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Grapevines and Pierce’s Disease: a xylem fluid mineral-status

dependent condition.

Breno Leite, PhDX-Ray Microanalysis

breno.leite@thermofisher.com

Geological Society of America: Portland 2009

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Outline

Problem and economical impactPierce's disease – How grapevines are affected?What is the disease mechanism?Why chemistry is so important?“Calcium Bridging” hypothesis, based on Energy Dispersive Spectroscopy results.Latest research resultsConclusion

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The wine industry: California

There are more than 2,800 wineries in California

The California wine industry has an annual impact of $51.8 billion on the state's economy, and an economic impact of $125.3 billion on the U.S. economy

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The wine industry: Florida

Florida has a $5 million viticulture industry.Number of Wineries: 33

Vitis rotundifoliaMuscadine grapes

Why is Florida Important?

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American grapes vs. European grapes

V. rotundifolia. “Muscadines”, “American grape”

“native American grapes”

V. vinifera. "Old world grape", "European grape“

“True grapes”

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Muscadine Florida Wines: Noble red and Carlos

Noble RedSemi-sweet Noble

CarlosSemi-sweet Carlos

Unique FL industry

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Muscadine grapevines are very “tolerant” to Pierce's disease

Some grape cultivars are very susceptible, usually dying within two years.

Leaf Scorching

Leaf petioles still attached after leaf drop

Bacterial Leaf Scorch on Red Oak

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Control of Plant Diseases: Fundamentals and PrinciplesControl of Plant Diseases: Fundamentals and Principles

Disease Triangle for Pierce's DiseaseDisease Triangle for Pierce's Disease

Amount of Disease

HostHostConditions favoring susceptibilityConditions favoring susceptibility

EnvironmentEnvironmentConditions favoring diseaseConditions favoring disease

PathogenPathogenTotal Virulence, abundanceTotal Virulence, abundance

Xylella fastidiosa

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XylellaXylella fastidiosafastidiosa living within the vectorliving within the vector

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XylellaXylella fastidiosafastidiosa living within the vectorliving within the vector

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Colonization Process “In planta”

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Xylem transport

Carrying water, minerals, amino acids and organic acids

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Typical Symptoms of Pierce`s Disease

nutritional-based disease

Disease Mechanism

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Pierce’s disease

Pierce’s diseaseNutritional deficiency

Nutritional deficiency

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The surface of XF cells is negatively charged

Leite, B et al. 2002. Genomics and X-ray Microanalysis Indicate that Ca++ and thiol mediatethe Xylella fastidiosa Aggregation and Adhesion. B.J.M.B. Sci. 36:645-650.

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Aggregation and adhesion Aggregation and adhesion X. X. fastidiosafastidiosa cells in xylem wallscells in xylem walls

SS((--))

++

((--)S)SSS

SS

SS

SSCaCa

SS

SSCaCa

HpSS

MgMg

SS

SS

MgMg

SS S

Xf

Xf

++++ SS S

SS((--))

((--)S)S

SSSS

SS

SSCaCa

SS

SS

CaCa

SSMgMg

SS

SS

MgMgXf

Xf

Hp

??COOHCOOHCOOHCOOH ??COOHCOOHCOOHCOOH

XSXS

The calcium bridging hypothesisThe calcium bridging hypothesis

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Xylem fluid form susceptible plants induce aggregation?

Cell aggregation >> Cell aggregation >> BiofilmBiofilm Formation >> More Disease?Formation >> More Disease?

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Pattern of aggregation of Pattern of aggregation of XylellaXylella fastidiosafastidiosa cells after being cells after being incubated with xylem fluid from different cultivars. incubated with xylem fluid from different cultivars.

Tolerant varieties

Susceptible varieties

Testing Aggregation “in vitro”

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[P]*

[cit]

/[Ca]

*[M

g][P

]*[c

it]/[C

a]*[

Mg]

a

b

d

e ededf

ef f f

Resistance of Grapes based on Xylem Chemistry

Most tolerant varieties

Most susceptible varieties

Grape varietiesGrape varieties

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Duncan's Multiple Range Test for [P]*[cit]/[Ca]*[Mg]

Duncan Grouping Mean N varieties

A 1.5613 3 V. rotundifolia cv. NobleB 1.2512 3 V. rotundifolia cv. CarlosC 0.9321 3 V. ruspestris cv. Saint GeorgeD 0.3670 3 V. ruspestris cv. Constancia

E D 0.3270 3 V. simpsoni cv. PixiallaE D F 0.2399 3 V. champini cv. DogridgeE F 0.0560 3 V. vinifera cv. Exotic

F 0.0235 3 V. vinifera cv. Chenin BlancF 0.0221 3 V. vinifera cv. Chardonnay

Xylem fluid collected in California (dormant season) Xylem fluid collected in California (dormant season)

Profiles separate resistant and susceptible plants

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Xylem vessel = works as an affinity column chromatography Xylem vessel = works as an affinity column chromatography

XF Bacterial cellsXF Bacterial cells

FLOW

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FLOWFLOW

Resistant/Tolerant plantResistant/Tolerant plant

XF Bacterial cellsXF Bacterial cells

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Xylem vessel = Column chromatography Xylem vessel = Column chromatography

Bacterial cells attracted byBacterial cells attracted bythe vessel wallthe vessel wall

FLOW

Susceptible/Diseased plantSusceptible/Diseased plant

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Bacterial cells attracted byBacterial cells attracted bythe vessel wallthe vessel wall

FLOW

Susceptible/Diseased plantSusceptible/Diseased plant

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FLOW

Susceptible/Diseased plantSusceptible/Diseased plant

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FLOW

Symptoms: Leaf scorching and Symptoms: Leaf scorching and chlorosischlorosis

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FLOW

Symptoms: Leaf scorching and Symptoms: Leaf scorching and chlorosischlorosis

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FLOW

Symptoms: Leaf scorching and Symptoms: Leaf scorching and chlorosischlorosis

= = FastidianFastidian gum/gum/glycoproteinsglycoproteins

BiofilmBiofilmCity of Cells = better nutrient uptake and City of Cells = better nutrient uptake and

toxins/enzymes productiontoxins/enzymes production

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Typical Symptoms of Pierce`s Disease

Disease Mechanism

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Pierce`s Disease of Grapevines: Lesion of leaf scorched areas accumulate silicon.This is the first time that PD symptoms are associated silicon accumulation.

Lesion areaLesion areaHealthy TissueHealthy Tissue

Silicon accumulation is severely affected grapevine leaves

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Is there a difference between healthy and diseased plant tissue?

Healthy

Diseased

Intensity map 3D display

Si rich area

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Silicon accumulation is severely affected grapevine leavesReduces drought stress

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Silicon accumulation questions

•How general is this response?

•What happens to PD symptoms in plants exposed tocalcium silicates?

Silicon helps plants to cope with drought.

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Why soil pH range?Why soil pH range?

TheThe effecteffect of of soilsoil pH is pH is greatgreat onon thethe solubilitysolubility of of mineralsminerals oror nutrientsnutrients. . FourteenFourteen of of thethe seventeenseventeen essentialessential plantplant nutrientsnutrients are are obtainedobtained fromfrom thethesoilsoil. . BeforeBefore a a nutrientnutrient cancan bebe usedused byby plantsplants it it mustmust bebe dissolveddissolved in in thethe soilsoilsolutionsolution..

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Precipitation of calcium phosphate in different pHsPrecipitation of calcium phosphate in different pHs

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Soil Climate Cultural Decision

Depth

Texture

Water and NutrientSupply

Radiation

Temperature

Humidity

Wind speed

Rainfall

Evaporation

Vine density

Rootstock

Fertilization

Irrigation

Pest and Diseases control

Pruning level

Smart (1985)Smart (1985)

Vigor stimulation

Foliar Characteristics

Canopy Management

Vine physiology Fruit Wine

Cultivated GrapevinesCultivated Grapevines

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California California –– Cucamonga ValleyCucamonga Valley Florida Florida –– Tallahassee / CVSFRTallahassee / CVSFR

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Acknowledgements

Peter Andersen, University of Florida

Maria Ishida, Florida Department of Food and Agriculture

Eduardo Alves, University of Lavras, Brazil

David Melton, Thermo Fisher Scientific

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References

Leite, B. & Andersen, P. C. 2009. Localized accumulation of silicon (Si) in grape leaves affected by Pierce’s disease. Microsc. Microanal. 15 (2):918-19).

Alves, E., Leite, B., Pascholati, S. F., Ishida, M. L. & Andersen, P.C. 2009. Citrus sinensis leaf petiole and blade colonization by Xylella fastidiosa: details of Xylem vessel occlusion. Scientia Agricola. 66:145-285.

Alves E. , Leite B., Marucci,R. C. , Pascholati S. F, Lopes J.R.S. & Andersen P. C. 2008. Retention Sites for Xylella fastidiosa in Four Sharpshooter Vectors (Hemiptera: Cicadellidae) Analyzed by Scanning Electron Microscopy. Current Microbiology. 56:531-538.

Andersen, PC., Brodbeck B. V., Oden S., Shriner. A., Leite, B. 2007. Influence of xylem fluid chemistry on planktonic growth, biofilm formation and aggregation of Xylella fastidiosa. FEMS Microbiology Letters Vol. 274: 210.

Ishida, M.L.., Andersen, P.C.& Leite, B. 2004. Cecropin B antimicrobial activity against Xylella fastidiosa: effect on colony formation. Physiological and Molecular Plant Pathology. 64:73-81.

Leite, B., Andersen, P.C. & Ishida, M.L. 2004. Colony aggregation and biofilm formation in xylem chemistry-based media for Xylella fastidiosa. FEMS Microbiol. Lett. 230:283-290.

Alves, E., Marucci, R.C., Lopes, J.R.S. & Leite, B., 2004. Relationship between the proportion of colonized vessels by Xylella fastidiosa in plum, coffee and citrus and the leaf symptomatology exhibited. J. Phytopathology. 152: 291-297.

Andersen, P.C., Momol, E.A., Leite, B., Momol, M.T Ishida, M.L. 2004. The Effect of Lytic Peptides on Xylellafastidiosa in buffer and in Xylem Fluid Grapevines. Vitis 43: 19-25.

Alves, E., Kitajima, E.W. & Leite, B. 2003. Interaction of Xylella fastidiosa with different cultivars of Nicotianatabacum: a comparison of colonization patterns". J. Phytopathology. 151:500-506.