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

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ISSR-PCR Differentiation Among Four G. morrilli Populations. Exact tests (simultaneous analysis) for population differentiation indicated that highly significant differences in marker frequencies existed among the G. morrilli populations (Table 1). Total genetic diversity (Ht) was high (35%), whereas the average genetic diversity within populations was low (3%). Table 1 also shows a comparison of other genetic differentiation estimates, G ST and θ, which evaluate the degree of genetic subdivision among populations. Excellent agreement was seen between G ST and θ values, 0.92 and 0.94, respectively. Theses values indicate that about 92 to 94% of the variance is distributed among populations. The indirect estimate of gene flow, Nm base on G ST , demonstrated a low value (0.04) among the geographic populations; this value indicates highly restrictive gene flow. Overall, genetic differentiation measurements (exact tests, G ST , θ, and Nm) indicate profound genetic divergence/structuring between G. morrilli populations from Texas and California. Genetic Relatedness Among G. morrilli Populations. Levels of genetic divergence among populations were also determined by calculating pairwise estimates for genetic distance by the procedures of Nei (1978) and Reynolds et al. (1983) (Table 2). Average genetic divergence (D) among populations was extremely high [Nei = 0.82 (0.89-1.07) and Reynolds = 2.79 (1.4-3.4)]. A dendrogram based on Nei’s genetic distance is shown on Fig. 2 with all G. morrilli geographic populations. Two clades are identified on the dendrogram with the California and Texas populations appearing on separate clusters. These two clusters are supported by strong bootstrap support values, 68 and 64%, respectively for the California and Texas populations. Amplification of the ITS-1 and –2 regions in G. morrilli Geographic Populations. Monomorphic patterns were demonstrated with amplification of the ITS-1 region in all of the populations from California and Texas (~850 bp) (Fig. 3); whereas, polymorphic or different DNA fragment sizes were detected within the ITS-2 region. The California populations were observed with an ITS-2 fragment size of about 865 base pairs and the Texas populations with a size of about 1099 base pairs. Good agreement is seen between the two molecular methods and they both suggest that cryptic species may exist. The results with ISSR-PCR demonstrating distinct banding patterns (no band sharing) between geographic populations typically is not found unless the populations are reproductively isolated. Similar results were obtained by Hoy et al. (2000) with two populations of Ageniaspis citrocola performed by RAPD-PCR. The following genetic differentiation parameters, extract test, G ST , θ, genetic distances, and gene flow (Nm) lend support to this observation. The extremely low value for gene flow between the populations from California and Texas lend support that these populations are isolated reproductively. Restricted gene flow usually leads to increased differentiation among populations as seen from the G ST and θ values (92 to 94% of the variance is seen among populations). In addition, the divergence (D) between these populations is also high. Methods incorporating SSR appear to be sensitive at detecting DNA polymorphisms in natural populations. Previously, we utilized ISSR-PCR to distinguish three species of Homalodisca sharpshooters (H. coagulata, H. liturata, and H. insolita) (de León and Jones 1994). Even though this method is sensitive, there are not many reports in the literature utilizing ISSR-PCR to study insect population genetics and phylogenetics. We have also had success determining the population genetic structure of H. coagulata representing 19 populations from through the U. S. (de León et al. 2004). The Internal Transcribed Spacer regions (ITS-1 and –2) have been used extensively to examine the taxonomic status of species and for diagnostic purposes, and success with this approach has been reviewed by Collins and Paskewitz (1996). Stouthamer et al. (1999) used ITS-2 DNA fragment sizes as taxonomic characters to develop a precise identification key for sibling species of the genus Trichogramma. In cases where species were observed with similar sized ITS fragments these authors suggested amplification, sequencing, and restriction digestion. Figure 1. Representative example of ISSR-PCR DNA fingerprinting of G. morrilli populations from California and Texas. Reactions were performed with genomic DNA from separate individuals and the 5’-anchored ISSR primer HVH(TG) 7 T (Zietkiewicz et al. 1994) as describe in the Materials and Methods. M: 1.0 Kb Plus DNA Ladder. - 319 -
These novel observations strongly suggest that G. morrilli may exist in nature as a species-complex. Results from our recent study with H. coagulata suggest that a subset of these insects have their origin in Texas (de León et al. 2004). Those results together with our present results with G. morrilli may suggest that this egg parasitoid from Texas may be a good candidate for the biological control efforts in California against H. coagulata, the causative agent of Pierce’s disease. Table 1. Nei’s analysis of gene diversity in populations of G. morrilli from Texas and California. Ten individuals per population (40 total) were subjected to ISSR-PCR DNA fingerprinting. Genetic variation was analyzed using the POPGENE 3.2 genetic software program and the program Tools for Population Genetic Analyses (TPFGA). X 2 , exact tests (simultaneous analysis) for population differentiation, df = degrees of freedom; Ht, total genetic diversity (SD), Hs, average genetic diversity within populations (SD); G ST (mean), coefficient of gene differentiation; θ theta (analogous to F ST ), and Nm, gene flow. *** P = 0000. ____________________________________________________ X 2 (df) Ht Hs G ST θ Nm __________________________________________________________ 400.8 (50) *** 0.35 0.03 0.92 0.94 0.04 (0.04) (0.00) (0.02) __________________________________________________________ Table 2. Nei’s unbiased (1978) genetic distance (below diagonal) and Reynolds et al. (1983) genetic distance (above) diagonal. Four geographic populations of G. morrilli, two from Texas (Hidalgo Co, Wes-2 and Wes-3) and two from California (OrCo, Orange county and SDCo, San Diego county). ______________________________________ Pop OrCo SDCo Wes-2 Wes-3 __________________________________________ OrCo *** undef 3.40 2.88 SDCo 0.00 *** 3.40 2.88 Wes-2 1.07 1.07 *** 1.40 Wes-3 0.89 0.89 0.20 *** _________________________________________ Figure 2. Dendrogram based on Nei’s genetic distance (1978) by the method of UPGMA. Relationships among the four geographic populations of G. morrilli performed by ISSR-PCR DNA fingerprinting. Genetic distances are indicated above the dendrograms and bootstrap support values are indicated at the nodes. Figure 3. Amplification of the Internal Transcribed Spacer regions (ITS). The ITS-1 and –2 regions were amplified with standard ITS-specific primers with genomic DNA from five separate individuals from each geographic population. Arrows indicate different ITS fragment sizes. M: 1.0 Kb Plus DNA Ladder. - 320 -
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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.
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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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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
Page 209 and 210: Figure 2. Individual leaves from a
Page 211 and 212: The state variables, process functi
Page 213 and 214: DEVELOPMENT OF A FIELD SAMPLING PLA
Page 215 and 216: Figure 1. Three main dispersion pat
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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
Page 229 and 230: Figure 2. Polymer disk accumulation
Page 231 and 232: CHARACTERIZATION OF NEONICOTINOIDS
Page 233 and 234: EVALUATION OF RESISTANCE POTENTIAL
Page 235 and 236: Project Leader: Doug Cook Dept. of
Page 237 and 238: Based on the in silico analysis, de
Page 239 and 240: CONTROL OF PIERCE’S DISEASE THROU
Page 241 and 242: Figure 1. Oleander ‘White’ afte
Page 243 and 244: RESULTS During the reporting period
Page 245 and 246: DEVELOPMENT OF AN ARTIFICIAL DIET A
Page 247 and 248: DESIGN OF CHIMERIC ANTI-MICROBIAL P
Page 249 and 250: Neutrophil elastase Cecropin B Chim
Page 251 and 252: and WTXb is very low (0.00-0.40%) a
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Page 255 and 256: GENETIC DIFFERENTIATION AMONG GEOGR
Page 257 and 258: Table 1. Single-populations descrip
Page 259: MOLECULAR DISTINCTION BETWEEN POPUL
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Page 265 and 266: Table 3. Pairwise sequence distance
Page 267 and 268: DEVELOPMENT OF MOLECULAR DIAGNOSTIC
Page 269 and 270: A. B. Relative Density of SCAR Band
Page 271 and 272: THE ALIMENTARY TRACK OF GLASSY-WING
Page 273 and 274: We have dissected and identified al
Page 275 and 276: REALIZED LIFETIME PARASITISM AND TH
Page 277 and 278: REPRODUCTIVE AND DEVELOPMENTAL BIOL
Page 279 and 280: Mean adult longevity (days) 25 20 1
Page 281 and 282: the proposed exploratory work; thes
Page 283 and 284: SEARCHING FOR AND COLLECTING EGG PA
Page 285 and 286: REFERENCES Hoddle, M. S. and S. V.
Page 287 and 288: some Gram(+) bacteria, but are inac
Page 289 and 290: 7. Hultmark, D., et al., Insect Imm
Page 291 and 292: Isolation of Fungal Pathogens Soil
Page 293 and 294: IDENTIFICATION OF MECHANISMS MEDIAT
Page 295 and 296: concentrations of 1.5 x 10 7 bacter
Page 297 and 298: anchor it in the outer membrane (sh
Page 299 and 300: SYMBIOTIC CONTROL OF PIERCE’S DIS
Page 301 and 302: MANAGEMENT OF PIERCE’S DISEASE OF
Page 303 and 304: pfF mutants are taken up by insects
Page 305 and 306: Simpson, A. J. G., F. C. Reinach, P
Page 307 and 308: Al-Wahaibi, A.K., Owen, and J. G. M
Page 309 and 310: 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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