U.S. flagAn official website of the United States government
                    Here's how you know **Official websites use .gov**
                            
                            A **.gov** website belongs to an official government organization in the United States.
 **Secure .gov websites use HTTPS**
                            
                            A **lock** (                            
                              A locked padlock                                                      
                            
                            ) or **https://** means you’ve safely connected to the .gov website. Share sensitive information only on official, secure websites.


Menu Agricultural Research Service
						
                    U.S. DEPARTMENT OF AGRICULTURE
 Temperate Tree Fruit and Vegetable Research: Wapato, WA
 close



    Research

                    Research Projects
                                    Publications
                                    Collaborations
                                    News
                People

                    Staff Listing
                                    Careers


        


                    Facilities
                        USDA Laboratory in Wapato, WA
                                    USDA Prosser Worksite
                                    USDA Moxee Farm
                    About UsSearch small
                    Search
            
            ARS Home
                                About ARS
                                Contact Us




ARS Home Pacific West Area Wapato, Washington Temperate Tree Fruit and Vegetable Research Research





- Cooper, William - Rodney
- Crop Protection & Quarantine (304)

Research Project: **Location:
                            Temperate Tree Fruit and Vegetable Research** Project Number: 2092-22000-022-024-S
                        
                        Project Type: Non-Assistance Cooperative Agreement Start Date: Jun 1, 2021
                        
                        End Date: May 31, 2024 **Objective:**
                        
                        The goal of our study is to identify viral pathogens that could be developed or propagated as biological control agents for potato psyllid. Objective 1: 1A: Examine the transmissibility of the novel BcPLV between infected and uninfected potato psyllids. 1B: Determine whether observed morphological abnormalities or colony crashes are associated with high rates of virus infection with colony populations. Objective 2: Screen wild and laboratory populations of potato psyllids for the presence of Bactericera cockerelli Picorna-like virus (BcPLV). Objective 3: Use a metagenomic approach to identify other viral pathogens potentially infecting potato psyllids. **Approach:**
                        
                        Objective 1:  We will examine how readily BcPLV is transmitted among psyllids and explore the possible effects of virus infection on potato psyllid. Objective 1A:  To examine the transmissibility of BcPLV, we will confine virus-infected and uninfected psyllids to potato leaves using sleeve cages. Initial experiments will include cLso-uninfected psyllids only. Treatment pairs will test whether the virus is transmitted by contact or close proximity, sexually, or via plant conduits. Objective 1B:  We suspect that the viral load will fluctuate within colony cages. Twenty psyllids from each colony cage will be collected from colony cages every three weeks (approx. 1 generation). The adults will be examined for the presence of morphological abnormalities, which will be photo documented. The psyllids will then be tested for the presence or absence of BcPLV using diagnostic RT-PCR. In addition to collecting psyllids, population fluctuations within cages will also be monitored to determine if colony “crashes” are associated with high rates of virus infection. Objective 2:  We will collect potato psyllid adults from potato fields and from the uncultivated host plant, matrimony vine, using a vacuum sampler. The psyllids will be examined for presence of morphological abnormalities and will be tested for the presence of BcPLV using RT-PCR. We will also screen potato psyllids captured in 3D printed traps and sticky card traps deployed in potato fields for another study involving beet leafhopper. Results from Objective 2 will be used to 1) determine BcPLV infection rates among field populations of potato psyllid, and 2) determine whether BcPLV can be readily detected in psyllids captured on sticky card traps, or if preservation of virus particles by use of the 3D printed traps is necessary for accurate detection of the virus. Objective 3. The three colonies of psyllids belonging to the central, northwestern, and western haplotypes will be subjected to the HTS analysis, using 10-20 pooled psyllids per colony. Total RNA will be extracted using a TRIzol reagent-based protocol. DNAse-treated, purified RNA will then be depleted of ribosomal RNA using the RiboMinus kit for RNA-Seq. Before proceeding further, quality of resulting ribo-depleted RNA will be checked by fragment analysis. The ribo-depleted RNA will then be submitted to the Genomics Resources Core at the University of Idaho for library preparation, multiplexing, and sequencing on the Illumina MiSeq platform. Bioinformatics analysis of the high-throughput sequencing data will be performed as described previously. Any matches to the custom-prepared virus sequence database entries will be confirmed through the RT-PCR or PCR verification using specific primers, with subsequent Sanger-based sequencing of amplified PCR fragments.


Return to top


Connect with ARS Sign up


ARS Home
                            USDA.gov
                            Plain Writing
                            Policies & Links
                            Civil Rights Statements
                            FOIA
                            Accessibility Statement
                            Privacy Policy
                            Non-Discrimination Statement
                            Quality of Information
                            USA.gov
                            WhiteHouse.gov