The world leader in serving science Grapevines and Pierce s Disease: a xylem fluid mineral-status dependent condition. Breno Leite, PhD X-Ray Microanalysis breno.leite@thermofisher.com Geological Society of America: Portland 2009
Outline Problem and economical impact Pierce'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 results Conclusion 2
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 3
The wine industry: Florida Florida has a $5 million viticulture industry. Number of Wineries: 33 Why is Florida Important? Vitis rotundifolia Muscadine grapes 4
American grapes vs. European grapes V. rotundifolia. Muscadines, American grape native American grapes V. vinifera. "Old world grape", "European grape True grapes 5
Muscadine Florida Wines: Noble red and Carlos Noble Red Semi-sweet Noble Carlos Semi-sweet Carlos Unique FL industry 6
Muscadine grapevines are very tolerant to Pierce's disease Leaf petioles still attached after leaf drop Bacterial Leaf Scorch on Red Oak Leaf Scorching Some grape cultivars are very susceptible, usually dying within two years. 7
Disease Triangle for Pierce's Disease Control of Plant Diseases: Fundamentals and Principles Xylella fastidiosa Pathogen Total Virulence, abundance Environment Conditions favoring disease Amount of Disease Host Conditions favoring susceptibility 8
Xylella fastidiosa living within the vector 9
Xylella fastidiosa living within the vector 10
Colonization Process In planta 11
Xylem transport Carrying water, minerals, amino acids and organic acids 12
Disease Mechanism Typical Symptoms of Pierce`s Disease nutritional-based disease 13
Pierce s disease Nutritional deficiency Nutritional deficiency Pierce s disease 14
The surface of XF cells is negatively charged Leite, B et al. 2002. Genomics and X-ray Microanalysis Indicate that Ca ++ and thiol mediate the Xylella fastidiosa Aggregation and Adhesion. B.J.M.B. Sci. 36:645-650. 15
Aggregation and adhesion X. fastidiosa cells in xylem walls The calcium bridging hypothesis (-)S + S (-) + Xf S Xf S S S Ca Mg S S S S S S Ca Mg S S Hp Xf S (-)S S Xf S (-) S S S Ca Mg + + S S S COOH COOH COOH? COOH? XS S Ca Mg S S S Hp 16
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Xylem fluid form susceptible plants induce aggregation? Cell aggregation >> Biofilm Formation >> More Disease? 18
Testing Aggregation in vitro Tolerant varieties Susceptible varieties Pattern of aggregation of Xylella fastidiosa cells after being incubated with xylem fluid from different cultivars. 19
Resistance of Grapes based on Xylem Chemistry a Most tolerant varieties [P]*[cit]/[Ca]*[Mg] b d e ed edf Most susceptible varieties ef f f Grape varieties 20
Profiles separate resistant and susceptible plants Xylem fluid collected in California (dormant season) Duncan's Multiple Range Test for [P]*[cit]/[Ca]*[Mg] Duncan Grouping Mean N varieties A 1.5613 3 V. rotundifolia cv. Noble B 1.2512 3 V. rotundifolia cv. Carlos C 0.9321 3 V. ruspestris cv. Saint George D 0.3670 3 V. ruspestris cv. Constancia E D 0.3270 3 V. simpsoni cv. Pixialla E D F 0.2399 3 V. champini cv. Dogridge E F 0.0560 3 V. vinifera cv. Exotic F 0.0235 3 V. vinifera cv. Chenin Blanc F 0.0221 3 V. vinifera cv. Chardonnay 21
Xylem vessel = works as an affinity column chromatography FLOW XF Bacterial cells 22
FLOW XF Bacterial cells Resistant/Tolerant plant 23
Xylem vessel = Column chromatography Bacterial cells attracted by the vessel wall FLOW Susceptible/Diseased plant 24
Bacterial cells attracted by the vessel wall FLOW Susceptible/Diseased plant 25
FLOW Susceptible/Diseased plant 26
FLOW Symptoms: Leaf scorching and chlorosis 27
FLOW Symptoms: Leaf scorching and chlorosis 28
Biofilm City of Cells = better nutrient uptake and toxins/enzymes production FLOW = Fastidian gum/glycoproteins glycoproteins Symptoms: Leaf scorching and chlorosis 29
Disease Mechanism Typical Symptoms of Pierce`s Disease 30
Silicon accumulation is severely affected grapevine leaves Pierce`s Disease of Grapevines: Lesion of leaf scorched areas accumulate silicon. This is the first time that PD symptoms are associated silicon accumulation. Healthy Tissue Lesion area 31
Is there a difference between healthy and diseased plant tissue? Intensity map 3D display Healthy Si rich area Diseased 32
Silicon accumulation is severely affected grapevine leaves Reduces drought stress 33
Silicon accumulation questions How general is this response? What happens to PD symptoms in plants exposed to calcium silicates? Silicon helps plants to cope with drought. 34
Why soil ph range? The effect of soil ph is great on the solubility of minerals or nutrients. Fourteen of the seventeen essential plant nutrients are obtained from the soil. Before a nutrient can be used by plants it must be dissolved in the soil solution. 35
Precipitation of calcium phosphate in different phs 36
Cultivated Grapevines Soil Climate Cultural Decision Depth Texture Water and Nutrient Supply Radiation Temperature Humidity Wind speed Rainfall Evaporation Vine density Rootstock Fertilization Irrigation Pest and Diseases control Pruning level Vigor stimulation Foliar Characteristics Canopy Management Vine physiology Fruit Wine Smart (1985) 37
California Cucamonga Valley Florida Tallahassee / CVSFR 38
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 39
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 chemistrybased 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 Xylella fastidiosa 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 Nicotiana tabacum: a comparison of colonization patterns". J. Phytopathology. 151:500-506. 40