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

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Thompson JD, Higgins DG, Gibson TJ. 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific gap penalties and weight matrix choice. Nucleic Acids Research 22:4673-4680. Swofford DL. 2002. PAUP*. Phylogenetic Analysis Using Parsimony (*and Other Methods). Version 4. Sinauer Associates, Sunderland, Massachusetts. Vickerman DB, Hoddle MS, Triapitsyn S, Stouthamer R. 2004. Species identity of geographically distinct populations of the glassy-winged sharpshooter parasitoid Gonatocerus ashmeadi: morphology, DNA sequences, and reproductive compatibility. Biological Control 31: 338-345. 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. Wu Z, Hopper KR, O’Neil RJ, Voegtlin DJ, Prokrym DR, Heimpel GE. 2004. Reproductive compatibility and genetic variation between two strains of Aphelinus albipodus (Hymenoptera: Aphelinidae), a parasitoid of the soybean aphid, Aphis glycines (Homoptera: Aphididae). Biological Control 31: 331-319. FUNDING AGENCIES Funding for this project was provided by the USDA Agricultural Research Service. - 313 -
GENETIC DIFFERENTIATION AMONG GEOGRAPHIC POPULATIONS OF GONATOCERUS ASHMEADI, A PRIMARY EGG PARASITOID 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 genetically comparing different populations of the same species of natural enemies is to identify the strain that is most adapted to the environment where it will be released. In the present study, Inter-Simple Sequence Repeat-Polymerase Chain Reaction (ISSR-PCR) was utilized to estimate the population genetic structure of Gonatocerus ashmeadi. Six populations from throughout the U. S. and a population from Argentina identified as near G. ashmeadi were analyzed. Four populations [California (CA), San Antonio, TX (SATX), Weslaco, TX (WTX-2), and Quincy, Florida (QFL)] were field collected and two [Louisiana (LA) and Weslaco, TX (WTX-1)] were reared. Three ISSR-PCR reactions were pooled to generate 41 polymorphic markers among the six U. S. populations. Nei’s expected heterozygosity values (h), including the reared population from Louisiana were high (9.0-14.3%) for all populations, except for a reared population from WTX-1 (2.9%). The total genetic diversity value (Ht) for the field populations was high (23%). Interestingly, the Florida population that was collected from one egg mass generated the greatest number of polymorphic markers (20) and was observed with the highest gene diversity value (14.3%). All populations, except WTX-2 generated population-specific markers. Comparison of genetic differentiation estimates, which evaluate the degree of genetic subdivision, demonstrated good agreement between G ST and θ values, 0.38 and 0.50, respectively for field populations, and 0.44 and 0.50, respectively for all populations. Average genetic divergence (D) indicated that the WTX-1 population was the most differentiated. Average D results from the Argentina population support the taxonomic data that it is a different species. The present results estimate the population genetic structure of G. ashmeadi, demonstrating extensive genetic divergence and restricted gene flow (Nm = 0.83) among populations. These results are of interest to the Pierce’s Disease/Glassy-winged Sharpshooter biological control program because the key to successful biological control may not be in another species, but instead in different geographic races or biotypes. INTRODUCTION Gonatocerus ashmeadi (Girault) (Hymenoptera: Mymaridae) is a primary egg parasitoid of Homalodisca coagulata (Say) (Homoptera: Cicadellidae), the glassy-winged sharpshooter (Huber 1998). A biological control program is currently in progress in California against H. coagulata because this xylem feeding sharpshooter is a serious economic pest that vectors a strain of <strong>Xylella</strong> fastidiosa (Wells), a bacterium that causes Pierce’s Disease in grapevines. Studies of allele or marker frequencies in naturally occurring parasitoid populations are important, not only for identifying genetic variation of potential benefit in the selection and screening of biological control organisms, but also for the detection of genetic markers indicative of specific biological traits or geographic origins. In addition, the recognition of intraspecific variation can be as crucial for the success of biological control programs as is sound species determination. Populations of parasitoids from distinct geographical regions may differ in relevant biological characteristics of importance to biological control (Powell and Walton 1989; Narang et al. 1993; Unruh and Woolley 1999). An aim of genetically comparing different populations of the same species of natural enemies is to identify the strain that is most adapted to the environment where it will be released (Messenger and van den Bosch 1971); in other words, the key to successful biological control may not be in another species, but instead in different geographic races or biotypes (Diehl and Bush 1984). Reliable methods are needed for distinguishing various exotic strains of these biological control agents from those indigenous to the U. S., including parasitoids from different states within the U. S. Release of unidentified and uncharacterized strains can make it difficult to document their establishment and dispersal. Therefore, genetic typing of strains prior to their release in the field is highly desirable (Narang et al. 1993). OBJECTIVES 1. Estimate genetic variation or gene diversity within and among populations. 2. Estimate the population genetic structure. 3. Determine whether ISSR-PCR was sensitive enough to identify diagnostic markers in geographic populations. 4. Confirm the species identification of a population of egg parasitoids from Argentina identified as near G. ashmeadi. RESULTS AND CONCLUSIONS ISSR-PCR Marker Heterozygosity and Genetic Diversity A total of 41 polymorphic markers were generated in the six populations of G. ashmeadi (163 individuals) from the U. S. with three pooled ISSR-PCR reactions. G 2 -contingency tests indicated significant heterogeneity of marker frequency across all U. S. populations for 31 of 41 markers and for 25 of 34 markers for the field populations (not shown). All populations, - 314 -
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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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Page 205 and 206: SPATIAL DATABASE CREATION AND MAINT
Page 207 and 208: Project Leader: Thomas M. Perring D
Page 209 and 210: Figure 2. Individual leaves from a
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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 257 and 258: Table 1. Single-populations descrip
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Page 261 and 262: These novel observations strongly s
Page 263 and 264: SEQUENCE DIVERGENCE IN TWO MITOCHON
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
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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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