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

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REFERENCES Collins FH, Paskewitz SM. 1996. A review of the use of ribosomal DNA (rDNA) to differentiate among cryptic Anophels species. Insect Molecular Biology 5: 1-9. de León JH, Jones WA. 2004. Detection of DNA polymorphisms in Homalodisca coagulata (Homoptera: Cicadellidae) by polymerase chain reaction-based DNA fingerprinting methods. Annals of the Entomological Society of America 97: 574- 585. de León JH, Jones WA, Morgan DJW. 2004. Population genetic structure of Homalodisca coagulata (Homoptera: Cicadellidae), the vector of the bacterium Xylella fastidiosa causing Pierce’s disease in grapevines. Annals of the Entomological Society of America 97: 809-818. Hopkins DL, Mollenhauer HH. 1973. Rickettsia-like bacterium associated with Pierce’s disease of grapes. Science 179: 298-300. Hoy MA, Ayyamperumal J, Morakete R, Lo MKC, Nguyen R. 2000. Genomic analyses of two populations of Ageniaspis citricola (Hymenoptera: Encyrtidea) suggest that a cryptic species may exist. Biological Control 17: 1-10. Huber JT. 1998. The species groups of Gonatocerus Nees in North America with a revision of the sulphuripes an ater groups (Hymenoptera: Mymaridae). Memoirs of the Entomological Society of Canada 141: 1-109. Löhr BA, Varela M, Santos B. 1990. Exploration for natural enemies of the cassava mealybug, Phenococcus manihoti (Homoptera: Pseudococcidae), in South America for the biological control of this introduced pest in Africa. Bulletin of Entomological Research 80: 417-425. Messing RH, Aliniazee MT. 1988. Hybridization and host suitability of two biotypes of Trioxys pallidus (Hymenoptera: Aphidiidae). Annuals of the Entomological Society of America 81: 6-9. Narang SK, Leopold RA, Krueger CM, DeVault JD. 1993. Dichomotomus RAPD-PCR key for identification of four species of parasitic hymenoptera. In: Narang SK, Barlett AC, Faust RM, editors. Applications of Genetics to Arthropods of Biological Control Significance, 53-67. CRC Press Inc, Boca Raton, Florida. Nei M. 1978. Estimation of average heterozygosity and genetic distance from a small number of individuals. Genetics 89: 583-590. Powell W, Walton MP. 1989. The use of electrophoresis in the study of hymenopteran parasitoids of agricultural pest. In: Loxdale HD, den Hollander J, editors. Electrophoretic Studies on Agricultural Pests, 443-65. Oxford, Clarendon. Reynolds J, Weir BS, Cockerham CC. 1983. Estimation of the coancestry coeffient: basis for a short-term genetic distance. Genetics 105: 767-779. Stouthamer R, Hu J, Van Kan FJPM, Platner GR, Pinto JD. 1999. The utility in internally transcribed spacer 2 DNA sequences of the nuclear ribosomal gene for distinguishing sibling species of Trichogramma. BioControl 43: 421-440. Triapitsyn SV, Mizell RF III, Bossart JL, Carlton CE. 1998. Egg parasitoids of Homalodisca coagulata (Homoptera: Cicadellidae). Florida Entomologist 81: 241-243. Turner WF, Pollard HN. 1959. Life histories and behavior of five insect vectors of phony peach disease. USDA Technical Bulletin 1188: 28 pp. Unruh TR, Woolley JB. 1999. Molecular Methods in Classical Biological Control. In: Van Driesche RG, Bellows TS, Jr., editors. Biological Control, 57-85. Chapman and Hall, NY. Zietkiewicz E, Rafalski A, Labuda D. 1994. Genomic fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics 20:176-183. FUNDING AGENCIES Funding for this project was provided by the USDA Agricultural Research Service. - 321 -
SEQUENCE DIVERGENCE IN TWO MITOCHONDRIAL GENES (COI AND COII) AND IN THE ITS2 RDNA FRAGMENT IN GEOGRAPHIC POPULATIONS OF GONATOCERUS MORRILLI, A PRIMARY EGG PARASITIOD OF THE GLASSY-WINGED SHARPSHOOTER Project Leader: Jesse H. de León USDA, ARS BIRU Weslaco, TX 78596 Cooperators: Walker A. Jones USDA, ARS BIRU Weslaco, TX 78596 David J. W. Morgan CDFA Mount Rubidoux Field Station Riverside, CA 92501 Russell F. Mizell, III University of Florida Quincy, FL 32351 Reporting period: The results reported here are from work conducted from fiscal year 2003 to fiscal year 2004. ABSTRACT The aim of the present study was to resolve the genetic relationships of geographic populations of Gonatocerus morrilli, a primary egg parasitoid of the Glassy-winged Sharpshooter. A phylogenetic approach was implemented by sequencing two mitochondrial genes (COI and COII) and the Internal Transcribed Spacer-2 (ITS2) region of several individuals per population. Two populations from Weslaco, TX (WTX) (collected at different times), one from Quincy, FL (QFL), two from California (CA) (Orange and San Diego counties), and an outgroup (G. ashmeadi) were analyzed. For all three sequence fragments, percentage sequence divergence (%D) (as measured by genetic distance), the results demonstrated that both the WTX and QFL populations were closely related; in constrast, the %D between WTX and CA fell within the range of the outgroup, G. ashmeadi. For all three sequence fragments, Nieghbor-Joining distance trees separated the CA and WTX and QFL populations into two distinctive clades (A and B). The topology of the clades in each case was supported by very strong bootstrap values, 100% in the three sequence fragments (COI, COII, and ITS2). The present molecular phylogenetics results provide strong evidence that G. morrilli from California may be a different species. The findings of the present study are important to the Glassy-winged Sharpshooter/Pierce’s disease biological control program in California. INTRODUCTION Gonatocerus morrilli (Howard) (Hymenoptera: Mymaridae) is an egg parasitoid of Homalodisca coagulata (Say) (Homoptera: Cicadellidae), the Glassy-winged Sharpshooter (Turner and Pollard 1959; Triapitsyn et al. 1998). This primary egg parasitoid species is common in the southern United States and Mexico (Huber 1988). A biological control program is currently in progress in California against H. coagulata, a xylem feeding leafhopper, which is a serious economic pest that transmits a strain of Xylella fastidiosa (Wells), a bacterium that causes Pierce’s disease in grapevines (Vitis vinifera L. and V. labrusca L.) (Hopkins and Mollenhauer 1973). Accurate identification of natural enemies is critical to the success of classical biological control programs. Lack of proper identification procedures has affected the early stages of several projects (Messing and Aliniazee 1988; Löhr et al. 1990). OBJECTIVES Determine the phylogenetic relationships of geographic populations of G. morrilli by sequencing two mitochondrial genes (COI and COII) and one rDNA spacer region (ITS2). RESULTS AND CONCLUSIONS Sequence divergence in the mitochondrial COI gene in G. morrilli geographic populations. Levels of genetic divergence in the mtCOI gene among populations were determined by calculating the pairwise estimates for genetic distance. Recently, we determined that populations of G. morrilli from California and Texas shared no ISSR-PCR banding patterns, indicating that these populations were reproductively isolated. In addition, we demonstrated that the ITS2 rDNA fragments varied in size between these geographic populations (de León et al. 2004). The percentage sequence divergence (%D) for mtCOI is shown on Table 1. In general, the intra-populational variation (0.0-0.6%) was small within each population and species, with the exception of the Quincy, FL population (QFL) (2.0-4.8%). The %D between Weslaco, TX (WTX) and QFL is 0.0-4.8%, which falls within the intra-populational variation of these populations; these results indicate that these geographic populations are genetically similar. In constrast, the %D of WTX and CA is 5.4-5.6%, falling within the range (5.4-6.9%) of the outgroup (G. ashmeadi). The Nieghbor-Joining distance tree in Fig. 1 demonstrates that the CA and WTX and QFL populations cluster into two distinctive clades (A and B). These clades are supported by very strong bootstrap values (100%). Sequence divergence in the mitochondrial COII gene in G. morrilli geographic populations. The percentage sequence divergence (%D) for mtCOII is shown on Table 2. Intra-populational variation is seen in both the WTX (0.0-4.5%) and QFL (0.0-3.2%) populations. The %D between WTX and QFL is 0.3-4.7%, these values fall within the intra-populational variation range and therefore these populations would be considered closely related. On the other hand, the %D between WTX and CA is 7.4-11.1%, these values fall within the range (7.4-11.5%) of the outgroup (G. ashmeadi). The Nieghbor-Joining distance tree in Fig. 2 demonstrates that the CA and WTX and QFL populations cluster into two distinctive clades (A and B). These clades are supported by very strong bootstrap values (100%). - 322 -
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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
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Figure 2. Individual leaves from a
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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
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Page 235 and 236: Project Leader: Doug Cook Dept. of
Page 237 and 238: Based on the in silico analysis, de
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Page 243 and 244: RESULTS During the reporting period
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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
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Page 295 and 296: concentrations of 1.5 x 10 7 bacter
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Page 299 and 300: SYMBIOTIC CONTROL OF PIERCE’S DIS
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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,
Page 311 and 312: 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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