Sponsoring Institution
Agricultural Research Service/USDA
Status
ACTIVE
Funding Source
USDA INHOUSE
Reporting Frequency
Annual
Project Director
YOKOMI R K
Accession Number
434468
Project Number
2034-22000-013-012T
Dates
2018-01-01 - 2020-09-30
Recipient Organization
AGRICULTURAL RESEARCH SERVICE
PARLIER,CA 93648
Keywords
(hlb)
clas
ddpcr
disease
greening
huanglongbing
qpcr
stubborn
Research Effort
Applied (40%)
Basic (50%)
Developmental (10%)
Classification Parameters
| Knowledge Area | Subject of Investigation | Field of Science | Percent |
|---|
| 212 - Pathogens and Nematodes Affecting Plants |
920 - Orange |
1170 - Epidemiology |
60% |
| 212 - Pathogens and Nematodes Affecting Plants |
910 - Grapefruit |
1160 - Pathology |
20% |
| 212 - Pathogens and Nematodes Affecting Plants |
930 - Lemon |
1160 - Pathology |
20% |
Goals / Objectives
The objective is to improve reliable diagnosis of "Candidatus Liberibacter asiaticus", presumptive bacterial causal agent of huanglongbing (HLB) (aka greening) and to more readily distinguish between HLB and Spiroplasma citri, the bacterial causal agent of citrus stubborn disease.
Methods (unparsed)
Deep sequencing using MiSeq PacBio platform to obtain additional full-length seqeunces of "Candidatus Liberibacter asiaticus" and Spiroplasma citri; conduct bioinformatic analysis, design new primers and probe (Taqman) with appropriate software and develop a new isothermal amplification assay using loop-mediated (LAMP) or Recombinase Polymerase Amplification (RPA) along with real time quantitative PCR, droplet digital PCR, high throughput robotic DNA extraction and purification of nucleic acids.
Methods
Deep sequencing using MiSeq PacBio platform to obtain additional full-length seqeunces of "Candidatus Liberibacter asiaticus" and Spiroplasma citri; conduct bioinformatic analysis, design new primers and probe (Taqman) with appropriate software and develop a new isothermal amplification assay using loop-mediated (LAMP) or Recombinase Polymerase Amplification (RPA) along with real time quantitative PCR, droplet digital PCR, high throughput robotic DNA extraction and purification of nucleic acids.