Disrupting transmission of Candidatus Liberbacter asiaticus with antimicrobial therapy

Disrupting transmission of Candidatus Liberbacter asiaticus with antimicrobial therapy

Report Date: 03/15/2020
Project: 18-018   Year: 2020
Category: ACP Vector
Author: Kirsten Pelz-Stelinski
Sponsor: Citrus Research and Development Foundation

 Objective 2. Determine the effect of antimicrobials on Las transmission.  Hypothesis: ACP will be less capable of transmitting CLas after feeding on antimicrobials because trees treated with antimicrobials are more likely to have lower CLas titers for acquisition. Eight-year old CLas-infected citrus trees have received six foliar applications (May-December) of streptomycin, oxytetracycline (Treatments), or receive no antimicrobials (Control). Ten CLas-free insects per plant from a laboratory colony were caged on young leaves (flush) of treatment and control trees to analyze ACP survival, CLas-acquisition in ACP P1 and F1 progeny, the total trees sampled consisted of 5 individual tree per treatment. In microcentrifuge tubes containing 1 mL of 80% ethanol, ACP adults were collected individually and then stored at -20°C for subsequent CLas detection using real-time PCR. Survival of ACP and CLas-acquisition were replicated twice from June 2019 to March 2020. During the first replicate, ACP P1 adults were collected on the 26th of June. Approximately two weeks later, five to ten ACP adults corresponding to the F1 progeny were collected. The adults collected on the first replicate in June showed higher CT means (> 38.5) and low copy numbers (< 3); indicating that ACP were unable to acquire the pathogen from treated trees.  The second (July), the third (September), fourth (October), the fifth (November), the sixth (January), and the seventh (March) replicates were collected using the same conditions previously described. Concurrently, the titer of CLas had been monitored at the same time-points using three leaves per tree to determine the CLas-infection rate. Currently, psyllids collected from late June through March are being processed to analyze the CLas-infection rate.  Objective 3: Determine the effect of antimicrobials on plant response and associated ACP behavior.  The objective of this experiment is to determine whether antimicrobial treatments applied to citrus plants affect behavior of Asian citrus psyllid that may change plant susceptibility to ACP infestation or pathogen inoculation. Two antimicrobial treatments are being investigated. These are Fireline (oxytetracycline HCL) and Firewall (streptomycin sulfate). Each is being applied to trees at label recommended rates with recommended adjuvants. To date, all treatments have been applied as foliar sprays; however, experiments are in progress. Treatments were applied to two-year-old Citrus sinensis L. Osbeck cv Valencia grafted onto US-812 rootstock. As described in our previous report, experiments comparing all uninfected (treated with antimicrobials versus untreated) versus all infected plants are ongoing using the T-maze olfactometer to determine whether Fireline affected ACP preferences for antimicrobial-treated versus untreated plants.  Experiments are still in progress with Firewall to determine whether application will induce an effect on plants that would cause a consequential change in the behavior of the vector to increase or decrease their preference for treated versus untreated trees.  


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