseases and disorders Serological, molecular, other tests
Laboratory tests
Bacterial isolations
Grind tissue in sterile water. Streak suspension on bacterial medium. Can use selective media Transfer single, isolated colony
Cultures of Ralstonia solanacearum

Used by permission of K. Ong
Isolate identification
Bacterial identification Biochemical tests Carbohydrate utilization (BIOLOG) Fatty acid methyl ester analysis (FAME) Molecular methods Polymerase chain reaction (PCR) Serological tests, e.g. ELISA
Identification of other pathogens

Non-culturable pathogens
Viruses, viroids Phytoplasmas Fastidious bacteria
Identification of non-culturable pathogens

Methods Serological tests, e.g. ELISA Electron microscopy Staining for virus-induced inclusion bodies Molecular tests, e.g. PCR Enzyme activity testing Host range testing
Identification of other pathogens

ELISA tests
Multi-well test for multiple samples
Individual ELISA kit

Identification of Nematodes
Extraction from Soil and Plant Material
Soil extraction using centrifugal flotation Baerman funnel techniques for soil and tissue Mist chamber extraction for nematodes in root or stem tissue
Direct examination of nematodes in tissue.
Laboratory tests
Diagnostic tests for abiotic causes
Soil, water pH testing Nutrient analysis Soluble salts analysis Analysis for chemicals Plant tissue tests Nutrient analysis Analysis for chemicals
Plant Disease Diagnosis Steps

13) Investigate pathogenicity.
Is identified organism a known pathogen - of host? - of hosts in same genus? Consult literature to determine
If not a host, Kochs Postulates may be next step A lengthy process, rarely used in routine diagnoses Problem arises when organism cannot be cultured
Kochs Postulates
Robert Koch German physician in late 1800s Proved that a bacterium caused anthrax in cattle Developed steps proving the bacterium was causal Same steps used to prove pathogenicity for plant disease organisms
Kochs Postulates
1. Note constant association of organism with diseased plants and consistent, observable symptoms. 2. Isolate and characterize organism in pure culture. 3. Inoculate healthy plant with organism, and observe the same disease and symptoms. 4. Re-isolate same organism.
Plant Disease Diagnosis Steps

14. Final Diagnosis
All information compiled and analyzed Organism identified, found to be pathogen of host OR Lab tests, other information reveal an abiotic cause
15. Develop control recommendations and present to grower.
Authors
Meg Williamson, Clemson University Melissa Riley, Clemson University Otis Maloy, Washington State University
This publication can be used for non-profit, educational use only purposes. Photographers retain copyright to photographs or other images contained in this publication as cited. This material was developed as a core training module for NPDN First Detector Training. The authors and website should be properly cited. Images or photographs should also be properly cited and credited to the original source. Publication Number: 0013 Publication Date: December 2006

Diagnosing Plant Problems-Plant Diseases

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National Plant Diagnostic Network

Diagnosing Plant Problems: Plant Diseases

NPDN Publication No. 0013

MODULE 4 Diagnosing plant problems: Plant Diseases

Adapted from Plant Disease Diagnosis by M. B. Riley, M. R. Williamson and O. Maloy.

MODULE 4 Diagnosing plant problems: Plant Diseases

Plant Problem Diagnosis

MODULE 4 Diagnosing plant problems: Plant Diseases

Plant Problem Diagnosis: Preliminary Considerations

Proper plant identification is critical.

Recognize healthy plant appearance.

MODULE 4 Diagnosing plant problems: Plant Diseases

Plant Problem Diagnosis Steps

Biotic living agent

MODULE 4 Diagnosing plant problems: Plant Diseases

Plant Problem Diagnosis Steps

MODULE 4 Diagnosing plant problems: Plant Diseases

Plant Problem Diagnosis Steps

MODULE 4 Diagnosing plant problems: Plant Diseases

Plant Problem Diagnosis Steps

When was the problem noticed?

MODULE 4 Diagnosing plant problems: Plant Diseases

Plant Problem Diagnosis Steps

MODULE 4 Diagnosing plant problems: Plant Diseases

Loblolly pine killed by southern pine beetle

MODULE 4 Diagnosing plant problems: Plant Diseases

Used by permission of M. Williamson

Cotton field with chemical damage

MODULE 4 Diagnosing plant problems: Plant Diseases

Plant Problem Diagnosis Steps

7. Review environmental conditions.

8. Check host specificity many hosts may mean an abiotic problem

MODULE 4 Diagnosing plant problems: Plant Diseases

Plant Problem Diagnosis Steps

MODULE 4 Diagnosing plant problems: Plant Diseases

MODULE 4 Diagnosing plant problems: Plant Diseases

Photo: J. Dunez, www.forestryimages.org

MODULE 4 Diagnosing plant problems: Plant Diseases

Photo: Joseph O'Brien, USDA Forest Service, www.ipmimages.org

Oak leaf blister, caused by Taphrina caerulescens.

Gall on stem of eastern redcedar caused by Agrobacterium tumefaciens.

Overgrowth of leaf tissue causes thickening and distortion.

Galls form on all plant parts, caused by many pathogens.

MODULE 4 Diagnosing plant problems: Plant Diseases

MODULE 4 Diagnosing plant problems: Plant Diseases

Used by permission of M. Willliamson

Oedema of camellia, a physiological disorder

MODULE 4 Diagnosing plant problems: Plant Diseases

Symptoms and Signs

Photo: Paul Bertrand, University of Georgia, www.ipmimages.org

Target spot of tobacco, caused by Thanatephorus cucumeris.

MODULE 4 Diagnosing plant problems: Plant Diseases

Symptoms and Signs

Photo: Volcani Center Archives, Agricultural Research Organization, www.ipmimages.org

MODULE 4 Diagnosing plant problems: Plant Diseases

Symptoms and Signs

Photo: Clemson University - USDA Cooperative Extension Slide Series, , www.ipmimages.org

Brown rot of peach, caused by Monilinia fruticola

MODULE 4 Diagnosing plant problems: Plant Diseases

Photo: Joseph O'Brien, USDA Forest Service, www.ipmimages.org

Sugar maple canker caused by Nectria spp.

MODULE 4 Diagnosing plant problems: Plant Diseases

Fire blight caused by Erwinia amylovora