Figure 2: Dynamics of tomato yellow leaf curl virus sense and complementary sense strand DNA molecules accumulation in Bemisia tabaci individuals.

Time-course analysis of the amount of tomato yellow leaf curl virus (TYLCV) virus sense (VS, dark grey bars) and complementary-sense (CS, light grey bars) strand DNA molecules present in individuals of the Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED) species of Bemisia tabaci that were given a 24 h acquisition access period (AAP) of TYLCV Israel (TYLCV-IL) (two replicated experiments, panels a,b) or Mild (TYLCV-Mld) (two replicated experiments panels c,d) strains virion purifications. Acquisition was done in an in vitro feeding (artificial diet) experiment and whiteflies were maintained on an artificial diet after virus acquisition until analysis. The amounts of CS and VS were quantified at 0, 24, 36, 48, 72, and 96 hours after the beginning of the AAP from DNA extracted from 50 whitefly individuals (values for three biological replicates per time point, for each B. tabaci genotype and virus combination are shown). The mean CS:VS ratio (±standard error) for each virus and whitefly genotype combination through each time-course study is indicated; CS:VS ratio in virion purifications used for virus acquisition is also indicated. Quantification of VS and CS molecules was performed on DNA extracts according to Rodríguez-Negrete et al.¹⁸. Data from insect extracts were normalized to the 18S ribosomal RNA gene. The mean and standard error of three independent qPCR technical replicates are shown. PCR efficiency values along a dynamic range of 10³ to 10⁹ molecules were 94.13% for TYLCV-Mld and 94.6% for TYLCV-IL for VS detection. Efficiency values for CS measurements were 96.48% for TYLCV-Mld and 95.15% for TYLCV-IL.