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We observed the health status of oak trees in a conservation seed orchard for over twenty years, focusing on characteristic virus-suspected symptoms. The orchard was established in 1992 in Kreuztal, North Rhine-Westphalia (Germany) with 1302 seedlings in 186 clusters. The number of seedlings showing chlorotic ringspots and mottle on leaves has fluctuated annually, but has increased from 3.3% to 12.1% in the last 20 years; the number of affected clusters has risen from 8% to 25.9%. A scientific breakthrough was the identification of a novel virus related to members of the genus Emaravirus in diseased oak by high-throughput sequencing (HTS). Screening of the oak seedlings in three consecutive years, using a newly established virus-specific diagnostic reverse transcription polymerase chain reaction (RT-PCR), confirmed the virus infection and revealed a close to 100% association between the observed leaf symptoms and the novel virus. As no other plant virus could be identified in the HTS-datasets, we assume the novel virus is primarily causing the symptoms. To reliably detect the novel virus in oaks, RT-PCR targeting the viral RNA3 or RNA4 should be applied in routine testing of symptomatic leaf tissue. It was obvious that most groups with virus-infected plants cluster, with only five out of the 42 affected groups being offside, not bordering on other affected groups of plants. There was no clear correlation between the detection of the virus and the overall vitality of the seedlings. In some groups, the strongest seedling emerged to be virus-infected, whereas in others virus-infected seedlings continued to grow. Forecasts on the future growth behavior of these virus-infected oak trees are therefore not possible.

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