Impact Of Host Plant Xylem Fluid On Xylella Fastidiosa Multiplication ...

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-5 with RST-31 and RST-33 was also examined, and found to detect 6.5×10P P µg/mL of DNA. In addition, an internal set of primers was developed so that nested PCR is now possible for samples appearing negative with traditional methods. Pathogen Strain A simple assay was conducted to categorize Xf by its common strains. Recent genetic and cross-inoculation studies showed that Xf had genetically distinct strains in different host plants (e.g., oak, oleander, grapes) (Almeida & Purcell 2003c, Chen et al. 1995, Henderson et al. 2001). Typically, Xf isolates from one plant species are genetically similar, despite different geographical origins. However, Xf isolated from almonds can be genetically separated into three distinct strains – with one ALS strain recovered in orchards in the northern San Joaquin Valley (ALS-XfSJV) that is genetically more similar to grape strains than the two other ALS strains (ALS-Xf1, ALS-Xf2). The few weeds samples that returned positive results in the winter and spring were analyzed using restriction enzyme digestion, and have so far been found to be all of the northern San Joaquin Valley (ALS-XfSJV). The lack of positive results for Xf in vineyard weeds after mid-March prevented us from analyzing any changes (new inoculations) of Xf strains. However, we were able to analyze the strain of Xf in the mouthparts of the GWSS tested, and found that these insects were also found to be carrying Xf of the PD type. These results are consistent with previous findings that strains of Xf tend to be host-specific (Almeida and Purcell 2003c). CONCLUSIONS The results of this study indicate that the winter and spring weeds may be the most important reservoirs for Xf in vineyards infected with Pierce's Disease. We recovered Xf from four species of weeds that have either not been studied in depth (Stellaria sp. and Capsella sp.) or would benefit from further investigation (Erodium sp. and Poa annua). We seem to have caught the tail end of the season where Xf is abundant in weeds, so the next season's sampling scheme will focus more heavily on vineyard groundcover during the winter months of December, January and February. Future research along these lines could illuminate the importance of previously overlooked alternate host plant species. One hypothesis for the importance of winter weeds for the persistence of Xf is that when symptomatic leaves senesce in late fall, they land directly on the groundcover, thus greatly enhancing the likelihood that any insect feeding there will transmit the bacteria to the weeds. Conclusive evidence of this hypothesis could provide a simple and low cost method for controlling the spread of PD. REFERENCES Almeida, RPP and AH Purcell. 2003a. Homalodisca coagulata transmission of Xylella fastidiosa to almond. Plant Disease 87: 1255-1259. Almeida, RPP and AH Purcell. 2003b. Transmission of Xylella fastidiosa to grapevines by Homalodisca coagulata. J Econ Entomol 96: 264-271. Almeida, RPP and AH Purcell. 2003c. Biological traits of Xylella fastidiosa strains from grapes and almonds. Appl Environ Microbiol 69: 7447-7452. Andersen, PC, B Brodbeck and RF Mizell, III. 1992. Feeding by the leafhopper, Homalodisca coagulata, in relation to xylem fluid chemistry and tension. J Insect Physiol 38: 611-622. Bextine, B, S Tuan, H Shaikh, MJ Blua and TA Miller. 2004. Evaluation of methods for extracting Xylella fastidiosa DNA from the glassy-winged sharpshooter. J Econ Entomol 97: 757 - 763. Chen, J, O Lamikanra, CJ Chang and DL Hopkins. 1995. Randomly amplified Polymorphic DNA analysis of Xylella fastidiosa Pierce’s disease and oak leaf scorch pathotypes. Appl Environ Microbiol 61: 1688-1690. Daane, KM and MW Johnson 2003. Biology and Ecology of the glassy-winged sharpshooter in the San Joaquin Valley In M. Athar Tariq et al. [eds.], Proc Pierce’s Disease Research Symposium. CDFA Digital Logistix, Sacramento, CA. Daane, KM, MW Johnson, T Ruiz and J Hashim. 2004. Research shows GWSS have their urban preferences. Kern/Tulare GWSS Update. March 5, 2004. Feil H and AH Purcell. 2001. Temperature-dependent growth and survival of Xylella fastidiosa in vitro and in potted grapevines. Plant Disease 85:1230-1234. Fukatsu, T. 1999. Acetone preservation: a practical technique for molecular analysis. Mol Ecol 8: 1935 – 1945. Goodwin, P and AH Purcell. 1992. Pierce’s disease, pp 76-84. In DL Flaherty et al. [eds.], Grape pest management. Univ Calif Div Agric Nat Res Publ 3343, Berkeley, CA. Hendson, M, AH Purcell, D Chen, C Smart, M Guilhabert and B Kirkpatrick. 2001. Genetic diversity of Pierce’s disease strains and other pathotypes of Xylella fastidiosa. Appl Environ Microbiol 67:895-903. Hoddle, M S, SV Trispitsyn and DJW Morgan. 2003. Distribution and plant association records for Homalodisca coagulate (Hemiptera: Cicadellidae) in Florida. Fl Entomol 86: 89-91. Perring, TM, CA Farrar and MJ Blua. 2001. Proximity to citrus influences Pierce's disease in Temecula Valley vineyards. Calif Agricul 55: 13-18. Purcell, AH 1990. Homopteran transmission of xylem-inhabiting bacteria, pp. 243-266 In: Advances in Disease Vector Research, vol., 6. Springer-Verlag, New York. Purcell, AH and NW Frazier. 1985. Habitats and dispersal of the leafhopper vectors of Pierce’s disease in the San Joaquin Valley. Hilgardia 53: 1-32. - 183 -
Purcell, AH and SR Saunders. 1999a. Glassy-winged sharpshooters expected to increase plant disease. Calif Agricul 53(2): 26-27. Purcell, AH and SR Saunders. 1999b. Fate of Pierce's disease strains of Xylella fastidiosa in common riparian plants in California. Plant Disease 83: 825-830. Redak, RA, AH Purcell, JRS Lopes, MJ Blua, RF Mizell and CP Andersen. 2004. The biology of xylem-fluid feeding vectors of Xylella fastidiosa and their relation to disease epidemiology. Annu Rev Entomol 49: 243-270. FUNDING AGENCIES Funding for this project was provided by the University of California Pierce’s Disease Grant Program for fiscal year 2002-03, and by the CDFA Pierce’s Disease and Glassy-winged Sharpshooter Board for fiscal year 2004-05. - 184 -
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IMPACT OF HOST PLANT XYLEM FLUID ON
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0.18 600 Optical density 0.16 0.14
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OPTIMIZING MARKER-ASSISTED SELECTIO
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esistant and susceptible genotypes
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MAP BASED IDENTIFICATION AND POSITI
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esistant genotypes are in process a
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BREEDING PIERCE’S DISEASE RESISTA
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Progeny from crosses of field resis
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a = (1=low, 4= high); b = (1=green,
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v.8) for differences due to the pla
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SHARPSHOOTER FEEDING BEHAVIOR IN RE
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correlated with all other findings
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EFFECTS OF FEEDING SUBSTRATE ON RET
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REFERENCES Bextine, B. and T. Mille
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will also provide insights into the
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OBJECTIVES 1. Determine glassy-wing
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GWSS per 30 s sweep (seasonal avera
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ELISA for the presence of GWSS egg
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Project Leader: Thomas P. Freeman E
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Freeman, T. P., R. A. Leopold, D. R
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pathogen, when they move into viney
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A NOVEL IMMUNOLOGICAL APPROACH FOR
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1.6 GWSS Adults That Fed on Protein
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Hagler, J.R. & S.E. Naranjo. 2004.
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RESULTS: Oviposition Survey Wild gr
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Host specificity testing: No-choice
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currently employed morphological ch
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increase our present understanding
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BIOLOGY AND MORPHOMETRIC ANALYSIS O
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Table 1. Mean a developmental durat
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EFFECTS OF USING CONSTANT AND CYCLI
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REFERENCES Gautam, R. D. 1986. Effe
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PARASITISM OF THE GLASSY-WINGED SHA
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Table 1. Parasitism by G.ashmeadi o
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Page 75 and 76: A more interesting analysis using t
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Page 79 and 80: POPULATION DYNAMICS AND INTERACTION
Page 81 and 82: No egg masses were recorded on olea
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Page 89 and 90: Table 2. Mortality and flight perfo
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Page 93 and 94: REFERENCES Brodbeck, B. V., P. C. A
Page 95 and 96: Cohorts H. coagulata neonates were
Page 97 and 98: REFERENCES Blua, M. J. and D. J. W.
Page 99 and 100: Figure 2 shows the average numbers
Page 101 and 102: REPRODUCTIVE BIOLOGY AND PHYSIOLOGY
Page 103 and 104: REFERENCES Blua, M. J., Phillips, P
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Page 109 and 110: RESULTS Some plant families had no
Page 111 and 112: Table 5. Winter, spring and summer
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Page 115 and 116: DNA MICROARRAY AND MUTATIONAL ANALY
Page 117 and 118: Inhibition of biofilm is BSA concen
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Page 121 and 122: Borneman, J., M. Chrobak, G. Della
Page 123: P (CFU P OBJECTIVE 1. Determine the
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Page 129 and 130: Project Leader: David Gilchrist Dep
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Page 133 and 134: RESULTS Construction of cDNA Librar
Page 135 and 136: CONCLUSIONS Genetic resistance and
Page 137 and 138: Mutagenesis of Xylella The EZ::TN T
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Page 143 and 144: outer membrane fractions using two-
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Page 147 and 148: DEVELOPMENT OF SSR MARKERS FOR GENO
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Page 153 and 154: wild-type cells was not conclusive
Page 155 and 156: DETERMINATION OF GENES CONFERRING H
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Page 159 and 160: MULTILOCUS SEQUENCE TYPING TO IDENT
Page 161 and 162: GENOME-WIDE IDENTIFICATION OF RAPID
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Page 165 and 166: EFFECTS OF CHEMICAL MILIEU ON ATTAC
Page 167 and 168: CONCLUSIONS Our overall objective i
Page 169 and 170: RESULTS Objective 1. We conducted t
Page 171 and 172: Redak, R. A., Purcell, A. H., Lopes
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PATTERNS OF XYLELLA FASTIDIOSA INFE
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% of vessels 100 80 60 40 20 Mugwor
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DOCUMENTATION AND CHARACTERIZATION
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Magnolia002 showed more identity (9
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PLASMID ADDICTION AS A NOVEL APPROA
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GENETIC VARIABILITY OF XYLELLA FAST
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probing activities). In preliminary
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the slow release valve was opened a
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12. Hopkins DL (1977) Diseases caus
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The almost complete absence of info
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QUANTIFYING LANDSCAPE-SCALE MOVEMEN
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Table 1. The mean (±SD) ELISA read
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EPIDEMIOLOGICAL ASSESSMENTS OF PIER
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multiply to relatively high (easily
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SPATIAL DATABASE CREATION AND MAINT
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Project Leader: Thomas M. Perring D
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Figure 2. Individual leaves from a
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The state variables, process functi
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DEVELOPMENT OF A FIELD SAMPLING PLA
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Figure 1. Three main dispersion pat
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OBJECTIVES 1. Track the movement of
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REFERENCES 1. Beard, C.B., Cordon-R
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cases, positive phloem samples were
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EXPLOITING XYLELLA FASTIDIOSA PROTE
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Figure 2. Polymer disk accumulation
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CHARACTERIZATION OF NEONICOTINOIDS
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EVALUATION OF RESISTANCE POTENTIAL
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Project Leader: Doug Cook Dept. of
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Based on the in silico analysis, de
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CONTROL OF PIERCE’S DISEASE THROU
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Figure 1. Oleander ‘White’ afte
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RESULTS During the reporting period
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DEVELOPMENT OF AN ARTIFICIAL DIET A
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DESIGN OF CHIMERIC ANTI-MICROBIAL P
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Neutrophil elastase Cecropin B Chim
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and WTXb is very low (0.00-0.40%) a
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100 100 0.056 North America 100 0.0
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GENETIC DIFFERENTIATION AMONG GEOGR
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Table 1. Single-populations descrip
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MOLECULAR DISTINCTION BETWEEN POPUL
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These novel observations strongly s
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SEQUENCE DIVERGENCE IN TWO MITOCHON
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Table 3. Pairwise sequence distance
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DEVELOPMENT OF MOLECULAR DIAGNOSTIC
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A. B. Relative Density of SCAR Band
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THE ALIMENTARY TRACK OF GLASSY-WING
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We have dissected and identified al
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REALIZED LIFETIME PARASITISM AND TH
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REPRODUCTIVE AND DEVELOPMENTAL BIOL
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Mean adult longevity (days) 25 20 1
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the proposed exploratory work; thes
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SEARCHING FOR AND COLLECTING EGG PA
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REFERENCES Hoddle, M. S. and S. V.
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some Gram(+) bacteria, but are inac
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7. Hultmark, D., et al., Insect Imm
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Isolation of Fungal Pathogens Soil
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IDENTIFICATION OF MECHANISMS MEDIAT
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concentrations of 1.5 x 10 7 bacter
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anchor it in the outer membrane (sh
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SYMBIOTIC CONTROL OF PIERCE’S DIS
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MANAGEMENT OF PIERCE’S DISEASE OF
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pfF mutants are taken up by insects
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Simpson, A. J. G., F. C. Reinach, P
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Al-Wahaibi, A.K., Owen, and J. G. M
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patterns differed among the lines,
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Punja, Z.K. 2001. Genetic engineeri
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mind, we conducted an additional st
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REFERENCES Toscano, N., Byrne, F.,
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RESULTS AND CONCLUSIONS The program
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COMPATIBILITY OF INSECTICIDES WITH
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More tests are in progress to addre
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Impact Of Host Plant Xylem Fluid On Xylella Fastidiosa Multiplication ...

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