Sponsoring Institution
Agricultural Research Service/USDA
Status
ACTIVE
Funding Source
USDA INHOUSE
Reporting Frequency
Annual
Project Director
HANDLER A M
Accession Number
432264
Project Number
6036-22000-030-16R
Dates
2016-10-01 - 2017-08-31
Recipient Organization
AGRICULTURAL RESEARCH SERVICE
GAINESVILLE,FL 30604
Keywords
candidatus
citri
citrus
conditional
diaphorina
grenning
insect
lethality
liberiabacter
population
replacement
transgenesis
Research Effort
Applied (20%)
Basic (70%)
Developmental (10%)
Classification Parameters
| Knowledge Area | Subject of Investigation | Field of Science | Percent |
|---|
| 211 - Insects, Mites, and Other Arthropods Affecting Plants |
999 - Citrus, general/other |
1130 - Entomology and acarology |
100% |
Goals / Objectives
The goal of this research is to develop methodologies for germ-line transformation of the Asian citrus psyllid (ACP), Diaphorina citri, which is a plant disease vector for the bacterium, Candidatus Liberibacter. Genetically-transformed strains will be used to replace or suppress populations of ACP in order to control citrus greening disease.
Methods (unparsed)
Methodology for transposon-mediated germ-line transformation of Diaphorina citri will be developed and transgenic strains of the plant disease vector will be created for either population replacement or suppression. Transposon vectors to be tested will include the piggyBac and Minos transposable elements marked with polyubiquitin-regulated flourescent proteins. Protocols will initially include embryonic microinjection of vector and helper plasmids, that will be extended to maternal abdominal injection, electroporation and biolistics delivery. Efficient transformation protocols will then be used to create vector-incompetent transgenic Asian citrus psyllid (ACP) having a synthetic "Media-like" element that will replace vector-competent strains in the field. Transgenic strains will also be created having a conditional tetracycline-suppressible embryonic lethality system that will result in inviable progeny after field release.
Methods
Methodology for transposon-mediated germ-line transformation of Diaphorina citri will be developed and transgenic strains of the plant disease vector will be created for either population replacement or suppression. Transposon vectors to be tested will include the piggyBac and Minos transposable elements marked with polyubiquitin-regulated flourescent proteins. Protocols will initially include embryonic microinjection of vector and helper plasmids, that will be extended to maternal abdominal injection, electroporation and biolistics delivery. Efficient transformation protocols will then be used to create vector-incompetent transgenic Asian citrus psyllid (ACP) having a synthetic "Media-like" element that will replace vector-competent strains in the field. Transgenic strains will also be created having a conditional tetracycline-suppressible embryonic lethality system that will result in inviable progeny after field release.