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

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REFERENCES 1. Davis, M. J., W. J. French, and N. W. Schaad. 1981. Axenic culture of the bacteria associated with phony diseaseof peach and plum leaf scald. Curr. Microbiol. 6:309-314. 2. De Feyter, R., Yang, Y., & Gabriel, D. W. 1993. Gene-for-genes interactions between cotton R genes and Xanthomonas campestris pv. malvacearum avr genes. Mole. Plant-Microbe Interact. 6:225-237. 3. Feil H, Feil WS, Detter JC, Purcell AH, Lindow SE. 2003. Site-directed disruption of the fimA and fimF fimbrial genes of Xylella fastidiosa. Phytopath. 93: 675-682. 4. Gaurivaud P, Souza LC, Virgilio AC, Mariano AG, Palma RR, Monteiro PB. 2001. Gene disruption by homologous recombination in the Xylella fastidiosa citrus variegated chlorosis strain. Appl Environ Microbiol. 68:4658-65. 5. Guilhabert, M.R., Hoffman, L.M., Mills,D.A., & Kirkpatrick, B.C. 2001. Transposon mutagenesis of Xylella fastidiosa by electroporation of Tn5 synaptic complexes. Mole. Plant-Microbe Interact. 14:701-706. 5. Kovach, M.E., Elzer, P.H., Hills, D.S., Robertson, G. T., Farris, M.A., Roop, R. M. and Peterson, K.M. 1995. Four new derivatives of the broad-host range cloning vector pBBR1MCS, carrying different antibiotic resistance cassettes. Gene 166:175-176. 6. Purcell, A. H. and Hopkins, D. L. 1996. Fastidious, xylem limited plant pathogens. Annual Review of Phytopathol. 34:131-151. 7. Simpson, A. J. G. et al. 2000. The genome sequence of the plant pathogen Xylella fastidiosa. Nature 406:151-159. FUNDING AGENCIES Funding for this project was provided by the University of California Pierce’s Disease Grant Program. - 187 -
Project Leader: David Gilchrist Dept. of Plant Pathology University of California Davis, CA 95616 ISOLATION AND FUNCTIONAL TESTING OF PIERCE’S DISEASE-SPECIFIC PROMOTERS FROM GRAPE Cooperators: Douglas Cook Dept. of Plant Pathology University of California Davis, CA 95616 James Lincoln Dept. of Plant Pathology University of California Davis, CA 95616 Reporting Period: This two-year project was initiated on October 1, 2004. Obviously, there are few results to report at this time. Only a discussion of the justification, objectives, and timetable will be presented per request by the Pierce’s Disease Symposium organizers. ABSTRACT Among the potential solutions to Pierce’s disease in grapes are approaches based on gene transfer technology that focus on understanding the underlying biochemical and molecular mechanisms regulating PD. One of the research priorities identified by the 2003 PD/GWSS project reviews and as indicated in the 2004 RFP was the need to identify, clone and characterize unique DNA sequences that specifically regulate the expression of grape genes in tissues that are infected with Xf. Emphasis was placed on the urgency and practical utility of isolating promoters of PD responsive genes. One of the major bottlenecks in using transgenes, either expressed as proteins or as inhibiting RNAs in grape (or any plant) is the lack of suitable promoters to specifically drive the expression of a transgene on a specific trait (susceptibility to PD) in particular tissues (e.g., vascular tissue) or in response to particular situations (e.g., sharpshooter feeding or Xylella infection). In the absence of tissue or response-specific promoters, transgenic strategies to either understand or control PD one can use only so-called constitutive promoters. The basic problem associated with the use of constitutive promoters is that the transgene is expressed in all cells all the time, not just in the tissue or cells where the gene is needed. Highly controlled induction is needed if the interest is in altering gene expression to avoid a cellular change (disease) that is initiated in one or a few isolated cells. The isolation and characterization of Xf-responsive promoters has immediate and direct application to several current PD projects that are studying the biochemical or molecular genetic basis of PD at the cellular and tissue levels in grape. It also is of practical importance that these promoters will be useful in either the up- or down-regulation of the expression of a specific gene-of-interest. The difference in presence or absence of the target gene product is determined by whether the promoter is used to drive a sense or an anti-sense construct of the gene of interest. INTRODUCTION The objective of promoter analysis is to identify and characterize cis-acting DNA (adjacent) sequences that, when induced, regulate PD-associated gene expression in grapes. Although regulatory sequences frequently occur just upstream of the transcription start site, they can also be found much further upstream. Transcript abundance can also be controlled posttranscriptionally, often by cis-acting sequences in the 3' untranslated region of a gene. Thus, the challenge in our studies is to demonstrate that the cis-acting sequences have a unique functional role in PD symptom development. It is not the goal of this proposal to understand mechanisms of transcriptional regulation, but rather to isolate and confirm sequences that are active in the regulation of gene expression when Xf is present as an inducer of a select set of genes. To test whether a particular DNA sequence, that lies adjacent to a gene of interest, is involved in the regulation of that gene, it is necessary to introduce such putative regulatory sequences into a cell and then determine if they are activated when the inducer (in our case, Xf) is introduced into the system. This is done by combining a regulatory sequence with a reporter sequence (in our case, GFP) that can be used to monitor the effect of the regulatory (promoter) sequences in the presence of Xf. We have identified a set of plant genes whose expression is correlated with infection by Xylella fastidiosa as part of a recent study of expressed sequence tags from Xf-infected and healthy V. vinifera plants in the Napa Valley. The genes are essentially off (silent) in plants that have not been exposed to the pathogen, but strongly induced in both natural field infections and greenhouse inoculated plants. Three of these genes are induced early during disease development, prior to the occurrence of symptoms, while the fourth gene is induced in symptomatic tissues only. In addition to their utility for engineering PD resistance in grape, the advent of Xf-induced reporter gene expression would provide an extremely powerful tool to examine other host responses in their intact cellular and tissue context. With such tools, it should be possible to examine the chemical and/or physical cues from the insect or pathogen that trigger host gene - 188 -
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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
Page 77 and 78: MYCOPATHOGENS AND THEIR EXOTOXINS I
Page 79 and 80: POPULATION DYNAMICS AND INTERACTION
Page 81 and 82: No egg masses were recorded on olea
Page 83 and 84: EXPLORATION FOR FACULTATIVE ENDOSYM
Page 85 and 86: Although Baumannia and Wolbachia we
Page 87 and 88: EFFECTS OF SUBLETHAL DOSES OF IMIDA
Page 89 and 90: Table 2. Mortality and flight perfo
Page 91 and 92: A NOVEL METHOD TO INDUCE OVIPOSITIO
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
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: Figure 1: Southern blot of tolC::ap
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 and 178: % 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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