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

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2. Utilize Bulk Segregant Analysis (BSA) with the AFLP marker system to saturate with markers the region around the previously mapped Xf resistance locus and eventually convert confirmed candidate markers to stable SCAR primers. 3. Confirm candidate marker linkage to resistance within families derived from resistant by susceptible crosses such as the ‘8909’ x V. vinifera and (‘8909’ x V. vinifera) x V. vinifera back-cross generations. RESULTS AND CONCLUSIONS Sub-objective 1. Initial mapping of the PD resistance locus PdR1 in the male parent F8909-17 of the 9621 family localized it to chromosome 14, and identified 6-8 SSR markers on the same linkage group. Marker placement on published SSR linkage maps of Vitis were used to preferentially target chromosome 14, bringing the total number of SSR markers on the linkage group up to 30. Approximately 9 SSR markers are localized within a 10 cM distance of the resistance gene. These SSR markers are reliable and are the easiest of the molecular markers to incorporate within a MAS breeding program. Correlation tests of these candidate markers to PD resistance when functioning within a V. vinifera genetic background are underway and described in sub-objective 3. The SSR marker analysis has allowed us to confirm that marker alleles linked in coupling to PD resistance alleles of the PdR1 locus in another PD resistant progeny of b43-17 (F8909-08) are different than the alleles linked in coupling the resistance alleles in F8909-17. It is apparent from these results that b43-17 is homozygous resistant for the PdR1 locus, and that F8909-17 inherited its resistance allele from one chromosome 14 and F8909-08 inherited its resistance allele from the homologous chromosome 14. In either case the markers linked to resistance will function for MAS, however, different alleles linked in coupling to the resistance alleles will have to be followed through the downstream MAS process. Placement of SSR markers to chromosome 14 via the comparative mapping strategy continue as the markers become available, however, the number of SSR markers that can be targeted to a specific chromosomal region via comparative mapping is limited. Figure 1 Resistant Variety Poor Fruit Characters R Resistant Variety Mix of Fruit Characters R Breeding PD resistant grapes Resistant Variety Excellent Fruit Characters R Sub-objective 2. For high density marker saturation within a narrow window around the PdR1 locus, a bulk segregant analysis (BSA) strategy (Michelmore et al. 1991) in concert with the AFLP marker system was chosen as the method of choice. Initial BSA was attempted within the 9621 family, however, confounding effects of the resistance loci within the D8909-15 parent made the attempt more difficult than expected. To avoid confounding affects from resistance inherited from other genetic backgrounds and focus the BSA procedure only on the PdR1 locus, work has begun within two segregating families from susceptible by resistant crosses. The first family, 99217 (C8909-07 x F8909-08) consists of 33 genotypes, has been screened for PD resistance (Krivanek et al. submitted) and segregates 1:1 resistant to susceptible (Table 1). DNA has been extracted from these genotypes, flanking SSR markers were run and a good correlation between resistance and resistance marker alleles has been established (Table 1). A bulk of the DNA from the 12 most susceptible and a bulk of the DNA from the 12 most resistant genotypes are in process and will be tested for AFLP polymorphisms utilizing florescent primers and visualized on a PE 3100 sequencer. The second family derived from a susceptible by resistant cross is a V. vinifera x F8909-08 family; it consists of 40 genotypes and has been designated as 0062. Testing of this family for PD resistance is currently underway via our standard greenhouse testing procedure (Krivanek et al. in press; Krivanek and Walker in press). It is expected that the progeny in this family will segregate in a 1:1 manner, and if so, DNA extraction and BSA procedures will be undertaken as with the 99217 family. Candidate AFLP markers will be converted to stable and more reliable SCAR primers before incorporation into the MAS program. Sub-objective 3. Work is progressing with two distinct breeding populations for testing of candidate resistance markers and initial application of those markers to MAS. <strong>On</strong>e family is a cross of the PD resistant F8909-08 to a female V. vinifera wine grape F2-7 (Cabernet Sauvignon x Carignane) and designated as the 0062 family. A second breeding population consists of a cross of F8909-08 to several elite V. vinifera table grape genotypes (the 500 series). A subset of the 500 series has been screened for PD resistance and screened for markers flanking the PdR1 locus. Five confirmed resistant genotypes have been utilized in the development of the first backcross generations BC1 (backcrossed to additional elite V. vinifera genotypes). The BC1 population (25000 series) consists of approximately 200 individuals and was planted in the field in 2003. Marker analysis for flanking markers to the PdR1 locus has been completed for the 25000 series and the marker information was utilized in selection of genotypes for the spring of 2004 crosses for the development of the BC2 generations. Subsets of candidate - 65 - X Marker Assisted Breeding Susceptible Variety Excellent Fruit Characters
esistant and susceptible genotypes within the 25000 series have shown improved fruit quality (Figure 2) and are currently being screened to confirm the correlation between the resistance markers and the PD resistance trait. We are also utilizing these populations to confirm the effectiveness and economics of the MAS relative to our greenhouse screening procedure. Table 1. Resistance classification and marker genotypes for the individuals of the full-sib family derived from the susceptible by resistant cross of C8909-07 x F8909-08. * = Genotypes selected for Bulk Segregant Analysis procedure. Genotype Overall resistance level to PD Mean natural log (cells/ml) Mean CMI score Mean % leaf scorch Alleles of SSR markers flanking the PdR1 resistance 99217-21 * Resistant 9.51 1.00 58.3 Rr / Rr 99217-40 * Resistant 9.70 1.33 75.0 rr / Rr 99217-18 * Resistant 9.77 2.75 95.0 Rr / Rr 99217-41 * Resistant 10.19 4.25 76.3 Rr / Rr 99217-35 * Resistant 10.55 1.33 100.0 rr / Rr 99217-19 * Resistant 11.08 2.50 76.7 rr / Rr 99217-01 * Resistant 11.52 2.25 90.0 rr / Rr 99217-23 * Resistant 11.57 3.00 87.5 Rr / Rr 99217-34 * Resistant 11.83 3.75 65.0 Rr / Rr 99217-46 Resistant 11.87 5.75 100.0 Rr / Rr 99217-27 * Resistant 12.20 4.25 100.0 Rr / rr 99217-22 * Resistant 12.29 4.00 100.0 Rr / Rr 99217-12 * Resistant 12.50 4.00 95.0 Rr / Rr 99217-38 ? 12.69 5.00 100.0 Rr / Rr 99217-36 ? 13.09 5.00 100.0 rr / rr 99217-50 ? 13.52 4.25 83.8 Rr / Rr 99217-14 Susceptible 14.06 5.50 88.8 rr / Rr 99217-07 Susceptible 14.87 5.50 100.0 rr / rr 99217-04 * Susceptible 15.42 6.00 100.0 rr / rr 99217-33 * Susceptible 15.59 5.75 100.0 rr / rr 99217-06 * Susceptible 15.80 5.25 68.3 rr / rr 99217-09 * Susceptible 15.81 5.75 100.0 rr / rr 99217-10 Susceptible 15.82 4.75 100.0 rr / rr 99217-13 * Susceptible 15.84 5.50 100.0 rr / rr 99217-42 Susceptible 15.85 4.25 75.0 rr / Rr 99217-15 * Susceptible 15.87 5.25 100.0 rr / rr 99217-32 * Susceptible 15.87 5.50 100.0 rr / rr 99217-28 * Susceptible 15.91 5.75 100.0 rr / rr 99217-05 * Susceptible 15.91 5.75 100.0 rr / rr 99217-37 * Susceptible 15.92 5.25 100.0 rr / rr 99217-26 * Susceptible 15.95 5.50 100.0 rr / rr 99217-24 * Susceptible 16.04 6.00 100.0 rr / rr Figure 2. Vitis arizonica PD Resistant poor fruit quality Hybrid BC1-25017 with flanking PD resistance markers Improved fruit quality Vitis vinifera PD Susceptible Excellent fruit quality - 66 -
Page 1 and 2: IMPACT OF HOST PLANT XYLEM FLUID ON
Page 3 and 4: 0.18 600 Optical density 0.16 0.14
Page 5: OPTIMIZING MARKER-ASSISTED SELECTIO
Page 9 and 10: MAP BASED IDENTIFICATION AND POSITI
Page 11 and 12: esistant genotypes are in process a
Page 13 and 14: BREEDING PIERCE’S DISEASE RESISTA
Page 15 and 16: Progeny from crosses of field resis
Page 17 and 18: a = (1=low, 4= high); b = (1=green,
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Page 23 and 24: v.8) for differences due to the pla
Page 25 and 26: SHARPSHOOTER FEEDING BEHAVIOR IN RE
Page 27 and 28: correlated with all other findings
Page 29 and 30: EFFECTS OF FEEDING SUBSTRATE ON RET
Page 31 and 32: REFERENCES Bextine, B. and T. Mille
Page 33 and 34: will also provide insights into the
Page 35 and 36: OBJECTIVES 1. Determine glassy-wing
Page 37 and 38: GWSS per 30 s sweep (seasonal avera
Page 39 and 40: ELISA for the presence of GWSS egg
Page 41 and 42: Project Leader: Thomas P. Freeman E
Page 43 and 44: Freeman, T. P., R. A. Leopold, D. R
Page 45 and 46: pathogen, when they move into viney
Page 47 and 48: A NOVEL IMMUNOLOGICAL APPROACH FOR
Page 49 and 50: 1.6 GWSS Adults That Fed on Protein
Page 51 and 52: Hagler, J.R. & S.E. Naranjo. 2004.
Page 53 and 54: RESULTS: Oviposition Survey Wild gr
Page 55 and 56: 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.
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Figure 1: Southern blot of tolC::ap
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Project Leader: David Gilchrist Dep
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III. Production of transgenic plant
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RESULTS Construction of cDNA Librar
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CONCLUSIONS Genetic resistance and
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Mutagenesis of Xylella The EZ::TN T
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ISOLATION OF BACTERIOPHAGES SPECIFI
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Project Leader: Michele M. Igo Sect
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outer membrane fractions using two-
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1995; Purcell and Saunders, 1999).
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DEVELOPMENT OF SSR MARKERS FOR GENO
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Strain Name Host of Origin County o
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ROLE OF ATTACHMENT OF XYLELLA FASTI
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wild-type cells was not conclusive
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DETERMINATION OF GENES CONFERRING H
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S1 S2 S3 S4 Nb123456789bb123456789b
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MULTILOCUS SEQUENCE TYPING TO IDENT
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GENOME-WIDE IDENTIFICATION OF RAPID
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Americas and since most of the plan
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EFFECTS OF CHEMICAL MILIEU ON ATTAC
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CONCLUSIONS Our overall objective i
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RESULTS Objective 1. We conducted t
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Redak, R. A., Purcell, A. H., Lopes
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In parallel, an “activating trans
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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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- 246 -
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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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- 278 -
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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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