Longevity of ingested mRNA transcripts in the gut of a homopteran (Bemisia tabaci): avoiding experimental artifacts

Abstract

Reverse transcription-polymerase chain reaction was used to detect the presence of ingested tomato transcripts in whiteflies (B. tabaci) collected immediately after removal from tomato and following a 72-h feeding period on either sucrose solution or cotton plants after removal from tomato. Primers selected for ribulose-bisphosphate carboxylase (Rubisco) and chlorophyll a/b binding protein transcripts were very specific and did not amplify their cotton counterparts. A 162-bp amplimer of Rubisco was observed only in plants and whiteflies allowed to feed on sucrose for 3 days after removal from tomato. This was observed with both the non-viruliferous and viruliferous whiteflies. The ampliner was not observed in whitefly fed with cotton for 3 days after removal from tomato. The results were similar for tomato chlorophyll binding protein, where the transcript was only successfully amplified in the total RNA from tomato and from whitefly feeding n a sucrose artificial diet. Plant transcripts persisted in viruliferous and non-viruliferous whiteflies after 72 h of feeding on sucrose.