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

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62.6 ± 9.1% (n = 98) and 44.6 ± 9.1% (n = 104), respectively. There was a significant difference in percentage hatch of 1- d-old GWSS eggs between the control eggs (83.0 ± 7.4%, n = 317) and those eggs stored for 25d (F = 3.939, df = 3,45, P = 0.014), but no significant differences were found between those groups stored in the cold for 15 or 20 days and the control. After storage for 80d under the daily cycled regime, no hatching was observed. To determine effect of cold storage during GWSS egg stage on nymphal development and adult reproduction, newly hatched nymphs from eggs stored at 12 ºC for 20 days, and at the daily cycled temperature regime for 15 and 20 days were reared on sunflower plants until they emerged as adults. When the characteristic patch of brochosomes was observed on the forewings of the adult females (brochosomes were considered as the sign that females had mated), they were then individually maintained on sunflower plants and their egg mass output recorded until death occurred. Our preliminary data (Table 1) shows that 50% of nymphs from eggs stored at 12ºC for 20 d and 50% and 40% of nymphs held under the stepwise temperature regime for 15 days and 20 days, respectively, successfully developed into adults. In comparison with the control groups, GWSS males and females from those eggs that had been exposed to either cold storage regime took significantly longer to complete their nymphal stages (Table 1). There were no differences in male and female developmental times among the nymphs that hatched from GWSS eggs that had undergone cold storage. The number of eggs produced/female and the ovipositional period was considerably greater for the control groups and approached 2-fold differences. Effects of Cold Storage of GWSS Eggs on Parasitism and Emergence by G. ashmeadi Following storage in incubators set at a constant 12 or 12.5ºC and also at the stepwise cycled regime as described above, GWSS egg masses were exposed to caged G. ashmeadi colonies for 2 days at room temperature (ca. 22 ºC) and under an 10L:14D photoperiod. Before statistical analysis, the data recorded for parasitism and emergence were square-root transformed to correct non-normality because the number of eggs/mass was not constant. After storage at 12ºC for 30d, 69.6 ± 11.7% of the 3-d-old GWSS eggs (n = 90) and 47.7 ± 11.7% (n = 106) of the 1-d-old eggs were successfully parasitized by G. ashmeadi. The percentage wasp emergence was 68.5 ± 11.3 for 3 day-old eggs and 35.3 ± 10.0% for the 1 day-old eggs. There were no significant differences in the incidence of parasitism, as determined by egg dissection, (F = 4.034, df = 1,14, P = 0.066) and emergence (F = 1.728, df = 1,14, P = 0.211). Further, G. ashmeadi successfully parasitized about 77% of the 4-d-old , 52% of the 5-d-old, and 45% of the 3-d-old GWSS eggs stored at 12.5ºC for 30d, and 46% of 3-d-old eggs stored for 50d. As above, there were no significant differences between parasitism and emergence in any of the comparable groups (data not shown). When stored under the cycled stepwise temperature regime (10-13 ºC), the parasitism (F = 14.934, df = 8,137, P < 0.001) and emergence (F = 13.661, df = 8,137, P < 0.001) of 1-d-old GWSS eggs by G. ashmeadi varied significantly with storage time (Table 2). More than 75% of GWSS eggs stored up to 25d were successfully parasitized and there was no significant difference in the incidence of parasitism between the control (92.1 ± 9.9%, n = 172) and the eggs stored for 15, 20 or 25d (F = 1.764, df = 3,35, P = 0.172). However, percentage emergence for the eggs stored for 25d was significantly lower than that for the control (91.7 ± 2.7%, n =172) (F = 3.250, df = 3,35, P < 0.033). Further, there were no significant differences in percentage emergence between the control eggs and the eggs stored for 15 or 20d (P = 0.099). After storage for 65d, < 44% eggs were parasitized by G. ashmeadi, and about 23% of wasps emerged, which was significantly lower than for eggs for stored for 25d or less. When stored for over 80d, the percentage parasitism and emergence were less than 12% and 7%, respectively. When these data were analyzed via a regression analysis, the percentage parasitism and emergence vs. storage time was found to be inversely correlated (Figures 1 and 2). Cold Storage of GWSS Eggs Parasitized by G. ashmeadi The experimental conditions for this study consisted of a constant 4 or 4.5ºC storage temperature and 2 daily cycled stepwise-increasing regimes (4, 6, and 8ºC or 4.5, 6, and 7.5ºC - each temp. changing at 8h intervals) under an 8 L: 16 D photoperiod. After the parasitized eggs were stored at 4 ºC for 10d, only 7.2 ± 5.0% (n = 85) of the wasps emerged, which was significantly lower than those parasitoids similarly stored at 4.5ºC (33.5 ± 7.2%, n = 280), 20 days (33.9 ± 6.9%, n = 114) or 25 days (21.7 ± 5.2%, n = 125) (F = 11.962, df = 4,66, P < 0.001). No parasitoids (n = 164) emerged from host eggs stored at 4ºC for 20d (Figure 3). When parasitoids were stored within hosts under the cycled stepwise temperature regime starting at 4 ºC, percentage emergence was 42% (n =126) at 10 d, 8 % (n = 420) at 20d and 0% (n = 184) at 25d (Figure 4). However, for parasitized eggs stored at the other cycled regime starting at 4.5ºC, the wasp emergence was at or above 60% throughout the 25d of storage. Thus, the percentage emergence for the parasitoids stored under the stepwise regime starting at 4.5ºC for 10-25d was significantly higher than that for the eggs stored for 15d under the regime starting at 4ºC (F = 48.237, df = 5, 114, P < 0.0001). Parasitoids within GWSS eggs did not emerge after storage for 80 days, but further research is needed to ascertain if maintenance of the Euonymus cuttings that bear the egg masses during the storage period is causing a problem. - 125 -
REFERENCES Gautam, R. D. 1986. Effect of cold storage on the adult parasitoid Telenomus remus Nixon (Scelionidae: Hymenoptera) and the parasitized eggs of Spodoptera litura (Fabr.) (Noctuidae: Lepidopetra). J. Entomol. Res. 10: 125-31 Iacob, M. and N. Iacob. 1972. Influence of temperature variation on the resistance of Trichogramma evanescens Westw to storage with a view to field release. Anal. Inst.de Cercetari pentru Protctia Plantelor 8: 191-199. Leopold, R. A. 1998. Cold storage of insects for integrated pest management. In: Temperature sensitivity in insects and application in integrated pest management. G. J. Hallman and D. L. Denlinger (eds.). Westview Press, Boulder. pp. 235- 267. Leopold, R.A., R.R. Rojas, & P.W. Atkinson. 1998. Post pupariation cold storage of three species of flies: increasing chilling tolerance by acclimation and recurrent recovery periods. Cryobiology 36: 213-224. Leopold, R. A, W. Chen , D. J. W. Morgan & G. D. Yocum. 2003. Cold storage of parasitized and unparasitized eggs of the glassy-winged sharpshooter, Homalodisca coagulata. pp 221-224. In M. Athar Tariq, S. Oswalt, P. Blincoe & T. Esser (eds). Proc. of Pierce’s Disease Research Symposium, Dec. 8-11, San Diego Liu, J.J. & Y. Tian. 1987. Cold storage of Encarsia Formosa Gahn. Chin. J. Biol. Cont. 36:4-6. FUNDING AGENCIES Funding for this project was provided by the USDA Animal and Plant Health Inspection Service and the USDA Agricultural Research Service. % Parasitism 110 100 90 80 70 60 50 40 30 20 10 0 0 20 40 60 80 100 120 140 160 Storage time (d) % Emergence 110 100 90 80 70 60 50 40 30 20 10 0 0 20 40 60 80 100 120 140 160 Storage time (d) Figure 1. Relationship of the % parasitism (y) of G. ashmeadi to storage time (x) of the GWSS eggs at stepwise temperatures (10~13ºC)(y = 5.10 + 1393.18/x, r = 0.58) (F =68.24, df =136, P < 0.001) Figure 2. Relationship of the % emergence (y) of G. ashmeadi to storage time (x) of the GWSS eggs at stepwise temperatures (10~13ºC) (y = -0.35 + 1286.50/x, r = 0.59) (F = 79.01, df = 136, P < 0.001). Table 1. Egg hatch, development time of nymphs and reproduction of adults for GWSS eggs stored under different temperature conditions (mean ± SE). Storage conditions Egg hatch Development time of nymphal stage Adult reproduction (%) % survival Male (d) Female (d) No. eggs/ female Ovipositional period (d) Control (25ºC) 82.9 ± 7.4 80.2 35.9 ± 0.5 a 35.3 ± 0.6 a 1068.8 ± 187.7 113.4 ± 49.6 12ºC for 20 d 52.7 ±10.2 50.0* 43.9 ± 0.9 b 42.5 ± 0.7 b 589.3 ± 81.9 65.7 ± 26.0 10-13ºC for 15 d 73.9 ± 5.4 50.0* 43.0 ± 0.7 b 41.0 ± 1.2 b 662.7 ± 111.1 65.0 ± 11.2 10-13ºC for 20 d 62.6 ±10.3 40.0* 43.0 ± 3.5 b 41.9 ± 0.4 b In progress In progress Only 1 replicate. Means within a column followed by different letters were significantly different at the significant level of 0.05 (SAS Proc GLM with LSD). Data for egg hatch were square-root transformed before analysis. - 126 -
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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
Page 15 and 16: Progeny from crosses of field resis
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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
Page 57 and 58: currently employed morphological ch
Page 59 and 60: increase our present understanding
Page 61 and 62: BIOLOGY AND MORPHOMETRIC ANALYSIS O
Page 63 and 64: Table 1. Mean a developmental durat
Page 65: EFFECTS OF USING CONSTANT AND CYCLI
Page 69 and 70: PARASITISM OF THE GLASSY-WINGED SHA
Page 71 and 72: Table 1. Parasitism by G.ashmeadi o
Page 73 and 74: Project Leader: Robert F. Luck Dept
Page 75 and 76: 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
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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
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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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