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Research Project: IPM TECHNOLOGIES FOR INSECT PESTS OF ORCHARD CROPS

Location: Subtropical Insects and Horticulture Research

Title: A new excised-leaf assay method to test the inoculativity of the Asian citrus psyllid with Candidatus Liberibacter asiaticus associated with citrus huanglongbing disease

Authors
item Ammar, Eldesouky
item Walter, Abigail -
item Hall, David

Submitted to: Journal of Economic Entomology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: November 9, 2012
Publication Date: February 12, 2013
Citation: Ammar, E., Walter, A., Hall, D. 2013. A new excised-leaf assay method to test the inoculativity of the Asian citrus psyllid with Candidatus Liberibacter asiaticus associated with citrus huanglongbing disease. Journal of Economic Entomology. 106:25-35.

Interpretive Summary: The Asian citrus psyllid (ACP) is the primary vector of the bacterium associated with huanglongbing, or citrus greening, the most devastating citrus disease worldwide. However, normally, only a low percentage of ACP can actually inoculate this bacterium into healthy plants after acquiring it from diseased citrus. Thus, to understand the epidemiology of this disease, it is important to assess the inoculativity of psyllids in any given population. Here, we developed a new ‘excised-leaf assay’ that can speed up inoculativity tests on ACP from the current 3-12 months (when using whole citrus seedlings for inoculation) to only 2-3 weeks. This new excised-leaf assay method saves considerable time, material and greenhouse space, and may hopefully enhance vector-relation and epidemiological studies on this and probably other bacterial species associated with huanglongbing disease.

Technical Abstract: The Asian citrus psyllid (ACP, Diaphorina citri, Hemiptera: Psyllidae) is the primary vector of Candidatus Liberibacter asiaticus (Las) associated with huanglongbing, or citrus greening, the most devastating citrus disease worldwide. Here, we developed a new excised-leaf assay that can speed up Las-inoculativity tests on ACP from the current 3-12 months (when using whole citrus seedlings for inoculation) to only 2-3 weeks. Young adults of ACP that had been reared on Las-infected plants were caged on excised healthy sweet orange leaves for 1-2 wk inoculation access periods (IAP), and both psyllids and leaves were tested later by quantitative PCR. When single adults were tested per leaf, percentages of Las-positive leaves averaged 2-6 percent using HLBaspr primers, and 10-20 percent using the more sensitive LJ900 primers. Higher proportions of Las-positive leaves were obtained with the following: 1) higher densities of inoculating psyllids (5-10 adults/leaf), 2) longer IAP, and 3) incubation of leaves for 1-wk post-inoculation before PCR. Logistic regression analysis indicated a positive correlation between Las titer in ACP adults tested singly and the probability of detecting Las in the inoculated leaves, which can be very useful in epidemiological studies. Comparison between excised leaves and whole seedlings, inoculated consecutively for 1 week each by one or a group of psyllids, indicated no significant difference between Las detection in excised leaves or whole plants. This new ‘excised-leaf assay’ method saves considerable time, material and greenhouse space, and may hopefully enhance vector-relation and epidemiological studies on Las and probably other Liberibacter spp. associated with huanglongbing disease.

   

 
Project Team
Lapointe, Stephen
Hall, David
Hunter, Wayne
Shatters, Robert - Bob
Patt, Joseph - Joe
 
Publications
   Publications
 
Related National Programs
  Crop Protection & Quarantine (304)
 
Related Projects
   REPELLENTS AND ATTRACTANTS FOR ASIAN CITRUS PSYLLID
   ASIAN CITRUS PSYLLID ATTRACTANTS AND REPELLENTS
   SPEEDY EVALUATION OF CITRUS GERMPLASM FOR PSYLLID RESISTANCE
   TARGETING THE ASIAN CITRUS PSYLLID (ASCP) FEEDING MECHANSIM AS A MEANS OF BLOCKING PSYLLLID FEEDING ON CITRUS
   DEEP SEQUENCING OF DIAPHORINA CITRI
   DEVELOPMENT OF CDNA MICROARRAYS FOR GENE EXPRESSION RESEARCH IN FLORICULTURAL CROPS
   COMBINATORIAL LIBRARY SCREENING FOR PSYLLID DISRUPTION MOLECULES
   ATTRACT AND KILL TECHNOLOGY TO CONTROL CITRUS LEAFMINER IN CITRUS NURSERIES AND ORCHARDS
   ASSESSMENT OF A MECHANICAL SAMPLING DEVICE FOR ASIAN CITRUS PSYLLID
   CHEMICAL ECOLOGY OF THE MEXICAN BROMELIAD WEEVIL AND ITS PARASITOID, LIXADMONTIA FRANKI
   COLLABORATIVE RESEARCH ON THE CITRUS LEAFMINER, PHYLLOCNISTIS CITRELLA
   SEMIOCHEMICAL-BASED TECHNOLOGY FOR CONTROL OF CITRUS PESTS
   DEVELOPMENT OF NOVEL INSECT CONTROL STRATEGIES BASED ON RNAI AND INSECT DETERRENT PROTEINS FOR INSECT PESTS OF CITRUS
   VIRUS OF HEMIPTERANS: LEAFHOPPERS AND PSYLLIDS
   PATHOGENS OF INVASIVE INSECTS
   SEMIOCHEMICALS FOR CONTROL OF CITRUS LEAFMINER AND CITRUS CANKER DISEASE WITH APPLICATION FOR CONTROL OF ASIAN CITRUS PSYLLID AND HLB
   INTERDISCIPLINARY APPROACHES TO ADVANCE CITRUS DISEASE RESEARCH & PRODUCT DVLPMT TO ENSURE THE SUSTAINABILITY OF THE NATL CITRUS INDUSTRY
   DEVELOPING A PHLOEM PENETRATION/ANTIMICROBIAL TREATMENT TO REDUCE/ELIMATE CANDIDATUS LIBERIBACTER FROM EXISTING CITRUS TREES
   DETERMINATION OF ATTRACTIVE HOST PLANT VOLATILES AND SEX PHEROMONES OF ASIAN CITRUS PSYLLID USING EAGS AND GC-EAD
   APPLICATION OF AN AGGREGATION PHEROMONE FOR MANAGEMENT OF THE DIAPREPES ROOT WEEVIL
   Rear and Release Psyllids as Biological Control Agents-An Economical and Feasible Mid-Term Solution for Huanglongbing (HLB) Disease
 
 
Last Modified: 05/22/2013
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