C I T R U S A D VA N C E D T E C H N O L O G Y P R O G R A M
QUARTERLY & FINAL REPORTS: Control of Citrus Greening, Canker & Emerging Diseases of Citrus
INSTRUCTIONS
Quarterly Report
Annual Report
Final
Required: What is the “headline” for this report (e.g. a one-sentence “newspaper headline” describing what you accomplished)
Evaluation of leafminer populations and movement
Proposal Title
Optimizing Spatial Distribution of Pheromone Traps for Monitoring Citrus Leafminer and Related Species
Today’s Date
4/15/15
Sponsoring Organization (drop-down)
Citrus Research and Development Foundation
Category (drop down)
Epidemiology and Cultural Control
REPORT UPDATE (500 words; summarize your accomplishments )
Citrus leafminer (CLM) is a major pest of citrus, causing direct damage and increased incidence/intensity of citrus canker.
Pheromone traps are valuable for monitoring CLM but need further evaluation to optimize use for management. The
objectives of the proposed project include: (1) assess species composition of trap catches, (2) optimize spatial and
temporal distribution of pheromone traps.
Laboratory Technician, Vincent Ficarrotta has continued to extract DNA from P. citrella samples. An additional 196
samples have been sequenced for COI barcoding to target the 658 bp region of COI gene. We recently switched
extraction methods from QIAGEN DNA Blood Tissue Kit to OmniPrep. The dataset was merged with our previously
published genetic dataset (Kawahara et al. 2013) generated from this project. The OK Slough had the highest number of
samples sequenced, and within these samples, P. citrella was the most common, followed by P. vitegenella. Other
species, such as the two new Phyllocnistis species, are lured to traps at a low frequency (≤11 specimens/year) in all years
sampled thus far. While the numbers show strong bias towards P. citrella, it is clear that in all years, multiple species of
Phyllocnistis readily come to CLM lures, following the initial finding of Kawahara et al. (2013).
We also generated a new ML tree with additional samples analyzed on UF’s High Performance Computing Cluster
(HPC). This tree shows largely similar results as Kawahara et al. (2013), where the majority of samples obtained grouped
with already-known specimens of P. citrella. Bootstrap values for clades were generally low for most groups (<50%),
which is comparable to prior results based on a single marker (e.g., DePrins and Kawahara [2012]). However, there were
some notable results, such as the strong sister group relationship of P. citrella (North American) to P. citrella (Asia).
Further, there is strong support for the monophyly of P. citrella (NA and Asia), P. insignis, and Phyllocnistis new sp. 1.
Other taxa that were included in the dataset, but not generated from this particular project, also show strong bootstrap
support for their monophyly, which will be relevant to future studies on non - P. citrella related species in the genus (e.g.,
P. hyperpersea (100% BP), P. longipalpus (100% BP), P. perseafolia (100% BP), P. populiella (93% BP), P. saligna
(100% BP)). We also see relatively strong genetic distance between each group (>2-4% in many cases), supporting the
hypothesis that these clades should be treated as separate species. Finally, 212 sequences of the 658 bp COI barcode
region that were recently sequenced were submitted to GenBank (www.ncbi.org) as proposed in the original project
proposal. The GenBank numbers are being acquired and verified by NCBI staff.
We are awaiting results of review and a decision on "Placement Density and Longevity of Pheromone Traps for
Monitoring of Citrus Leafminer, Phyllocnistis citrella (Lepidoptera: Gracillariidae)" by P. Vanaclocha, M. M. Jones, C.
Monzó and P. A. Stansly.
Mark-release-recapture work using fluorescent powders has indicated that male moths are capable of flying 140 m in
24 h. We determined that release immediately after application of powders is essential to capturing marked moths. We
have replicated traps at the distances over four release dates and are preparing to deploy traps at 280 m and 420 m from
the release site. We are also preparing to sample Rb-spiked citrus to evaluate Rb uptake and detection in CLM for
mark-recapture applications.
PI First Name Philip
Organization SWFREC - University of Florida
PI Last Name Stansly
Contract Number 00089614
Email [email protected]
Project Duration (years) 3
Phone 239-658-3400
Total Funds (current year) $103,436.04
Year of Project 3
SUBMIT REPORT