Comprehensive Assessment of Reference Gene Expression within the Whitefly Using RT-qPCR.

The citrus whitefly, , is a destructive pest that infests citrus plants. It is a major vector in transmitting plant viruses such as citrus yellow vein clearing virus (CYVCV), which has caused severe economic losses worldwide, and therefore efficient control of this pest is economically important. However, the scope of genetic studies primarily focused on is restricted, something that has potentially limited further study of efficient control options. To explore the functionalities of target genes, screening for specific reference genes using RT-qPCR under different experimental conditions is essential for the furtherance of biological studies concerning . The eight candidate reference genes were evaluated by dedicated algorithms (, , and Δ method) under five specific experimental conditions (developmental stage, sex, tissue, population and temperature). In addition, the software, a comprehensive evaluation platform integrating all of the above algorithms, ranked the expression stability of eight candidate reference genes. The results showed that the best reference genes under different experimental settings were and at different developmental stages; and in both sexes; and in different tissues; and under different populations; and and in different temperatures. The validation of selected reference genes was further identified using heat shock protein () 70 as a reporter gene. Our study, for the first time, provides a detailed compilation of internal reference genes for that are suitable for RT-qPCR analysis, which is robust groundwork for comprehensive investigation of the functional target genes of