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Body of Text Huanglongbing (HLB) is the most serious disease of citrus, putatively caused by the bacterium ‘Candidatus Liberibacter asiaticus’ (CLas), transmitted by the Asian citrus psyllid (Diaphorina citri Kuwayama) in the USA. Most commercial citrus cultivars are susceptible to the disease, particularly conventional sweet oranges and grapefruit, decline severely in health and productivity (Bové 2006, Gottwald 2010). Early detection and prompt response are key factors in the eradication or suppression of HLB epidemics in California. Real-time PCR is the standard regulatory method for CLas detection and it can detect as little as a single copy of bacterium in a sample. However, due to lack of visible symptoms, low titer and uneven distribution of CLas in a tree, selecting the best sample is a major hurdle for early detection by qPCR. Regulatory testing of CLas by qPCR stipulates sampling of mature leaves. Such leaves are good for visual inspection of HLB symptoms but can be unreliable tissue for early detection and low titer infections. Symptoms of HLB develop slowly and without symptoms, collecting that right leaf/shoot amongst >200,000 estimated leaves per mature tree is practically impossible for early diagnosis.

Reliable Sampling Tissue and Seasonality for Consistent Detection of ‘Candidatus Liberibacter asiaticus’ by qPCR