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

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Hendson, M., Purcell, A. H., Chen, D., Smart, C., Guilhabert, M., and B. Kirkpatrick. 2001. Genetic diversity of Pierce’s disease strains and other pathotypes of Xylella fastidiosa. Applied and Environmental Microbiology 67:895-903. Pooler, M. R., and J. S. Hartung. 1995. Genetic relationships among strains of Xylella fastidiosa from RAPD-PCR data. Current Microbiol. 31:134-137. FUNDING AGENCIES Funding for this project was provided by the CDFA Pierce’s Disease and Glassy-winged Sharpshooter Board. - 217 -
MULTILOCUS SEQUENCE TYPING TO IDENTIFY RESERVOIRS OF XYLELLA FASTIDIOSA DIVERSITY IN NATURAL HOSTS IN CALIFORNIA Project Leader: Robert Luck Dept. of Entomology University of California Riverside, CA 92521 Cooperators: Richard Stouthamer Dept of Entomology University of California Riverside, CA 92521 Len Nunney Dept. of Biology University of California Riverside, CA 92521 Don Cooksey Dept of Plant Pathology University of California Riverside, CA 92521 Danel Vickerman, Dept. of Entomology University of California Riverside, CA 92521 Reporting period: The results reported here are from work conducted from July 2004 to October 2004. ABSTRACT INTRODUCTION The ability to identify accurately and track the strains of an important infectious agent causing a plant disease is fundamental to its surveillance and management. It is also fundamental to the recognition of future changes in strains of the disease that result from 1) the invasion of exotic strains or 2) the recombination and evolution of known strains, including recombination with native strains that are as yet unrecognized. Unambiguous identification of Xylella fastidiosa (Wells) (Xf) strains and clones is of vital importance in understanding 1) the epidemiology of this bacterium, 2) the relationships between the different Xf strains and their host plant species, and 3) the geographic distribution of the “ancestral” strains in California. In the case of Xf, this is all the more critical because the introduction of the Glassy winged Sharpshooter, Homalodisca coagulata (Say) (GWSS), has changed the population dynamics, epidemiology, and the potential virulence trajectory of these bacterial pathogens. GWSS allows for frequent transmission between hosts not normally or as frequently visited by the native Xf vectors. GWSS adults feed on a wide variety of plants, and they are known to acquire multiple strains of the Xf (Costa et al. 2003). This observation takes on added significance when it is combined with the recent research findings of several recombination events between different host strains (Nunney et al. 2003, Scally et al. In Prep). Thus, the emergence of new strains that can infect new hosts or become more virulent on their traditional hosts is to be expected. To this, we can add two additional concerns. First, the identified strains in California consist of only those that are associated with a syndrome in an agricultural or ornamental host plant. We do not know how many asymptomatic indigenous strains exist in California, especially in native or naturalized alien plants because they have not, as yet, given rise to a recognizable syndrome. Second, the possibility of invasions by novel strains from other parts of the Americas cannot be ignored. Therefore, it is critically important that we characterize the diversity of X. fastidiosa strains present in California especially those presumed to be the ancestral strains, i.e., those in native and naturalized alien plant hosts as a benchmark. This information is essential for fully understanding the potential for recombination and the generation of new strains. In both central and northern California, the incidence of Xf in commercial vineyards is associated with the occurrence of the blue green sharpshooter (BGSS), Graphocephala atropunctata (Signoret) (Freitag 1951, Purcell 1975, 1976). BGSS inhabits riparian areas and has been documented as feeding on at least 16 riparian host species sequentially through the season (Purcell 1976). However, the principal species on which it feeds are the native grape, Vitis spp., blackberry, Rubus spp., Elderberry, Sambucus spp., stinging nettle, Urtica spp., Mugwort, Artemesia douglasiana, and cocklebur, Xanthium strumarium (Purcell 1976). These species occur in riparian habitats both in northern (Purcell 1975, 1976, Purcell and Saunders 1999) and southern California (Hickman 1993, B. Boyd and M. Hoddle pers. comm.). Inoculations of these species with PD Xf-infected BGSS in a controlled experiment showed that the inoculated plants maintained populations of Xf (Purcell and Saunders 1999). A similar inoculation experiment showed that Xf overwintered in a subset of these plants (Purcell and Saunders 1999) but they mostly manifested asymptomatic infections that were only detectible by culturing. It is highly likely that other nonculturable, asymptomatic forms exist in these and other plants as well (Cooksey and Costa 2003, Costa et al. In Prep). These riparian habitats harbor Xf which is spread from them to cultivated grapes by infected BGSS as they move from the riparian vegetation in late spring - early summer into the vineyards and plant communities adjacent to the riparian areas (Purcell 1975). Presumably GWSS acquires the inoculum from the infected plants in these areas, yet we know precious little of the variety of strains that reside in these riparian habitats. It is these ancestral strains that we seek to characterize and to associate with their host plant species and geographic locations. This information underpins the work on strain diversity and - 218 -
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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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Page 109 and 110: RESULTS Some plant families had no
Page 111 and 112: Table 5. Winter, spring and summer
Page 113 and 114: 16S rDNA is by far the most sequenc
Page 115 and 116: DNA MICROARRAY AND MUTATIONAL ANALY
Page 117 and 118: Inhibition of biofilm is BSA concen
Page 119 and 120: CULTURE-INDEPENDENT ANALYSIS OF END
Page 121 and 122: Borneman, J., M. Chrobak, G. Della
Page 123 and 124: P (CFU P OBJECTIVE 1. Determine the
Page 125 and 126: 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
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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 and 178: % of vessels 100 80 60 40 20 Mugwor
Page 179 and 180: DOCUMENTATION AND CHARACTERIZATION
Page 181 and 182: Magnolia002 showed more identity (9
Page 183 and 184: PLASMID ADDICTION AS A NOVEL APPROA
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Page 189 and 190: probing activities). In preliminary
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Page 193 and 194: 12. Hopkins DL (1977) Diseases caus
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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
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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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