RNAi Feeding Bioassay: A Protocol for dsRNA Screening Against Asian Citrus Psyllid and Related Hemipteran Insects.

RNA interference (RNAi) comprises a natural mechanism of gene regulation and antiviral defense system in eukaryotic cells, and results in sequence-specific degradation of RNAs. Recent scientific studies demonstrate the feasibility of use RNAi-based strategies to control pest and pathogens in plants. A key step in developing RNAi-based products is a reliable method to appropriated screening of selected dsRNAs.Herein presented are a bioassay for screening dsRNAs to control the Asian citrus psyllid (ACP), Diaphorina citri, vector of citrus Huanglongbing (HLB) and other hemipterans. The RNAi feeding bioassay, called in plant system (iPS), uses vegetative new growth citrus flush to deliver double-strand RNA (dsRNA ) to ACP during natural feeding .

Infestation dynamics of Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae) in citrus orchards as affected by edaphic and climatic variables.

Brevipalpus phoenicis (Geijskes) is a cosmopolitan and polyphagous mite that transmits important phytoviruses, such as coffee ringspot virus, passion fruit green spot virus and Citrus leprosis virus C. To characterise the dynamics of the probability and the rate of B. phoenicis infestation in response to edaphic and climatic factors, monthly inspections were performed in nine orchards in a citrus region of the State of Bahia, Brazil, for 35 months. Three fruits per plant were examined using a magnifying glass (10×) on 21 plants distributed along a "W"-shaped path in each orchard. Meteorological data were collected from a conventional station. To determine the correlations among the climatic variables, the data were analysed using Spearman correlations. Variables were selected by principal component analysis, and those that contributed the most to differentiate the groups were evaluated via a Mann-Whitney test. Using the quantile-quantile method, the limit values for the following climatic variables were determined: temperature (24.5 °C), photoperiod (12 h), relative humidity (83%), evapotranspiration (71 mm) and rainy days (14 days). The combination of longer days, higher temperatures, lower relative humidity levels and lower evapotranspiration increased the probability of B. phoenicis infestation, whereas successive rain events decreased that risk. Infestation rates were negatively affected by relative humidity levels above 83% and were positively affected by a decreasing available soil-water fraction and increasing insolation and photoperiod.