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ARS Home » Pacific West Area » Salinas, California » Crop Improvement and Protection Research » Research » Research Project #436486

Research Project: Identification of Additional Viruses Contributing to Lettuce Dieback Disease

Location: Crop Improvement and Protection Research

Project Number: 2038-22000-018-05-T
Project Type: Trust Fund Cooperative Agreement

Start Date: Apr 1, 2018
End Date: Mar 31, 2020

Objective:
1. Identify lettuce samples with lettuce dieback-like symptoms that test negative for tomato bushy stunt virus (TBSV) and Moroccan pepper virus (MPV), but positive for a virus-like agent through mechanical inoculation of test plants. 2. Conduct RNA sequencing (RNA-seq) analysis and compare to lettuce genomic and virus-like sequences to identify virus sequences. 3. Develop primers for reverse transcription-polymerase chain reaction (RT-PCR) detection of newly identified targets and evaluate against additional field isolates.

Approach:
Lettuce plants with symptoms of lettuce dieback will be collected from growing areas and tested by mechanical inoculation and RT-PCR for tombusvirus infection. Mechanical positive, but RT-PCR negative samples will be subjected to RNA sequencing (RNA-seq) analysis to compare total RNA content (which should include most viruses) with those of the lettuce genome. The goal is to identify additional unknown viruses associated with lettuce dieback disease. Virus genomic sequences identified using this method will be used to develop new RT-PCR primers, which will be used to identify virus infected lettuce from the field.