Tomato plants at the 7–8 true-leaf stage that had been infected witheither TYLCV or ToCV for 14 days were used for this procedure. Newly emerged (0–24 h) non-viruliferous female adultwhiteflies were first placed in small self-sealing Petri dishes (10-cm diameter) containing moistened filter paper for the 3-h preacquisition starvation period. They were then moved onto TYLCV-infected, ToCV-infected or ToCV+TYLCV mixed infected tomato plants for a 6-, 12-, 24-, or 48-h timed acquisition feeding periods. Each viral acquisition test was conducted using 60 adult whiteflies. When the acquisition efficiency of TYLCV by whiteflies was tested, the whiteflies were placed on TYLCV-infected and ToCV+TYLCV mixed infected tomato plant upper leaves (the second leaf from the top). When the acquisition efficiency of ToCV by whiteflies was tested, the whiteflies were placed on lower leaves(the second leaf from the bottom) of ToCV infected and ToCV+TYLCV mixed infected tomato plants. Thereafter the adults were collected, stored at −70°C and later assayed individually for detectable TYLCV or ToCV using PCR methods with primers specific for ToCV (ToCV-F1/ToCV-R1) or TYLCV (TYLCV-F1/TYLCV-R1) (Supplementary Table 1). Finally, viral acquisition rates (no. infected/no. inoculated) from singly or doubly infected tomato plants were calculated.

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