Grant Information
| Knowledge Area | Subject of Investigation | Field of Science | Percent |
|---|---|---|---|
| 202 - Plant Genetic Resources | 1050 - Date | 1080 - Genetics (excludes breeding) | 20% |
| 202 - Plant Genetic Resources | 910 - Grapefruit | 1080 - Genetics (excludes breeding) | 20% |
| 202 - Plant Genetic Resources | 920 - Orange | 1080 - Genetics (excludes breeding) | 20% |
| 202 - Plant Genetic Resources | 930 - Lemon | 1080 - Genetics (excludes breeding) | 20% |
| 202 - Plant Genetic Resources | 999 - Citrus, general/other | 1080 - Genetics (excludes breeding) | 20% |
Establish reliable real-time PCR detection of graft transmissible pathogens of citrus including but not limited to Pospiviroids, Hostuviroids, Spiroplasma citri, Dweet mottle virus, citrus tatter leaf virus. Establish protocols for rapid extraction of pathogen nucleic acid from target tissue. Develop genomic information of graft transmissible pathogens of citrus including pathogens which are presently uncharacterized.
This project is related to objectives 1C, "Develop, validate, and apply alternative means, such as cryotherapy, to free citrus germplasm from graft transmissible pathogens", of the parent project. Regulatory requirements are changing for citrus nurseries in California as a result from the increasing spread of the Asian citrus psyllid and huanglongbing (HLB). As nurseries are moving operations under protected conditions, there is an increased need for laboratory testing for presence of graft transmissible pathogens as the facilities for the traditional biological indexing will not meet the increased demand. A 10-plex assay for RNA pathogens and a 3-plex assay for DNA pathogens has been developed utilizing Luminex-based technology. This assay allows for the testing of 10 RNA pathogens or 3 DNA pathogens in one reaction well simultaneously with quantitative results. Application of this methodology in the citrus nursery industry would reduce the need to do multiple sample collection for the different graft transmissible pathogens and the cost per pathogen would be less than real time PCR.