SUB00004552S
STATEMENT OF WORK: IN THIS PROJECT, WE WILL DISTRIBUTE AND DISSEMINATE SURVEYS AND EDUCATIONAL MATERIALS. I WILL PROVIDE INFORMATION ON ACREAGE, NUMBER OF GROWERS, AND MANAGEMENT APPROACHES USED IN CA TO FULFILL THE OBJECTIVE INVOLVING RISK ASSESSMENT. I WILL BE FACILITATING AND WORKING WITH THE UF TEAM IN ORGANIZING AND DELIVERING EXTENSION/OUTREACH ACTIVITIES IN CALIFORNIA. I WILL ORGANIZE A TRIP TO CA FOR THE UF TEAM DURING THE GRANT PERIOD, IN WHICH THE UF-TEAM WOULD DELIVER INFORMATION ON THE PROJECT AND ON LAUREL WILT DISEASE. THE UF TEAM WILL BE SCHEDULED TO VISIT THE UNIVERSITY, THE CDFA AND MEET WITH RESEARCHERS AND REGULATORY PERSONNEL. I WILL ALSO HOST (AND ADVERTISE) HYBRID EVENTS WHICH CA GROWERS/RESEARCHERS/AGENCIES WILL ATTEND ONLINE. I WILL HELP THE UF TEAM SPREAD THE NEWS OF THE EVENTS SUCH AS ONE HYBRID WORKSHOP/YEAR.
Regents Of The University Of California At Riverside
Project Grant 20245118143302
$250.0k 9/24/24 SUB00004554S
THE CO-I AT UC RIVERSIDE (UCR) WILL ASSUME RESPONSIBILITY FOR GENERATING ALL BEETLE DATA FROM CALIFORNIA. THIS WILL INCLUDE SEASONAL FIELD-TRAPPING AND COLLECTION OF INFESTED LOGS, FOLLOWED BY GENETIC IDENTIFICATION/CHARACTERIZATION OF THE AMBROSIA BEETLE COMMUNITIES ASSOCIATED WITH AVOCADO ORCHARDS IN THE STATE, AND ACROSS THE US. FIELD TRAPPING & COLLECTION OF INFESTED LOGS. AT LEAST TWO COMMERCIAL AVOCADO ORCHARDS WILL BE SAMPLED IN EACH OF THREE COUNTIES; VENTURA CO, SAN DIEGO CO, AND RIVERSIDE/SAN BERNARDINO CO. EACH ORCHARD WILL BE VISITED TWICE IN THE SPRING, SUMMER, AND FALL, AND SAMPLING WILL BE PERFORMED IN TWO CONSECUTIVE YEARS. ON THE FIRST VISIT, OF EACH SEASON, A SERIES OF LINDGREN FUNNEL TRAPS WILL BE PLACED IN THE ORCHARDS, AND VARIOUSLY BAITED WITH ONE OF THREE KNOWN AMBROSIA BEETLE ATTRACTANTS (HIGH-RELEASE ETHANOL, ALPHACOPAENE, OR QUERCIVOROL). THE COLLECTION CUP AT THE BASE OF EACH TRAP WILL BE FILLED WITH ALCOHOL-FREE RV ANTIFREEZE (E.G., YODRVAFPREM-06) TO PREVENT ESCAPE AND PRESERVE THE DNA OF INSECTS ENTERING THE CUP. CONTENTS OF THE TRAPS WILL BE COLLECTED AFTER 2 WEEKS. TRAPS WILL BE RENEWED AND LEFT IN PLACE FOR A FURTHER 2 WEEKS. THE CONTENTS OF EACH TRAP WILL BE RETURNED TO UCR IN A COOL BOX. ON EACH VISIT, WE WILL ALSO WALK THROUGH THE ORCHARD LOOKING FOR EVIDENCE OF ANY AMBROSIA BEETLE INFESTED BRANCHES. SUCH BRANCHES WILL BE REMOVED USING A PRUNING SAW (STERILIZED WITH 100% ETHANOL BETWEEN EACH CUT), CUT TO A CONVENIENT SIZE, DOUBLE-BAGGED INSIDE HEAVY DUTY TRASH CAN LINERS), AND TRANSPORTED BACK TO UCR INSIDE SEALED PLASTIC TOTES. THIS PROTOCOL IS CURRENTLY REQUIRED TO ALLOW THE MOVEMENT OF WOOD THAT IS POTENTIALLY INFESTED WITH KUROSHIO SHOTHOLE BORER INTO RIVERSIDE COUNTY AND THE UCR INSECTARY & QUARANTINE FACILITY (I & Q), IN ACCORDANCE WITH CALIFORNIA PLANT PEST MOVEMENT PERMIT 3976. ON ARRIVAL AT UCR, THE SEALED TOTES WILL ENTER I & Q. ONCE INSIDE, INDIVIDUAL LOGS WILL BE PLACED IN EMERGENCE JARS AND MONITORED DAILY FOR 6 WEEKS. ALL EMERGING ARTHROPODS WILL BE COLLECTED, AND IMMEDIATELY EUTHANIZED IN >95% ETHANOL, BEFORE BEING REMOVED FROM I & Q. AFTER 6 WEEKS, THE LOGS WILL BE PLACED IN DOUBLE LAYER AUTOCLAVE BAGS, AUTOCLAVED AT 121O C FOR 30 MINUTES (TO KILL ANY REMAINING INSECTS OR FUNGI), PRIOR TO DISPOSAL. INSECTS IN ETHANOL WILL BE STORED AT -20O C FOR GENETIC CHARACTERIZATION. SPECIMEN IDENTIFICATION. AT UCR, THE CONTENTS OF EACH FIELD TRAP WILL BE SORTED INTO PARA-TAXONOMIC Â SPECIESÂ (MORPHOLOGICAL GROUPS), UNDER A STEREO DISSECTING MICROSCOPE. REPRESENTATIVE SAMPLES OF EACH GROUP, FROM EACH ORCHARD, WILL BE TRANSFERRED TO >95% ETHANOL AND STORED AT -20O C FOR GENETIC CHARACTERIZATION. SUBSEQUENTLY, DNA WILL BE EXTRACTED FROM THESE SPECIMENS USING A NON-DESTRUCTIVE HOTSHOT METHOD, AND WE WILL AMPLIFY AND SEQUENCE TWO GENE REGIONS (THE MITOCHONDRIAL CYTOCHROME C OXIDASE SUBUNIT I (COI) AND THE NUCLEAR RIBOSOMAL 28S). BOTH GENES ARE WIDELY USED IN TAXONOMIC AND PHYLOGENETIC STUDIES AND AS SUCH ARE VERY WELL REPRESENTED IN PUBLIC DNA SEQUENCE REPOSITORIES. AS SUCH, ATTEMPTS TO IDENTIFY SPECIMENS WILL BE INITIALLY MADE USING BLAST SEARCHES OF THE NCBI AND BARCODE OF LIFE SEQUENCE DATABASES. THE BEAUTY OF THE HOTSHOT EXTRACTION METHOD MEANS THAT IF A SPECIES ID CANNOT BE OBTAINED, THE SPECIMEN CARCASS REMAINS, AND WILL BE IDENTIFIED USING MORPHOLOGICAL KEYS. REPRESENTATIVE SAMPLES FROM FLORIDA, HAWAIIAN, AND PUERTO RICAN COLLECTIONS WILL ALSO BE SENT TO UCR FOR SEQUENCING. TOGETHER, THIS GENETIC INFORMATION WILL BE USED TO DOCUMENT AND COMPARE SPECIES COMPOSITION, AND ASSESS INTRASPECIFIC GENETIC VARIATION, BETWEEN ORCHARDS, COUNTIES, AND STATES. ALL EXTRACTED SPECIMENS AND THEIR DNA WILL BE MAINTAINED AT -20O C, AND A SUBSET OF THE SEQUENCED SPECIMENS WILL BE RETRIEVED AND DEPOSITED IN THE UCR ENTOMOLOGY MUSEUM.
Regents Of The University Of California At Riverside
Project Grant 20245118143302
$217.6k 9/18/24