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

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7. Hill, B.L. and A.H. Purcell. 1995a. Acquisition and retention of Xylella fastidiosa by an efficient vector, Graphocephala atropunctata. Phytopathology 85: 209-212. 8. Hill, B.L. and A.H. Purcell. 1995b. Multiplication and movement of Xylella fastidiosa within grape and four other plants. Phytopathology 85: 1368-1372. 9. Hill B.L. and A.H. Purcell, 1997. Populations of Xylella fastidiosa in plants required for transmission by an efficient vector. Phytopathology 87: 1197-1201. 10. Newman, K.L., Almeida, R.P.P., Purcell, A.H. and S.E. Lindow, 2003. Use of a green fluorescent strain of analysis of Xylella fastidiosa colonization of Vitis vinifera. App. and Env. Micro. 69: 7319-7327. 11. Purcell A.H., Finlay, A.H. and D.L. McLean, 1979. Pierce’s disease bacterium: mechanism of transmission by leafhopper vectors. Science 206: 839-841. 12. Purcell, A.H., 1981. Vector preference and inoculation efficiency as components of resistance to Pierce’s disease in European grape cultivars. Phytopathology 71: 429-435. 13. Raju, B.C.,and A.C. Goheen, 1981. Relative sensitivity of selected grapevine cultivars to Pierce’s disease bacterial infections. Am. J. Enol. Vitic. 32: 155-158. 14. Stafford, S.G. and T.E. Sabin, 1999. Statistical Analysis for the Natural Resources and Ecological Sciences, 1-22 to 1-25. Oregon State University. 15. Tyson, G.E., Stojanovic, B.J., Kuklinski, R.F., DiVittorio, T.J.and M.L. Sullivan, 1984. Scanning electron microscopy of Pierce’s disease bacterium in petiolar xylem of grape leaves. Phytopath. 75: 264-9. 16. Wistrom, C.W., and A.H. Purcell. 2003. Fate of Xylella fastidiosa in alternate hosts. Proc. Pierce’s disease research symposium, California Dept. of Food and Agriculture. FUNDING AGENCIES Funding for this project was provided by grants from the Viticulture Consortium, the California Agricultural Experiment Station (at College of Natural Resources, University of California, Berkeley), and the CDFA Pierce’s Disease and Glassywinged Sharpshooter Board. - 237 -
DOCUMENTATION AND CHARACTERIZATION OF XYLELLA FASTIDIOSA STRAINS IN LANDSCAPE HOSTS Project Leaders: Frank Wong Dept. of Plant Pathology University of California Riverside, CA 92521 frank.wong@ucr.edu Donald A. Cooksey Dept. of Plant Pathology University of California Riverside, CA 92521 donald.cooksey@ucr.edu Heather S. Costa Dept. of Entomology University of California Riverside, CA 92521 heather.costa@ucr.edu Cooperators: James Downer, UCCE, Ventura County Mike Henry, UCCE, Riverside County John Kabashima, UCCE, Orange County John Karlik, UCCE, Kern County Michelle LeStrange, UCCE, Tulare County David Shaw, UCCE , San Diego County Reporting Period: The results reported here are from work conducted from October 15, 2003 to October 12, 2004. ABSTRACT To better understand the impact of Xylella fastidiosa on the urban environment and the potential for ornamental hosts to serve as reservoirs for agronomically important diseases caused by the bacteria, a survey project was initiated to document and characterize strains of the bacteria harbored in landscape plants. Targeted sampling of 122 landscape species either symptomatic for bacterial scorch or testing positive for X. fastidiosa by ELISA in 2003 was performed. Of the 830 samples, 321 tested positive by ELISA (representing 77 of the 122 species tested). X. fastidiosa was also detected in 23 species by PCR-amplification using X. fastidiosa specific primers. Twenty-seven isolates from 13 host species were obtained from samples testing positive by ELISA. Isolates from plants not previously reported as hosts in southern California urban environments included mulberry, heavenly bamboo, magnolia, day lily, western redbud, jacaranda and peach. Genetic characterization of these isolates by 16S-23S rDNA sequencing distributed these isolates amongst previously characterized strain groups: almond leaf scorch (crape myrtle, ornamental plum, liquidambar, gingko, olive), Pierce’s disease (magnolia, peach, western redbud), mulberry leaf scorch (mulberry, heavenly bamboo), and oleander leaf scorch (magnolia, jacaranda, day lily). The role of some X. fastidiosa strains in their ability to cause disease is presently being tested by fulfilling Koch’s postulates in glasshouse experiments. The data collected from this study strongly suggest that X. fastidiosa is causing a number of scorch diseases in the urban landscape, and that strains of agronomic importance may be harbored in this environment. INTRODUCTION Xylella fastidiosa (Xf) is a xylem-limited, insect-vectored, plant pathogen that can cause severe damage to a wide range of host plants. Diseases caused by this pathogen include Pierce’s disease of grapevine (PD), oleander leaf scorch (OLS) and almond leaf scorch (ALS). In 2003, a survey of landscape plants in five urban locations in southern California was initiated to document the incidence of the Xf infection in landscape ornamental hosts and to characterize strains existing in these hosts that may prove a threat to landscape ornamentals or crops of agronomic importance. Two hundred twenty one samples (29%) representing 48 species tested positive by ELISA. Ten isolates of Xf were obtained from eight plant species (Fatsia japonica, Ginkgo biloba, Lagerstroemia indica, Liquidambar styraciflua, Morus alba, Nandina domestica, Olea europea, and Prunus cerasfiera) not previously described as hosts of X. fastidiosa in southern California. Based upon the results of the first year, targeted sampling of host species testing positive by ELISA was performed primarily in the Riverside and Redlands areas in order to obtain additional isolates for characterization. To prove the role of Xf in causing disease in previously identified hosts, test plants were inoculated in glasshouse experiments to fulfill Koch’s postulates for these isolates, and to determine if they were able to cause disease in grapevine and oleander. OBJECTIVES 1. Use laboratory methods to identify landscape host species that are infected with X. fastidiosa. 2. Secure isolates from these hosts to document infection and provide material for genetic characterization of the X. fastidiosa strain(s) involved. 3. Genetically characterize the strains of pathogen in landscape plant species. 4. Confirm pathogenic infection through inoculation studies with specific isolates. 5. Test ability of new strains to infect agricultural crops including grape, olive, and almond. RESULTS Objective 1 In 2004, 830 samples from 122 landscape plant species were collected. Sampling focused on plant species that were symptomatic or had tested positive by ELISA in 2003 surveys. Three hundred twenty one samples (39%), tested positive by ELISA. At least one sample from 77 of the 122 species tested was positive by ELISA (63%). Attempts to isolate the - 238 -
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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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Project Leader: Robert F. Luck Dept
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A more interesting analysis using t
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MYCOPATHOGENS AND THEIR EXOTOXINS I
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POPULATION DYNAMICS AND INTERACTION
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No egg masses were recorded on olea
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EXPLORATION FOR FACULTATIVE ENDOSYM
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Although Baumannia and Wolbachia we
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EFFECTS OF SUBLETHAL DOSES OF IMIDA
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Table 2. Mortality and flight perfo
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A NOVEL METHOD TO INDUCE OVIPOSITIO
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REFERENCES Brodbeck, B. V., P. C. A
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Cohorts H. coagulata neonates were
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REFERENCES Blua, M. J. and D. J. W.
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Figure 2 shows the average numbers
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REPRODUCTIVE BIOLOGY AND PHYSIOLOGY
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REFERENCES Blua, M. J., Phillips, P
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RESULTS Some plant families had no
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Table 5. Winter, spring and summer
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16S rDNA is by far the most sequenc
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DNA MICROARRAY AND MUTATIONAL ANALY
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Inhibition of biofilm is BSA concen
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CULTURE-INDEPENDENT ANALYSIS OF END
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Borneman, J., M. Chrobak, G. Della
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P (CFU P OBJECTIVE 1. Determine the
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Purcell, AH and SR Saunders. 1999a.
Page 127 and 128: Figure 1: Southern blot of tolC::ap
Page 129 and 130: Project Leader: David Gilchrist Dep
Page 131 and 132: III. Production of transgenic plant
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
Page 139 and 140: ISOLATION OF BACTERIOPHAGES SPECIFI
Page 141 and 142: Project Leader: Michele M. Igo Sect
Page 143 and 144: outer membrane fractions using two-
Page 145 and 146: 1995; Purcell and Saunders, 1999).
Page 147 and 148: DEVELOPMENT OF SSR MARKERS FOR GENO
Page 149 and 150: Strain Name Host of Origin County o
Page 151 and 152: ROLE OF ATTACHMENT OF XYLELLA FASTI
Page 153 and 154: wild-type cells was not conclusive
Page 155 and 156: DETERMINATION OF GENES CONFERRING H
Page 157 and 158: S1 S2 S3 S4 Nb123456789bb123456789b
Page 159 and 160: MULTILOCUS SEQUENCE TYPING TO IDENT
Page 161 and 162: GENOME-WIDE IDENTIFICATION OF RAPID
Page 163 and 164: Americas and since most of the plan
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
Page 173 and 174: In parallel, an “activating trans
Page 175 and 176: PATTERNS OF XYLELLA FASTIDIOSA INFE
Page 177: % of vessels 100 80 60 40 20 Mugwor
Page 181 and 182: Magnolia002 showed more identity (9
Page 183 and 184: PLASMID ADDICTION AS A NOVEL APPROA
Page 185 and 186: GENETIC VARIABILITY OF XYLELLA FAST
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Page 189 and 190: probing activities). In preliminary
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Page 195 and 196: The almost complete absence of info
Page 197 and 198: QUANTIFYING LANDSCAPE-SCALE MOVEMEN
Page 199 and 200: Table 1. The mean (±SD) ELISA read
Page 201 and 202: EPIDEMIOLOGICAL ASSESSMENTS OF PIER
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Page 205 and 206: SPATIAL DATABASE CREATION AND MAINT
Page 207 and 208: Project Leader: Thomas M. Perring D
Page 209 and 210: Figure 2. Individual leaves from a
Page 211 and 212: The state variables, process functi
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Page 221 and 222: OBJECTIVES 1. Track the movement of
Page 223 and 224: REFERENCES 1. Beard, C.B., Cordon-R
Page 225 and 226: cases, positive phloem samples were
Page 227 and 228: 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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